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Flavonoids in Citrus Hybrids

a a a
Shintaro Kamiya , Sachiko Esaki & Fukuko Konishi
a
Food Chemistry Laboratory, Department of Foods and Nutrition, Shizuoka Women’s University, Shizuoka City, Yada 409, Japan
Published online: 09 Sep 2014.

To cite this article: Shintaro Kamiya, Sachiko Esaki & Fukuko Konishi (1979) Flavonoids in Citrus Hybrids, Agricultural and Biological Chemistry, 43:7, 1529-1536
To link to this article: http://dx.doi.org/10.1080/00021369.1979.10863651

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 Agric. Bioi. Chern., 43 (7), 1529-1536, 1979 1529

Flavonoids in Citrus Hybrids*

Shintaro KAMIYA, Sachiko ESAKI and Fukuko KONISHI

Food Chemistry Laboratory. Department 0/ Foods and Nutrition,

ShizlIoka Women's University, Shizuoka City, Yada 409, Japan
Received February 5, 1979

The occurrence and distribution of flavanone glycosides in the leaves and fruits of many
kinds of artificial citrus hybrid plants were investigated by polyamide thin-layer chromato-
graphy. The citrus hybrids can be divided into two broad categories, a) those containing
rutinosyl glycosides, b) those containing neohesperidosyl glycosides in accordance with the
case of natural citrus species. The flavonoid patterns of rutinosyl glycosides are classified
into the following groups, a) hesperidin, b) narirutin, c) hesperidin and narirutin, d) didymin
and narirutin, e) hesperidin, narirutin and eriocitrin and f) hesperidin and eriocitrin, while
the pattern of neohesperidosyl glycosides fall into six groups, a) naringin, b) neohesperidin and
naringin, c) neohesperidin, naringin and neoeriocitrin, d) neohesperidin and neoeriocitrin,
e) naringin and neoeriocitrin, and f) poncirin, neohesperidin, naringin and neoeriocitrin.
It is worthy of note that a hybrid (accession number 1088) between C. unshill and C. hassaku
contains only narirutin. Among the ninty-four hybrids examined, fifty-three varieties were
obviously different from female parents in their flavonoid pattern and could be judged as
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true hybrids by flavonoids but the others could not.

Additionally, a survey of f1avonoids in newly found natural pummelo- and Daidai hybrids
were carried out in connection with their origin.

It is important for citrus breeding to create O

new cultivars which are resistant to disease,
insects and climate extremes, which produce
fruit with high quality and good flavor. How-
ever, since nucellar embryony is of regular
occurrence in many species of citrus, it is
necessary to discriminate between nucellar and
zygotic seedlings produced by hybridization.
It is also interesting to study inheritance of
flavonoid glycosides in citrus by hybridization
from the view point of chemicogenetics. V ntil
present, various chemical, physical and enzy-
matic tests have been tried for identifying zygo-
tic versus nucellar seedlings. Namely, Furr
et al. ll and Nishiura et aU) used modifications
of root stock color reaction test, Pieringer and
3
Edwards ) used infrared spectroscopy and
Pieringer et al!) tried to apply gas chromato-
graphy to analysis of essential oils of hybrid
leaves as a basis of distinction. None of these
tests are entirely satisfactory. Recently, Tatum
* Flavonoids in Citrus and Related Genera. Part
VlI. For previous paper, See Agric. Bioi. Chern., 38,
339 (1974).
et al. ) studied flavanones and coumarines in
leaves of hybrids resulting from a cross between
two oranges by thin-layer chromatography and
obtained good results. Iglesias et al. BI and
7
Veno et al. ) have reported the possibility of
distinction of zygotic seedlings by analysis of
enzymatic patterns of leaf peroxidase and
isoesterase, respectively. From the chemico-
genetical standpoint, Kefford BI and Horowitz 9 )
intended to correlate specific flavonoids with
certain species of citrus. Also, Albach and
Redman 10 ) have surveyed the composition and
inheritance of flavanones by thin-layer chro-
matography and discussed the species problem
of citrus in connection with the pattern of
flavanone glycosides. In previous papers ll • 121
we reported the flavonoid patterns in some
citrus and poncirus hybrids and obvious dif-
ferences were observed in the chromatogra-
phic pattern of flavonoids between the female
parents and their hybrids and we considered
that flavonoid survey may be useful means for
judging the citrus plants as true hybrids.
In the present paper we further surveyed the
 1530 S. KAMIYA, S. ESAKI and F. KONISHI

distribution of flavonoid glycosides contained glycosides in the leaves of parents of citrus hybrids are
in a number of hybrid leaves and fruits by given in Table I.

