VoLv~ XXX MAI¢CH, 1935 No.

T H E GI" E NETICS
" q I
A N D CHEMISTI~Y OF F L O W E R
COLOUI~ I N D A H L I A : A N E W T H E O R Y OF
SPECIFIC PIGMENTATION.
BY W. J. (J. LAWI~ENCE
(John Innes Horticultural Institution)
AND ROSE SCOTT-~iIONORIEFF
(Dyson Perrins Laboratory, Oxford, and John Innes Hortioult~trcd
Institution).
(With Plates VII and VIII, Six Text-figures and One Folder.)
CONTENTS.
rAGE
Infll'OdUOfiion 156
Terminology 157
Part I. Genetics 159
(i) Flavones 159
(a) Yellow (Y) 159
(b) Ivory (z) 162
(ii) Anthoeyanin 166
(a) "Light" ~nthocyanin pigmenfi~fiion: the A factor 166
(b) "~eavy" ~nfihoey~uin pigmentation: the B factor t68
Off) Interaction of factors 168
(a) Interaction of Y and I . 168
(b) Infieraetion o f A ~nd I . 173
(e) Interaction of B and I . 178
(d) Interaction of Y with A and B 181
(c) l!)~fiflerns 185
(f) Cumulative effects 186
l)~rfi If. Chemistry . 187
(i) The flower pigments of Dahlia . 187
(c0 Fla,vones 187
(b) Anthocyalfins 190
(ii) GeneticM ~speet of ehemicM results 196
(a) Dahlic~ sI)eeios ~md species hybrids
(b) D. variabilis . 197
(1) ]J'lowers with non-yellow grounds ]97
(2) Flowers ~dth yellow grounds 198
(3) ]~'[Uto,tions 20O
(c) CqenerM observations 2O2
J o u r n . of Genetics x x x Ii
156 Genetic8 (end Che~rdst'ry of Flowc'r Colour i n D a h l i a
P~rg I i [. A new theory of t'ae~orial balance and specific pigmentarY,ion in Dahlia, 203
(i) GcnerM . 203
(if) Pigmen~ bahmec in '~he A I ~ypes 205
(iii) Pigment balance in the B Y t;ypes 207
(iv) Pignmng balance in the ]3AYI types 208
(v) SpEcific a,nghocyanin production 209
(vi) VaeCorial conditions governing SpECific pigmenC,~tAon in Dahlia 213
(vii) A biochemical bheory ibr specific pigmental)ion in Dcttdia . 216
I~ecapigula~ion 220
(i) Gcnegioal 220
(if) Chemical 221
(iii) TheorelAcal 222
I~eferences . 224:
Explanation of Pl~ges VII and VI.tl. 225

INTI:~OD UCTION.

IN two previous papers (Lawrence, 1929, 1931) accounts were given of

the gpneties of flower coleus in the garden dahlia, De&fig va~'~;abilis. It
was shown that, with the sole exception of D. vc~'ictbilis, Dc~hlia species
can be divided into two distinct groups for flower colour, viz. Group I,
pale to deep magenta with ivory varieties; Group II, orange to scarlet
with yellow varieties. Both series occur within D. sc~'iabilis. Further it
was shown that of the six species in cNtivation four were tetraploid
(2n = 32), one was hypertegraploid (2~ = 36) and the sixth, D. sc~'4c~bilis,
oetoploid (2~z= 65). The genetic studies on D. vc~'iabigis showed that four
main factors governed flower colour, namely I and Y producing ivory and
yellow fig+one, and A and B "light" and " h e a v y " gnthocyanin pigmen-
tation respectively (see p. 158). No pigments are produced by the
recessive gllelomorphs of these four factors. A fifth factor, now called I-I,
progressively inhibits the production of yellow flavone. The anthocyanin
may be either cyanin or petargonin, or a mixture of both, but ~here are
~zo specific fa, ctors for these 2igments.
The flower-colour range results from the mLxture of th.e flavoue and
anthocyanin pigments in varying proportions and intensities. Thus,
anthocygmn with yellow flavone appears orange to scarlet according t~o
the intensity of the anthocygnin. Similarly gnthocyanin with ivory
flavone gives magenta and purple colom's. Owing to the cumNative action
of the inhibibor, I-I, of yellow flavone; the colom:s cream, primrose and
pale yellow are commonly found, and with these intermediate flgvone
eolours the anthoeyanin gives intermediate flower colours (e.9. crimson)
(see Table XVII).
 W . J . C. LAW.R,ENO~,~AND _ROS:E Soo~r~L'-MoNo~I:~7~ ~ ]57
The chief eolours resulting from the combination of the fern" factors
are as follows:
bayi =white baYi =yellow
b a y I =ivory baYI 1-- yellow
b A y l = magenta BAYi = aprico g
B a y I =purple B a Y i = scarlet
Th.e factor Y was shown definitdy to be tetrasomie, giving the character-
istic ratios 5 : 1, 11 : 1 and 35 : 1 in the appropriate crosses. A similar
constibatioa was assmned for the factors A and. B. Th.e results presented
in this paper substantiate the conclusions previously reached, with one
exception. Although a number of plants had[ been tested, no evidence
had previously been found to indicate that I was tetrasomie, disomic
ratios onIy occurring. Improved methods of testing and the requisite
progenies for determining the constitution of this factor now show that I
too is tetrasomie, but with a modified expression.
The combined morphologicM, eytologioM and geneticM data strongty
suggested that the garden dahlia was a hybrid octoploid derived from
the crossing of two hybrid tetraploid species, one belonging to Group I
and the other to Group II, hybridisation having been accompanied by
doubling of the chromosome nmnber. It was concluded thab the tetra-
ploid dahlias hays probably descended from a diploid ancestral stock,
now extinct; and that during this descent differentiation occurred giving
rise, among other differences, to the two flower-eolour groups. Text-fig. 1
1?resents the scheme for the origin of the garden dahlia. It will be seen.
that D. w~'ic~bitis combines the products of specific differentiation with. a
high degree of polyploidy.
Further breeding experiments with the garden dahlia have had to be
postponed. In view of this, certain data will be presented and discussed
which normally would have been withheld until furbher results were to
hand.
The complexity arising from the combination of four tetrasomio eolour
factors is eonsiderM)le and of peculiar interest, especially in relation to
interaction effects. An attempt isnow made to state the relationship of
the specific flower-colour pigments in D. vc~'~abitis as elueidaSed by
genetioal and ehsmicM experiments.
TEI~NINOLOGY.
The flower eolours Mways refer to the pigmentation as seen on the
~l)l)e~" side of the ray floret, since differences of intensity, and even of
colour, not infrequently occur between the upper and lower surfaces.
a y is epis~atiogo I.
11-2
158 Genetics a n d C h e m i s t r y of F l o w e r Colour i n D a h l i a
In describing the various colom'ed forms, "light" and "heavy" will
be used to denote the an~hocyanin hl~ensity normally associated[ with
~he A and B factors respectively. Fro'Chef disflhlctions of intensity in
be Ch ligh~ and heavy forms will be indicated by "pale," "medium" and
"deep." By comparison of known genotypes and apart from interaction

Diploid ancestral stock

;t= 8

I
Hybrid diploid species

uroup II
Magenta or ivory flowers Scarlet or yellow flowers

D. dissects D. cocdnea (n = 16)

D. excelsa
D. imperialis (n= 16) D. coronata (n-- 16)
D. Lehmanni
D. Maximiliana? (n = 16) D. gracilis
D. Maxoni (n = 16)
D: Merckii (n= 18) D. tenuis
D. platylepis
D. pubeseens
D. scapigera

Hybrid oetoploid,
D. variabiliS
.n = 32

Flowers scarlet, magenta, yellow, ivory, etc.

Texd-fig.1.

the deepes~ light anthocyanin pigmentation is much paler fihan the

palest het~vy one (see p. 182), so that the comparative value of the germs
pMe, medium and deep is restricted to differences of intensity within the
light and heavy groups and not bet'wee~ them. It should be especially
noted that the germs "light" and "heavy" do not necessarily indicate
particular anthocyanins but only differences of intensity characteristic
of the A and B factors.
 . . . . . . ~ ~ P

(f[~3" i oii "

× x × x x x x x × x x x x × × x x x x z x x x x x x × x x x x x × x. x x x x x x x

~ ~ o~ ~ ~ o~ .2' ~ ~.~

o A

0~ " Very fah~fly

t~ ~ . ~ ~ ~, t~ % ~ - ~ . ~ • • . *- . . . . ~ ~ . t~ ~ • t~ to . . . . ~ . . . . ~[edium ~t)rleob

• . ~, - • ~ • • ~ . . . . . . . . . Or&nge

Bleached ~carle~-

ta . ~ . . . . . . . . . . . . . . , . . . . . . . t~ ~ . . . . . . . . . . . ]][e~hed crimson

° L
• ~ ~ ~ ~ ~ ~ t~ ~ t~ ,2 . . . . . . . . . Sctu'leborimson

~o ~ , . . , ~ • . . • ,~ ~Z ~ ~ ¢~ t~ . . . . . . . . . Crimson

e~ . . . . . . . . . . ~ . ~ ~ . . . . . t~ ¢* • • o~ . . . . . . . . . . . . I~urplish.orhn~m~

"4 . . . . . . . . . . . . . . . ~ ~ • s~ f . . . . . . ~n . t~ . . . . . . . . . 1)eep llm.plJ~h.

ot ~ ~ ~o to ~ ~ ~ . . . . . . . . . Pale pm'plo

~, Very dsep
v-d . . . . . . . ~ . . . . . . . . o: . • . • . . . . . . . • ' " ~ . . . . . . . . . . mngenLf~

4.

V~ry fldn~ly
. . . . . . ~ . . . . . . . . . . ~ . . . . . . . . . . . . ~ t~ c~ • t, i n g e d nmgenta

,~ . . . . . . . . . . . . . . ~ ~ - " . ~ . . . . . . . ~ . . . . . ~ . . . . . ~ Whii,e

tb tb g~ rb dl r3~ e;~ e}L 6t ~ ¢~ Ch tb Cb £1 al r3L l"~

........ '~"~ ...... ~ ...... ~" £'" ............. ~" ..... !~'" I 1 .............................................................................................. Ivm'yorwhi[e
 W. J; C. LAWI~ENCE AND I~OSE SCOTT-I~{ONC]{IEFF 159
Forms with anthoeyanin in their ray florets will be distinguished from
those without anthoeyanin by the terms cyanic and acyanic.
Finally, in order to avoid a cumbersome and confused symbolism
cje'netic constitutions will be denoted by dominant fa, ctors only ~, except in the
case of the bottom recessive a,,b,~y,~i,~. This abbreviated symbolism is
ilLtstrated below.
Factorial
Genotype Written abbreviation
xxxx N~flliplex --
Xxxx Simplex Xi
XXxx Duplex X~
XXXx Triplex Xa
XXXX Quadriplex X,t
E.ff. A a a a b b b b Y Y Y y l I i i - - A 1 Y ~ Ia .

When the number of dominants is unknown the factor will be represented

without a suffix.

PAI~T I. GENETICS.
New results are given in Table I. In the majority of eases the
factorial constitution of the parents has been derived from the analysis
of two or more different progenies. Although most of the families are of
quite moderate size, if cross-reference be made in the Tables I-VI it will
be seen that the interpretations adopted are substantially in agreement.
Unless otherwise stated, the genetic constitutions of the individuals
mentioned in this paper have been determined by their ancestry or
ot~spring, or both, the data for which will be found in the tables or in
earlier papers.
(i) F~Avo~s.
(a) Yellow (Y).
The inheritance of Y has been studied in families raised (a) fl'om
crossing forms with yellow ray florets together or with ivory or white,
and (b) fl:om the crossing of cyanic forms having yellow grmmds, with
each other or with recessive forms (Table II). In the case of (b) it is
usually quite easy to distinguish yellow from non-yellow grounds when
anthocyanin is present but, until chemical methods were used, in certain
deep cyanics it was impossible to decide whether the ground was yellow
or not.
i Since all analysis of inheritance in polyploids necessitates frequent reference to reces-
sive forms, it is simpler to denote the dominant factors only. Tiffs is opposite to the practice
of the Drosophila workers, who give the recessive but omit bhe dmninant (wild type)
factors.
].60 Genetics and Chemistry of Flower Uolour i~ Dahlia
TABLE II.
Inherita~ce of the J'actor Y.
Parents Observed Calculated
_ _ A
f h c-
Family ~ ~ Y y Y y
Y:~ × Y~ (and reciprocal)*
-- 14/26 M5 246 0 246 0
26/28 14/26 228/27 86 0 86 0
17/29 14/26 27~/27 24 0 24 0
21/29 27~/27 14/26 19 0 19 0
60/30 14/26 1"-/25 69 0 69 0
Total 444 0 44,1 0
Ya × Yl
231271
38/28 J' 14/26 Union Jack 175 0 175 0
6/28 14/26 ::deal 4-9 0 49 0
18/29 ] 4/26 27~/27 35 0 35 0
Total 259 0 259 0
"Y~ x Y,l (and reciprocal)
2/28 14./26 Whii~e St;at 79 51" 84 0
4/28 14/26 36/26 78 i"I- 79 0
29/30 32/26 14126 69 0 69 0
50/30 1,i/26 h'Is 152 1"l 153 0
Total 378 7I 385 o
Y.. x Y2
26/291 27~/27 27~/27 107 3 106'9 3-1
24/31 J
Y2x Yi (and reciprocal)
26/27 Union Jack 34/26 191 14
3/20 J 187.9 17.1
34/27 i~/25 Union Jack 134 14 135.7 12.3
40/30 2a/28 41i/28 83 9 79.3 7.7
54/30 2a/28 31~/27 72 8 72.3 6.7
66/30 Amy ]~arilet 31i/29 33 5 34.8 3.2
Total 513 50 516 46.9
38/29 22'J/27 Amy Barflet c a . 79 13 84-3 7-7
:~8131 42~I29 4D/28 ca. 50 8 53,2 4.8
Y. x Y,i (and reciprocal)
27/27 ~{~ M i 29 6 29.2 5'9
36/27 12/25 ii/25 16 16.7 3"3
37/27 1"/25 27/24 49 3 43'3 8.7
40/28 35/26 229/27 41 9 41.7 8"3
i2/29 36/26 ~{5 33 7 33'3 6-7
37/29 229/27 35/26 21 3 20 4
9/30 2:V28 32/26 38 8 38'3 7'7
14/30 34/26 White Star 22 5 22'5 4"5
] 5/30 3~/26 Everest 11 4 12'5 2.5
17/30 23/28 35/26 32 12 36.7 7'3
] 8/30 2a/28 Whi~c Star 21 8 24.2 4.8
20/30 10'/28 2:1/28 68 16 70 14
22/30 2'~/28 :Everest 35 11 38'3 7'7
26/30 lO/29 2:~/28 42 13 45.8 9'2
30/30 38~/29 Everest 76 10 72-5 14.5
45/30 White Star M~ 12 5 14,2 2.8
13/31 22s/27 Everest 18 4 18.3 3-7
14/31 22s/27 White Star 26 12 31.7 6'3
.I9/31 27'~/27 White S~ar 26 5 25.8 5.2
22/31 27~/27 Everest 30 12 35 7
37/31 42i/29 Everest 62 12 61.7 12.3
Total 708 169 730.8 146-2
31/31 27~/27 Everest c a . 25 6 25.8 5"2
 W . J . CJ. LAW]KENCE AND I~OSE SCOTT-M.ONCI~IEFF 161

TABLE II (co,~lt.)
Lnhe'rita'nce of l]~efactor Y .
Parents Observed Calculated
A

F~mity ? ~ Y y Y y
Yi x Y l
22127 Union Jack Glenshee 274: 79 264,8 88.3
32/27 Union Jack Ideal 7 1 6 2
6/29 41J/28 4.1~-/28 42 13 ~1..3 13.8
33/3o 41L/28 2"-/28 14 5 14.3 4.8
55/3o 31n/27 2~/28 38 12 37.5 12.5
67/30 Amy .Barilet 38"-/29 28 6 25.5 8.5
3~t/3I ,~2~/29 4:11/28 8 2 7.5 2.5
To,at 411 118 396.8 132.3
Yi × Y,i Gm d reciprocal)
30/27 Union Jack 32/26 33 27 30 30
17/28 Me ~'Vhi~e Sgar 21 22 21.5 21.5
4:[/28 32/26 31 ~/27 28 27 27.5 27.5
7/30 White St,~r Ideal 31 33 32 32
11/30 2-"/28 32/26 23 16 19.5 19.5
13/30 Ideal 32/26 ,£ 4: ,l: 4
16/30 35/26 2~/28 20 17 18.5 18.5
23/30 Everest 2"-/28 15 22 18.5 18.5
24/30 2~"/28 10/29 34~ 34 34: 34
31/30 Colgness Gem Everesg 56 -50 57"5 57.5
56/30 Glenshee Wlfite S~ar 102 92 97 97
61/30 32/26 Union Jack 32 21 26.5 26.5
17/31. 27a/27 White S~ar 7 ]4 10.5 10.5
32/31 20/29 42e/28 10 10 10 10
33/31 42"/29 White S~ar 14~ 17 15.5 15.5
~t3/31 38~/29 White Star 58 57 57'5 57"5
To~al 488 ~72 480 480
* Differences ill reciprocal pollination have never been observed in regard ~o the four
eolom' factors.
"~ Probably due to ehromagid segreg~gion.

This difficnlty regarding the identity of the ground oolour has been
encountered in three of the families analysed in Table I (XIII, XVI and
XXXIX), but only in the last case is the constitution of the parent
(27"/27) for the factor Y doubtful. It will be noted that 279/27 has
purplish-crimson rays, a colour normally associated with a white or ivory
ground. 279/27 is known to have a yellowish ground from the occm'rence
of a mutant sector revealing the ground colour.
With the exception of these three families segregation of yellows and[
non-yellows is beyond dispute, the coincidence of heavy anthooyanin
aud pale yellow being rare in these experiments.
In families V and X X I X (Table I), the plants tabNated as "Yellows"
carry :factors for anthooyanln (in the flowers), bug for reasons presented
later (p. 184) the anthoeyanin is produced in the stems and leaves only
and not in the ray floregs. These" pseudo-yellows" therefore are included
with the cyanic forms iu column 7 of Table I. Table II summarises the
162 Genetics and Uhemist~'y of Flowe~' Coleus" in Dahlia
total results for inheritance of Y, which is completely dominant itt the
simplex condition.
Attention has previously been drawn (Lawrence, 1929, 1931) to the
appearance of reeessives in Dc&l'ic~,where none was expected on ]~{uller's
theory (].914.) t h a t four chromosomes mated and were distributed at
random in gamete formation. Nine such "unexpected" reeessives have
been recorded in these experiments an/[ in, each case the parent 14./26 was
directly involved..Reference to Table II will show t h a t 14./26 must be
triplex or quadriplex for Y. From indirect evidence it had hitherto been
assume/[ t h a i ' i t was triplex, an assumption which is now confirmed by
the fact t h a t 4.2"/29 (14./26 x 288/27) and 2"/28 (14-/26 x White Star) prove
to be simplex lot Y.
I t is probable t h a t these unexpected recessives arose from chromatid
segregation. Two of the recessives were fl:om 15~/28 × 14./26 (Y1 x Ya), the
other seven from 14./26 x Y4.
I t is noteworthy t h a t although 14-/26 has been used as a parent in five
different crosses with plants duplex for Y, yet no recessives have ap-
peared in a total of 4.4.4. individuals. On the random segregation of
chromatids the chances against a recessive appearing in the cross
Y a x Y 2 are 130: 1, whereas they are only 27 : 1 in the cross Ya xy~.
No evidence has been found for chromatid segregation in regard to
the factors A, B and I.

