Tabelle 1. P r o z e n t u a l e V e r t e i l u n g der 14C-Aktiviti.

t a u f die J u v e n i l e H o r m o n e and Caste D e t e r m i n a t i o n

A s s i m i l a t e der 16slichen F r a k t i o n n a c h 10 rain P h o t o s y n t h e s e in a S t i n g l e s s Bee
Assimilat P. convolutae P. tetrathele ~ Sex D e t e r m i n a t i o n in Bees, V I I
L. A. de Oliveira C a m p o s
Oligomere 1,1 1,1 Depto. de Gendtica, Fac. Medicina, 14100 Ribeir~o P r e t o S. P.
Phosphatester 2, 7 11,7 Brazil
Asparaginsgure 5,4 4,4
Glycin 3,0 t, 5 F. M. V e l t h u i s - K l u p p e l l a n d H. H. W. V e l t h u i s
Serin 1,1 2,1 Lab. of C o m p a r a t i v e P h y s i o l o g y , U t r e c h t , T h e N e t h e r l a n d s
Alanin 2, 5 22, 3
l~brige A m i n o s g u r e n 3, t 3,6 D i f f e r e n t i a t i o n in q u e e n a n d w o r k e r caste in h o n e y b e e s d e p e n d s
Malat 6,0 10,2 on a food factor. A q u e e n l a r v a c o n s u m e s a m u c h g r e a t e r
Citrat 0, 5 0, 5 q u a n t i t y of food t h a n a w o r k e r larva. A special c o m p o n e n t
Glycerat 3,t 1,0 in t h e r o y a l jelly m a y in a d d i t i o n d e t e r m i n e q u e e n d e v e l o p m e n t
Fumarat 0, 5 3,2 [t]. Q u e e n d e v e l o p m e n t is d e t e c t a b l e as a raised juvenile-
Fructose 4,6 t4,0 h o r m o n e (JH) t i t r e in t h e h a e m o l y m p h of a 2- to 3-day-old
Glucose t 4,2 4, 5 l a r v a [2]. Topical a p p l i c a t i o n of J H to 3-day-old h o n e y b e e -
Mannit 51,4 14,2 worker larvae i n d u c e d queenlike c h a r a c t e r s in t h e e m e r g i n g
a d u l t s [2]. Caste d e t e r m i n a t i o n s e e m s to be u n d e r t h e control
a S t a t u r e 161-2b S a m m l u n g G 6 t t i n g e n . of differential J H p r o d u c t i o n as i n d u c e d b y different food
regimes; t h i s conclusion h a s n o t been accepted, however, b y
R e m b o l d el aL [3].
I n t h e stingless bee Melipona quadrifasciata Lep. it is n o t o n l y
s t r u k t u r e l l [4, 5] g u t u n t e r s u c h t . Die IZenntnis der stoffwechseI- t h e food factor t h a t influences q u e e n d e v e l o p m e n t . T h i s
p h y s i o l o g i s c h e n I n t e r d e p e n d e n z der beteiligten P a r t n e r ist species h a s 2 q u e e n - d e t e r m i n i n g genes which, w h e n food is
d a g e g e n w e n i g e r weft f o r t g e s e h r i t t e n , obwohl y o n a n d e r e n a b u n d a n t , c a u s e 25 % of t h e e m e r g i n g bees to be q u e e n s [4].
S y m b i o s e n viele D e t a i l i n f o r m a t i o n e n v o r h a n d e n s i n d [t, 6]. Lower a m o u n t s of food p r o d u c e o n l y workers, 25 % of w h i c h
Die S y m b i o s e zwischen Convoluta u n d P l a t y m o n a s w u r d e stoff- are genetically q u e e n s b u t f u n c t i o n as workers.