simple thin-layer chromatography and intended TABLE I. FLAVONoms IN THE LEAVES OF

to discuss the inheritance of flavonoids in con- THE PARENTS OF CITRUS HYBRIDS
nection with the origin of citrus species and Common name Scientific name Flavonoids
also to distinguish zygotic from nucellar
Citrus
seedlings. Archicitrus
Limonellus
Mexican lime c.aurantifolia Swingle 3
EXPERIMENTAL
Citrophorum
Materials and methods. Hybrid samples (leaves Genoa lemon C.limon Burm. f. Genoa 3, 7
and fruits) were collected from the trees planted in the Cephalocitrus
orchards of Shizuoka Citricultural Experiment Station Hirado-buntan C.grandis Osbeck f.Hirado 6
and Okitsu Branch of Fruit Tree Experiment Station. Kao Panne C.grandis Osbeck f.Kao Pann 6
The trees have been judged as true hybrids by their Mato-buntan C.grandis Osbeck f.Mato 6
tree behavior and fruit characters. Mature leaf samples Grapefruit C.papadisi Macfad. 6
were harvested from trees in June. Each sample con- Kinukawa C.graberrima Tanaka 4,6,8
sists of ten leaves. Several immature fruits of each Hassaku C.hassaku Tanaka 4,6,8
hybrid plant were collected from September to October. Yamamikan C.intermedia Tanaka 6
Procedures for isolation of flavanone glycosides from Aurantium
the leaves and fruits were described in previous Sanbokan C.sulcata Takahashi 3,5,7
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papers,1l ,12) A crystalline mixture of flavanones from Natsudaidai C.natslidaidai Hayata 4,6,8
peels was separated with Woelm polyamide column Sour orange C.aurantium Linn.f.Kabusu 4,6,8
using benzene-methanol as an eluent. Individual Joppa C.sinensis Osbeck f.Joppa 3
flavanones were identified by mp and IR spectra in Maltese Blood C.sinensis Osbeck v.Sanguinea
comparison with those of authentic samples. In ad- Y.Tanaka f.Maltese 3
dition, the taste properties of crystals and also ethyl Parson Brown C.sinensis Osbeck
acetate extracts of leaves were examined, because the f.Parson Brown 3
flavanone rutinosides and neohesperidosides can also Trovita C.sinensis Osbeck f.Trovita 3
be distinguished by their taste properties, the former Meiryuto C.sinensis Osbeck f.Meiryuto 3
being tasteless while the latter intensely bitter. Detec- Mizunawa C.sinensis Osbeck f.Mizunawa 3
tion of the isomeric rutinoside and neohesperidoside Temple C.temple Y.Tanaka 3
pairs of flavanones was accomplished by polyamide Funadoko C.FlInadoko Y. Tanaka 4
thin-layer chromatography using nitromethane-metha- Tankan C.tankan Hayata 3
nol (5: 2) as a developing solvent and sodium boro- Hyiiganalsu c.tamllrana Tanaka 3,5,7
hydride-hydrochloric acid as a chromogenic reagent. Metacitrus
Spot colors and Rf values of flavanones are: didymin, Osmocitrus
red (0.67), poncirin, red (0.62), hesperidin, purple (0.57), Yuzu c.junos Sieb ex Tanaka 3,5
neohesperidin, purple (0.43), narirutin, red (0.38), Kunenbo C.nobilis Lour. val'
naringin, red (0.32), eriocitrin, blue (0.22), neoerio- Kunep Tanaka 3
citrin, blue (0.18). The chemical structures of the King C.nobilis Lour. 3
above flavanones are listed in Figure I. Flavanone Satsuma C.lInshili Marc. 3
Dancy tangerine c.tangerina Tanaka 3
Ponkan C.reticlilata Blanco 3
x -O~::::::::- .J" 0 ......
CH Mankitsu C.tardiferax Tanaka 3
I ' Clementine
C,..CH z C.clementina Tanaka 3
II Tankan C.tankan Tanaka 3
OH 0
Fortunella
X Rutinose Neohesperidose Kinkan F. Swingle 2
R , =OCH 3 ,R 2 =H Didymin Poncirin Choju Kinkan F.obovata Tanaka 2
R 1 =OCH 3 ,R 2 =OH Hesperidin Neohesperidin Poncirus
.
R 1 =OH ,R,,=H Narirutin Naringin Trifoliate orange P.trifoliata Rafin. 2,6
R 1 =OH,R,=OH Eriocitrin N eoerioci trin 1, didymin; 2, poncirin; 3, hesperidin; 4, nea-
FIG. 1. Chemical Structures of Citrus Flavanone hesperidin; 5, narirutin; 6, naringin; 7, eriocitrin;
Glycosides. 8, neoeriocitrin.
 Flavonoids in Citrus Hybrids
RESULTS AND DISCUSSION
The present authors surveyed the occurrence
and distribution of flavanone glycosides in the
leaves and partly fruits of ninty-four varieties
of artificial citrus hybrids by polyamide thin-
layer chromatography and also isolated flava-
none glycosides from the fruits of some hybrids