(b) Ivory.
When the tetrasomie inheritance of Y was first elucidated it was
thought t h a t I too would prove to be inherited in a similar manner. With
one exception, however, only 3 : I and 1 : 1 ratios were obtained from
crosses between ten different ivory and white individuals. The exception
was family 9/28 (Lawrence, 1929) which gave 5 ivory : 30 white, but the
issue was obscured in this ease by the discovery t h a t one of the two white
parents (viz. 35/26) occasionally sported ivory sectors and even whole
ray florets. The appearance of the ivory individuals in 9/28 was attributed
to the accidental use of a capitulum mosaic for ivory and white. Such
mutations are relatively common in ~he garden dahlia, and in this case
mutation of an inhibitor of I was considered responsible. Since no
tetrasomic ratios were fotmd, the inl/eritance of I was assumed to be
disomic and due to the allosyndetic pairing of four chromosomes.
Improved methods of testing for the presence of ivory flavone and the
requisite progenies now show t h a t the tetrasomie inheritance of I was
not observed owing to the fact tha~ simplea~ cts well ~ts n~dli2Iexforms ct~'e
 W. J. C. LAWr~ENOE AND I%OSE 8COTT-~VfONC]~IEFF 163
virtually ~vhite, i.e. the factor I is incompletely dominant, ivory pigment
being produced in quantity only when two dominant factors are present.
TABLE III.
Inheritance of the factor I.
Parents Observed Calculated
¢ h % r h ~ r-______A__~
Family -9- d' Ivm'y I'ghite Ivory White
Ia
I,i x
26/3o 10/29 26/28 13 0 13 0
I,l x 12 (and reeil?rocal)
24/30 2"128 10/29 34 0 34 0
56/30 Glenshee White Star 55 0 55 0
Total 89 0 89 0
Ia x I,~
18/30 2:~/28 White Star 8 0 7-3 0"7
~2/30 2~/28 Everest 10 1 10.l 0.9
i0/30 23/28 'Iii/28 8 0 7"3 0'7
54/30 2,~/28 31~/27 8 0 7"3 0'7
13/31 228/27 Everest~ 3 0 2.8 0.3
14/31 228/27 White S~ar 10 0 9.2 0-8
Total 47 i 44 4
Ia X II
9/30 2a/28 32/26 4 ,1 6 2
12 x 12
6/30 Everes~ White S~ar 10 0 7.5 2.5
7/30 White Star Ideal 27 6 24 8
23/30 Everest 2°"/28 17 5 16.5 5.5
33/30 411/28 2"/28 3 2 3.8 1.3
55/30 315/27 2"/28 12 0 9 3
48/31 30t/30 Everest 60 22 61.5 20.5
Total 129 35 123 41
Ie x 11 (and reciprocal)
10/28 32/26 Whi~e Star 16 15 15.5 15.5
40/28 35/26 229/27 5 4 4.5 4.5
4=1/28 32/26 316/27 12 i5 13.5 13.5
6/29 ) ~11/28 412/28 4 9 6.5 6.5
35/3O
5/30 Everest 32/26 9 3 6 6
11/30 22/28 32/26 8 8 8 8
13/30 Ideal 32/26 1 3 2 2
16/30 35/26 2"-/28 16 13 14.5 14.5
29/30 32126 14126 5 8 6.5 6.5
8/3I 41/29 Whi~e Star 34 33 33.5 33.5
9/31 4:4/29 ~Vhi~e S~ar 26 19 22.5 22.5
53/31 32/26 31"/30 23 31 27 27
55/31 Colbness Gem Everes~ 16 26 21 21
Total 175 187 181 I81
I i x Ii
9/28 35/26 32/26 5 30 8.8 26.3
10/3l 41/29 44/29 14 47 15'3 45'8
11/31 4~/29 4~/29 5 21 6-5 19-5
51/3]. 32/26 31i/30 8 31 9'8 29'3
Total 32 129 40'3 120'8
Simplex forms may develop a little ivory pigment, but this is always
much less in amount than in any normal ivory (I2_,l).
 164 Genetics and Che~nist,ry of Flower Colour in Dahlia
The calculated ratios resulting h:om the incomplete dominance of I are
given below. The remaining combinations give dominants only. It will be
nobiced that only one characteristic a,u~otebrasomie ratio occurs (11 : 1).
Ivory : White
i,~ x i a -- : ~1
I, x i. t -- : All
11 x 11 1 : 3
L x %, 1 : 5
xI 1 1 : I
I~ x I~ 3 : i
I a x i.~ l : l
I a xI~ 3 : i
Ia x Iz 11 : ]

The first evidence for the conclusions stated above was from :family
8/31 (4i/29 x White Star) and family 9/31. (44/29 x White Sta'r). 4~/29 and
4'1/29 are sister seedlings with magenta-colom:ed florets, and bhe majority
of their respective progenies also had magenta florets. Finned with
ammonia these magentas were found to turn a bright green if ivory
flavone was present, or a medium blue if it was gbsent. The reliability of
these colour reactions was further tested by the ethyl-acetate method to
be describe/[ later. In both of the families 8 and 9/31. approximately
equal numbers of ivory and white grounds were obtained, which on the
scheme already mentioned was taken to indicate t h a t one parent in each
family was simplex and the other duplex for I. " W h i t e Star" was already
known to have ivory florets, therefore both 41/29 and 44/29 were expected
to give a blue reaction with ammonia, and were found to do so.
On this assumption, if 41/29 (A211) and 4"~/29 (AzIi) were crossed to-
gether then the/~i should give 1 ivory : 3 white. The numbers actually
obtained were 14 ivory : 47 white. Seedling 4z/29 is evidently simplex
for I, since it too gives a 1 : 3 ratio when crossed with 44/29.
]~evision of families raised earlier now gave the constitution of other
parents (TM)le III). For example, 32/26 (white)x }Vhite Star (I~) gave
16 ivories: 15 white individuMs, thereby showing 32/26 to be simplex
for I, and on crossing this white seedling to a purple seedling (31/30)
without flavone, a 1 : 3 ratio was again obtMned according to eN?eetation.
It now seems unnecessary to attribute the ivory flowers obtained in
family 9/28 to the use of ~ mutant ~nd mosMc ivory capituhtm, since the
ratio of 5 ivory : 30 white is a fMr approximation to expectation on the
tetrasomic basis (i.e. 8.8 : 26.3). Further, the explanation of the m u t a n t
ivory sectors in 35/26 can now be brought into line with other colour
m.ntations in Da,hgG as arising from irregular mitoses in a plant simplex
for I so that two I-chromosomes are distributed to one ceil and its lineal
descendants.
 W. J. C. LAWRENCE AND ~,OSE SOOTT-MONORIEFF 165
Except when anthocyanin pigmentation is very light, it is usually
possible to distinguish at sight magentas with ivory grounds from those
with white grounds, the respective flower colours being bluish-magenta
and[ rosy-magenta. Similarly iu deeply pigmented[ flowers with uon-
yellow grounds those forms urith ivory flavone are some shade of purple
(i.e. reddish-b/us), whereas when flavone is absent the eolour is a purplish-
crimson to "chocolate" (i.e. bluish-~'ed). In other words the p,ress'nce, of
ivo~'y flavone ha.s el, bh~eing effect ufoon a'nthocyc~nin flower colou~' (see
Plate VII, figs. 1-4). Prof. and Mrs Robinson's (1931a,~1932 a) very
interesting discovery of the existence in many flowers of "co-pigments"
which have a surprisingly big effect in modifying flower colour threw light
upon this observation, it being apparent that the ivory flavone in Dahlia'
is acting as such a substance (Lawrence, 11932). By taking advantage of
the l~obinsons' observation that extracts of co-pigments will have a
blueing effect upon un-co-pigmented anthocyanin solutions in sit~'o, it
has been possible to estimate the amount of ivory flavone present not
only in flowers containing this pigment Mone, but also in those con-
raining yellow flavone and anthocyanin.
The blueing effect of ivory flavone is seen on turning to the records of
families (1) 8/28, and (2) 4- and 5/29, where it was found that there were
(1) 36 bluish: 3 rosy-magentas (expectation 35.8:3.3), and (2) 53:10
(expectatioi~ 4.7.3 : 15.8) respectively. These resuRs clearly show 36/26
to be triplex for I. Similarly 32/26 (white)x Union Jack (crimson-
scarlet) gave 26 purplish forms in the proportion 11 pm~plish-crimson :
3 crimson-purple:9 purple, thereby indicating that Union Jack is
probably duplex for I. That it is at least duplex for I is certahl from the
fact that loss mutations of the yellow flavone in this variety result in the
l)u~'ple flower colour associated with co-pigmentation.
In these experiments every hldividnal scored for ivory or white has
been ~ested with ammonia or by the ethyl acetate or co-pigment method.
Parents with anthoeyanin on ivory or white grounds have all given the
reactions expected from a knowledge of their constitution for I as shown
by the breeding experiments (see Table III).
No nulliplex forms have been used, the eight parents wRhout ivory
pigment all proving to be simplex for I. Nevertheless, nulliplex forms
must occm', since the ratios obtained show no deficiency in the white
class as would be exl~eeted if reeessives were appreciably less viable than
dominants. Fm'ther breeding work is being done to confirm these results
for the inheritance of i.
To a certain extent the factor I is cumulative in its effect on file
166 Genetics and Chemistry of Flower Colour in Dahlia
p h e n o t y p e . Nulliplcx forms are almost certainly devoid of flavone,
simplex forms usually h a v e a little, a n d duplex, triplex and quadriplex
individuals all p r o d u c e flavone in q u a n t i t y . The evidence suggests t h a t
the s a t u r a t i o n point is r e a c h e d in t h e duplex condition.
Certain of the simplex individuals h a v e been f m m d to be devoid of
flavone, and more t h a n one grade of i n t e n s i t y has been observed in
duplex forms. I n view of this it seems probable t h a t an inhibitor of i v o r y
flavone c u m u l a t i v e l y suppresses this pigment.

TABLE IV.
Inheritauce of the factor A.
P~ren~s Observed Calculated
I,'amfly 9 c~ A a A a

A~? x a.~ (and reciprocal)

4/29 36/26 32/26 4o o ~10 o
5/20 36/26 35/26 23 0 23 0
8/28 36/26 White Star 39 0 39 0
4/28 14[26 36/26 78 0 78 0
Tottd 1.80 0 180 0
A. x A 2
10/31 4'~/29 41/29 59 1 58-3 1.7
11/31 42/29 44/29 26 0 25.3 0.7
Total 85 1 83.6 2.4
A 2 x a a (~nd reciprocal)
17/29 14/26 27~/27 21 3 20 4
18[29 14/26 273/27 30 5 29.2 5"8
8/31 4~/29 White St~r 54 13 55.8 11.2
9/31 46/29 White Star 39 6 37.5 7.5
t7/31 276/27 White St~r 15 6 17.5 3.5
19/31 274/27 White Star 27 4 25.8 5'2
22[31 27n/27 Everest 35 7 35 7
Total 221 44 220.8 44.2
A 1 × a 4
56-59/30 Glenshee White St~r 90 104 97 97
32/31 20/29 412/28 9 11 10 10
Total 99 115 107 107

(ii) k ~ e ~ o o Y A m N .
(a) "Light" anthocyanin pigmentation: the A factor,
As previously s h o w n (Lawrence, 1931) two i n d e p e n d e n t factors,
A and B, control the p r o d u c t i o n of a n t h o c y a n i n in the garden dahlia.
A governs light p i g m e n t a t i o n , B heavy. D a t a are presented in Tables I
a n d I V which clearly establish the tetrasomie n a t u r e of A.
The expression of A is cumulative in m o d e r a t e degree. T h u s t h e
extremes of intensity a m o n g the r o s y - m a g e n t a s in families V I I I and I X
(Table I, A 2 x A2) were n o t far removed, a l t h o u g h simplex, duplex a n d
ti'iplex forms m u s t h a v e c o m m o n l y occurred. The deepest types in these
 W. J. C. LAWIRENCE AND R, OSE SCOTT-MONCI~IEFF 167
f a m i l i e s w e r e m u c h p a l e r ~ha.n t h e p i g m e n f a t i o n p r o d u c e d b y t h e f a c t o r
B , a n d i t is u s u M ~o find t h i s d i s t i n c f i o n q u i t e c l e a r l y e x p r e s s e d .

T A B L E V.
Pla.nts scored Jbr anthoeya,nin in seedling stage.
Parents Observed CMculated Compare witch
c- ~ u - - , r~ A , r--~--~ Table I
Family ~ d~ Cyanic Acyanic Cyanic Acyanio ]~ef. nos.
12/31 20/29 Evcres~ 20 34 27 27 XLI
15/31 27~/27 411/28 37 3 33.3 6.7 XXXI, XXXVI
20/31 27'1/27 Everest 27 4 25.8 5.2 XXXIV
21/3 l 27'~/27 41~/28 17 3 16.7 3.3 XXXIV
23/31 27,~/27 4tl/28 18 4 18.3 3.7 XXXV, XXXVt
39/31 Everest 601/30 35 8 35.8 7.2 XL
40/31 Everest 60:/30 4.8 6 45 9 XL
41/31 Everest 603/30 66 l.O 63"3 12.7 XL
42/31 Wlfite Star 60~/30 7 2 7'5 l'5 XL
45/31 38~/29 Everes~ 57 15 60 12 XIX, XXII
46/31 38~/29 411/28 6 1 5'8 1'2 XIX, X X I I
54/31 ~rhite St~r 313/30 16 3 15.8 3.2 XXIif, XXV

T A B L E VI.
Iqzheritance of the factor 13,
Parents Observed OMeul~ted
*

Family 9 6' ]3 b ]3 b
138 or ]3,i × ]32
66/30 Amy ]3~rflet 38i/29 38 0 38 0
67/30 ±buy ]3arileb 382/29 34 0 34 0
Total 72 0 72 0
B a or 13.I x b I (and reoiproeM)
38/29 22"/27 Amy Barilet 92 0 92 0
] 3 2 × b 4 ( a n d reciproca, l)
30/30 38i/29 Everesb 70 16 71.7 14.3
68/30 34/36 381/29 27 3 25 5
43/31 382/29 White Star 99 16 95.8 19.2
53/31 32/26 312/30 41 9 41.7 8.3
Total 237 44 234.2 46.8
B i x b,i (~nd reoiproeM)
-- M5 14/26 123 123 123 123
3/29 151/28 14/26 24 33 28.5 28-5
33/31 42"-/29 ~Vhit e St~r 16 15 15-5 15'5
34/31 42~"/29 411/28 7 3 5 g
35/31 391/28 411/28 15 12 13.5 13"5
37/31 421/29 Everest 37 37 37 37
38/31 421/29 41.1/28 29 29 29 29
48/31 301/30 Everest 37 46 ,11-5 41'5
51/31 32/26 311/30 21 16 18-5 18.5
Total 309 314: 3] 1.5 311.5

As will be shown later, however, the interaction of flavone and an.tho-

eyanin factors may modify fhe intensify and quality of fl.ower colour. For
f h i s r e a s o n t h e figures given in T a b l e s I V a n d V I a r e h ' o m crosses i n
w h i c h e i t h e r o n e or t h e o t h e r of t h e t w o f a c t o r s A a n d B is i n v o l v e d , so
168 Genetics and Chemistry of Flowe+' Colour in Dahlia
that no doubt should remain as to their identity. From Table IV, 36/26
could be triplex or quadriplex for A. It is probably triplex, since mutant
acyanic sectors have been observed in its progeny from crossing with
nulliplex forms, and these loss mutations have been proved in Dahlia
nearly always to occur in forms simplex for an anthocyanin :factor.
Plants which have anthocyanin in their ray florcts always have it in
their foliage and stems in addition. Hence, unless the pigmentation is
very faint, the inheritance of anthocyanin can be scored in the seedling
stage.
In Table V are given the results of scoring cyanic pigmentation in
seedling families. With the exception of the first family all faintly tinged
and green-stemmed individuals were grown on. to maturity to ensnre
certainty in recording the ratio of pigmented to non-pigmented forms.
The excess of recessives in 12/31 is almost certainly due to the inclusion of
faintly tinged (flowers) forms in this class, since comparison with
family XLI (Table I) shows that these are commonly derived front
20/0.9.
I~eference to Table Iwill show that the results from the remainder of
the crosses in Table V are confirmatory. The figures obtaflled from these
seedling families are not included in the total results in Tables IV and VI.

(b) "Heavy" anthoeyanin pigmentation: the B factor.

The results from the inheritance of the B anthocyanin factor are
presented in Table VI. Like the factor Y, B is completely dominant in
the simplex condition, and no difference of intensity can be detected
between the dominant forms. It was thought at first (Lawrence, 1931)
that B was cumulative in its phenotypic effect. Differences were found
which seemed to be correlated wit]] the number of B factors; but recent
work has shown that these differences are dne to the interaction of the
factors for flavone and anthocyanin.
The results for the inheritance of the four factors Y, I, A and B are
summarised in Table VII.

(iii) INTERACTION OF THE :FLOWEI{-OOLOUI¢ FACTORS.

(a) Interaction of Y and I.

Although the inheritance of the colour factors in Dahlia is inde-
pendent, this is not the case as regards the production of the pigments
which the factors control, increase of one pigment being accompanied by
decrease of another.
The interactions are quantitative, two types being found, (1) between
 W . g. C. LAWRENCE AND :~0S:B ~COTT-MONCR,:[EFF 169
the flavones, (2) between the flavones and anthoeyanins. The interaction
of (1) cannot be detected on casual :inspection as in the case of (2), but
will be dealt with first in order to ]laVe a surer basis for tile discussion of
the second type.
The detection and estinaation of ivory flavone in aeyanie flowers
depended upon the presence and degree of the blueing co-pigment effect
by the flower extracts upon a standard cyauin solution (see p. 189). Since
TABLE VII.
Tet.rasomic inhe.rita.'~ce i'~ tM octoploid Dahlia variabilis.
N,i,ngle:factor ,ratios.
Observed CMculaged
N o . of , - - ~ - - - - - - ~ , *
Combination families Dora. : Recessive Dora. : l%eeessive Derivation ~Dov,/g
A a x a, t ,t 180 :, 0 180 : 0
A~ x A~ 2 85 : i 83.6 2.4 - 1.4 0.9
A 2xaa 7 221 : 44 220.8 : 44.2 - 0.2 0.03
A1 xa4 2 99 : 115 107 : 107 + 8 1.1
a 4 x a 4 47 0 : 2327 0 2327
::Bs or ]3 a x B ~ 2 72 : 0 72 : 0
B S o r B , t x :b4 i (32 : 0 92 : 0
]3 2 x:b~ ,4 237 : 44 234-2 46.8 - 2.8 0.5
B j . x:b,~ 9 309 : 314 311"5 : . 311..5 + 2.5 0.2
b,~ xb,~ 46 0 : 2004 0 : 2004
Y~ xY~ 5 444 : 0 444: : 0
Y3 xYt 3 259 : 0 259 : 0
Y~xy4 ,i 378 : 7* 385 : 0
Y~ x Y~ I 107 : 3 106-9 3.1 - 0.1 0.03
Y~ xY1 5 513 : 50 516 : 46"9 + 3 0"47
Ys x y,~ 2l 708 : 160 730"8 146"2 +22.8 2-1
Y1 x YI 7 411 : 118 396"8 : i32"3 -14-2 1-4:
YI xY,l 16 488 : 472 480 : 480 - 8 0-5
Y,~ x Y4 14 0 : 558 0 : 558
I,~xI~ 1 13 : 0 13 : 0
I a x Is 2 89 : 0 89 : 0
Is x Is 6 47 : 1 44 : 4 - 3 1.6
Ia x 11 i 4: 4 6 : 2 + 2 1.0
12 x I s 6 129 : 35 123 : 41 - 6 i.I
13 175 : 187 181 : 181 + 6 0'6
11 x 11 4 32 : 129 40,2 120.8 + 8.3 1.5
* l l e e e s M v e s prob~flJly ~ r i s i n g f r m n o h r o m ~ t i d s e g r e g a t i o n .

yellow flavone exerts no such effect, it is possible by this method to

eliminate the masking of ivory pigment in ),allow forms.
While white forms gave no reaction, ivory extracts turned the reddish
anthocyanin solution to a bluish-red, deep ivory individuals having an
even stronger effect than pale ivory.
Apart from a slight dtange towa,rds orange tile addition of yellow
flavone extract to the standard solution had no appreciable effect, but if
the extract also contained ivory flavone the blueing reaction occurred as
with pure ivory forms (see Table VIII).
170 Genetics and Chemistry of Flower Colour in Dahlia
Various yellow-flowered varieties whose constitution for Y and I was
known from breeding experiments were tested, along with other seedlings
which had given mutant sectors reveMing white and ivory grounds. The
results are given in Table VIII.
As expected, 61~/28 gave a completely negative reaction. Each of the
rem Mning seven plants was expected to give a definite positive reaction,
since six of them were duplex and one triplex :for I. As will be seen,
however, four of the 12 plants had only a trace of ivory at the most.
Further, the yellow pigment was suppressed in both of the two individuals
giving good positive reacgions. It is especially noteworthy tlhat the only
one of the four YII~ plants giving a positive reaction had c~'eamflowers.
"IdeM" apart from breeding results is known to be at least duplex for I,
TABLE VIII.
Yellow-floweredplants testedfor ivoryflavone content.
Flower colour Constitution Ivory flavone reaction
41"/28 Yellow ¥111 Nfl
411/28 Yellow YlI2 Trace
3 i~/27 Yellow YII2 Truce
IdeM Yellow YII~ Practically nil
22/28 Cream Y212 Positive +
229/27 Yellow Y2L. Practically nil
2a/28 I)rimrose Y~Ia Positive
14/26 Yellow YaI2 -Weak posigivo
291/30 Yellow White sectors Nil
29:/30 I)rimrose White sectors ~Practically nil
29a/30 Yellow Whi~e sectors Practically nil
295/30 Yellow Whi~e sectors Nil
]3 32 a Yellow Wlfi~e sectors Nil
13 32 ° PMe yellow Ivory sectors Positive + +
A 221° Yellow Ivory sector 1)osigive
]3 g:3G Cream Ivory sector Positive +

loss mutation of yellow flavone having shown that this variety produces
ivory in the absence of yellow flavone. The results from testing the
mutating seedlings are in agreement with the observations on the
mutant sectors.
As previously reported (Lawrence, 1929, p. 142; 1931, p. 263) there
is ample evidence to show that the diminution of yellow as observed in
cream and primrose forms is not due to the presence of ivory but to the
action of the inhibitor H. In view of this, the results of these preliminary
tests on the parents listed in Table VIII suggested that the p~'oduction of
ydlow flavone i~ quantity was at ~he expense of ivory flavone.
A series of yellow-flowered seedlings was next tested in order to
determine whether yellow flavone in quantity masked or inhibited the
blueing reaction. The results are presented in Table IX. From this gable
we see that one deep and four moderately deep (i.e. good) ydlows in
 W. J. C. L2~WIgENOE AND I~OSE SCOTT-IVioNcI%IIflF]~ ~ 171

family 37/31 gave positive reactions. At the other extreme, while none
of the five creams in Table IX was iouud to be without ivory, three
primrose forms gave negative results. That cream forms can occur
without ivory flavone being present is quite certain from numerous tests
made in 1930. In certain families the inhibition of yellow was extreme
(Lawrence, 1931, p. 262), as much as half of the petals being white, a,nd
on fuming with ammonia the reactions showed t h a t no ivory flavone
was present, i~![uta~lt sectors similarly fumed also gave negative results.
Others of these creams gave positive results for the presence of ivory.
We see, then, t h a t ivory may occur with any degree of yellow, and the
TABLE IX.
Reactions for l)resence of ivory,flavone i'~ yellow-flowered seedlings.
l~eacgion
To~al
t>osi- ]?osi- ]?rac- g -J'-- %