w e c h s e l p h y s i o l o g i s c h b i s l a n g k a u m u n t e r s u e h t [7, 8]. Zur Stingless bees are a d v a n t a g e o u s for a s t u d y of t h e relation
A n a l y s e der p h o t o s y n t h e t i s c h e n 14COe-Assimilation w u r d e n b e t w e e n J H a n d n a t u r a l caste d e t e r m i n a t i o n . W h i l e h o n e y b e e
I n d i v i d u e n y o n Roscoff (Bretagne) v e r w e n d e t . Bet der P h o t o - l a r v a e are fed a b o u t 150 t i m e s d u r i n g larval d e v e l o p m e n t ,
s y n t h e s e lagern die E n d o s y m b i o n t e n I~C a u s H14CO~ sehr r a s c h stingless bees h a v e m a s s p r o v i s i o n i n g a n d need no g r o o m i n g
ein. Die D u n k e l f i x i e r u n g s r a t e b e t r g g t n u t < 0, 5 % der P h o t o - b y n u r s e bees. T h i s allows b e t t e r c o n t r o l of t h e food factor a n d
s y n t h e s e l e i s t u n g . D a s legt die A n n a h m e n a h e , d a b die E n d o - m o r e a c c u r a t e e s t i m a t e s of t h e a m o u n t of active m a t e r i a l
s y m b i o n t e n ihren CO~-Bedarf i m D u n k e l n n i e h t a u s d e m u m - a c t i n g on t h e developing larva.
g e b e n d e n M e d i u m , s o n d e r n a u s der R e s p i r a t i o n ihres W i r t e s T h e effect of topical a p p l i c a t i o n of several J H a n a l o g u e s was
decken. D e r tierische P a r t n e r l~lat keine ins G e w i c h t fallende s t u d i e d b y one of us (L.O.C.). I n t h e s e e x p e r i m e n t s fullgrown
CO2-Fixierung e r k e n n e n . D u r c h D C - T r e n n u n g der 16sliehen l a r v a e t h a t h a d n o t y e t defecated were t a k e n f r o m t h e c o m b s
F r a k t i o n n a c h 10 rain P h o t o s y n t h e s e k o n n t e n ca. 30 A s s i m i - a n d , after a p p l i c a t i o n of a k n o w n a m o u n t of t h e m a t e r i a l to be
late e r k a n n t w e r d e n (Tabelle t). U n t e r d e n M a r k i e r u n g s p r o - t e s t e d [5], placed on filter p a p e r in P e t r i dishes. T h e i r f u r t h e r
d u k t e n k o m m t d e n A m i n o s g u r e n m i t e t w a t 5 % der eingelager- d e v e l o p m e n t w a s s t u d i e d in a n i n c u b a t o r (29 ~: I ~ C a n d 80 %
t e n 14C-Aktivitgt der gr6gte A n t e i l zu. Auffallenderweise fehlt RH).
d e m A s s i m i l a t m u s t e r dieser e n d o s y m b i o n t i s c h e n Griinalge T h e o t h e r two (F.V. a n d H.V.) s t u d i e d t h e effect of v a r i o u s
l~C-Saceharose. Daftir t r i t t laC-Mannit m i t e i n e m M a r k i e r u n g s - c o n c e n t r a t i o n s of t h e J H p r e p a r a t i o n after R611er [8]. L a r v a e
anteil v o n > 50 % n e b e n laC-Glucose a n d 14C-Fructose auf. Die were reared in vitro b y g r a f t i n g l a r v a e less t h a n one d a y old
M e n g e n dieser K o h l e n h y d r a t e , die i n s g e s a m t die a m s t ~ r k s t e n into artificial cells c o n t a i n i n g k n o w n a m o u n t s of food. T h e
m a r k i e r t e F r a k t i o n stellen, v e r h a l t e n sich wie e t w a t 0 : 3 : t . l a r v a e were t r e a t e d w i t h a droplet of a c e t o n e c o n t a i n i n g t h e
I m U n t e r s c h i e d zu E r g e b n i s s e n y o n a n d e r e n ein- u n d m e h r - q u a n t i t y to be tested. I n t h e e x p e r i m e n t r e p o r t e d here, all
zelligen Algen konnte 14C-Glykols{~ure n i c h t n a c h g e w i e s e n l a r v a e o b t a i n e d t 70 m g of food a n d were t r e a t e d while s p i n n i n g
werden. M6glicherweise s t e h t d a m i t der relativ niedrige G e h a l t t h e i r cocoon.