for more rigorous chemical proof. As the
result, it became apparent that qualitative as
well as quantitative differences existed between
the hybrids and cross parents in many cases.
The citrus hybrids can be divided into two cate
gories, a) those containing rutinosyl glycosides,
b) those containing neohesperidosyl glycosides
in accordanoe with the case of natural citrus
species. 12 ) It was not observed that when a
species containing only flavanone rutinosides
is crossed with another containing only neo-
hesperidosides, the progeny contains both types
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of flavanone glycosides. The patterns of
rutinosyl glycosides fall into the following
groups: a) hesperidin, b) narirutin, c) hes-
peridin and narirutin, d) didymin and nari-
rutin, and e) hesperidin, narirutin and erio-
citrin, and f) hesperidin and eriocitrin, while
the patterns of neohesperidosyl glycosides fall
into six groups, namely, a) naringin, b) neo-
hesperidin and naringin, c) neohesperidin,
naringin and neoeriocitrin, d) neohesperidin
and neoeriocitrin, e) naringin and neoerio-
citrin, and f) poncirin, neohesperidin, naringin
and neoeriocitrin. A hybrid of the lemon
with yuzu is a very curious variety because its
leaves contain only narirutin through a year,
•
but fruit contains hesperidin predominantly
and minor amounts of narirutin and eriocitrin.
This fact deduces that narirutin introduced into
the fruit from leaves changes biochemically into
hesperidin. Among four hybrids between the
Hassaku and Mato-buntan (pummelo), three
varieties contain flavanone neohesperidosides
but the remaining one(23~31) flavanone
rutinoside, hesperidin. Since both of parents
contain only flavanone neohesperidosides, the
appearance of rutinosyl glycosides in a progeny
is very interesting. This fact suggests that any
citrus species which contains flavanone rutino-
1531
sides might be concerned with the production
of both of the parents in the past time. Among
the ten hybrids originated by crossing the
Daidai (Sour orange) with each of the man-
kitsu, Ponkan and sweet orange (Mizunawa),
nine varieties contain neohesperidosyl glyco-
sides while the other one (23 ~ 140) rutinosides.
In survey of flavonoid distribution in the leaves
of twenty-two kinds of hybrids of the Satsuma
with the pummelo, twenty-one individuals
contain flavanone neohesperidosides and only
one individual (29 ~ 36) contains rutinoside,
hesperidin. The composition of flavanone
neohesperidosides in the artificial pummelo-
and Daidai hybrids mentioned above can be
divided into five groups, namely, a) those con-
taining only naringin, b) those containing neo-
hesperidin and naringin, c) those containing
naringin and neoeriocitrin, d) those contain-
ing neohesperidin and neoeriocitrin, and e)
those containing neohesperidin, naringin and
neoeriocitrin. Many natural species belong-
ing to the Cephalocitrus and Aurantium sec-
tion have been considered to be natural hy-
brids which have pummelo or the Daidai as
one of the parents. Since these species con-
tain one of the five flavonoid patterns men-
tioned above, they might be recognized as
pummelo or Daidai hybrids from the stand-
point of chemosystematics. Among the hy-
brids between the Hassaku and sweet orange
(Joppa) four varieties contain neohesperidosyl
glycosides and the one rutinosyl glucosides,
hesperidin, while all the hybrids of the Natsu-
daidai with sweet orange (Maltese blood)
produce rutinosyl glycosides. Very interesting
artificial hybrids are two varieties (accession
number 5857, 1088) created by crossing the
Satsuma with the Hassaku. Their flavonoid
patterns in the fruits are a) hesperidin and
narirutin, and b) narirutin, respectively. Es-
pecially it is worthy of note that the hybrids
(1088) contains only narirutin. Among more
than two hundred kinds of natural citrus plants
examined thus fur, the Jagatarayu (c. pseudo-
gulgul Shirai) and the Tengu (c. tengu Tanaka)
have been found to contain narirutin alone.
The hybrid (5857, or Okitsu No. 20), a variety
 1532 S. KAMIYA, S. ESAKI and F. KONISHI