live t i r e Posi- tically Posi-

Family Parents Ground colour + -F -F give Trace nil Nil give Trace Nil
43/31 38'-'/29 (B~.YJ.) h{edium + yellow I 1
x W h i t e Star N e d i u m yellow 2 1
(I.,) N e d i u m - yellow i I i I 4
37/31 ,~2~/29(B~¥~I~) Deep yellow 1 i 4
x E v e r e s t (I~) Miedium + yellow 4 2
l~iedium yellow I i 2 l
N e d i m n - yellow 2 i I
Primrose yeltow 1 i i1 I6
4/31 Ideal (Yile) Medium yellow 2
x White Star Prhm'ose 8 i
(I~) Pate primrose i 1
Cream I8
40/31 E v e r e s t (L) Deep yellow 3
x 60'/30 Pale yellow I
(BAYI)
42/31 ~'Vliite Star (I~) Cream 1 l
x 60'1/30)
(BAYI)
blueing reaction can be observed even when ivory occurs with deep
yellow flavone.
The results given by the eight inedimn yellows in family 43/31 are
particularly interesting. The aeyanic forms in this family comprised eight
yellows and eight ivories. A ratio of 58 Y: 57y showed t h a t the yellows
must be simplex for Y. The cyanic forms with non-yellow grounds were
scored as 11 blmsh-purple, 22 pm:ple and 16 crimson-purple (see Table I,
family X I X ) . One parent was duplex for I, so t h a t the possible propor-
tions of ivory to white were I : 5, i : 1, 3 : 1 or 11 : I according to the
constitution of the other parent, 38"'/29. As will be seen the results indi-
cated t h a t 38a/29 was duplex for I, since 11 : 22 : 16 is a fair approxima-
tion to expectation (i.e. 12.3 : 2~.5 : 12.3) for I~ x 12.
J o u r n . of Genetics x x x I2
172 Genetics and Chemist~'y of Flower Coleus" in Dahlia
011 testing the eight yellows only one was found to give a positive
reaction (weak), whereas the other eight acyanie individuals were all
ivories giving strong reactions. Thus, instead of getting a ratio of
7.2 Yellow+Ivory : 2.~ Yellow : 7-2 Ivory : 2 4 White, the actual num-
bers observed were 1 : 7 : 8 : 0 (X°"=19.3, P = l e s s than 0.01)L The
cMculation for independence of yellow and ivory gives X~= 13.0 with P
agai~ less than 0.01.
These figures show that it is highly iml)robaJ)]e that the results re-
corded above would be obtMned i[ there were no interaction between N~e
yellow and ivory flavone factors. It should be noted that none of N1e
yellows was deep. In four of them a trace of blueing was observed in the
colonr reaction test, and it seems possible ~hat these four plants might
have been duplex for I, but that interaction had resulted in the sup-
pression of ivory.
I n the next family examined (37-31) the ratio of 62Y : 12y showed
one parent (421/29) to be duplex for Y. The constitution of the other was
I~. Out of eight acyanie forms in the progeny without yellow flavone six
were ivory and two white. Twenty-seven of the yellow-flowered plants
were tested for the presence of ivory, eleven giving positive and sixteen
negative results, which means that 421/29 must be at least simplex for I.
On this assumption the values obtained for goodness of fit are %2=,1.62,
and P=0.20. We have seen that the Y~I2 forms are apparently on the
border line with respect to the production of ivory flavone and Y~I2
individuals produce none whatever. Applying these results to an analysis
of family 37/31 we find that the expected proportions of yellows giving
positive and negative reactions for ivory are 20.25 positive : 6.75 negative
assuming Y~I~ is without ivory flavone, or 11.25 positive : 15.75 negative
if Y~I~ produces ivory flavone. The actual numbers were 11 positive : 16
negative.
In family 37/31 ~herefore ~here is no appreciable suppression of ivory
in YII~ forms. Compared with 43/31 this difference is almost certMnly
due to the action of the yellow inhibitor I-I, since examination of the two
families in their entirety shows that there is a wider variation in the
intensity of yellow in 37/31--partiM suppression of yellow permitting a
greater production of ivory flavone.
This conclusion is supported by the evidence from :family ~1/31
(Table IX) in which no suppression of ivory flavone is apparent. In this
family the intensity of the yellow flavone is distinctly lower than in the
1 The constitution of 38"'/29 tbr I m'usl be duplex or simplex. E v e n if ig were simplex
t h e n ~:" w o u l d equal 13.0 a n d P less t h a n 0.01.
 W . J. C. LAWRENCE AND I~,OSE SCOTT-1V]]ONCIAIEFF 173
other two families. The figures for the inheritance of I, viz. 27 ivory : 6
white, probably indicate a 3 : 1 ratio, in which case "IdeM" is duplex
for I as postulated from other data. We may note in passing that when
the two groups of yellows (Table IX) giving positive and negative re-
actions are compare([ for average intensity of yellow, taking the value
of cream as 1 and deep yellow as 5, the figm:es obtMned are 2.8 for the
positive and 4-.5 for the negative group, i.e. the paler the yellow tile more
ivory pigment is present.
Considering these results together the following points emerge:
(1) From Table VIII it appears that only a trace of ivory flavone is
produced in yellow-flowered plants of tile constitution YIIa, and practic-
ally none at all in a Y,I~ individual. If these examples are characteristic
then it seems that the et~ect of Y on the action of I is decidedly pro-
nmmced.
(2) [['he eight yellows in family 43/81 were simplex for Y. Two of
them should have been Ia, foul: 12 and two I i. The ratio therefore of
1 positive : 4 trace : 3 nil in the reactions for ivory supports the evidence
from (1) that some YiI2 plants develop a trace of ivory only.
(3) In families with pale yellow flowers (due to the action of the
yellow-inhibitor I-I) Iz forms give the normM positive reaction for the
presence of ivory flavone.
(4) Ivory pigment occurs more frequently in the paler than hi the
deeper yellow flowers.
We see therefore that though further study of the interaction of I and
Y is desirable, it has yet been shown that (a) some yellow-flowered plants
duplex for I do not produce any ivory flavone at all, or only in slight
degree, and (b) that the production of ivory flavone approximates go
normal when the amount of yellow flavone is considerably diminished.
(b) I%te~'acgon of A and I.
Qbservations on interaction in the production o f " l i g h t " anthocyanin
(i.e. the product of the factor A) and ivory flavone, can be made with
facility owing to the blueing effect of ivory on anthoeyanin in ~he ray
florets, and (2) because A is cumNative in its expression, increase or
decrease in the amount of anthocyanin being directly observed (see
Plate VII, fig. 1).
A magenta seedling, 36/26, was crossed with ivory and white forms
and was found to give no acyanic individuals in the progenies. Three
deep rosy-magenta seedlings, 41/29, ~2/29 and ~1'~/29, were selected from
.Fi and were crossed with an ivory variety and with each other. The
12-2
174 Genetics and Chemistry of l~lower Colour i~ Dahlia
results (Tables I, III and IV) showed that the three magentas were each
duplex for A and simplex for I. Tests with ammonia had shown that
A individuals without ivory flavone were rosy-magenta, whereas the
flower colour was bluish-magenta when ivory was present. The F2
families (8 and 9/31) were accordingly divided into rosy- and bluish-
magenta groups and the groups analysed and compared for intensity of
anthocyanin coloration. The results are given in Table X. The division
into pale, medium and deep groups, etc., is mainly arbitrary. As will be
seen from Text-fig. 2, where the results are set out graphically, and in
Plate VII, fig. 1, there is a considerable decrease i~ the intensity of
anthocyanin in the bhfish-magenta group of individuals with ivory
grounds.

TABLE X.
Showing variation in anthocyanin intensity (a,) in the absence, (b) in the
presence of ivory flavone.
/~i, Cyanie intensity
Magenta f h
Family Parents eolour Pale B{edinm D e e p
8/31 41/29(magenta) x White Star (ivory (a) rosy 11 9 8
(A2.Ii) (I2) (b) bluish 9 8 9
9/31 44/29(magenta) x Whi~e Star (ivory) (a) rosy 6 8 5
(A~I~) (I,,) (b) bluish 5 7 8
Anthocyanin intensity
grades, pale to deep

10/31 4'1/29(magenta) x 4i/29 (magenta) (a) rosy 4 5 14: 17 5 2

(~I~) (/L2I~) (b) bluish 3 5 5
11/31 4"/29 (magenta) x 44/29 (magenta) (a) rosy 3 9 5 3 1 0
(AaI1) (A2Ii) (b) bluish 2 i 2

In contrast with these figures are the results from 4i/29 x 4~/29 and
42/29 x 44/29. The intensity of anthooyanin in these two families was
noticeably deeper than in 8 and 9/31, owing, undoubtedly, to the greater
number of dominant factors and the small number of forms with ivory
grounds. It is noteworthy that the cumulative effect of A is much more
pronounced in families 10 and 11/31 in the absence of I, than in families 8
and 9/31 with an inverse proportion of A and I factors (Table X and
Texbfig. 2).
Considering these results together, we see that anth,ocyc~nin ~ntensity
is diminished when @cry flavone is present, a diminution which without
doubt corresponds with an actual decrease in the amount of pigment.
Since in these families the ratios of ivory to white and cyanic to aoyanic
are iu good agreemenb with expectation, we may also conclude that the
interaction between the factors for these pigments in the majority of the
 W. J. C. LA~rI~ENCE AND P~OSE SCOTT-MONCIaIEFF 175
/~1 g e n o t y p e s is n o t e x t r e m e e n o u g h t o s u p p r e s s e n t i r e l y t h e f o r m a t i o n of
either.
l~osy magenta (A.~ll)x Ivory (I2)
46 Bluish magentas = 47 :gosy magentas . . . . . .
I%osy magenta (A2Ii) x Rosy magenta (A~Ii)
18 Bluish magentas . . . . . . . . . 68 igosy magentas . . . . . . . .
25

'20 / ,
r l..~-. /
t5 " "-
'~ \\ \
I0 \ \

®
~o 5
/
I/
/ ........... ?
/
t

,
\
,,\
X,.
, •\
m~O
, ¢:/ , ,/ , \ ' -..

I 2 3 4 5 6 7 8 9 10 II
Anthoeyanin intensity grades, pale --~ deep
Text-fig. 2. Showing variation in cyanic intensity in relation to the presence
or absence of ivory flavone.

TABLE XI.
£'howing interaction of factors for flavone and light anthocya,nin pigmentation
(see text). (a) = ingivic~uals with ivory grounds, (b) those with yellow grounds.
Observed Calculated
fro c 2,,__ _.~ [ A__~
Constitution pale FMntly Faintly
~ef. Family ¢ z ~ No. of cyanic Flavone Full tinged Full tinged
no. no. Parents ~ c~ plants forms index coloured or nil coloured or nil
I 10/31 4't/29x 41/29 A,I 1 ~Ii 60 5 0'25 58 2 60 0
(magenta x magenta)
II 11/31 42/29 ×4'1/29 AaI 1 A~I1 26 12 0.25 26 0 26 0
(magenta × magenta)
III 8/31 41/29 × White Star /k~I1 12 54 17 0.58 51-53 1-3 51 3
(magenta × ivory)
IV 9/31 4'V29 x White Star /k~I1 i. 39 13 0'58 34-38 1-5 37'8 2'2
(magenta × ivory)
V 4/29 36/26 x 32/26 AaI ~ I~ 40 33 1 34 6 30 10
(magenta × white)
VI 5/29 36/26 × 35/26 AaI a I~ 23 26 1 21 2 17.4 5'6
(magenta × wtfite)
VII 8/28 36/26 × Whi~e Star AaI ~ I,a 39 44 1.5 28 11 27.6 11.~
(magenQ~ × ivory)
\ r l l I 56/30 Glenshee × White Star Y1AII,I I~ (a) 36 69 2 11 25 5 30
(apricob xivory) (b) 54 100 2.5 1 53 0 54
The interaction of A and I may be examined further by comparing
the proportions of plants above and below a fixed degree of intensity in
eight families derived from parents differing in constitution as regards
the nmnber of A and I factors (Table XI).
176 Genetics and Che~rdd'ry q[ Flowe~' Coleus" in Dahlia
The results strikingly confirm the effect of interaction between I and
A. First, the proportion of pate forms is seen to increase Ft~'i 'Ftssu with
the nmnber of I factors (see column 7). Secondly, b y making the assump-
tion t h a t each I factor (after the first) potenticdly governs the production
of equal and additional amounts of ~avone, we can predict the average
cyanic intensity of a given family, as :follows. If we assign the vNues 1, 2
and 3 to Ie, I a and I~ respectively, we can calculate the average amount of
flavone per fatuity for any given cross. This value, referred[ to as the
"flavone index," is given in colmnn 8 of Table XI. Plotting the flavone

I00 -

m
8o
;s X

.v 60
b
o9
c8

~8 40

X X

X
I
0-5 I'0 1"5 2.0 2"5
l~lavone index
Textfig. 3. Showingthe inverse correlation begweenthe production of ivory flavone
and light anthoeyanin in the eight families given in Table XI.

indices against the percentage of cyanic forms of low intensity, the points
so obtained form approximately a straight line (Text-fig. 3), i.e. in the
A cyctnics there is ctppctrently an inverse correlcttion between the production
of ivory flc~vone c~nd ctnthocyc~nin.
Six individuals in family VII (Table XI) were entirely devoid of
anthoeyanin in the flowers, although their stems were tinged. On the
assumption that this is due to a certain balance of I over A factors we
m a y a t t e m p t to establish the precise nature of this balance. 86/26 is
probably triplex for A (see p. 168), therefore o~ crossing to White Star the
progeny would be duplex or simplex for A. I~orms qugdriplex for 1
should be most e:l~eetive in suppressing anthocyanin formation, but it is
 W. J. C. LIW:[%ENCE AND I%OSE SCOTT-~V[ONCIIIEFF 177
hardly probable that the six seedlings referred to were all 1,t, since only
1 in 12 of the progeny would have had this constitution. The actual
intensities scored for this family were 3 deep, 19 medium, 6 pale, 5 very
faintly tinged and 6 with no anthocyanin. The medium and pale classes
graded insensibly, but the other classes were fMrly distinct. Summing the
last two classes there are 11 seedlings with very little or no anthoeyanin.
The expected proportions of top dominants are J:f (A~I,~), s~ (A2Ia),
JT (AlI,~) and XT ~ (Alia). I-llence to fit expectation we must assume
that little or no anthocyanin is formed in the first class and the last two
classes, ~.e. in a family of 39 individuMs l l - t would be expected to be
without anthoeyanin. We may no~e in passing that it is probably signifi-
cant that the number of pale individuals recorded was six, since the
nmnber of A~!s forms expected (i.e. the next highest class in regard to
I factors) is 5.
If the above analysis is approximately correct then very little or no
anthocyanin will be produced in A~I4, AIIa and A~I,~ forms, all A~
individuMs with fewer I factors producing a variable but moderate
amount of anthocyanin.
This hypothesis can be tested by analysis of the other families in
Table XI. The results are presented in the last four columns of the table.
The yellow-ground forms in family VIII were praeticaIly uniform in
intensity of yellow, so that little or no variation in anthoeyanin produc-
tion can be attributed to variation in the amount of yellow. The occur-
rence of two faintly tinged forms in family I (Table XI) suggests that
factors other than the colour factors may sometimes modify anthoeyanin
intensity. Notwithstanding, the agreement between observation and
expectation in the eight families is sm'prisingly good considering the
many variables which might affect the issue.
If the scheme outlined above is correct, the important point emerges
tha~, in their governance of pigment production, the interaction of the
I and A factors is reciprocal bug ~mequal, the ei~ect of I on A being more
pronounced than that of A on I. Also we have seen that an A~Ia form
has more anthocyanin than an AIIa form.
A provisional classification of the grades of anthocyanin intensity
arising from interaction between A and I is given below.
Little or no Anthocyanin
atighocyoolfin r x
A:~I,~? /~.tiIi?
Alls AIL " A~I a Aal ~ A.:Ia
All,i All i A~I 2 A~I. A,~I,,
i%~l.i A i i~l i A~I i A.ili
178 Genetics and Chemistry qf Flower Uolour in Dahlia
The data for the A i and A~ types ~re from the results presented in the
previous pages. Since 36/26 (Aala) is a medimn magenta we may suppose
that all A a and A,i forms with three or less I :factors will also develop a
:fair alnotmt of anthoeyanin. This view is supported by the evidence from
the four tetraploid Dc~hlia species, each presmnably with four dominant
factors :for flavone attd four for anthocyanin. In these species the great
majori V oi the individuals are of intermediate intensity in their respec-
tive light and heavy classes. If, however, the range of cyanic intensities
found in D. variabilis is taken as the scale of reference, the predominating
cyanic intensities of the species are certainly nearer the pale than the

TABLE XlI.
Heavy cyanic plants tested for ivory flavone. (a) cyanies teste[l by ethyl
acetate method, (b) aeyanies tested with ammonia.
Cotour reactions
A
h
Parents Very Expectation
Family ( "t pale r--~'--~
c~ Yellow yellow Nil Ivory White
51/31 82/26 (white) 311/30 (deep put- (eL) 5 -- 18 -- - -

I1 plish crimson) (b) 3 -- 13 -- --

Bili
Total 8 -- 31 9.8 29.3
53/31 32/26 (white) 31~"/30 (purplish) (a,) 20 3 22 -- --
Ii B212 (b) 3 - - 6 - - - -

Total 23 3 28 25.5 25.5

48/31 301/30 (p~le purple} Everest (ivory) (a) 27 3 7 -- --
B~L 12 (b) 33 - - 1 3 - - - -

Total 60 3 20 61.5 20.5

55/31 Coltness Gem Everest (ivory) 22 16 21.1 21.1
(scarle~ crimson) I.
B~.A2YIli

deeper end of their respective scales. From this observation it seems that
forms with equal numbers of not only A and I (excepting the Aili forms)
but also B and Y factors are usually of medium intensity, and that inter-
action between the flavones and anthocyanins is in each class to the
advantage of the flavones.

(o) Interaction o r b and I.

In addition to many individual forms, foul: families with heavy
anthocyanin on non-yellow grounds were tested for ivory flavone. The
results from the four families are given in Table XlI. As will be seen, the
ratios obtained are in good agreement with the postulated constitutions
of the parents. Apparently tlde interaction of B and I on B is not strong
 W. J. C. LAWI:tENCE AND I-%O8E SCOTT-I~ONCI~IEFF 179
enough to disturb the ratio of dominants to recessives in the families
examined, i.e. in any combination from BiIi to BeIa.
It is well known that the cyanic flowm: colours in the garden dahlia
are often pater towards the end of the season. Apparently this is not
associated with, or confined to, any particular colour, since we have
observed it in the case of the yellow flavone also. It would be difficult to
detect in ~he case of the ivory flavone, and we have no evidence that it
occurs here. It is not very evident in the light cyanic (A) forms, but is
conspicuous in many heavy cyanic varieties.
In the four families just mentioned a number of individuals were
observed ~o'pro&me less anthocyaniu late in the season, a change usually
associated with an increase in the blueness of the flowers. In certain
individuals the change in colour was sufffeient to turn a crimson-purple
to purple or even (rarely) to bluish-purple. We have already shown that
blueness is mainly determined by the amount of ivory flavone, and it
seems reasonable to suppose that we may ascribe the colour changes to
an increased production of this flavone together with a corresponding
fall in cyanic intensity. That this conclusion is correct is supported[ by the
evidence from family 48/31. Thirty-seven cyanic individuals in this
family were tested for ivory flavone in August and again in October, with
the following results:

Flgvone
rl~{uoh Some Trace Nil
August 8 ii I 17
0ctober 23 4 3 7

The decrease of anthocyanin pigmentation cannot be due to direct action

of light, which has no such effect oll dahlia flower colours as far as can be
ascertained. It is almost certainly due to a change in the metabolism of
the plant in such a way as to modify the actual production of pigments.
Family 55/31 was highly productive of somatic mutations (which
usually arise when Everest is used as a parent). Some of these mutations
involved a change from ivory to white and vice verscb, thus comparative
tests could be made with normal and mutant sectors and petals. In every
ease the presence of ivory was retold to confer blueness and to diminish
the intensity of an~hocyanin pigmentation. Similar mutations in other
families also gave like resNts (see Table XVIII).
It is dear from these data that increase of ivory flavone diminishes
the cyanic intensity of B forms. In consequence it is important to know
if, for example, a BlI,~phenotype will be a ligh~ or a heavy form.
180 Genetics and Chemistry of Flower Uolour iv, Dahlia
The most direct evidence is obtained from the flower colour intensity
of the following B individuals of known constitution.
311/30 (BlI1) has deep purplish-crimson flowers, 30i/30 (BiI2) ])ale
purple aud 31~/30 (B~I2) rosy purple (i.e. deeper than 30~/30 and with a
reddish suffusion due to the presence of pelargonldin instead of cyanin).
311/30 and 30i/30 produce pure cyanin. The difference between 30'/30
and the other two individuals is very striking, the presence of ivory
flavoue considerably diminishing the cyanic intensity (see Plate VII,
figs. 2-¢).
Further evidence may be obtained from the progeny of these tkree
plants on crossing to aoyanic ivory and white forms. Thus the evidence
from family XX (Table I), the progeny of which were all simplex for B,
suggests that B,I a phenotypes are pale heavy cyanles or on the border
line between light and heavy forms, since 8 B,I a and 1 BiI,, were expected
and the palest forms observed were 3 deep and 1 medium magenta.
The effect of an additional I factor is particularly noticeable on com-
paring families XXIV (I i × BiI,) and X X V (I 1 x B2I~) in Table I. In
family XXIV the five pale purple individuals were, on testing (see
Table XII), found to be the only plants with ivory flavone, and these,
from the constitution of theh' parents, must all have been I~. By contrast,
family XXV shows a higher percentage of pale forms, viz. 74 per cent. as
compared with 28 per cent. in family XXIV.
The expected numbers of cyanic genotypes in family XXV were
0.52 (B~Ia), 2.07 (B~Ia), 2"58 (B2I~), 10"33 (B,I2) plus 14"98 individuals with
no flavone. The colour classes observed comprised 3 very deep magentas,
12 pale purple, and 8 more or less purple. On testing these 8 purples,
3 were found t.o be without flavone. The deep magentas and pale purples
all gave good flavone reactions. The cyanic forms with flavone therefore
comprised 3 very deep magentas, 12 pale purple and 5 more or less purple,
in addition to 11 individuals without flavone. Comparing these colour
classes with the calculated numbers of the expected genotypes it seems
probable that the very deep magentas were BlI a forms, the pale purple
BiI.~ and the more or less purpIish individuals B~I~ types. In other words
BlI ~ forms are usually medium heavy eyanics (pale purple) and BlI a
forms are usually pale heavy cyanics (very deep magenta).
In view of these results it seems probable that BlI,i genotypes are
pale heavy eyanles. However, the possibility of other factors diminishing
cyanic intensity should not be overlooked.
The action of 13 on I is more difficult to analyse and fewer data are
available. We have noticed, however, that traces, as opposed to definite
 W . ,]. C. LAWRENCE AND I~OSE SCOTT-){ONC1RIEFF 181
quantities, of flavone are usually lnu~h more common in B cyanies than
in acyanic forms. Further evidence is obtained from am~lysis and com-
parison of 37 cyanics and 46 aoyanics in family ,18/31,as follows:
Fl~vono
A
r "1
Miuch Some Trace Nil
% % % %
Cyanics 62 I0 8 19
Acyatlics 72 i5 II 2
The increase ill blueing observed in purple forms at the end of the season
is doubtless the result of a weakened interaction between B and I, the
suppressing effect of B upon ivory flavone being greatest at the com-
mencement of the flowering season, when anthoeyanin colours are
particularly intense.