a n 14C-Glycin u n d 14C-Serin in Z u s a m m e n h a n g . A n d e r e r s e i t s T h e two e x p e r i m e n t a l p r o c e d u r e s u s e d led to p a r t i a l l y different
k 6 n n t e dies u n d der i m Vergleich zur freilebenden Platymonas results, a n d a detailed a n a l y s i s will be p r e s e n t e d elsewhere.
tetrathele (Tab. 1) w e s e n t l i c h geringere G e h a l t a n P h o s p h a t - Here we r e p o r t t h e m a i n conclusion we h a d in c o m m o n .
e s t e r n die Folge ether A u f n a h m e y o n Stoffwechselintermedi~- E v e n d u r i n g t h e early p u p a l s t a g e e x t e r n a l c h a r a c t e r i s t i c s
t e n d u r c h d e n tierisehen P a r t n e r sein. enable one to d i s t i n g u i s h b e t w e e n q u e e n s a n d workers. T h e
H i n s i c h t l i c h V o r k o m m e n a n d A r t der n a c h K u r z z e i t f i x i e r u n g m o s t p r o m i n e n t a s p e c t is t h e relative size of t h e eye, w h i c h is
in der e n d o s y m b i o n t i s c h e n P. convolutae faBbaren Assimilate s m a l l in q u e e n s a n d large in w o r k e r s ; t h e s t r u c t u r e of t h e h i n d
b e s t e h e n keine U n t e r s c h i e d e zu d e n E r g e b n i s s e n y o n Parallel- leg is slender in q u e e n s b u t a d a p t e d to pollen collection in
v e r s u c h e n m i t der freilebenden P. tetrathele (Tab. t), soweit workers. Several o t h e r m o r p h o l o g i c a l differences exist.
deren C h e m o k o n s t i t u t i o n b i s l a n g a u c h a n d e r w e i t i g u n t e r s u c h t T h e r e s u l t s of s o m e of o u r e x p e r i m e n t s are s u m m a r i z e d in
w u r d e [9, 10]. Die tiefgreifenden m o r p h o l o g i s c h e n U m g e s t a l - Table 1. T h e a n i m a l s o b t a i n e d in t h e e x p e r i m e n t a l series all
t u n g e n n a c h E t a b l i e r u n g als E n d o s y m b i o n t [4] f i n d e n i m
I n t e r m e d i g r s t o f f w e e h s e l y o n P. convolutae offenbar keine
Parallele. I n s g e s a m t dflrften die S t o f f w e c h s e l b e z i e h u n g e n Table 1. Effect of s o m e J H a n a l o g u e s a n d of s y n t h e t i c J K on
zwischen Alge a n d Convobuta rosco//ensis s e h r k o m p l e x a n d caste d i f f e r e n t i a t i o n
n i c h t n u t a n t d e n lDbertritt eines einzigen A s s i m i l a t s [6, 8]
beschr~inkt sein. Origin Material Quantity Number Number
of l a r v a e studied per bee of of
E i n g e g a n g e n a m t 7. O k t o b e r a n d 26. N o v e m b e r 1974 [[xg] queens workers

t. Taylor, D. L. : A n n . R e v . Microbiol. 27, t 7t (1973) Fullgrown Altozar TM 9 9 4

2. Keeble, F., G a m b l e , F. W . : Quart. J. Microscop. Sci. ~1, Fullgrown Altozar TM t8 28 0
t67 (t907) Control t0 42
3. Provasoli, L., et al.: J. m a r . biol. Ass. lJ. K. 48, 465 (1968)
Fullgrown Altosid TM t8 8 t3
4. O s c h m a n n , J. L.: J. P h y c o l . 2, 105 (1966)
Control 2 17
5. P a r k e , M., M a n t o n , I.: J. mar. biol. Ass. U. K. 47, 445
(t967) Fullgrown Hoffmann-L. 2' t 0 -a 23 0
6. S m i t h , D., et al.: Biol. Rev. 44, 17 (1969) Control 2 17
7. Gooday, G. W . : J. m a r . biol. Ass. U. K. 50, t99 (1970)
Reared Hoffmann-L. 2" t 0 -a 22 0
8. M u s c a t i n e , L.: Proc. Roy. Soc. L o n d . 187, 221 (t974)
in vitro Hoffmann-L. 2" 10-7 tI 11
9. Craigie, ~[. S., etal.: Can. J. Bot. 45, I327 (1967)
Control 3 17
10. Suzuki, H . : P h y i o e h e m i s t r y 13, t 159 (1974)

98 N a t u r w i s s e n s c h a f t e n 62 (t975) 9 b y Springer-Verlag 1975

 could be classified as either queen or worker, for neither the
pupal (experiments of L.O.C.) nor the adult forms (experiments
of F.V. and tt.V.) showed any intermediate stages. This con-
trasts with experiments with honeybees [2, 3]. Moreover,
within the proper concentration range of a given substance, no
morphological differences were observed as between experi-
mental and control animals. This is suggestive of natural
development of both castes in the experimental insects.