TABLE II. FLAVONOIDS IN THE LEAVES OF CITRUS HYBRIDS

Accession Cross parents Flavonoid content Judgement

number
C. aurantifolia x C. limon f.
Genoa (Perrine lemon) 3 (+++)
23~68 C. limon x C. junos 5 (+++) +
23~70 C. grandis f. Mato x C. sinensis (. Joppa 4 ( +), 6 ( + +), 8(+ + +) +
23~31 C.hassaku x C.grandis f. Mato 3 (+++) +
23~41 /I 4 (tr), 6 (+ + +), 8 (+ + +) +
23~ 102 /I 6 (+++) +
23~ 103 /I 6 (+ + +), 8 (±) +
23~72 C.hassaku x C.sinensis f. Joppa 3 (+++) +
23~73 /I 4 (+ + +), 6 ( + + +) +
23~74 /I 4 ( + + +), 6 ( +), 8 ( + )
23~75 /I 4 (±), 6 (+ + +), 8 (tr) +
23~76 /I 4 (+ + +), 6(+ + +), 8 ( +)
23~201 C.natsudaidai(Kawano)
x C.sinensis f. Maltese 3 ( + + +), 5 ( + ++-), 7(+) +-
23~202 /I 3 (+++), 7(+++) +
23~203 /I 3 (+ + +), 5 ( + + +), 7 (tr) +
23~204 /I 3 (+++), 5 (+++),7 (tr) +
23~35 c.aurantium x C.tardiferax 4 (+ + +), 6 ( + + +), 8 ( + ++)
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23~47 11
4 (+ + +), 6(+ + +), 8(+ + +)
23~65 /I
4 ( + + +), 6 ( + + +) +
23~ 140 /I
3 (+++) +
23~ 199 /I
4 (+ + +), 6(+ + +) +
23~97 c.aurantium x C.reticulata 4 (+ + +), 6(+ + +), 8(+ + +)
23~93 c.aurantium x C.sinensis f. Mizunawa 6(+ + +), 8(+ + +) +
23~94 /I
4 (+ +), 6(+ + +), 8(+ + + )
23~96 II
4 (+ + +), 6(+ + +), 8(+ + +)
23~205 6 (+ + +), 8(+ + +)
/I
+
23~54 C.llnshill(Sugiyama) x C.grandis f. Hirado 4 (+), 6(+ + +), 8(±) +
23~55 6(+++)
23~58
/I
+
/I
4 (+), 6(+ + +) +
23~59 /I
4 (±), 6(+ + +) +
23~61 4 (±), 6(+++)
23~62
/I
+
/I
4(±),6(+++) +
23~63 /I
4 ( +), 6 ( + + +), 8 ( +) +
23~82 /I
4 (tr), 6 (+ + +), 8 (±) +
23~83 /I
4 (+), 6(+ + +), 8 (±) +
23~84 4 (tr), 6(+++)
29~32
/I

C.unshiu(Tomono) x C.grandis f. Hirado

+
4 (+ + +), 6(+ +), 8(+ +) +
29~33 /I
6(+++) +
29~34 II
4 ( ++ +), 8 ( + +) +
29~36 II
3(+++)
23~34 C.unshiu x C.grandis f. Kao Pann 4 (±), 5 (±), 6 (+ + +), 8 (+) +
23~
128 11
6(+++)
23~ 130
+
/I
4 (±), 6 (+++),8 (+) +
23~ 131 /I
4 (±), 6(+++),8 (+) +
23~ 132 /I
4 (±), 6 (+ + +), 8 (±) +
23~ 133 /I
4 (tr), 6(+++) +
23~191 11
4 (+ + +), 6(+ + +), 8(+ + +) +
23~ 192 /I
4 (+ + +), 6 (+ +), 8(+)
23~60 C.llnshill X C.glaberrima
+
4 (tr), 6 (+ + +), 8 (tr) +
29~82 C.unshiu x (C. paradisi x c.tangerina) 3(+++)
23~145 1/
6(+++) +
 Flavonoids in Citrus Hybrids 1533