(d) In~eracgon of Y with A c~nd B.

Text-fig. 4 shows the diminution in cyanic intensity caused by ~he
presence of yellow flavone in the F I from Glenshee (apricot flowers,
A1YlI4) x White Star (ivory, I2). The effect of the single Y factor is
clearly seen.

40 - 7#~
I
[
t I
|
¢
m t
"~ 30 t
I It

20 / L A
o

t I

io- I ,

0 "~f 1..... I
1 2 3 4 5 6 7 8
Anthoey~nin intensity grades, pale --~ deep
=Aprieo~ (36) - = M ~ g e n t ~ (54)
Text-fig. 4. Showing the effect of Y on cya,~fic intensity in the ~1 from A~Y~I,~ × 12.

Evidence of interaction between Y and A is also provided by co]our

mutations in the ray florets. The complete loss of yellow flavone fl'om a
narrow sector of a petal is comparatively common, and with ligh~
anthocyanin pigmentation the change in the cyanic intensity of the
sector is conspicuous (Lawrence, 1929, Plate XII, fig. 9). Numerous
182 Genetics and Chemistry of Flower Uolour i~ Dahlia
examples have been recorded ill these experiments (e.g. Lawrence, 1931,
pp. 266, 276-7). It will be noticed that, on mutation, the qualitative
change in flower colour is due, not only to the increased anthocyanin
intensity which occurs in the absence of yellow flavone, but also to the
lack of flavone ground colour which plays such an important part in
Dahlia colour variations.
Loss mutations of yellow flavone in heavy cyanic forms produce
changes of intensity similar to those described for light cyanic forms, b a t
on a more intense scale (Lawrence, 1929, Plate XII, fig. 16; 1931, Plate IX,
figs. 9-11). Loss mutations of yellow in pale forms of the heavy cyanic
class a:e particularly interesting. Reference to Table I, families X X X V I I
and X X X I X , will show that a far higher proportion of pale cyanic forms
occmTed in 1~'1 when 27"/27 was crossed with 4-11/28 than when Everest
was used as male, i.e. the presence of yellow flavone in addition to ivory
lind a pronounced diminishing effect on anthoeyanin intensity.
The interaction of Y and I with B may modify anthocyanin
intensity in the heavy cyanic forms to such an extent that B genotypes
come to resemble and even be indistinguishaMe from A forms. In
estimating the degree of this interaction it is necessary first of nil
go examine the cyanic intensity of normM A and B forms in which
interaction has little or no effect. Data from earlier work (Lawa'ence,
1931) suggested that the deepest of the light cyanics (e.g. A a and A~, etc.)
approached and even equalled the heavy cyanic intensity, and an inter-
pretation of the results was based on this assumption. Observations on a
wider range of material and with plants of known constitution now show
that, apart from interaction, the light and heavy cyanic types are widely
separated. Thus the deepest magentas (As_~, Plate vii, fig. 1) are much
paler than any B~ phenotype (e.g. Plate VII, figs. 2 and ~). Similarly
the deepest light cyanics with a yellow ground are never deeper than
apricot (Lawrence, 1931, Plate IX, fig. 6) whereas B1Y~ individuals are
an intense scarlet (Lawrence, 1931, Plate IX, fig. 10). In other words
AB genotypes are always magenta, apricot or an intermediate colour,
all other types being heavy cyanics (aB or AB).
l~evision of the families raised from crossing Union Jack and
22s/27 (Lawrence, 1931) indicates that the constitution of the former
should be B~¥~I 2and of the latter BpA1YpIa or ,t. The proportion of light and
h.eavy cyanics and acyanics in the above families together with. some new
results are given in. Table X I I I below (of. Lawrence, 1981, pp. 266-269).
Six different heavy cyanic forms were crossed with (a) ivory or wJaige,
(b) yellow coloured individuals, and in every case there was an fl~crease in
 W . J. C. LAWIgENCE AND ~OSE SCOTT-I~0NCI%IEFF 183
~he total number of ligh~ cyanics and acyanics on crossing to the yellow
forms. More significant is the occurrence of light eyanies where none
was expected, since four of the heavy cyanic pa.rents used in nine crosses
were nulliplex for A (viz. Union Jack, 42~/29, 42"/29 and 38~/29). In

T A B L E XIII.
Cyanics
]?arenM r - - ' L ----~
No. ? ~ Heavy Light Acyanios
I 32/26 (I~) x Union Jack (B2YII2) 51 I? 8
CMc, 52 0 i0
II 34/26 (Y-fl2-s) x Union Jack (B~YII'Z) ht9 4 52
Calc. 170'8 0 34.2
III 14/26 (Yal2) x Union Jack (B2YII,) 140 3 32
CMc. 145.8 0 29.2
IV Glonshco (A1YlI,l} x Union Jack (B,zYlI2) 285 24 44
CMc. 294.2 29.4 29.4
V 421/29 (B1Y.I2) x Everest (I~) 37 0 37
Calc. 37 0 37
VI 42~/29 (B~Y.I.) "<'111/28 (Xi~) 25 4 29
CMc. 29 0 29
VII 42~/29 (B1YlIa_~) x Whi~c Star (I.) 16 0 15
CMc, 15'5 0 15.5
VIII 42z/29 (B~Y~IsJ x 41~/28 (Y~I,) 3 ,l 3
CMc. 5.5 0 5.5
IX 381/29 (B2Y2I) x Everest (L) 62 i0 16
Gale. 71"6 0 14'3
X 34/26 (Y.L_a) x 381/29 (B.Y.I) 19 8 3
Gale. 25 0 5
XI 27o/27 (BY21) x Everest (12) 22 7 i
XII 27u/27 (BY.I) x 41z/28 (Yzl¢.) 30 27 20*
XIII 22s/27 (B~AIY~I3_a) x Everest (I~) 12 9 1
Calc.
18.3 1"8 1.8
XIV 22s/27 (BoA1Y2Is_a) X White Star (I.) 22 12 4
f.Jalc. 31-6 3'2 3"2
XV 14/26 (Y~I~) x 22s/27 (B2A1Y.Ia~) 50 21 15-~
CMc. 71'6 7'2 7.2
* Including fern" pseudo-yellows.
1" Including five pseudo-yellows.

eight families there is a decided increase in the number of light cyanics

when a heavy cyanic parent is crossed with a yellow individuM as opposed
to a white or ivory. The actuai figures from crossing to ivory and white or
yellow respectively were as folIows:
Percentgge of ligh~ cyanics from
heavy oyanics crossed with
Ivory or white Yellow
Families % %
V and VI 0 7
VII and VIII 0 40
IX and X 22 27
XI and XII 23 35
184 Ge~etics a~,d Che'mistry of Flower Uolour i~ Dahlia
In three other families, XIII, XIV and XV, the same phenomenon is
seen, except that interaction is so strong as to result in twice the expected
number of acyanics through the total suppression of anthocyanin in
certain of the progeny from crossing 22s/27 to a yellow form.
The case of the seedling 22s/27 is particularly interesting, since the
ratios observed in its progenies were unconformable until the interaction
of flavone "rod anthocyanin factors was realised. The :flower colour of
22a/27 is scarlet-orange, l~esults from crossing 22s/27 and 14/26 (yellow)
were given in the 1931 paper, when the ratio of 71 cyanic : 15 acyanic
forms was reported. This was interpreted as showing that 228/27 was
duplex :for A, the ratio of 71 : 15 being Mmost exactly 5 : 1, or expecta-
tion from crossing together duplex and nNliplex forms.
On crossing 22s/27 to the ivory forms Everest and White Star, only
five acyanic individuals appeared in a total of sixty plants. The propor-
tions of light and heavy forms were also different. The records were
therefore re-examined with a view to elucidating the cause of the
discrepancy.
One of the individuMs scored as a "yellow" in the progeny of 14/26
× 228/27, was found to be described as having tinged stems--a character
now known never to be associated with true aoyanic forms. Fern' other
"yellows" were recorded as being unusually deep-ooloured. Since this
family was rMsed a nmnber of these "pseudo-yellows" have been ob-
served and there is now no doubt that they are extreme examples of the
snppression of anthocyanin due to interaction with flavones, lVor
example, twelve plants in family 4/28 and four in family 8/28 were
entirely without anthocyanin in the ray florets. The stems, however, were
varyingly tinged and in family 4/28 the florets were the typical "pseudo-
yellow" colour. The normal yellow of the dahlia is a lemon yellow; the
pseudo-ydlows are usually nearer chrome yellow, and in extreme cases
the stems may be without pigment and the flowers a clear (not chrome)
yellow.
The true number of cyanic to acyanic genotypes, therefore, in the
cross 22s/27 x 14/26 is approximately 76 : 10 and, considered along with
figures from the sister families, it seems highly probable that the total of
131 cyanics to 15 acyanics is an 11 : 1 ratio (expectation 133.8 : 12.2) or
expectation when an individual, duplex and simplex for two independent
factors respectively; is crossed with the double recessive. Since the
parents of 22s/27 were B~ and A I respectively, it follows that this
seedling must be BzA I in respect of the anthocyanin factors.
Thus the results presented in Table X I I I suggest that the addition of
 W . J . C. LAW1%ENCE AND I{OSE SCOTT-1V~ONC:RIEFF 185
Y factors or, more probably, the ratio of flavone to anthocyanin factors,
in heavy cyanic pareuts is correlated in some measure with the average
cyanic intensity of the progeny, increase in the nmnber of flavone factors
contributed by the parents leading to diminution of anthocyanin
iuteusRy in F I B genotypes so that they come to resemble A forms, or,
more extremely, have no anthocyauin at all. The results are probably
confused to some extent by the action of the yellow flavone iuhibRor H,
since any suppression would tend to minimise interaction.
We see therefore that flower-colour intensity in cyanic forms is the
result of a complex balance between the actions of the four tetrasomic
eolom: factors.
Finally, we have to examine the evidence for the interaction effects
of A and B upon Y. No ease has been found where A has caused diminu-
tion of yellow-flavone intensity, b u t t h e r e are indications that a further
suppression of yellow flavone is caused by B factors if the yellow
inhibitor tI has already diminished the intensity to primrose or cream.
Further, on testing a mosaic individual 1)~ (B1Y2I~ 4) more flavone was
found in the lightest mosaic as compared with the self-coloured purplish-
crimson areas. To a rough approximation twice as much flavone was
found in the mosaic parts. The ground colom' of 1~ is near primrose,
fading slightly towards the tips of the petals where ivory flavone makes
itself evident by its blueing effect on the anthocyanin.

(e) P~ttte,r'~s.
The evidence from mutation effects in respect of interaction between
Y and A is supported by the general observation that the deepest light
cyanic forms with yellow grounds are definitely paler than those with
ivory grounds. In particular, the effect of Y on cyanic intensity is clearly
seen in those forms with differential distribution of yellow flavone between
the upper and lower surfaces of the petal.
In the ease of (i) many of the delicately shaded named varieties owe
their patterns to the strong suppressing action of yellow flavone, l~'[any
instances have been noted in these experiments (e.g. Lawrence, 1929,
p. 1~I2, Plate XII, figs. 7 and 8) in which the yellow ground diminishes m
intensity from base to tip of the ray florets. For example, in petals con-
gaining anthocyanin and both yellow and ivory flavone, the flower colour
is seen to change from scarlet at the base to purplish-crimson at the tips
(Lawrence, I929, Plate XII, fig. 8). If no ivory is present the colour at
the tips is deeper and more of a chocolate shade. In the ease of light
cyanic forms with shaded yellow grounds it is common to find no antho-
186 Genetics and Chemist~'y of Flower Coleus' in Dahlia
cyanin except at the tips or distal portion of the petals, anthoeyanin
intensity increasing as yellow intensity decreases (Plate VIII, fig. 9). The
same phenomenon may be seen in the edges of t]~e petals, the flavone
distribution shading off across the petals instead of along its length. In the
case of (ii) differences in colour between the upper andlower surfaces of the
ray florets "~lso constitute a common pattern in named varieties. In such
instances the tinder sides of the petals have less yellow flavone than the
upper sides, cyanic intensity being inversely correlated. When ivory is
also present the under surface is usually bluer, sometimes s~rikingly so
(see Plate VIII, fig. 5).

(f) Cumulative @cts.

We have shown that (1) i and A are incompletely dominant, I being
cumulative up to the duplex condition, A being cumulative from
simplex to quadriplex; (2) Y and B are completely dominant in ~he
simplex condition--duplex, triplex and quadriplex forms being in-
distinguishable from simplex. Thus in theh: phenotypic effects I and A
are cumulative whereas Y and B are non-cumulative. But we have also
shown (Table XI) that the interaction of I with A is strictly proportional
to the nmnber of I factors, i.e. I is potentially cumulative from I~ to I~
and aetually shows this in interaction effects. The evidence from inter-
action also supports the cumulative nature of A, as seen in the pheno-
types.
The fact that I, though associated with maximum flavone production
when in the duplex condition in aeyanics, is, nevertheless, potentially
eumNative from I 1 to I,t necessitates examination of the Y and B forms
to see if these too are potentially cumulative. In the ease el Y the clearest
evidence is from comparison of families 56/30 (Glenshee AiYi%i x White
Star I~) and ,t/28 (1~/26 YaI2x 36/26 AaIa). The eolour of the yellow
ground in both families was uniformly good, therefore the effect of tt can
be disregarded. No eyanies were found in 56/30, but twelve pseudo-
yellows occurred ill 4/28. This result is almost certainly due to the
greater number of Y factors in family 4/28, especially as the flavone
index for I is of higher value in the ease of 56/30.
Y therefore is potentially cumulative in effect. The difference in
intensity of BY types might be due either to the cumulative effect of Y
upon B, or to the eumula.tive nature of B independently of Y, or to both.
It will be shown later that 31"/30 (B~I2) produces pelargonin as com-
pared with 30~/30 (BlI,) producing cyanin alone, a :fact which suggests
that B also is potentially cumulative.
 W. J. C. LAWI~ENCE AND I%OSE SCOTT-MONCI~IEFF 187

The hypothesis that the difference between potential and actual

cumulative effects may be due to a limited pigment source is discussed
later.

PAI:~T Ii. CHEIVIIST1%Y.

(i) T~ ~5owmz rlc~NTS OF D~rLIX.
Flower colonr in the garden dahlia depends upon the presence in
varying proportions of soluble sap pigments of the anthocyanin and
anthoxanthin (flavone or flavonol) type.
The occurrence of these pigments and their association with each
other from a genetieal and chemical standpoint has been investigated by
means of a series o~ quick qualitative colonr tests upon crude or semi-
pm'e sap extracts, based upon careful preliminary identifications and
standardisations.
Owing to the slight texture of the florets, the pigments were extracted
with the greatest ease by macerating the petals with a glass rod in a test-
tube containing 3 c.c. of aqueous 1 per cent. hydrochloric acid. The base
of the petal was always removed to prevent contamination from a yellow
" e y e " which frequently occurs. Only fresh flowers were used, and the
extracts were examined as soon as possible. When both flavones and
anthocyanins were present, the flavones were removed by exhaustive
extraction with ethyl acetate, in which solvent the anthocyanins are
completely insoluble.
It was necessary in some cases to keep petals for further investigation.
These were air dried at about 30° C. and kept in a dry, dark cupboard.

(c~) Flc~vones.
Two pigments of the anthoxanthin type are present in the dahlia.
Both the ivory and yellow pigments have been isolated from the yellow
flowers by Schmid and his co-workers (1928, 1932 and 1933), who state
that the former is the ivory flavone apigenin (Text-fig. 5 e) and the latter
an isomeric trihydroxy compound, from which they obtained para-
hydroxybenzoie acid on degradation. Their inability to methylate more
than two of the three hydroxyls points to one of these being at positon 5.
The third hydroxyl must therefore be at 3, 6 or 7. In the last of these
papers Schmid has published the surprishlg fact that on acetytation the
purified pigment forms a triacetyl product identical with b:iacetyI
apigenin, which on de-acetylation gives apigenin itself. This yellow
pigment therefore appears to be an isomer of apigenin. The exact nature
Journ. of Geneticsxxx 13
188 Ge~etics a'nd Che~nist~'y of Flower Colorer i~ Dahlia

o 8 ~ o -~

o "~ ~

a~ d ga

~
~ ~~
~ ~ ~o® i

~ 0

q~o ~. ~.~
0 ,O-4

d~

© ©

0 0 °

.~
e~
o ~

N
 W. J. C. LAWRENCE AND 1%OSE ~COTT-I~.ONCRIEI?'F 189
of this anthoxanthin is therefore still uncertain (Text-fig. 5 b), but for
the sake of simplicity the pigment is rein'red to throughout this papm" as
the yellow "flavone."
According to Sehmid and his collaborators both the ivory and yellow
pigments are non-glycosidic.
The ivory flavone behaves as a strong co-pigment according to the
qualifications laid down by the l%obinsons (1931 cb, ].932 cb) for this
interesting class of compounds, Extracts have a strong blueing effect
upon cyanin solutions and also upon pdargonin ones to a smaller degree.
This effect disappears on boiling the co-pigmented anthocyanin solution,
but returns on cooling. On repeated extraction with ethyl acetate the
colour of this solution changes from bluish red to crimson scarlet, btlt the
original colour can be recovered by the addition of a water solution of the
extracted flavone or by addition of a crude extract of the petals of any
ivory dahlia.
Pale purple and bluish-magenta fowers were strongly co-pigmented by
the ivory flavone which they carried, whereas the corresponding choco-
lates and rosy-magentas were comparatively flavone-free an([ uneo-
pigmented. The lowering effect of this co-pigment upon the distribution
number 1 with amyl alcohol of the crude anthocyanin extracts was very
apparent in these flowers. While extracts of rosy-magentas, and especially
the chocolates, had a fairly high distribution due to the monoglueoside
present, this was reduced almost to zero in the bhtish-magentas and pale
purples. The yellow flavone showed no co-pigment effect.
Experims~tcd. The presence in the crude dahlia extracts of one or
both flavones was determined by the following methods. Apigenin was
easily identified in the ivories and magentas by the yellow or green reac-
tion on fuming the petals with ammonia. When much anthocyanin was
present the crude extract was shaken in a test-tube with an equal
quantity (3 c.c.) of ethyl acetate. After standing a few minutes the latter
was decanted off and shaken with 1 c.e. of saturated Na2COa solution,
and any development of colour in the aqueous layer observed. Total
absence of colour indicated absence of both flavones, while the ivory
pigment gave a lemon yellow, and the yellow one a golden yellow rapidly
turning orange and then intense golden brown. In the absence of antho-
cyanin these alkali colour reactions could be made directly upon the acid
extracts.
Since in the presence of the yellow pigment, ivory flavone could not be
The "distribution number" of an anthocyaninrefersto its specificdistributionratio
between iso-r~mylalcoholand one per cent. aqueoushydrochloricacid.
13-2
190 Genetics and Chemistry of Flower Uolour in Dahlia
recognised by these methods, its presence in yellow flowers was detected
by making use of tile strong co-pigment effect obtained on addition of an
acid flavone extract to an equal vohme of a standard eyanin solution
(see p. 169). The standard solution was prepared by extracting about
fifty petals from rosy-magenta (flavone-free) flowers with 100 c.c. of
1 per cent. aqueous hydrochloric acid. This effect reached a maximum
in a :few minutes and remMned stable.
White (no flavone) flowers tested were found to be of two kinds,
(1) those which gave an entirely negative reaction, except for the
faintest suggestion of pink, and (2) those which gave a pale straw or
mMze colour, quite distinct from the lemon-yellow reaction of ivory.
This second effect was also observed with certain pale ivory forms, whose
weaker reaction was not strong enough to mask the maize colour.
Apparently the substance which gives the maize reaction is independent
of ivory, with which it may or may not occur. It gives a green reaction
with ferric chloride and is probably tannin. It was also found in rosy-
magenta (A) and in some purple (B) forms of the garden dahlia, and in
D. Merdcii and D. gmperialis, both with magenta flowers.