As Table t shows, some of the substances tested produced a
ratio of queens to workers t h a t differs statistically from the
control series. We may thus conclude t h a t induction of queen-
caste formation was caused by the artifical presence of J H
(analogue).
Most probably in this stingless bee natnral caste differentia-
tion is the result of differential J H production. Our results
further support the opinion t h a t the two developmental
pathways for the honeybee are under the control of differen-
tial J H production. This might well be the basic mechanism
for caste determination in the social Hymenoptera in general.
Financial aid was provided by the State of S~o Paulo Researct~
Foundation (FAPESP), the Organization of American States
and the Brazilian National Research Council (CNPq). Prof.
Dr. Warwick E. Kerr is heartily thanked for the invitation
to work in his department in Ribeir~o Preto, Brazil
Received September 27 and November 2~, 1974
t. Rembold, H., Lachner, B., Geistbeck, I.: J. Insect Physiol.
20, 3O7 (1974)
2. Wirtz, P.: Meded. Landbouwh. WageDingen 73, t (1973)
3. Rembold, H., Czoppelt, C., Rao, P. J.: J. Insect Physiol.
20, t193 (1974)
4. Kerr, W. E.: Evol. Biol. 3, 119 (1969). Above 155 mg a 3: t
segregation occurs, see Camargo, C. A. de: Diss. RibeirRo
Preto, 1972
5. Altozar (ZR 5 t 2, ethyl 3, 7,11-trimethyl-2,4-dodecadienoate)
and Altosid (ZR 5t5, ethyl isopropyI tl-methoxy-3,7,11-
trimethyl-2,4-dodecadienoate) were kindly given by Zoecon
Corp.
6. The isomer mixture of Hoffmann-La Roche, kindly given
by Prof. D r . J. de Wilde, Entomological Department,
Wageningen, was used.
I n s e n s i t i v i t y of B a c t e r i a l N u c l e i c Acid
B i o s y n t h e s e s to a M o r p h i n e - l i k e N a r c o t i c
A. D. Wolfe and F. E. Hahn
Department ok Molecular Biology, Walter Reed Army In-
stitute of Research, Washington, D.C. 20012, USA
Growth inhibition of E. coli by levorphanol [t ] or levallorphan
[2] is accompanied by failures of RNA and DNA biosyntheses.
We have made analogous observations ~.3] with N I H 7591,
a synthetic analgesic which is 30 times more potent than
morphine E4].
Treatment of E. coli with toluene renders the bacteria per-
meable to the nueleoside triphosphate precursors of nucleic
acids and permits the organisms to perform replicative
biosynthesis of chromosomal DNA [5] as well as RNA bio-
synthesis [6]. We treated suspensions of E. coli B with toluene
and measured incorporations of l~C-adenosine triphosphate
into RNA [6] and of aH-thymidine triphosphate into DNA
[5] in drug-free control experiments and in the presence of
l-5"10-aM N I H 7591 which is 2.5 times the concentration
which completely inhibits growth of E. coli B [3]- The time
course of RNA biosynthesis in the presence and absence of
N I H 759t is shown in Fig. t.