TABLE II. (Continued)

Accession Cross parents Flavonoid content Judgement
number
23~140 C.unshill X C.hassakll 4 (+ + +), 6 (+), 8 (+ ) +
23~ 107 C.unshiu x c.tamurana 3(+++)
23~ 189 C.unshill (Miyagawa)
x C.natsudaidai (Kawano) 3 (+++)
139 C.uns!till(Beni) x C.natusdaidai(Kawano) 4 ( + + +), 6 ( + + +), 8 ( + + ) +
23~81 C.lIns!tiu X C.natsudaidai 3(+++)
23~115 3(++ :-)
23~144
"
C.unshill x (C.natslldaidai
x C. sinensis f. Maltese) 3 (+++)
14 • C.unshiu(Sugiyama)
x C.sinensis f. Meiryuto 3 (+1-+)
18 3(+++)
"
C.lInshiu x C.sinensis f. Joppa
26 3 (+++)
38 C.lInshiu(Sugiyama)
X C.sinensis f. Meiryuto 3 (+++)
C.unshill(Miyagawa)
x C.sinensis f. Meiryuto 3 (+++)
C.unshiu(Sugiyama)
x C.sinensis f. Mizunazwa 3 (+++)
32~95 3 (+++)
"
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29~ 134 (C.unshiu x C.sinensis f. Parson Brown)

x C.sinensis f. Parson Brown 3 (+++)
(C.unshiu x C.sinensis f. Parson Brown)
x C.sinensis f.Parson Brown 3 (+ + +)
32 ~ 19 C.unshiu(Sugiyama) x C.temple 3 (+ + +) -
32~20 " 3 (++-+-)
32~29 " 3 (+++)
32~30 1/ 3 (+++)
32~32 " 3 (+++)
32~42 1/ 3(+++)
32~74 1/ 3 (+++)
23 ~ 28 C.unshill x C.sulcata 3 (+), 5 (+ + +), 7 (+ + + ) +
23 ~ 136 C.unshill x C.fllnadoko 3 (+ + +)
23 ~ 1 3 7 " 3 (+ ), 4 (+ + + ) +
29 ~ 62 1/ 3 (+ + + )
29 ~ 63 1/ 3 (+ + + )
23 ~ 150 C.lInshiu X c.junos 3 (+ + +), 5 ( + + +), 7 (+ + + ) +
29 ~ 96 C.llnshill X C.nobilis val' Kunep 4 (+ + +), 6 (+ + +), 8 ( + ) +
23 ~ 10 C.lInshiu X P.trifoliata 2( + + + ),4( + + + ),6( + + + ),8( +) +
23 ~ 69 C.tankan X C.intermedia 3 (+ + + )
29~59~9 Fortllnella sp. x c.allrantifolia(Sugiyama) +
23~109 Fortunellasp.xC.limon 3(+++),5(+++),7(+++) +
For arabian numerals under the item of flavonoid, See Table I. ±, +, + +, and + + + signs are
used to indicate approximately the relative proportions of the constituents in the sample. tr, trace.
In the item of judgement, + and - signs show that the citrus plant can and can not be judged as true
hybrids by flavonoid, respectively.
 J534 S. KAMIYA, S. EsAKI and F. KONISHI