(b) Anthocyanins.
Wiltstittter and 5{allison (1915) isolated cyaniu (Text-fig. 5 c) from
the deep brown-red garden dahlia iu which it was present to nearly
20 per cent. and staged that certai~ scarlet flowers were pigmented by
4-6 per cent. of pelargonin (Text-fig. 5 d), while a deep violet variety
contained this pigment together with a suM1 amount of cyanin. Both
these pigments are 3:5 diglueosides (I~obinson and Robinson, 1931 b).
We have found that one or both of these two anthocyanins occur through-
out the numerous colour varieties discussed in this paper, and have also
detected the presence of two other anthocyanins. In cyanin (pelargonin-
free) flowers small amounts of a cyanidin 3-monoside (Text-fig. 5 e) are
often present. Iu the magentas the quantity is negligible, but in the
deeply pigmented ell ocolate and purplish-crimson flowers it is present in
sufficiently large amounts for its separation on a suM1 scale and subse-
quent identification (see p. 194). It may be noted in this connection that
I£obinson and Todd (1932) remarked upon the presence of traces of a
monoglycoside in the specimen of Dahlia cyanin sent to them by Prof.
Willstgtter. It is Mso of interest that the purple disc florets of D. Me~'ekii
cont~ain large amounts of this monoglycoside, while the ray florets of
D. Me~'ckii and those of D. 4mpericdis are both pigmented by cyanin.
We have also found that flowers of D. coronctta contain a pelargonidin
 W. J. C. LAWRENCE m\TD I%OSE SCOTT-MONCI%IEFF 191
3-monoside (Text-fig. 5f). They were a slightly duller orange-scarlet
than those of D. coccinea and D. va~'iabilis, which are pigmented by
pelargonin.
The presence in flower extracts of pelargonin or cyanin or mixtures
of these two pigments could generally be detected by means of their
eolour reactions with sodium carbonate. When necessary the flavones
were first removed by means of ethyl acetate.
A certain "liveliness" of colour generMly distinguished the petals of
pelargonin-pigmented fowers, such as the rosy-purples, while cyanin ones
were slightly duller in appearance (see Plate VII, figs. 3 and 4). When
yellow flavone could be detected with the eye, some pelargonin was
always found to be present. Although in most cases it was thus possible
to distinguish between flowers contMning cyanin and pelargonin res-
pectively, those of the purplish-crimson, purple and very deep magenta
shades were often indistinguishable.
The pigment changes occurring in severM somatic mutations have
also been studied.
Prof. and ~Irs I~obinson (1932 cb) state flint in three di~erent dahlia
flowers taken at random, they came across 3:5-diglyeosides of pelargo-
nidin, cysnidin and delphinidin. The lat~er pigment was detected on one
occasion only, in the mauve-pro:pie dahlia "George Ireland," which they
have now found normally to be pigmented by cyanin. Since its occm'-
reuse here is unique, the presence of delphinidin in D. vctr4abigs is still
sub indict. This pigment has certainly not occurred in any of our families.
The stem pigments of Dcd~ic~ have not yet been identified.
Ex2)c~'imcntcd. In the case of flavone-free flowers, cyanin extracts were
a more crimson scarlet than those of pelargonin, while co-pigmented
cyanm solutions were markedly bluer. When yellow flavone was present
the extracts were orange-red. Acid alcoholic solutions of pelargonin
show a green fluorescence. When yellow flavone is absent the specific
anthoeyanins may readily be identified by observing the colour reactions
obtMned on adding aqueous sodium carbonate to the dilute acid extracts.
Cyanin gives a rich pure blue coloration in the absence of flavones. In
the presence of ivory the colour is green-blue to emerMd green according
to the amolmt of flavone. Similarly petargonin gives a red-violet reaction
(green-blue on addition of acetone) ill the absence of flavones, and a
greenish-violet if ivory is present. Yellow flavone gives an intense brown-
orange reaction which masks that of an accompanying anthocyanin.
]?Iowever, both ivory and yellow flavones may be remSved by repeaSed
extraction with a large excess of ethyl acetate. This can be done quite
 192 Genetics and Uhe'~nis/,ry of Flowe~" Uolou~" i~ DaAlia
simply by successive shakings in a test-tube and decantation of the upper
layer. Ivory, even in its greatest concentration, is removed quickly as
compared with yellow flavone, which evidently occurs in much greater
quantity. This observation is of considerable interest in regard to the
pronounced suppressing effect of yellow upon the co-production of ivory
flavone and anthoeyanin, since it supports the view that this action of
yellow is mainly due to its tendency to monopolise the available supply
of pigment-forming material.
When extracts of flowers of D. eo~'onata were, with great dill~oulty,
cleared of yellow flavone, the anthocyaNn solution gave a plum-red
reaction with carbonate, as might be expected from pelargonidiu
3-monoside. This plum reaction, however, was also obtained in certain
other cases with pelargonin solutions which were still contaminated by
traces of yellow flavone. The colour obtained was not such a clear red,
but it was necessary to guard[ against confusion. The pigment of
D. co,ro~uaa could be distinguished from pelargonin by its high distribu-
tion (monoglycosidic) in amyt alcohol.
While, on treatment with sodium carbonate, flavone-free cyanin
extracts give pure blue and pelargonin ones a red-violet, mixtures of
these two pigments give intermediate shades such as violet-blue, blue-
violet and violet, according to the proportions of each pigment present.
This range of colours being obtained on testing our series of crude
petal extracts (flavone-fl'ee), it was surmised that eyanin and pelargonin
were present in these flowers in varying proportions. 3{ore elaborate
tests however were made on several of these extracts, to ensure that the
blueness of some of the alkali reactions was due to the presence of cyanin
and not to tannin or some other modifying substance.
The purification and identification of the glucosidal and hydrolysed
pigments were based upon the methods described in Prof. and 3'h's t~obin-
son's "Survey of Anthooyanins. I and I I " (1931 c~, 1932 a). These
methods, adapted and modified to some extent to suit our requirements,
were invMuable for our purpose. We are also greatly indebted to these
authors for their help and advice.
Before hydrolysis the extracts were shaken twice with a large excess
of amyl alcohol to remove most of the flavone. Ethyl acetate was not
used, since any remaining trace upsets the normal extraction with the
"cyanidin reagent." Oyanin and petargonin are hydrolysed to cyanidin
and pe]argol~din respectively.
After hydrolysis of these extracts and subsequent thorough purifica-
tion of the sugar-free pigments, a violet ferric chloride reaction (in amyt
 W . ,~[.G. LA%VR]]]NCE A N D ]~OSE ~COTT-MONCI~,IEFF 193
alcohol over-saturated sodium acetate solution)was obtained when both
eyanidin and pelarg'onidin were present. Small amounts of these pigme~lts
were finally obtained uncontaminated by each other by lh:actional pre-
cipitation with benzene from amyl alcohol into 1 ]?el: cent. hydrochloric
acid. In the first precipitation cyanidin predominated, and this was
finally freed from pelargonidin by extraction with igobinson's "cyanidin
reagent." The aqueous layer then gave satisfactory reactions for cyanidin,
including a good[ blue eolour with ferric eNoride. Solutions of pure
pelargonidin were Obtained in the final precipitations into 1 per cent.
acid. These gave no ferric chloride reaction and resembled pelargonidin
in every way.
From the extracts which were thus examined it was found that small
amounts of cymlin tended to modify the pelargonin alkali reactions out
of proportion to the quantity present. Extracts giving violet and blue-
violet reactions contained only small amounts of cyanin. Those giving
violet-blue contained larger amounts, but pelargonin still predominated.
It was therefore concluded[ that alkali reactions showing even a tinge of
violet indicated the definite presence of pelargonin, while extracts giving
blue-violet and violet reactions contained a large proportion of this
pigment.
In apricot-magenta flowers in which Y is partly inhibited by I-I,
cyanin was often present in sut~cient amount to mask the pelargonin
reaction entirely, but after hydrolysis it was possible to isolate sufficient
pelargonidin for identification.
It is important to test extracts for cyanin as soon as possible after
removal of the ivory flavone, since its stability depends largety upon the
presence of this co-pigment. Cyanidin solutions are even less stable and
should be tested qmckly or kept in the dark. When testing mixtures of
cyanin and pelargonin with sodimn carbonate, the blue of the cyanin
reaction tends to fade out first, leaving the more stable purple of the
pelargonin.
Flowers containing very large amounts of yellow flavone together
with the anthocyanin were best cleared by frequent extraction, first with
ethyl acetate and then with amyl alcohol. When the flavone had{ been
removed, an extraction with butyl alcohol was made after saturating
the aqueous solution with sodium chloride. On washing back some of the
anthocyanin from the butyl alcohol into 1 per cent. hydrochloric acid,
good reactions for pelargonin were obtained. At tiffs stage of purity it
was possible to detect the fluorescence of the pelargonin in the butyl
alcohol layer.
194 Genetics and Chemistry of Flower Colour in Dahlia
Identification of cyanidin 3-monoside.
The separation and identification of the cyanidin 3-monoside from
the deeply-pigmented chocolate flowers was carried out in the following
Izlanner.
The dried petals wore left to stand overnight in 1 pro" cent. hydro-
chloric acid and the deep red extract was then filtered off. A second
amount of acid was added to the petals and the extraction repeated.
The combined concentrated extracts were extracted three times with a
large excess of amyl alcohol, l~{ost of the cyanin remained in the
aqueous layer but some was carried into the alcoholic layer along with
the monoghmoside. To renmve this, the pigment was precipitated into
dilute acid by means of petrol ether, taken up into butyl alcohol in the
presence of salt and well washed with a saturated salt solution. After
several precipitations into aqueous acid by petrol ether and re-extraction
with butyl alcohol, ~ho aqueous pigment solution was extracted three
times with ethyl acetate in the presence of picric acid, and reprecipitated
into 0.,5 per cent. acid by petrol ether. After three more extractions with
butyl alcohol (in the absence of salt) and reprecipitation with petrol
ether, a few c.c. of concentrated pigment extract were obtained giving
none of the reactions for cyanin but only those of a cyanidin 3-monoside
(Text-fig. ,5 e). A comparison with a pure sample of chrysanthomin
kindly supplied by Prof. Robinson showed no divergence. The somewhat
dull red acid solution gave a blue-violet soIution with sodium carbonate,
changing to pure blue with sodium hydroxide. With sodium acetate a
violet colour was obtained. The distribution with amyl alcohol was nor-
real for a monoghcoside, the high distribution when using very low
concentrations of pigment differentiating it from a rhamnoglueoside.
On hydrolysis and purification, the sugar-free pigment gave the colonr
reactions of eyanidin--inchding a good blue with ferric chloride. Oom-
parison of the anthocyanin with a sample of the cyanidin rhamnoglucoside
from crimson Antirrhinum majus (Scott-hfoncrieff, 1930) showed definite
divergence in behaviour. It was therefore established that a definite
though small proportion of cyanidin 3-monoside is present in the
chocolate-colom'ed dahlia, together with a large amount of eyanin.
A preliminary experiment on the purpte central disc florets of the
blue-magenta flowers of D. Merclcii showed that these were chiefly
pigmented by cyanidin 3-monoside. Since the pigment of the ou~er disc
florets is cyanin, the co-production of these two anthocyanins in Dahlia
should have a definite significance from the point of view of pigment
metabolism.
 W . J . O. LAWIgENCE A N D ~OSE SCOTT-NONCllIEFF 195
Identgficatgon of pda~'gonidin 3-monoside.
Unlike the orange flowers of D. va~'iabilis and D. coccinea, those
of D. co~'onata contain a 3-monoside of pslargonidin together with
abundant yellow anthoxanthin. The monogtyeosidic nature of the
anthocyanin was easily recognised by its high distribution in amyl
alcohol, even in dihte solution (distinction from rhamnoglycoside).
Great diflieNty was experienced in removing all traces of yellow flavone,
but eventually small amounts of uncontaminated anthocyanin solution
were obtained which gave clear and satisfactory reactions for pelargo-
nidin 3-monosids. Repeated extraction with 1 per cent. hydrocNoric
acid extracts of fresh flowers with large quantities of ethyl acetate
eventually gave a flavone-free solution, but the method was tedious and
extravagaut. ~'fethods involving repeated extraction with butyl aleohot,
amyl alcohol or ethyl acetate and[ pierie acid, and precipitation with
petrol ether, were mlsatisfactory, owing to much of the yellow flavone
being carried along with the anthocyanin through all these purifications.
Finally use was made of the slight but definite solubility of pelargonidin
monoside pierate in ether. 1 per cent. hydrochloric acid[ methyl alco-
holic extracts of the dried flowers were precipitated with ether, the
precipitate dissolved in acid methyl alcohol, reprecipitated, well washed
with further quantities of ether, and finally dissolved in I per cent.
aqueous hydrochloric acid. After filtering and shaking twice with larger
quantities of ether to remove more of the yellow flavone, solid pieric acid
was added, and the solution shaken three successive times with twice its
v o h m e of ether. A slight but definite extraction of the anthoeyanin was
obtained in the ether layers, which were then united and the pigment re-
moved by shaking with three successive amounts of 10 c.e. of concentrated
hydrochloric acid. The acid aqueous layers were qtfickly poured off, and
repeatedly shaken with benzene to remove the picrie acid. The solution
was then diluted with an equal quantity of water, saturated with salt,
and the pigment extracted with amyl alcohol, washed with salt solution
and reprecipitated with petrol ether into 1 per cent. hydrochloric acid.
This process was repeated three or fear times until the aqueous layer
gave the clear and unmistakable reactions recorded for a pelargonidin
3-monoside (Text-fig. 5f) by !gobsrtson and l%obinson (1928). The
pigment was scarlet-yellow in aqueous acid and slightly pinker in amyl
alcohol, in which it had a high distribution both in strong and weak
solutions, as is normal for a monoglyeoside. In the presence of an excess
of sodimu chloride it was almost quantitatively extracted by the alcohol.
Unlike pslargonin, the pigment showed no fluorescence. A characteristic
196 Gene~ics a~~d C h e m i s t r y Of F~owe'r Colour i n D a h l i a
plmn colom: was obtained with sodium acetate and sodium carbonate,
the colour in the latter test being unchanged by the addition of alcohol,
dilute caustic soda or acetone.
The nature of the aglucone was further established by hydrolysis and
purification of the crude flower extracts, which then gave the normal tests
for pelargonidin.
Owing to the difficulties of purification, the possibility of the presence
of other glycosides of pelargonidin cannot be ignored, but the 3-glycoside
is the only anthocyanin present in any qua,ntity. Beyond the fact that
it is a monose, the nature of the sugar residue has not been determined.
The pigment is probably identical with the monoglucoside oallistephin,
but may be the galactoside or a pentoside.

(if) G~NE'rIOaL ~SP~OT O~~ O~EM~O~La~SUL'rS.

In the course of these experiments we have tested either by quick
qualitative tests, or, when necessary, by more accurate methods of
identification, upwards of 500 flowers including species, hybrids and a
large range of D. varia,bilis of known geneticM constitution.

(a,) Dahlia species and species hybrids.

The pigments of the species fell rote two groups corresponding to the
two eolour series. D. Metric,if contained cyanin in the petals and the
corresponding 3-monoside in the d~rk central disc florets, much apigenin
co-pigmenting the flowers. D. impe~'icdis also contained cyanin and rather
less co-pigment. Pelargonidin glycosides and yellow flavone were iotmd
in D. coecinea and D. coronata, the former having %he diglucoside pelar-
gonin and ~he latter the corresponding 3-monoside.
Six species hybrids have been examined with a view to tracing the
inheritance of the different parental pigments in these offspring (see
Table XVI). Three of the four D. coccinea x D. variabilis plants were
natural hybrids, while 3/31 was from White Star (apigenin) x D. coccinec~.
In each case the pelargonin and yellow flavone type was inherited.
These pigments were also found in the two YD. coronata hybrids,
showing that the pelargonidin type is inherited in preference to the
cyanidin. The 3-monoside found in D. coronata is not present in any
other species, nor is it found in its hybrids, tt is noteworthy that the
orange hybrid 2/29 obtained on crossing a magenta D. variabilis flower
36/26 (cyanin and apigenin) wi~h D. coroneta should be, unlike either
parent, s~rongly pigmented with pelargonin, the aglucone being inherited
from the former and the diglucosidic sugar residue from the latter.
 W . J. C. LAWI~ENOE zkND I%OSE SCOTT-~{ONCI%IEFF 197

(b) D. va~'iabilis.
The distribution of the yellow and ivory flavones, their interaction
effects an d the effect of I-I, the yellow fl~vone inhibitor, are futly discussed
in Part I.
Investigation of the pigments in the cyanic flowers may be taken in
two sections: those with an ivory or whi~e ground, and those carrying
yellow flavone.
(1) Fl,owe,rs with non-yelbw g~'ounds.
In this section 290 individuals, with ivory or white ground, coin-
prising ].30 light and 160 heavy cyanics, have been tested for their specific
anthocyanins. The resNts will be found in Table XIV. The plants arc
divided into five classes according to their reactions with sodimn
TABLE XIV.
Showing the ~'elation of facto,re A a,~d 13 to the .productio~ of cya~zi,~ and
pela'rgonin i.~kforms with ivory or white g,rounds.
Colour reactions with sodium carbonate

J31ue -
tinged- Violeb Red
~Flower c o l o u r J31ue violet blue Violet violet
L i g h t c y a n i e s (A)
Pale magent~ 18 . . . .
Medium magen~ 62 1 (13) -- -- --
Deep magenta 48 1 (B) -- -- --
t I e a v y e y a n i e s (13)
Very deep magen~ -- 1 2 1 --
P,Mo p u r p l e 29 1 3 -- 1
Purple 20 12 10 3 --
Crimson-purple [
Purplish-crimson J 40 10 3 2 --
Deep pm'plish-crinmon 13 5 2 1

carbonate, which ranged from blue (pm'e cyanin) to red-violet (pure

pelargonin). With two exceptions--one fi:om family 48/31, the other
from family 53/31--all the light cyanics with non-yellow g,rounds gave the
pu~'e blue q'eaction fo~' cya~zin. The two exceptions were both known to
carry B factors (see Table gI) and gave btue-tinged-violet~ reactions.
While in the A cyanics all those with non-yellow grounds were found
to carry cyanin, in the I3 forms tested only 63 per cent. contained cyanin
alone while the remainder contained chiefly pelargonili together with
sm~ll ~mounts oi eyanin in varying proportions.
Three heavy cyanic families with non-yellow grounds were therefore
examined in order to ascertain this distribution of pelargonin and cyanin
in. B forms. The results are given in Table XV, and from these it will be
198 Genetics and Chemistry of Flower Colour in Dahlia
seen that there is no clear evidence as ~o the inheritance of the capacity
to form pelargonin. It is surprising to find that the majority of the
offspring from crossing 31~/30 (pure pelargonin) with a white-flowered

TABLE XV.
Showi~g the i'M~eritanceof cyanin and 2elargonin (in crossesbetween
cyanic and acyanicparents).
F I an~hocyanin reactions

]Bluedieged
Parents, ~nthoeyanin ]?ure blue violet to
~Parents alld flower colour reactions ~nd s~p- violet (somo
c - - ~ - - ~ plfire blue or all
Family ? d~ ~ • (cyanin) pelargolfin)
32/26 (white) 3] 1/30 (deep Nil Blue 22 1
I1 purple-erim- (pure eyanin)
son) B I I 1
53/3I 32/26 (whi~e) 312/30 (rosy- Nil i~ed-violet 33 8
I1 purple) B a l ~, (pure pel~rgonin)
48/3] 301/30 (bhm- Everest (ivory) ]Blue Nil 28 9
purple) BII ~ 12
55131 Coldness Gem Everest (ivory) ]31ue-violeg Nil ]9 18
(crimson- In (pelargonin
SCal'let ) + cyanin)
B2A.~YII1

form contain pure cyanin. The fourth family, 55/31, is included in the
table, since a fair number of purplish segregates were tested for their
anthocyanins.

(2) Flowers with yellow grounds.

The anthocyanins associated with yellow flavone were identified in
seven species and their hybrids and in tMrty-seven garden forms. Their
colour reactions are given in Table XVI. The plants are arranged in
order of diminishing yellowness of ground colour due to increasing effect
of the inhibitor t t upon the factor Y. It is at once apparent that this
arrangement results in the grading of the specific anthocyanin denomina-
tions in a sequence, ranging from pure pelargonin in the scarlets through
mixtures of pelargonin and cyanin to almost pure cyanin in the apricot-
magentas with pale cream grmmds. It will be noticed that 299/27 and
39~/28 are out of series. Owing to the large number of factors in these
types, excessive pelargonin formation might be expected on our theory
of specific pigmentation (see io. 21@ Conversely, B 414, 22s/27 and A431°
have bluer carbonate reactions than expected[, but since they are all three
related, it is probable that their case is exceptional.
In general, however, the productio~'~of yellowflavone c~ndof peh~,rgonin
is q,ua,ntitatively correlated.
 W. J. C. LAWr~E~aE Amid I~OSE SCOTT-MONc[RIEFF 199

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° ~,e .~ .o . z ° ~ .,~ ~, .E e '~ ~

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g
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~,~ ~ -,~.~.~. ~- ~. . ~ ~ ~ °l -~ ~ - ~ , ~ ~ ~ "~ ~
o~ ., ~ . ~<<I<~, ! ~ o. i G i ~ . ~,~ ,.~,~ ~H~ i -~;4,~,~ - I~ I I I I ,o, }'4
~ h~.~! ~ ? ~, ~~ ~I~ ~:~,~ ! E ~ ~ h ~ % / . :~ °l °,'~ ~, ~, ~,~,~, "i'

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~0 ~ °~ ~ o ~ .o~ ~
o ~ ~ ~ o ~0 ~ ~ ~' ~0 ~, ~~ o
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o
o ~
m o
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m
bg bg o
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4 ~ ~
~:~ ~ .,~
~ "~ ~ ~0~'-~ "~
o ~Oce I~ ~

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~×x ~.,× ~ "~

~ o ~ ¢o¢0¢0 ~ ~ (:~<::~o~O(:~C:, ~ ~ ~ l~q ~o 0~ OC,~ c.O~ 1.~ b-- ~ L~ b'- L~ 1~

200 Genetics and Chemistry of Flozoer Colour in Dahlia
In spRe of this correlation, each anthocyanin may occur independently
of the presence of flavones. Of the types in Table XIV six B ivory and
two 13 plants gave a pelargonin reaction. I-Iere we have the production of
pelargonin in the absence of yellow flavone and with or without the
presence oI ivory flavone. AgMn 57B Ivory and 45B plants gave the
blue cyaniu reaction. Here we have the production of cyanin with or
wNiout the presence of ivory flavone. The remMning 179 individuals
tested gave intermediate cyanic reactions, and here again a llumber of
them were devoid of flavone. In no flowers containing much yellow
flavone was cyanin found withou~ a much greater proportion of pelar-
T A B L E XVII.
~Flower colour Constitution" Anghoxanthin pigment Anthocyanin pigment
White to ivory Ii_,, Little or much ivory --
Yellow Y (and YI) Yellow (and ivory) --
Yellow to cream YHi_,~ Yellow
Apricot AY Yellow MMnly pehu'gonin
Apricot AYI~ Yellow and ivory MMnly pelargonin
Magenta-apricot to J AYHi_ l
apricot-magenta ( Ayi~I_ii_.l Much or little yellow Pclargonin and cyanin
Bhdsh-magenta AI~ Little or much iyory Cyanin
l~osy-magcnta A Nil Cyalfin
Scarlet BY Yellow Pelargonin
Scarlet BYI:_ a Yellow and ivory Pelargonin
Crimson-scarlet to BYI-Ii_,~ M.uch or little yellow MMnly pelargonin
purplish-crimson
Crimson to purplish- BYI,~tHI_. 1 Much or little yellow Mainly pehu'gonin
crimson and ivory
Purplish-crimson to 1312_~ Ivory ]
pMe-purple or rosy- t Mainly cyanin or
pro'pie mMnly pel~rgonin
Chocolate or crimson- B Nfl
purple
IYotc. A may be present in B forms without appreciable effect. I i may be present in any
form without additionM effect.