DNA biosynthesis in toluenized E. coli B was slightly in-
hibited (~-~t8 per cent) by N I H 7591 but high radioactive
counts in blank samples in which trichloroacetic acid was
present rendered the numerical estimates of inhibitions
somewhat uncertain. The important point is t h a t concentrations
of NIH 7591 which completely inhibit bacterial growth and
preclude RNA and DNA biosyntheses in intact E. coli B
have little influence on nucleic acid biosyntheses in toluene-
treated cells. Concerning RNA biosynthesis, Simon [7] stated
t h a t numerous attempts to demonstrate inhibitions of the
cell-free RNA polymerase reaction by " v e r y high levorphanol
concentrations" have been unsuccessful.
Naturwissenschaften 62 (t975) 9 by Springer-Verlag 1975
2Boo
X Xl,Sx IO-3M
2600 .x / 7591
2400
2200 CONTROL
2000
:E.,<
a- I 1800
(3o
1600
1400
1200
1000
800
600
400
2O0
I , I , i f ~ r 1 , , I
15 30 45 60
MINUTES
Fig. I . Time course of incorporation of 14C-adenosine tri-
phosphate into RNA of toluene-treated E. coli B in the absence
or presence of N I H 7591. Method is described in [6]
We conclude t h a t chromosomal DNA replication and RNA
transcription in E. coli are not directly affected by morphine-
like bacterial-growth inhibitors and that the failures of these
polymerizations in intact ceils may result from drug effects
on those portions of bacterial membranes which are dis-
organized by treatment with toluene. Nucleic acid bio-
syntheses in E. coli which were plasmolyzed by a sucrose-
EDTA mixture remained sensitive to levallorphan [8].
Received December 12, t 974
t. Simon, E . J . , Van Praag, D.: Proc. Nat. Acad. Sci. US
51, 877 (t964)
2. Greene, R., Magasanik, B.: Mol. Pharmacol. 3, 453 (t967)
3. Wolfe, A.D., Olenick, J.G., Hahn, F. E. : Antimicr. Agents
Chemoth. (in press)
4. Mellett, L.B., Woods, L.A.: Progr. Drug Res. 5, t55 (1963)
5. Moses, R.E., Richardson, C.C.: Proc. Nat. Acad. Sei. US
67, 674 (t970)
6. Peterson, R.L., Radcliffe, C.W., Pace, N.R.: J. Bact. 107,
585 (1971)
7. Simon, E . J . , in: Narcotic Drugs, Biochemical Pharma-
cology, p. 330 (D.t-I. Clouet, Ed.). New York-London:
Plenum Press 1971
8. Ben-Hamida, F., Gros, F.: Bioehimie 53, 71 (I971)
O u i n a c r i n e (Atebrin) :
I n t e r a c t i o n s w i t h R N A and w i t h R i b o s o m e s
Anne K. Krey and F. E. Hahn
D e p a r t m e n t of Molecular Biology, Walter Reed Army Institute
of Research, Washington, D.C. 20012, USA
Quinacrine binds to duplex DNA by intercalation [I] and
strongly inhibits DNA replication in vivo [2]. An ancillary and
lesser effect of the drug is an inhibition of protein biosynthesis
[2] and of the polycondensation of phenylalanine in a ribosome-
poly U system E3]; this latter effect is reversed by increasing
concentrations of ribosomes [3]. Quinacriue Iabilizes ribosomes
in vilro to h e a t [4]. We report optical observations on the
interaction of quinacrine with RNA and with ribosomes.
Fig. t a shows effects of t-RNA, poly rG and poly rA on the ab-
sorption spectrum of quinacrine. The major absorption bands
were decreased in intensity and exhibited bathochromie shifts.
Binding of the drug to DNA causes similar phenomena [5]-
Fig. t b compares the Cotton effect in the ORD spectrum of
quinacrine, induced by binding to t-RNA, with those induced
by binding to double- and single-stranded DNAs ~5]. The mole-
cular amplitude of the effect with t-RNA, centered around
350 nm, is smaller than that induced by duplex DNA but
greater than t h a t induced by single-stranded DNA; t - R N A
possesses extensive double-stranded regions.
Fig. 1 e shows that the thermal denaturation profile of t-RNA
was shifted by quinacrine to a higher temperature and rendered
99