TABLE III. FLAVONOIDS IN THE IMMATURE PEELS OF CrRUS HYBRIDS

Artificial hybrids Fresh Flavonoids Mother

Accession Cross parents weight Crystals yield liquid
number (g) (%)
c.aurantium f.Kabusu
x C.sinensis f.Mizunawa 320 6 3.86 0.89 4, 6,~8
23~68 C.limon x C.junos 130 3 0.35 0.27 3, 5, 7
23~62 C.unshiu(Sugiyama) x C.grandis f.Hirado 460 6 4.43 0.96 6
23~63 Q 750 6 13 .5 1.80 4,6,8
23~82 Q 460 6 5.12 1.11 4,6
23~ 132 C.unshiu x C.grandis f. Kao Pann 590 6 2.39 0.41 6
29~36 C.unshiu(Tomono) x C.grandis f.Hirado 270 3 4.77 1. 77 5
1088 C.unshiu(Suzuki Wase) x C.hassaku* 285 5 2.90 1.02 5
5 C. unshiu(Ueda) x C.hassaku* 286 3 5.30 1.85
5 0.26 0.09 5
C.unshiu x C.sinensis f.Parson Brown 140 3 4.1 2.93 5
C.unshiu(Sugiyama) x C.sinensis f.Meiryuto 170 1 0.45 0.26
5 0.50 0.29 I, 5
23~22 C.unshiu x C.sinensis f. Maltese 80 5 0.20 0.25 1, 5
14 C.unshiu(Sugiyama) x C.sinensis f.Meiryuto 51 3 0.20 0.39 5
18 Q 40 3 0.20 0.50 5
26 C.unshiu x C.sinensis f. Joppa 27 3 1.0 3.70 5
38 C.unshiu(Miyagawa) x C.sinensis f.Meiryuto 90 2.40 2.66 5
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3
29~ 145 C.unshiu(Miyagawa) x C.sinensis f.Trovita 110 3 1.40 1.27 5
32~91 C.unshiu(Sugiyama) x C.sinensis f.Mizunawa 55 3 1.20 2.18 5
32~95 Q 79 3 1. 70 2.15 5
29~96 C.unshiu x C.nobilis var. kunep 120 6 1.50 1.20 4'6
,-
C.unshiu(Owari) x C.nobilis[Kara mandarine] 200 3 5.5 2.75 5
32~19 C.unshiu(Sugiyama) x C.temple 48 3 0.80 1.64 5
32~20 Q 43 3 1.20 2.79 5
32~29 Q 45 3 1.20 2.66 5
32~30 Q 33 3 1.00 3.00 5
32~32 n 37 3 1.10 2.97 5
32~42 n 41 3 1.00 2.43 5
32~74 Q 40 3 1.20 3.00 5
C.nobilis x C.deliciosa
Ten. [Honey mandarine] 150 3 3.40 2.27 5
C. paradisi x Sampson tangelo(C. paradisi x
C.tangerina) [Wekiwa tangelo]* 180 3 2.50 1.34 5
Natural hybrids (not yet classified)
Akaogon 21 6 0.35 1.65 6
Chosen Daidai 9.5 6 0.26 2.77 2,6
Kaneko pomelo' 185 6 0.80 0.43
4 0.39 0.21 2,4,6,8
Murcott orange* 100 J 0.20 0.20
5 0.30 0.30 1, 5
Ojagatara e 28 6 0.26 0.92 2,6
Onatsu e 5.0 6 - 6
Wakayama shishiyuzu f 15.0 5 0.30 2.00
1 0.12 0.80 1, 5
Wheeny grapefruit* 170 4 0.15 0.26
6 0.67 0.38 4,6
The fruits of Japanese natural hybrids were collected from the following places by Dr. Y. Tanaka.
a Takuma Cho, Innoshima Shi, Hiroshima Prefecture
b Kizu, Kisawa Mura, Naka Gun, Tokushima Prefecture
c Iwata Shi, Shizuoka Prefecture
 Flavonoids in Citrus Hybrids
d Aid Shi, Kochi Prefecture
• Katsuura Cho, Tokushima Prefecture
f Fruit Tree Experiment Station, Wakayama Prefecture
* Obtained from Okitsu Branch of Fruit Tree Experiment Station
Flavonoid numbers are the same as those described under the Table II.
currently of increasing importance because of
its high quality, has the same flavonoid pattern
13
as that of the Satsuma. ) In the hybrids
between the Satsuma and pummelo-hybrids
such as the Kinukawa, Hassaku, and tangelo,
three varieties contain neohesperidosyl glyco-
sides while three varieties rutinosyl glycosides.
Of the ten hybrids obtained by crossing the
Satsuma with the Daidai-hybrids such as
Hyilganatsu, Natsudaidai, Sanbokan, and
Funadoko, rutinosyl glycosides are contained
in all except one sample which contains neo-
hesperidosyl glycosides. In survey of the ten
hybrids of the Satsuma with sweet oranges
such as Joppa, Trovita, Meiryuto, Mizunawa
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and Maltese blood, eight varieties contain
hesperidin only but the two ones contain
didymin and narirutin but hesperidin lacks
completely although both of the parents con-
tain hesperidin. Seven hybrids between the
Satsuma and Mandarines such as King man-
darine and temple, eight varieties contain
hesperidin only but interestingly the Satsuma x