gonin. Some pelargonin is always present if Y is present. No pelargonfil

is found in A types unless Y is present.
The co-pigment and interaction effects of the flavones upon ~he colom:
and intensity of the anthocyanin pigments have already been described
in Part I. In the A types the colour variations in the magentas (cyanin
with or without apigenin) and in the apricots (pelargonin and yellow
flavone with or without apigeuin) are produced entirely by these two
effects. Table XVII shows the flower colour and pigments present in the
series of genotypes.
(3) Mutations.
The distribution and character of flower-colom' mutations in Dahlia
were fully dealt with i~ an earlier paper (Lawrence, 1931). The evidence
 W. J. C. LAWI~ENOE AND ~OSE'SCOTT-MONCI~IEI~F 201
strongly suggests that the mutations arise from irregular distribution at
mitosis of the chromosomes bearing tlle colour factors. Published and
unpublished data clearly show that in the great majority of cases the
factor involved in mutation is in the sinaplex condition, the mutation
usually being a "loss mutation" to ~he nulliplex condition. Since no
colour is produced by any of the recessive forms Y4, i,~, a 4 and b~L, direct
comparison can be made between normM and mutant areas in regard to
the nature and interaction of the specific pigments. The individuals
listed in q/able XVIII a/ll gave large mutant sectors or entire ray floregs
in which the changes in pigmentation could be investigated. In each
instance it was the flavone ground which mutated, the change involving
toss or gain of ivory or yellow flavone, accompanied by an inversely
proportionM change in anthocyanin con~ent (see 1°"/31 on Plate VIII,
fig. 8) and sometimes also in its nature.
In the fi~:st six oases the mutation was from white or very wea,k ivory
go ivory. In one of these plants (C21.a) the cyanic reaction of the mutant
sector was different from the normal, and gain of ivory flavone was
accompanied by an increase in the proportion of pelargonin to cyanin.
The next four mutations involved loss of yellow flavone, which in the
case of B 40 a and B 42 ~"revealed a white grotmd. The presence of ivory
flavone in B 414 was demonstrated by its purple (co-pigmented) flower
colour. The mutation from yetlow to ivory ground in B 41 ~, 62"/29, and
probably B 40 s, was accompanied by a slight increase in ~he proportion
of cyanin to pelargonin.
In each of the last three plants the mutation was from non-yellow to
yellow, and in two cases (C 24 ~2 and C 207) ~here was a slight increase in
the amount of pelargoniu. No such difference was detected in the third
individuM, but in the apricot-magentas small amounts of pelargonin are
easily masked by the large proportion of eyanin.
The pattern A 139 (see Plate VIII[, fig. 5) maybe classed alongside these
mutations. Here the darker crimson-purple edge contains very little
yellow flavone as compared with. the lighter orange central portion. The
larger proportion of pelargonin in the latter is demonstrated by a change
in the carbonate reaction from violet-blue to bhe-violet. A further point
of interest concerning this flower is the substitution of ivory for yellow
flavone on its under side, where the central section is strongly co-pigmented.
The deep purplish-crimson flower 31r'/30 with a paie magenta-purple
edge pattern may also be mentioned here, since ~he decrease in cyanic
intensity was found to be accompanied by an increase in a,pigenin by four
and a half.
202 Genetics a n d C h e m i s t r y of _Plower Colour i n D a h l i a
Variation in the amounts of flavones and of anthocyanins present in
comparable genotypes was s~udied with a view to ascertaining the limits
of pigment formation required as a basis for the theory put forward[ in
Part III, and is referred to in the later section (pp. 205-209).
TABLE XVIII.
Flavone and eye,nit all,all colour ~'eactions ~n normal (a) and mutant (b)
.flowers. An aslerisl~ denotes.fiavones cleared, 5eJbre testi,ngfor oyanio reaction.
t~am. Seedling No. Flower eolour Flavone reaction Cyanic reaegio~
C13 a (a) Purplish-crimson Trace of yellow Blue
(b) 13{co:intomagenta Yellow and maize Blue*
53/31. C17 a (a) Crimson-purpie Nil BNe
(5) Blue-purple Yellow Blue-green
55/31. C 21a (a) Deep purplish-crimson Nil Violet-blue
(b) Purplish-crimson Pale yellow Blue-violet
55/31. 026 ll (a) Deep purplish-edmson Nil :Blue
(b) Purple Very pale yellow Blue
55/31. C261 (a) Purplish-crimson Very pale yellow Violet
(b) Pale magent)a Yellow Green, slighl~ly
muddy (violet?)
301/30 (a) Pale purple Pale yellow :Blue
(b) Magenta Yellow Green
3s/31. B,i0 ~ (a) Orange Orange Brown-tinged green
(b) Mageng~ Very pale maize? Pale violet blue
38/31. B41 ~ (a) :Deep orange Orange Violeb<red violet*
(b) Purple -- Violet < slightly
more blue
38/31. B42 ~ (a) Crimson<magenta Orange Green
tips
(b) :Deepmagenta Nil Sapphire-blue
42"/29 (a) Crimson scarlet Orange Violet*
(b) Purple Yellow Blue-violet*
55/31. 024 l" (a) Pm'ple Very pale yellow Violebblue <pale
blue-violet*
(b) Seadeberimson Orange Violebb/ue <violeb*
55/31. C207 (a) Livelyerimson-purple Yellow Viotebblue*
(b) Crimson-scarlet 0r~nge Violebblue, slightly
more violet*
55/31. C253 (a) 3'[agenta Yellow Blue*
(b) Tinged ~prieo~ Orange :Blue*

(o) genera~ observations.

Considering the results on the mutants together with those on the
normal ill regard to the relations of flavones and anthocyanins, it is quite
clear that the differences in flavone content are also accompanied by
differences in the amount and natm'e of cyanic content. In the first
place, where only one anthocyanin is concerned, gain of apigenin is
correlated with decrease in eyanin, and gain in yellow flavone with
decrease in pelm:gonin. Introduction of yellow flavone into pure cyanin
flavone-free types will cause the production of pelargonin partially or
wholly in place of the eyanin, together with a general decrease in cyanic
intensity. Secondly, gain of yellow flavone in flowers containing both
 W . 3. (I. LA}YRENCE AND I%OSE SCOTT-~[ONCI%IEFF 203
anthocyanins is accompanied by a general decrease in cyanic intensRy at
the expense of the eyaniu rather than the pelargonin. Conversely, loss of
yellow due to mutation or to the presence of tt is accompanied by decrease
in the proportion of pelargonin in the deeper pigmented flower. In one
instance (G 21 a) gain of ivory flavone in a mutant sector of a B I plant
is accompanied by a decided increase in the proportion of pelargonin
to cyanin. This has an important bearing npon the tlleory of pelargonin
formation which is to be discussed later (see p. 211).
It will be seen from Text-fig. 5 that the pigments of Dcd~Zic~ fall into
two series. Apigenin is associated with the two cyanidiu glycosides in the
bluish-magenta flowers of 1). Me~'clcii and D. i,m~)c~'ic~Zis, while the yellow
flavone is associated with pelargonidin in the orange flowers of D. eo,ro~a,tc~
and D. cocci~ec~. Combinations of the same two colour series are indicated
in ~he numerous types of D. va~'iabi~is. In this garden dahlia the ivory
and yellow flavones may be found together, as are also cyanin and
pelargonin. There is generally a balance in favour of one or the other
anthocyanin.

PAI%T III. A NEW TItEOI~Y OF FACTORIAL BALANCE AND

SPECIFIC PICSMENTATION IN DAHLIA.
(i) G~N~A~.
From a study of the genetical and chemical facts concerning flower
pigmentation in Dc~hlic~, three phenomena stand out clearly and demand
an attempt at biochemical explanation.
First there is the phenomenon of factorial eumulative action, demon-
strafed either by proportional increase in specific pigmentation or, where
pigment production is limited, by the second phenomenon of factor
interaction, which involves a proportional suppression in the co-produc-
tion of the other pigments involved. Thirdly, the production of cyanin
and pelargonin respectively does not appear to be controlled by specific
genetieal factors in spite of the presence of two general factors for non-
specific anthocyanin formation.
In this last respect Dcbhgc~ is quite unlike other flowers such as
A~zti~'~'hi~n m@~s (0nslow, 1925; Scogt-3{oncrief% 1930), Phcl,~'bil,is
Ni~ (Hagiwara, 1932), Pelc~go~,i~tm zo~cde 1 (Scott-Moncriett, 1931),
P~'im~&~ si~e~s~;s (de Winton and HMdane, 1933)and Pc~2)a,ve~" Rltoects,
67wi~'c~tt~,s ct~ei~'i and the polyantha rose (Scott-Noncrieff, 1935) in which
x The l%obinsons(1931c~)have sinceshown thM,the anthocyanins of Pcla.rfto~i.wm,zeal.ode
are neffpelargonin and cyaninbugpelargoninrindmMvin. This doesnot alter ~,heurgument.
Journ. of Genetics xxx 14-
204- Genetics and Chen~ist,ry of iFlower Colour i~ Dahlia
i~ has been found that individual factors control specific anthocyanin
formation.
]in Dc~h~4c~~he subble bMance in the formation, of both fiavones and
anthocyanins pomts to these pigments all being produced through some
common fundamental chemicM reaction, or from some limited, common
source from which all four pigments are at least in part derived, and for
which they MI compete. The supply of this hypothetical source must be
so limited that the quantity of each pigment derived from it can only
increase at the expense of the others, and depends upon the nnmber,
natm:e mid balance of MI the dominant pigment factors present, and
upon the relative clMms to existence made by the pigments formed, as
demonstrated by their specific interaction effects. Since this finding is
the basis of the theory we wish to advance, it must be examined and
tested carefully.
In D. sg,r~;abi~ispigmentation derived from the supposed AI magenba
ancestor is less intense, both qualitatively and quantitatively, than that
derived from the BY orange-sen:let one. This is clear not oNy from the
relative cyanic intensities of comparable A and B types (see p. 208), but
also from the existence of degrees of intensity in ivory and pale antho-
eyanin pigmentation as opposed to the maximum pigmentation found in
even simplex Y and B forms. It is also indicated by relative interaction
egects such as the relative suppression of A and B anthoeyanin pigmen-
tation by Y and I respectively, and the tendency of Y to suppress the
ivory flavone completely.
Except in the case of I and heavy anthocyanin, the interaction
effects of B, A, Y and I upon each other are roughly proportional to the
intensity of the respective pigments. At the same time it is also dear
that the interaction between flavone and anthocyanin factors resNts in a
far greater modification of anthocyanins than of flavones.
While yellow flavonc tends to secure a monopoly of the chromogen in
faetoriM bManees which include A and/or I, in the absence of Y ivory
pigment is formed from a cumulative but limited source at the expense of
A anthooyanin. In certain BY and B I types the effect of mntuM intm:-
action is apparent, the competition of both B and Y, or B and I, for the
same limited source causing a definite reduction in the amount of each
pigment produced, as compared with B, Y or I genoVpes.
Taking as a starting hypothesis the suggestion that the anthocyanin
and flavone pigments are produced from a common source of which at a
given moment there is only a limited supply avMlable, an examination
of ~he resNts obtMned wibh various genotypes shoNd give some indica-
 W . J . C. LAWRENCE AND [ROSE SOOTT-I~IONCRIEI~'F 205
[ion of the limits imposed, upon both the source and the pigments pro-
duced from it.

(ii) PIG-?c[ENT [BALANCE IN TEll A I TYPHUS.

The factors A and I are cumulative in action and produce, even in the
quadriplex condition, small amounts of pigment in comparison with the
orange-scarlet BY series.
The question arises whether the quantity of common source is con-
stunt for all A, AI and I flowers whatever their constitution, the cumula-
tive effect of th.e factors ])sing proportional rather than additive, or
whether [h.e source varies in amount according to ~he nature and number
of f~ctors presen¢.
To decide this point a study must be made of the relative concentra-
tions of ivory flavone and eyanin in the ivories and the rosy- and bluish-
magentas. In flavone-free flowers, light anthoeyanin production is
proportional to the number of A factors present, and is strictly additive
(see Plate VII, fig. 1). In the ivories, production of flavone is very slight
in simplex forms, while in duplex forms much pigment is produced[. This
quantity, however, is not increased in triplex or quadriplex forms,
pigment production apparently being limited to an amount equal to
that produced by I z. On combining the factors A and I it is possible to
get magenta flowers of varying depth of anthoeyanin colour and blueness
depending upon the number and relative proportion of these factors. The
cumNative nature of I in triplex and quadriplex forms is here shown by
eumNative interaction effects (Plate VII, fig. 1). In a certain AII 4 flower
anthocyanin was found to be entirely suppressed.
The possibiIity of a variable pigment source cannot be tested by a
comparison of cyanic intensities in A and AI genotypes, since any decrease
in anghoeyanin in the latter could be aeoouufied for by the known sup-
pressing effect of I. If, however, one compares AI and ! plants for ivory
flavone concentration, this shoNd be found to increase in AI plants if the
ehromogen source is dependent upon factorial contributions, and to fa.ll
if the source is constant, since in the AI plant it must be shared by the
two pigments present. To avoid complications due to the b~rely appre, ci-
able production of flavone by simplex forms, the duplex types AI, and
I~ were compared.
Equal. weights (0.5 gin.) of equal numbers of fresh petals (bases
removed) from flowers of the same age were macerated in 10 c.e. of
1 per cent. I{C1 a~d extracted with an equal volmne of ethyl acetate.
Two c.c. of this extract were shaken with an equal amount of saturated
1.4-2
206 Genetics and Chemistry of Flower Coleus" i n Dahlic~,
sodium carbonate solution, and the initial intensity of the colour reactions
compared. Confusion due to the later development of the maize reaction
in certain flowers was thereby eliminated. Two AIs bluish-magentas
A/321~ (double) and A/39 G (single) were compared with two 12 flowers
White Star (doable) and Everest (single) respectively. The flowers were
matched carefully for age since the ikovone content was found to fall
considerably as the flowers grew older. In no case was any decided
difference in ivory content found, if a slight margin was allowed[ for error
due to faulty matching. The possibility of the presence of an inhibitor of
ivory flavonc comparable to H also made it difficult to come to definite
conclusions, but the evidence was in favour of ivory I~ flowers containing
as much, if not more, ivory flavone than the corresponding AI~ magentas.
It is therefore suggested that the maximum pigment source avaih&le ~n tim
AI genoty2~es is ~'e(lui~'edfor apigeni,~ productio,~z by an 12 plant. Addition
of an aathocyanin factor therefore cannot increase this source, but
necessitates the division of the limited source between flavone and
anthocyanin, the former taking much the larger share. Since the pro-
duction of anthocyanin in the rosy-magentas is strictly additive, one may
deduce that &man& upon the source by the A factor do not exceed the
maximum limit (e~uivalent to that in I J even when A is in the ~ua&'@lex
state.
It may therefore be assumed that the factors A and I, when present
singly, have a cumulative effect upon the quantity of available chromogen
up to a limit represented by the production of flavone by I~ or antho-
cyanin by A,. When present together the proportion of each pigment
formed from this limited source depends upon the relative numbers of
each dominant factor present and their respective interaction strengths.
Since 11 has practically no effect upon pigment production, the case of
interaction in an AI, type, where pigment production should be below
the fixed maximum, does not arise. The fact that a single I factor in an
acyanic flower is nob sufficient to liberate enough source for appreciable
apigenin formation is also demonstrated by the fact that the interaction
in an h i 2 genotype is insufficient for the suppression of M1 anthocyanin
(see p. 177), while the plant A~I,1has much less cyanic colour. Since iu each
case the first I factor is without effect, the effective flavone-produeing
factors are one and three respectively, and we thus find less anthocyanin
suppression by this effective I factor upon one A factor in AtI ~ than that
of the three effective I factors upon the two A factors in A~I,~instead of
similar suppression in each ease.
 W. J. C. LAWlCENCE AND I~OSE SCOTT-!V~ONCI~IEFF 207

(iii) PZG~iENTBALANOEZN THE BY TYPES.

A study may now be made of the corresponding BY series. Although
iu the absence of the inhibitor t t there is no apparent difference in the
production of yellow flavone however many Y factors are present, the
cumulative effect of these factors is demonstrated by the quantitative
interaction of an increasing number of Y factors upon anthocyanin pro-
dnction; for example, Y~ has a greater suppressing effect upon cyanic
intensity than Y1. Similarly, the factor B produces a maximmn anaount
of heavy anthocyamn even in the simplex condition, and only shows its
cumulative character when in competition with other factors.
As in the AI series, the question arises as to whether the common
source competed for by B and Y has a maximma unchangeable value for
all B and/or Y genotypes, or whether the source is increased in the
presence of both factors. We have seen that maximum pigment produc-
tion is reached in the simplex B or Y plants respectively, and that Y has
a large interaction effect upon B anthocyanin as demonstrated by the
reduction of cyanic intensity from chocolate to scarlet through the
introduction of a single Y factor (see Plate VIII, fig. 6). In this connection
it is interesting to recall that Willstgtter Iotmd nearly 20 pet" cent. of
cyanin in chocolate dahlia petals as compared with 4-6 per cent. of
pelargonin in scarlet (BY) forms. Since Y has such a sh'ong interaction
effect upon B it is impossible to judge from any change in anthocyanin
intensity whether the total pigment som'cs available in BY flowers is
similar to that in B or Y flowers, or whether each factor contributes
separately to the common source, thereby augmenting the total pro-
duction of pigment. However, if the common som'ce is invariable in all
B, Y or BY genotypes there shmfld be more yellow flavone in Y than in
BY plants in which both factors come into competition for the same
limited source. Owing to mutual interaction, both pigments would[ then
be present in lower concentration than when occurring unaccompanied
by the other, the suppressing effect of If upon B probably being greater
than that of B upon Y.
Comparison of the yellow flavone content of comparable yellow and
scarlet flowers was carried out as in the case of the ivory flavone, eolour
reactions with saturated scdium acetate being compared as well as those
with sodimn carbonate. The former gave yellow reactions which were
easier to grade than the deep orange of the latter. As before, care was
taken in choosing comparable flowers since the petals lost both pigments
with age. Flowers were avoided in which ~he inhibitor t t was likely to be
208 Genetics a n d C h e m i s t r y of F l o w e r Colour i n D a h l i a
present. It was found that the BY types--4-2i/29 (Bi¥~Ii_~,) aud 42~/29
(B,YiIa_,l)--both had less flavone than petals from the full yellow B 331
(Yi-2). I~eductiou of flavone in the presmlce of heavy anthoeyanin was
as much as 50 per cent. in. the mosaic flower iV[~ (B'Y'~I~) ]}om which
s~ric~ly comparable normal scarlet and yellow mosaic petals could be
compared.
A comparison of crude acid solutions of equal fresh weights of petals
from purple (Bi_4) and deep rosy-magenta (A,l), of bluish purple (Bi_,iI~)
and deep bIuish magenta (A,iI~) and of orange (BY) and apricot (A¥)
respectively, showed that the cyanic intensity of all the B plalrbs was
about three times that of the deepest corresponding A types.
Therefore in all BY types both pigments are producecl from a constant
limited co~nmo'~t source, which is about th~'ee times as great as the q~zaxi~u~rt
source iu the AI types.

(iv) PIGMENT BALANOE IN THE BAYI TYPES.

In flowers carrying all four factors the AI and BY sm~ies are com-
bined, and we are dealing with two factors each capable of producing
only relatively small amounts of chromogen and pigment, ~ogether with
two factors which, even in the simplex condition, make use of as large a
pigment source as the plant is capable of supplying, however many
factors are present.
Since in any B or Y plant the maximum source is already in use, the
introduction of further factors cannot increase total pigment formation.
All the factors present therefore compete for a share in tiffs limited source,
according to their number and specific interaction capacity.
The in'~eraetion effect of I~_~ npon B is apparent in the remarkable
decrease in anthocyanin in the pale purples (BI=_,,) as compared with the
B or B I chocolates (see Plate VII, figs. 2 and 4), while the lesser effect of
B upon I is demonstrated in such families as ~18/31 (p. 181) by the Weaker
flavone reactions given by the B I cyanies as compared with the com-
parable series of ivory flowers. The mosaic ~{ 8 (B2Aa_~I2_~) also shows an
increase in ivory flavone in the ivory sectors as compared with the
normal ]?ale purple petals from the same flower, in which competition
for the limited source results in mutual snppression.
There is no evidence that B A Y I genotypes have a maximum pigment
source equal to the sum of the limit values found iu the AI and BY series
respectively. If this were so, no interaction could occur in BiI ~ forms in
which sufficient source should be available for the production of the full
amount of both these pigments. The cyanic intensil V of the pale purple
 W. J. C. LAWI~EN(]E AND I~OSE ~COTT-~ONC:RIEFF 209
BiI ~ type is, however, muoh less than that of the B i and BiIi purplish-
crimsons, while the flavone content is not appreciably greater than in an
12 type.
The effect of A in the presence of B is diglcult to gauge, since B is
epistatic, but it will be suggested later that this factor may i~ffluenee
pelargonin production in certain B 1 factorial balances.
The action of the inhibitor H is to reduce the productive and com-
petitive power of Y, probably at the source. Its effect appears to be
cumulative, and it may inhibit Yl-~t to such an extent that the production
of flavone is well below the usual Yi limit. In flowers combining the AI
and BY series, an increase in both ivory co-pigmentation and cyanic
intensity and a decrease in pelargonin may result from ~he action of I-I in
reducing the severiby of interaction of Y upon I, B or A.
It appears that in genotypes in which the maximum source for AI
types is increased by the presence of B or Y, the production of pale
anthoeyanin pigmentation and ivory fl.avone is still comparatively low.
This limib/tion is either due to the greater competition and suppressing
effect of B and Y or to the inability of A and I to produce more than a
limited amount of pigment whatever the quantity of source available.

(g) ~PEOIFI0 ANTtIOOYANIN PI~ODUOTION,

We now come to the question of factorial control in specific antho-
cyanin production. Throughout this paper both eyanin and pelargonin
have hifiherto generally been referred to under the common term
"anthocyanin." Light or heavy cyanic pigmentation might be clue to
either or bo{h of these pigments. Although {we factors A and B conb:ol
anthoeyanin production, these factors are not specific for any particular
anthocyanin. Just as the presence of any pigment in Dahlic~ has been
shown ~o depend firstly upon the presence of a specific factor and
secondly upon the degree of competitive interaction effects by the other
factors preseug, so in cyanic flowers we can show that the production of a
specific anthoeyanin depends for its presence upon an A or B autho-
cyanin factor, and for its specific nature upon the stun total of all the
factors present.
We have shown that in the light A eyanies, pdargonin is always
produced when Y is present, i~s relation to cyanin production being
inversely proportional ~o the degree of inhibition of Y by I-I. In ~he
absence of Y cyanin alone is formed.
Again in the .heavy B series, pelargonin is produced in proportion to
the amount of yellow fiavone presen~, but it is also produced in very high
210 Genetics and Chemistry of Flower Colour in Dahlia
proportion in the rosy-purples in the entire absence of Y. Pelargonin is
never found in the absence of both B and Y.
A careful examination of the records of specific anthoeyanin pigmen-
tation in fifteen B genotypes with non-yellow grounds, whose factorial
constitution was more or less certMn, showed the following interesting
classification. In the first column are all those genotypes in which any
pelargonin was found, whether with or without accompanying cyanin.
The possible presence of A in the pure cyanin genotypes is not excluded.

Forms produciW pdargoni~.

Constitution a n d colour Alkali reaction
a

31a/30 B=Ie, rosy-pro'pie :F~ed-violet

M 8 B~Aa_aI2=l, pm'ple :Blue-violet
C 18~ B,_.I~, v e r y deep magent~ Blue-violet
A46 ta Bl_2-&~I2_4, very dee]? magent~ Violet
]~ 40 ~ BzI2_l, v e r y deep magentt~ I~ed-violet
C 26 t B~_2(kl_e~)I2_a, pm'plish-crimson Violet
C 20 ~ BI_~(AI_~)I~ a, rosy-pro'pie Violet-blue
0 2 4 ~" BI_~(At_~7,)I1, purple Violet-blue
Forms producing pure cyanin.
30~/30 BlIe, pale purple
C 13 a B,I,a, pale pm'ple (violet-blue)
3D/30 BII], chocolate
3P/30 B I I 1, chocolate
013 ~ B 1, chocolate
C 13 ~ BlI0_ 1, pm'plish-erimson
C 16~" Bl_~Io_l, pm~plish-crimson

From this classification it appeared that exclusive cyanin production

is only found in B 1 forms accompanied by small numbers of I or A factors.
With increase in the number of B factors to two or more, or of I to three
or more, pelargonin was formed to a lesser or greater extent. Increase in
A factors also appeared to influence pelargonin production in certain
C~SeS,
l~eferring to the cyanin-pelargonin ratios obtained in the four
families recorded in Table XV, the following comparison was made
between these values and those calculated for specific pigmen~ produc-
tion on the assumption that pure cyanin is present in B~, B I l I _ 2 (~d
B~AI_~I~forms while 2)dargonin is Jbrmed in varying 2ro2ortions in B~,
tg~Ia_,~, tl~Aa_aI~ and B~A~_,~I~_~genotypes.
Pure cyanin Some or all pelargonin
_ _ A A

Family Parents Found Calculated Found Calculated

5]/3t It x BIII 22 22'0 i 0
5a/31 11 x B2I ~ 33 30'3 8 11'1
~s/31 BI~ x I,a 28 27'75 9 9"25
55, 56/31" B~A,,YtI 1 x I. 19 20'1 18 10'9
* Yellow-fl'ee forms.
 W . J. C. LAWRENCE AND ROSE SCOT~-MONCl~IEFF 211
A further ease examined was that of seven B plants with non-yellow
grounds in the progeny of two crosses of 42~/29 (B1YIla_~0 with White
Star (I~) and 411/28 (YII,,) respectively. All these were found to contain
pelargonin, the calculated ratios for B1YII,1being 7 : 1 and for B1YIIa being
1 : 1. The constitution for ~22/29 is as likely to be B~YII,t as B~YII3.
Considering the fact that the exact factorial limits controlling specific
anthocyanin formation have been postulated somewhat arbitrarily, and
that many of the anthoeyanin extracts were tested without preliminary
clearance of the flavones (under which circumstances the presence of
pelargonin might be obscured), a very satisfactory fit was found in these
ratios. The validity of the assumption was therefore tested further by
studying the ivory flavone concentration in B 1 pelargonin types, and the
pigment changes in certain mutant petals. The question of flavone con-
tent in B, genotypes was complicated by the small but definite inter-
action effect of B upon ivory flavone formation (see p. 181). It is also
possible that pelargonin production had a different interaction effect
upon apigenin than had cyanin.
Comparison of all the eyanin and pelargonin forms, with and without
ivory, which have been tested showed 52 ivory forms to 58 flavone-free
eyanin forms, as compared with ~I1 ivory to ]7 flavone-free pelargonin
forms. These figures are in favour of the suggestion that, generally
speaking, pelargonin forms are more often accompanied[ by ivory flavone
than are eyanin forms; several flavone factors being required for pelar-
gonin formation in types simplex for B. In the B~I 2 form 31~/30, men-
tioned above, pelargonin production would be independent of I, and
interaction effects upon I would not be encountered.
Evidence was next taken from the somatic mutations recorded in
Table XVIII. The first case of interest is the deep purplish-crimson CJ21s
which mutated to purplish crimson with increase both of ivory flavone
and of the proportion of pelargonin. Unlfl~e C14-1~ and 0207, no yellow
flavone was present in the mutant sector to account for increase in
pelargonin. The probable factorial constitution of 012 s is B~_oA,_ofl.
Since on our scheme this genotype would be at ].east on the border line
for pelargonin formation, we predict that addition of an I factor should
increase the proportion of this anthocyanin.
On the other hand iu the flowers C 13 s, C 17s and 02611, pigmented by
pure cyanin and probably simplex for I, an increase of one ivory factor
in the mutant petals would be insufficient to cause pelargonin production,
the factorial constitution still being below the B! a limit. The addRion of
an ivory factor to 301/30 (BI~) would be expected to give pelargonin in
212 Gene~ics and Chemistry of Flower Colour in Dahlia
the mutant sector, but tmfor~una~ely the green reaction obtained is
ambiguous. Since 0261 (Bi_~Ai_2Ia_4) Mready contained pelargonin the
increase in flavone in the paler mutant shows little extra effect.
Loss mutations for yellow flavone were found in purple and magenta
sectors on the orange flowers of B l0 a, B ~tl4 and zt22/29. There the cyanic
reactions in all cases showed a fall fit the proportion ot! pelargonin present.
I t is possible, and in the case of 42~/29 certain, that these B i plants had a
constitution for I of more than two factors. This w e n d account for the
reactions of the mutant sectors showing the presence of some pelargonin
in spite of the loss of Y. On the other hand the crimson flowers of
B ~122 (B[Y,,II_~H.0 had a deep magenta sector giving a reaction for pure
cyanin. The presence of pelargomn in the tinged apricot sector of the
magenta @25a is probably masked by the larger proporbion of cyanin
present--a situation which often occurs in the deep apricots, hi which
the proportion of pelargonin is small owfl~g to the yellow flavone being
partly inhibited by If.
It thus appears that pelargonin production depends upon the
presence of A or B in one of the following factorial balances:
(1) A or B together with Y.
(2) Two or more B factors.
(3) One or more B factors together with three I factors.
(4-) One or more B factors together with (a) two or more I factors and
at least one A factor or (b) one I factor and at least three A faefiors.
It remains to deduce what similar influence upon pelargonin metabol-
ism all these different factorial balances can have. Why should AY, BY,
B~_4, BlI a and EiAiI ~ flowers all apparently be capable of producing
pelargonin while AIo_~, BiAo_~, Bili_ 2 and BiAlI z genotypes can only
produce cyanin ?
We have ah'eady concluded that the pigment source, dependent up to
its maximum limit upon the dit~erent factors present, is competed for by
them all, and subsequently goes towards the proportional formation of
the specific pigments which they represent. We now see from the gone-
types above that when an anthocyanin factor is present pelargonin is
formed e~bher in the presence of Y, or when comparatively large numbers
of factors are present. We have also ah'eady noted that cumulative pig-
ment production is apparently only possible up to the limits of maximum
source available. A only reaches this maximum in the quadriplex state,
whereas I does so in the duplex and both B and Y even in the simplex
condition.
Thus in AY, BY and B~_~ plants the ~)ole~tia,l abiliV for pigment
 W . J. C. LAWI~ENCE AND I~OSE SCOTT-MONOI{IEFF 213
formabion must be greater than t h a t act~tal[y required to liberate the
maximmn amount of source the plant is capable of supplying. A sugges-
tioll as to the signific,~uee of bhis l)ote~tial ability and the nature of this
source will be discussed later (see p. 216).

(vi) ~AOTOI~IAL 001gI)YrIONS GOVEI~NINC¢ SPEOIFI0 PICI-NE1WrATION

IN Dahlia.
The following scheme to explain interaction and specific anthoeyanin
formation is based upon the deductions for which we have given evidence
in-the foregoing pages.
Let us suppose tha~ the limit of pigment source which can be made
available in Dahlia whatever factors are present can be expressed am six
act~tal units. All the values to follow are based upon this arbitrary
:figure. In the BY series these six units are made use of even in simplex
forms. Since cyanic intensity in the A series is approximately one-third
t h a t of the B types, the maximum source available in the A I series must
be limited to only bwo units, maximmn production of light anthocyanin
only reaching this value at A 4 and t h a t of ivory flavone at 12.
Let us also suppose that the factors B, A, Y and I are each capable of
contributing a specific number of 2)otentiaZ units, the maximum quantity
of source aca~ally available being limited to six or two. These specific
values based upon the arbitrary number six for maximum source pro-
duction, can be deduced from the following facts:
(1) B and Y make use of the maximmn sb: lmits of source available
when in the simplex condition. Therefore B and Y must each contribute
six or more potc~tia~ units, t t apparently has a direct reducing effect
upon the potential unit value of Y.
(2) A has a cumulative potential unit v a h e which in the quadriplex
condition is less than or equal to the maximmn value for available source
in the A I series ( = 2).
(3) While 11 has a potential v a h s below this m a x i m u m value for All
plants, t h a t of L, is eqnal to it, and those of I a and 14 exceed it.
(4-) All A and AI types produce pure cyaniu and AY and BY and Be
types pure pelargonin. Whereas BII ", and B1A~I1 types produce pure
cyanin, in Blla, BIAII 2 and B1A3I 1 plants pelargonin predominates. The
number of potential units essential for pelargonin production must
therefore lie between the values of the contributions of B~I2 and B~A~I~
and those of BII a and B1AaI~, B~A~I~, and these last two genotypes must
contribute less than AY, BY or B~, while A,II4 must contribute no more,
or less, t h a n B~I~.
214 Genelics a n d Chemist~'y of F l o w e r Colou,r i n D a h l i a
It will be fotmd that the unit Values which fit satisfactorily with the
above conditions are Y = 9 or more, B = 6 , I = l and A=½, the critical
value for pdargonin production being 8 (see Text-fig. 6). Thus:
A,~Ij = 6 units ]
B1 = 6 " t Can o n l y p r o d u c e c y a n i n
BIL " = 8 ,,
B1/LII= 8 ,,
A I Y 1 = 9½- or m o r e u n i t s
B1Y 1 = 1 5 ,,
B I A z l ~,= 8½ m i l t s
B 1 A a I I = 83c ,, Can p r o d u c e p e t a r g o n i n
BI 3 = 9 ,,
B., =12 ,,

Except for the two relative values for the limited source, these units
have been calculated independently of pigment concentrations and inter-
action -values, but it is significant that they appear nevertheless ~o bear a
close relationship to these as well. For instance, the specific maximum
suppression of the anthocyanins by the flavones found experimentally can
be expressed by the relative potential values of our scheme. Thus
Y/A=9/½-, I/A=e/I, Y/B=3/2, I/B=l/6.
Although we can only determine the minimum possible potentiM
value for Y, it is unlikely that it is much higher than nine if the inde-
pendent evolution of these interaction vMues has any significance. The
m a ~ m u m possible value must be well below twelve, since this figure
would give Y/B = 2/1 and we know that the suppression of B by Y is not
equal to but definitely less than that of A by I.
This theory of specific pigment production in Dahlia may be sum-
marised as follows:
(1) The potential unit values of the fou~" coleus' factor's are: Y = 9 o~'
,mo~'e, B = 6, I = 1, A = ½.
(2) A sum total of six units is su~cient to p~'oduce the maximum pigment
sou~'ce which plants ca~'~'ying B o~' Y a~'e capable of supplying. In AI plants
this maximum is fu~'the~" limited to two units.
(3) Each dominant factor' can cont~'ibute towa~'ds the 2~'oduction of this
common sou~'ce acco~'ding to its potentiM unit value; the supply of sou~'ee
being limited to two o~"six actual units whatevc,r the ~n~,mbc~'of potential ~ d t s
available.
(4-) Each dominant factor competes for the co'tureen sou~'ce i~ te,rms of
potential units, the amount of each pigment p~'oduced depending upon the
amount of sou~'ce available, and upon the balance a~'~dpower of i'r~te~'aetion
of all the factor's conce~'ned. Total pigment 2~'od,actio,~ cannot exceed the
limit of 2 or 6 actual units.
 = g ~- ~, ~- ~.
~.~ ~' ~ ~
°~'~~ . ~ ~ .
nr- •

U,
gl t ! i ~~ t ' '1" ; t 1' * ~ " '~""1 ,.~1 I I
~N N
r~
g
g I.~ •
a?. 4.
~.~
W > ~,'icdi u m rosy-magenta
.,~
p ~ Deep rosy-magettta
,n •

? g Very pale ivory

c

o
--I ='
t~
Deep ivory
c~

I--I "~
m
Pale bluish-magenta

o
5 > --I Pseudo-ivory
>~.

~/ Medium bluish-mageata

../), > Yellow

g ;-/ '-'-'///-'/-'-*~F~¢¢"/" "5~5"~552552~, --> Pseudo-yellow

g
g ,///~.////,
///'// // ~. ~ prieot

i~,./~.. __
~;: :
~ ? ;-< ~5 I~ '

> prieot-magenta
n

~, - .- > ~? ? / ' > 7 -"/~ . . . . . . "~

~g :-J -/.- .~ ".5, I > Apricot

NNNN > Deep crimson-purple

o ~5 Deep purplish-crlmson.
e~
go
b~
Pale tmrple

I
Rosy-purple
m- m
i=z,

Rosy-purple

cd
~ .?, i55~fS~5"./111/N _ z ~ Scarlet

o
L~ --

/~#~
~
.

. / /// / i ///// . /, /. ./ t/ ~/ / / / I / / //~//~/ i.~,

: / ~ l ~ . * s e ~ .~.l:.x. ~ l _ e ~c; /..,. z . / _ . / _ / . ~ . . . . . 7" --
i --2~
.~
~ Scarlet

7~ ~ ] Crimson
>
.k~ctlow
. . . . .4.

~.//.~//.~ . . . . . .. ~,.,~>~ _ _ ~ ~, . > ~,-,~

..- ~ _ _ . . ~ _ _ _
,,_~ Yellow

o ,-,, ~ 5", N
]Potcn linl lllllt~ .hctuul tlnits

Total pote~*tialfactorial eontribul.lonn Actnal'specillc lfign~,ent Flower eolour

production
 W . J. C. LAt,VICENCE AND I~OSE ~COTT-~J[ONCR,IEFF 215
(5) w govc~'ns the productio~ of yellozv flavone, cbnd I the production of
the ivory flavone apigcnin. The i~hibitor I-I prog,ressively reduces the potential
unit value of Y,
(6) In the presence of A or B.
(a) Pu~'e cya~in is produced when the sum total of potential units is
less than, or effua~ to, eight;
(b) Pda~'gonin is produced exclusively or in part when the sum total of
potential units exceeds eight.
Thus, in effect,, pelargonin production is only possible in the presence
of 13 or Y, since it depends upon an excess of more than two pote'nticd
units over an actual unit value of six. While Al_,lI0_~, BlIo_2, B1h0 ~I1
can only produce cyanin, Al_4Yl_,I, Bi_,iYl_,~, BlI3_,~, Bzhl_,iI2_,~,
B1Aa_,II1 and B2_,tI0_~ can produce pelargonin.
Text-fig. 6 represents the potential unit contributions and the aetucd
resulthlg proportions based upon the direct observation of specific
pigments formed in twenty-three different geuotypes. The specific
factorial contributions of A, I, B and Y are represented at the beginning
of the list. Any excess in the sum total of potential over the actual units
of limit in available source (2 or 6) is represented in broken line. When this
excess occurs factors compete according to their respective ability, and
the mutual interaction which results is demonstrated by the relative
production of each of the pigments concerned.
These diagrams shoNd be compared with the list of cyanic intensities
in AI genotypes listed on p. 177, and with the accruing of interaction
between Y alad A or 13 on p. 181.
The first seven genotypes demonstrate thelarge suppression of A antho-
cyanin by I2_4, a suppressioll which is complete in AtI ~ plants. Decrease
in flavone production as compared with an I~ ivory is slight.
I~_4 also has a high interaction value in BI~_~ types, but never sup-
presses 13 anthocyanin formation by much more than half. Ivory
flavone suppression by B is comparatively small. Since the exact inter-
action values in BI forms are nncertain, the actual pigment values given
for these forms are subjec~ to correction.
In BY tyl)eS the results of mutual interaction are very extreme owing
to the large excess of poten~icd unit values over the limit of six actual
units of source.
When more than two potential units in excess of six are present, the
anthocyanin is chiefly pelargonin; the small proportion of cyanin which
remains depel~ding upon the number of these excess units. In such gone-
types as AY, 1357 and B 2 pelargoniu formation is practically complete.
216 C~enetics a~,d Chemist~'y of Flower Uolou~" i'~ DaAlia,
The exact quantitative effect of tt upon Y is unknown. As the effect
of l~I in reducing the amount of yellow flavone increases, the proportiot~
of pelargmfin to cyanin falls. I-I therefore apparently reduces the
potential unit contribution of Y sufficiently to bring the total potential
units of the :flower to a value near or below the critical value 8, and
mixtures of the two anthocyanins result.
The suppression of ivory flavone by Y is also demonstrated.
The unusual phenomena concerned with pigment formation in DahZic~,
can therefore be expressed by means of this scheme. When the potential
unit values exceed the actual values of available source, competition mush
occur with subsequent interaction and suppression. The limitation of i

cumulative effects upon pigment production to those A and I genotypes

whose total potential, ~mit contribution is equal to or less than the ackt~a.1
milts of available source is also explained. Furthermore we are able to
express the correlation between A and[ I and between B and Y, the fall
in pelargonin content whml the potential value of ¥ is suppressed, by l~I,
and, finally, the fact that the A and B factors in contributing to mass
pigment production may automatically control specific anthocyanin
formation though not themselves specific factors.
Further breeding experiments are being undertaken with a view to
tenting this new theory.
In regard to the facts of interaction and the specificity of anthoeyanin
production, we would draw attention to the physiological theory of
heredity advanced by Goldschmidt (1927), who regards the precise
characteristics of the phenotype an being determined by differential
reaction processes originating in the genes but carried on at different
rates.

( v i i ) A BIOCHEivIIGAL THEOI~Y FO]g SPEOIFIO PIC-~I]gNTATION IN .Dc&]~$f&.