Kunembo, a variety of King mandarine,(29 ~
96) contains neohesperidosyl glycosides, name-
ly, naringin predominantly and small amounts
of neohesperidin and neoeriocitrin, and naringin
can be isolated as crystals from its fruit. This
fact indicates that gene which controls the neo-
hesperidose formation might be present in both
of the parents. A hybrid of the Satsuma with
the trifoliate orange contains all of neohes-
peridosyl glycosides, namely, poncirin, neo-
hesperidin, naringin and neoeriocitrin. As
already reported '2 ), we found the same flava-
none pattern in the Rusk, Troyer citranges and
lyo x Poncirus.
Of the ninty-four hybrids examined, fifty-
three varieties were obviously different from
each female parent in their flavonoid pattern
and could be judged as true hybrids by flavo-
noids but the others could not. Among fifty-
nine varieties resulting from the crosses of
1535
species containing flavanone rutinosides with
another species containing flavanone neo-
hesperidosides, forty-six varieties could be
judged as true hybrids, whereas among thirty-
three varieties resulting from the crosses be-
tween the species which have the same sugar
moieties of flavanones twelve varieties were
judged as true hybrids. Accordingly, this
technique can employ as supplemental means of
recognizing hybrids in addition to the survey
of their morphological properties such as
tree behavior and fruit characters.
Akaogon, Chosen Daidai, Kaneko pomelo,
Ojagatara, Onatsu which are natural hybrids
and also Wheeny grapefruit which is an Austra-
lian variety of considerable importance in New
Zealand are considered to be pummelo- or
Daidai hybrids by morphological characters.
Flavonoid patterns of them support this as-
sumption. Wakayama shishiyuzu which con-
tains didymin and narirutin is said to be
identical with Jagatarayu which contains only
narirutin. Accordingly both citrus plants may
not be identical from the view point of flavo-
noids. Murcott orange, which is considered
to be natural tangor currently of increasing
importance in Florida lacks hesperidin com-
pletely and has didymin and narirutin in ac-
cordance with that of two Japanese artificial
tangors (Accession number 23 ~ 14, 23 ~ 22).
Acknowledgment. We wish to express our sincere
thanks to Mr. E. Shikano, Shizuoka Citricultural Ex-
periment Station, and Dr. L Veno, Fruit Tree Research
Station, for their kind supply of citrus hybrid samples.
We are also grateful to Dr. Y. Tanaka for his kind
supply of several newly found citrus fruits.
REFERENCES
1) J. R. Furr and P. C. Reace, Froc. Amer. Soc.
Hart. Sci., 48, 141 (1946).
2) M. Nishiura, J. Matsushima and N.Okushiro,
Tokai Kinki Nashi Hakoku, 4, 19 (1957).
3) A. P. Pieringer and G. J. Edwards, Proc. Amer.
 1536 S. KAMIYA, S. EsAKI and F. KONISHI
Soc. Hart. Sci., 86, 226 (1965).
4) A. P. Pieringer, G. J. Edwards and R. W. Wol-
ford, ibid., 84, 204 (1964).
5) J. H. Tatum, R. E. Berry and C. J. Hearn, Proc.
Fla. State Hart. Soc., 87, 75 (1974).
6) L. Iglesias, H. Lima and J. P. Simon, J. Heredity,
65, 81 (1974).
7) 1. Ueno and M. Nishiura,Bull. Fruit Tree Res.
Sm. B., 3, 1 (1976).
8) J. F. Kelford, Adv. Food Res., 9, 285 (1959).
9) R. M. Horowitz, Symposium of biochemistry of
Downloaded by [Florida State University] at 02:20 24 December 2014
plant phenolic substances, Colorado State Univ.,
Ft. Colline, Colorado, pp. 1 ~ 8 (1961).
10) R.M. Albach and G.H. Redman, Phytochemistry,
8, 127 (1969).
11) M. Nishiura, S. Kamiya and S. Esaki, Agric. Bio/.
Chern., 33, 1109 (1969).
12) M. Nishiura, S. Kamiya and S. Esaki, ibid., 35,
1691 (1971).
13) M. Nishiura, S. Kamiya, S. Esaki and F. Ito, ibid.,
35, 1683 (1971).