Such a scheme as this cannot be put forward without at leant ven-
turing a plausible biochemical explanation for our theory of a limited
common pigment source and dependence of specific anthoeyanin produc-
tion upon a general factorial balance.
Very little is known of the source of ant]]ocyanin pigments. The earlier
view that flavones were their precursors has been mtled cubafrom lack of
evidence of suitable relationships between correlated pigments. Our
evidence for the emstence of a common pigment source points to the
production of specific pigments being simultaneous rather than sequential.
The lace Nrs Onslow (1931, 1932) made the interesting suggestion thal~
condensation of deaminated amino acids might be responsible for pigment
 W. J. C. LAW]~]~NCE AND ~OSE ~COTT-IV[ONC~IE:~:_~ 217
production; and she also suggestedthat oxidation in the side phenyl ring
might depend upon the hydrogen ion concentration of the cell sap, since
acidi~y is known to inhibit oxidation in the or~hoposition in such com-
pounds. The l%obinsons (1933 (b)) state that they have fmmd a progres-
sive rise in the/)I-I of the flowers, calyx and leaves of a certain nasturtimn,
these being pigmented by glycosides of pelargonidin, cyanidin and
delphinidin respectively. In certain cases oxidation of the phenyl ring
may therefore be dependent upon the reaction of the cell-sap.
In Dcbh~icbwe have shown that in many genotypes the potentiality to
produce pigment is greater than the possibility of chromogen production.
One at least of the materials required for pigment synthesis must be
common to both flavones and anthocyanins, and must be limited in
quantity. The other constituent is mllimited, its quantity, variation and
such structural modification as affects the nature of the condensation to
glucosidal anthocyanin (pyrane) or to either of the sugar-free flavone
(pya'one) types depending on the specificity, number and balance of the
various factors. Cumulative pigment production is found in A~_,~ and
Ii_ ~ genotypes, where it is dependent upon these additive factorial con-
tributions which in these plants never exceed the maximum available
supply of the limited precursor. In most other types this supplyis exceeded,
hut further pigment production is curtailed by the absence of a sufficient
equivalent of the limited component. The maximum available supply of
the lagfier is three times as great in the BY as in the AI series, but is
otherwise unaffected by the factors present. With anthocyanin produc-
tion, accumtflation of sufficient excess of the unlimited component
appears ill. some way to cause, either before or after condensation, an
inhibition of oxidation of the side phenyl ring in the ortho position 3',
pelargonidin instead of cyanidin glucoside being formed.
We suggest that when the pote~tia~ unit value exceeds an aca~cd
value of six by more than two units, the accumulation of this surplus
precursor might affect the reaction of the cell sap in such a way as to
inhibit oxidation of the phenyl ring. An investigation of p]~I variation in
these flowers is being undertaken. Although in some cases pelargonin
pigmented flowers appear to have a lower/)I-I, the evidence ag present is
by no means completeL
A study of the structure of the six pigments occurring in the Dcddia
species (Text-fig. 5) shows that, apart from the uncertain formulae given
:for the yellow flavone, the left half of each molecule is similar throughout
a ~,eeent l)II estimations on Dahlia, genotypcs reveal no connection between acidity
of the cell s~p ~md polarg'onin formation.
218 ~e~etics a,~d Ul~emistrg of F_lowe'r Uolo~cr i~ .Dcd~lic~
~he series, while ghe righ~ half only varies in the presence or absence of a
hydroxyl at 3'. Either half of the molecule may therefore be the one
which is supplied by the limited precursor common ~o all the p~gments,
oxidation in the ortt~o position in cyanic types depending upon the
absence of a large excess of the other unlimited component, wlfichever
and whatever this may be.
We pug this suggestion forward as a purely hypothegieal explanation,
which at least demonstrates a plausible line of attack upon this com.pli-
ca~,ed biochemical problem.
Some metabolic significance may underlie ~he fact that the ~wo
flavones in Da,h~ic~, are both non-ghcosidal, while the anthocyanins are
ghleosides. This absence of a sugar residue at positions 3 and 5 may have
some connection with a different form of condensation which results in
t,he formation of a pya'one instead of a pyrane ring. The non-glycosidal
nature of these pigments might also be connected with ~he greater inter-
action effects of I and ¥ as compared wi~h A and B.
The fact that monoglycosides of pelargonin and cyanin can also occur
in DcdaZicb may be significant. In each case they occur alongside or in
place of the corresponding diglycoside, which is the more usual form.
The chemical relationship between co-existing pigments shows no
evidence of their production being sequential. Pigment specificity is
probably determined at a very early stage. The maize rea.etions recorded
are possibly connected with the presence of a tannin, which may also be
concerned in pigmen~ production. Karrer and Schwartz (1928) mention
a high moIeeNar tannin in ~hs flower of a salmon-coloured Dc~h~ic~.
Proof has at last been obtained of the existence of "leuco-antho-
eyanins." The 1Zobinsons (1933 c~) have extracted them fi'om various
plants and seeds, and by treatment i~ ~it~'o with strong acid have pro-
duced from them true anthocyanidins. They have not yet de~ermined the
nature of this leuco compound, nor decided what r61e, if any, it plays in
anthocyanin production in the living plant.
In this connection it is worthy of note that while working on the
pigments of D. eo~'o~zcttawe obtained interesting ether-alcoholic solutions
from which all anthocyanin had been precipitated. They contained
yetlowflavone and other unidentified substances and formed a red pigment
after standing for some time in bright sunlight. The pigment appeared
to be a glycosidal flavyllium pigment of simple anthocyanin type. On
hydrolysis ig yielded a sugar-free pigment. We hope to investigate this
phenomenon further.
We have postulated (Text-fig. 1) that Dc~h~ic~ ~;e~'iabigis is a double
 W . J . C. LAW~,ENCE ~ D ROSE SCOTT-I~([ONCI%IEFF 219
autotetraploid, each o[ the 8 chromosome types consisting of two
quadrivaleut sets which, although similar, are too unlike to pair with one
another as a rule. Thus both A and B produce anthocyanin but B in
much greater quantity. I and Y both produce flavone but the two
pigments are qualitatively distinguished. It is also probable that the
yellow inhibitor, I-I, is represented in the other quadrivalent chromosome
set by an inhibitor of ivory (see p. 166). In other words the chemical
differences between the similar genes (c. 9. I and Y) and the processes
they govern, are an expression of the structural differentiation which
prohibits chromosome pairing between the homologous quadrivalent
sets.
It is possible that the genes I and Y and A and B respectively have
differentiated one from the other or from a common ancestral gone.
We suggest however that a simpler view not at variance with the facts
would be that the differences between cyanic1 and pelargonin, alid
bet~yeen the ivory and yellow flavone, in addition to the unique pheno-
menon of specific anthocyanin production, can be explained on the basis
of a single mutation very early in the evolution of Dc&l~c~, this mutation
involving the decrease of the actual value of the common source from
6 to 2 units, or vice versc~.
In the first place the synthesis of the pigments in Groups I and II
respectively must be along identical or very closely related lines, since
our ex-idence clearly shows that they are derived from a common source.
Secondly the fact that the dilute pigments A and I and cyanin produc-
tion are characteristic of Group I, and the intense pigments Y and B and
pelargonin production (which is dependent upon an excess potential unit
value) are characteristic of Group II, strongly suggests that this paral-
lelism arises from a single basic difference between the groups.
In regard to the phenomena of specific anthocyanin pigmentation
described in this paper, it is probable that D. variabilis is an exceptional
case, the peculiar conditions for which arise fl'om the high degree of
polyploidy and hybridity found in this species.
The occurrence of various types of bManced pigment production from
a common source is now being recognised in other genera (Pc~pavc~" Rhoec~s,
Pela~'goni~r~ zoncde, P~'imulc~ si~ensis, ~St~'el)tocr~'~)~s, etc. (Seott-Moncrieff,
1935)). There is no doubt that this situation is the rtfle rather than the
exception.

Journ. of Gono~icsxxx 15
220 Genetics and Chemist~'y of Flowe~" Coleus" in Dahlia
1RECAPITULATION.
In view of the complexity of the data presented, and the length of the
account; we have thought ifi advisable to recapitulate the leading facts as
briefly as possible.
(i) GE~E'rIOA~.
With one exception, Dc~hl'ic~species may be divided into two distinct
groups for flower colour, Group I with magenta or ivory flowers,
Group II with orange-scarlet or yellow flowers. The exception, the garden
dahlia D. vc~'iabilis, combines both colour series within itself. Cytological
examination of the six species in cultivation shows that five of these are
allo-tetraploids and the sixth, D. va~'iabilis, an allo-octoploid. The above
facts in conjunction with the evidence from breeding strongly suggest
that D. v~'ia3ilis arose as a hybrid between members of each of the two
flower-colour groups, one with magenta and the other with orange-
scarlet flowers.
Flower colom: in Da,ldia, is the expression of two series of soluble sap
pigments, flavones and anthocyanins. In Group I these pigments are of
light intensity and are the ivory flavone apigenin and a glucoside of
cyanidin respectively. In Group II the pigments are of comparatively
heavy intensity and comprise a yellow flavone and an anthocyanin of
the pelargonidin type. All four pigments may occur in D. va~'iabilis, in
varying proportions and degrees of intensity, in both light and heavy
forms.
~'our main factors govern flower colour in D. vc~'iebiIis, two for fiavone
and two for anthocyanin production. A is necessary for the production
of light anthocyanin pigmentation by either cyanin or pelargonin; B is
necessary for the production of heavy anthoeyanin pigmentation by either
cyanin or pelargonin; I produces ivoryflavone; Y produces yellow flavone.
Each factor is tetrasomie and gives the characteristic ratios arising from
the random pairing of four homologous chromosomes. The expression of
these four factors, however, is di~erential. Y and B are completely domi-
nant in the simplex forms. A is cmnulative from simplex to quadriplex.
I simplex produces very little pigment and is usually in.distinguishable from
the nulliplex form. I~L is saturated and of the same intensity as I3i~ and I~.
In consequence of the incomplete dominance of I, modified tetrasomie
ratios 1 : 5, I : 3, 1 : 1, 3 : 1 and 11 : 1 are obtained. A fifth tetrasomic
factor I-I is cumulative in effect and progressively inhibRs the formation
of yeltow flavone, titus giving rise to the intermediate cream and primrose
colours. There is evidence also for an ivory flavone inhibitor.
 W. J. C. LAWICENCE AND ~OSE SCOTT-MONOICIEFF 221.
Pronounced interaction occurs between the flower colour factors and
results in partial or complete snppression of the pigments, the precise
degree of suppression in all cases depending upon the total number and
proportion of the fower colour factors present. In general the flavones
suppress the anthocyanins much more strongly than site verscb.
The action of Y upon I or A is very strong, complete suppression of
ivory flavone and pate anthocyanin respectively occurring in certain
genotypes, while Y and I together are associated with complete sup-
pression of heavy anthocyanin (B) in given genotypes. The action of I
upon A is also strong, complete suppression of light anthooyanin occur-
ring in at least one genotype; its greatest effect upon heavy (B) antho-
cyanin pigmentation however is to reduce it to the intensity of light (A)
anthocyanin pigmentation. I has no apparent effect upon yellow flavone
unless the latter is already diminished by It.
Though B and Y are completely dominant in the simplex condition,
and I2i2 is saturated, these factors are seen, from their interaction effects,
to be potentially cumNative from simplex to quadriplex in the case of
B and Y and from duplex to quadriplex in the case of I.

(if) OnE~mAL.
The chief anthocyanins of Dc~hlia are the 3 : 5 diglucosides cyanin and
pelargonin (Willst5tter and ]Kallison); the flavones are apigenin and a
yellow isomeric tri-hydroxy compound not yet satisfactorily identified
(Schmid, etc.).
It has now been established that two monoglycosides of cyanidin and
pelargonidin also occur, though less fl:equently. Cyanidin 3-monoside is
present in the disc florets of D. Merc/cii and also often accompanies
cyanin in the chocolates, etc. Pelargonidin 3-monoside is only found in
D. co,ronata, where it is the sole anthocyanin present.
These six pigments have been identified, and their occurrence,
whether alone or together, has been studied over a large range of geno-
types by means of quick qualitative tests.

The pigment distribution in the species, varieties and mutant sectors

of Dah~ic~ is found to fall into two distinct colour groups. Although either
anthoeya,nins or anthoxanthins may occur alone, the presence of yellow
flavone in the apricots, oranges and scarlets is cdways accompanied by a
pelargonidin derivative, while ivory flavone alone is associated with the
two oyanidin glycosides in the magenta, purple and chocolate series.
An important exception was found in certain B rosy-purples which are
i5-2
222 Genetics and Che~nist'ry of Flower Colour in Datdia
deeply pigmented with pelargonin in the entire absence o~ yellow pigment.
Predominance of cyanin over pelargonin in the presence of yellow
flavone only occurs when this pigment is considerably suppressed by the
inhibitor It. An inverse correlation has been found in the co-production
of eyaniu and apigenin and also of pelargonin and yellow flavone.
Ivory flavone acts as a strong co-pigment in flowers pigmented with
cyauin, which it makes bluer in tone. Its action on pelargonin is con-
siderably weaker. Yellow flavone has no co-pigment effect on either
anthocyanin.
There is no evidence of the existence of specific factors for the
particular anthocyanins.

(iii) TmEO~m'IOAL.
The following three phenomena have been observed in our study of
Dahlia :
(1) ]Factorial cumulative action.
(2) Interaction effects involving pigment suppression.
(3) Specific anthocyanin formation by non-specific factors.
Balanced pigment production points to a limited common sotrrce for
bo~h anthocyanins and flavones, while interaction and suppression
indicate that this source is limited in such a way that it is competed for
by all the factors present, the actual proportions of pigments produced
depending upon the specific claims of the various factors.
The relative mtensity and interaction values of the flavones and
anthocyanins were estimated from a study of the limits of additive
pigment production by a single factor, and of proportional pigmentation
when interaction had occurred. In the AI types pigmentation is less
intense than in the BY. Each A and I factor contributes to the con-
version of common source into light anthocyanin, or ivory flavone,
respectively, until the limits of available source are reached at A,~ or
I2i~, factorial cure,dative action being represented by interaction a~id
subsequent pigment suppression when the potential ability for pigment
formation exceeds the available source. In the BY types, heavy antho-
cyanin or yellow flavone production is at a maximum even in B~B3 and
¥1Ya forms. Here proportional cumNative interaction alone occurs, both
flavone and anthocyanin intensity being less in a scarlet ]BY flower than
in the corresponding purple By and yellow bY types.

The case of the rosy-purples has shown that it is not only in the
presence of Y that pclargonin is produced in preference to cyanin.
 W . J . C. [LAWI:~ENCEAND ][~OSE SCOTT-MOlgCI%IEFF 223
A genedco-ohemical study of the a.n~hocyanins in fifteen known
By genotypes, together with that of certain somatic mutations and the
pigment distribution ratios in fern: B y families, shows that pelargonin
production in these forms depends upon the presence of more than one
B factor, or one B factor together with a sufficient number of A or I
factors. Thus BB3, BbaIi3, BbaI2i.z, BbaAaalia and BbaAoaeIi~ can only
produce eyaniu, while pelargonin oecm:s in such types as B~b~, BbaIai ,
BBaA~!~i,) an([ BbaAaaIiia.
If the maximum limit of available source in BY types is given the
arbitrary value of 6 units, and that of the AI types the value 2, then from
a study of the limits of actual pigment formation and specific anthocyanin
production in known genotypes we can deduce the specific l)otenticd
factorial contribution of each factor, a value which is found to resemble
their relative interaction values. These unit values, calculated from
practical data, are: Yi = 9 or more, B 1= 6, I~--1 and A i = ½. Pelargonin
replaces cyanin pigmentation to a lesser or greater degree whenever the
y)otential unit value exceeds an actual source value of six by more than
two units, a situation which can only occur in the presence of B or Y.
The scheme in Text-fig. 6 expresses a series of genotypes in terms of these
~)otential factorial contributions and shows the interaction values, limits
of available source, and the critical value for specific anthoeyanin produc-
tion, together with their influence upon the nature and proportions of the
acaeal pigments found in these genotypes.

We suggest that the pigments occurring in the Dahlia species (Text-

fig. 5) are each produced by condensation of two plant substances. One
of these precursors is strictly limited in cluantity and takes part in the
synthesis of all the pigments, while the other is ~mlimited and is dependent
upon the specific action of the various factors for its quantity, variation
and the nature of its condensation with the limited component to form
flavone or anthoeyanin. The amotmt of each co-produced pigment is thus
proportional to the specific factorial competition.
In cyalfic flowers accumulation of a certain excess of the unlimited
over the limited component appears to cause pigmentation by glucosides
of pelargonidin instead of cyanidin, through inhibition of oxidation of
the side phenyl ring at 3'. The nature of this inhibition and the identity
of the two precursors can only be conjectured.
An interesting comparison can be made between these resNts and
Goldsehmidt's physiological theory of heredity.
224 Genetics a n d C h e m i s t r y of F l o w e r Colour in D a h l i a

Our ghanks are d u e ~o Prof. a n d h{rs P~obinson for t h e i r iageresg a n d

h e l p f u l advice, ~o Prof. J. B. S. H a ] d a n e for v a l u a b l e criticism, a n d be
t h e D . S . I . : g . for a g r a n t go one of us w h i c h e n a b l e d ~he c h e m i c a l side of
t h i s w o r k ~o be underta,ken.
F o r ~he coloured d r a w i n g s we are m u c h i n d e b t e d go i~r H. C.
Osters~ock.

REFERENCES.

iBrJ~l~]~sL~]~,A. F, B~LLINC~',J. and ]LU~Nmt~r,]K. E. (1923). "Inheritance of totraploid

Dataras." Bet. Gaz. 76, 329.
BI~IDC'F~S,C. B. and ANm~RSON, E. G. (1925). "Crossing-over in the X-chromosomes
of triploid females of Drosophila meh~nogaster." Genetics, i 0 , 418.
D~ ]¥I~'J'oN, D. and HALDA!giE,J. ]~. ,S. (1933). "The genetics of Primula si~ensis.
Segregation and interaction of factors in ~he diploid." J. Genet. 27, 1.
C40LDSOm~IDT, I~. (1927). Physiologischs Theorie der Vererbang. Berlin.
HAGrW~-~, T. (1932). "On the gonetieo-physiological studies of the colom'-dovelop-
monb of flowers in Pha.rbitis Nil." Prec. imp. Acad. Japa~t, 8, 54.
H~J~DA~E, J. B. S. (1930). "Theoretical genetics of autopolyploids." J. Goner. 22,
359.
I(Anrcn~, P. and SOmVaX~TZ,I(. (1928). "Usher Pflanzonfarbstoffo. IX." Holy. chim.
Acta, t i , 917.
L ~ w ~ o ~ , W. J. C. (1929). "The gene~ies and cytology of Dahlia species." J. Gcnet.
21, 125.
- - (1931). "The genetics and cytology of Dahlia variabilis." Ibi& 24, 257.
- - . (1932). "Interaction of flavones and anthoeyanins." Nature, Lond.,i29, 834.
hivnLEg, K. ft. (1914). " A new mode of segregation in Gregory's tetr~ploid primtflas."
A'mer. Nat. 48, 508.
ONSLOW, 15{. W ~ L D ~ E (t925). The Anthocyani~ Pig~Tzenls of Plants. 2nd edition.
Cambridge.
-- (1931). "The chemical effect of a mendetiah factor for flower colom'." Nature,
Lend., 128, 373.
-- (1932). "Genetical and chemicaI aspects of anthocyanin pigments." Ibi& 129,
601.
~EDFIELD, H. (1930). "Crossing-over in the third chromosomes of triploids of
Drosophila melanogaster." Genetics, i 5 , 205.
EonEIVrSON, A. and l%o~r~sog, 1%. (1928). "A synthesis of 3fl glueosidyl peIargonidin
chloride (which is believed to be identical with eallistephin chloride)." J. chem.
Soc. 1~t60.
1%OBINSO~r, G. M. and ~O]3rNSON, 1~. (1931 a). " A sm'vey of an~hoey~uins. I."
Biochem. J. 25, 1687.
(1931 b). "Constitution of anthoeyanins." Arat.a~.c,Lend., 128, {13.
(1932 a.). "A survey of anthoeyanins. II." Biochcm. J. 26, 1647.
(1932 b). "Developments in the Chemistry of the Anthoeyanins." Nalu~'e,
Lend., t 3 0 , 21.
 W. J. O. L A W I ~ E N C E A N D ~OSE SCOTT-MONC1%IEFF 225
I~O~INSON, G. M. a.nd ROBINSON, ]~. (1933 a). "A survey of anthocyanins. III. Notes
on the distribution of louco-mlthocyanins." Biod~em. Y. 27, 206.
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address to the British Association: "Anthocyanixm as indicators and the colours
of flowers." Na/ure, Lend., t 3 2 , 625.
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cyanin chlorides." J. chem. ~goe. 2~195.
Som~no, L. and H i s o ~ K , L. (1933). " D e r gelbe Dahlieltfarbstoff." Mh. Chem.
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6 0 , 32.
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SCJOT'r-I){ONCJI~IEFF,]-~. (1930). "Nagm'M Anflhocy-min Pigments, I. The magenfi~
flower pigment of Antirrhinum majus." Biocb.em. J. 24, 753.
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Liebigs Ann. 408, 147.

EXPLANATION OF PLATES VII AND VIII,

I?Li'rE VII.
Fig. i. Cumulative and interaction effects in i - a I - i phenotypes of D. variabilis. A is
cumulative from Ai to A,I; I is saturated at 12but Imtentiallycumulative from I2 to i,i .
Note the strong co-pigmentblueing ei~eet of I upon i and the progressivesuppressionof
authoeyanin with the addition of each I factor for apigenin. The natm'M co]ours
are brighter than those shown in the plate.
Fig. 2. Seedling 31i/30 (BiIi), deep pm'plish-crimson (pm'e cyanin, no flavone).
Fig. 3. SeedYng 31"'/30 (B~), rosy-pro'pie (pm'e pelargonin plus apigenin).
Fig. ,L Seedling 301/30 (BiL), pMe purple (pure cyanin plus apigenin).
In the above tln'ee figures (2-4), note (I) the decrease in cyanic intensity in Figs. 3 and 4
compared with Fig. 2, owing to the presence of the ivory flavone (apigenin), (2) the stronger
co-pigment effect of apigenin upon cyalfin (Fig. 4) than upon pelargolfin (Fig. 3), and (3) the
more lively "rosy" colom' of the pelargonin pigmented flower.

PLATE VIII.
Fig. 5. Seedling A 13K [['he left hMfshows the lower surface, the right hMftlie upper surface
of the petM (for explanation see ~ext, p. 186).
Fig. 6. Crimson petal in hMf of which.the yellow ground has changed to ivory owing to the
loss of the Y factor. Note increase of cyanic intensity in absence of Y.
Fig. 7. P~rattel sectorial mutations due to (1) loss of the B factor in both mutant sect,ors
and (2) loss of the Y ihctor in the magenta sector. Note that the light (A) anfihocyanin
pigmentation is not developed in the presence of yellow flavone.
226 (/enetics and Chemist~'y of Flowe~" Co lou~' in Dahlia
t~ig. 8. Seeclling 1~/31. B~u~gion showing suppression of heavy anghoeyanin pigmengagion
on gain of yellow flavone. The large ~pricog mugang secgor h~d more flavone ghan ghe
normal scaz'le~-orange ])et~als. Cf. Fig. 6.
Fig. 9. A p~t~gern~rising from inger~egion effect)s. The yellow ground p~les ~owards the l~ips
of ghe pel~als. Anghocy~nin is produced only in ghe tips of t~he pe~ls, owing go ghe
sgrong supl?rcssing acgion of the yellow fi~vone. Ag ghe exgremigies of t)he peg~ls,
where yellow fla~rone is pr~ci,ic~lly ~t)senl), t~he presence of ivory flavone is indicated
by igs co-pigmeng (bhming) effect upon ghe anghocy~nin.
JOURNAL OF GENETI08, voL. x x x , NO. 2 PLATE Vll

i4 Ii I2 13 I4

8'4

, iii,iliS!iif
A2 t i

2 ,-, 4
JOURNAL OF GENETICS, VOL XXX, NO. 2'. PLATE VIII