B I O L O G I A P L A N T A R U M (PRAHA)

1 ( 4 ) : 3 2 8 - - 3 2 9 , 1959

BRIEF COMMUNICATION

P r o b l e m s o f Autosterility in Nicotiana alata LINK et OTTO

(Formation o f the inhibitory factor in the pistil)

~IARMILA TOM~OVX
D e p a r t m e n t of Genetics, Faculty of Biology Charles University, P r a h a

Received June I, 1959

Souhrn: PrSco je zam6~ena zvl~w n a }e~ieni ot~zky, zda je u autosterflniho d r u h u Nicotiana

alata LINx et OTTO var. grandiflora C o ~ s inhibiSni slo~ka v y t v ~ e n a nezg~visle n a opylenl,
nebo vznik~-li a~ vlivem pflsobenl prorfistajicich inkompatibilnich pylov~eh lhSek. Ot~zka byla
~e~ena pomoci um~16 kultivace pylu n a a g a r o v ) c h filmech zShsti pokryt:~ch rozt~ry pestikov~eh
pletiv. Pestiky byly p}ed preparaci b u d opyleny kompatibiln~ nebo inkompatibfln~, nebo po-
ncchgmy bez opyleni. Mezi pestlkovl~mi pletivy (v nich~ sterilJtnl reakce b u d prob~hla, nebo
neprob6hla) a kliSicim pylem, byla vys6v~nim pylu b u d t6ho~i nebo rozdfln6ho genetiek~ho
pfivodu n a preparhtech improvlsovgma obdoba inkompatibilnich nebo kompatibilnich vztah~.
Z v~sledkfi vyvozuje autorka p~edb6~.n:~ zSv~r, ~e je inhibiSnl slo~ka v pestlku pravd~po-
dobni~ p~itomna ji~ p~ed opylenlm.
Summary: F r o m the partial results of artificial cultures in the presence of pistil tissue-pulp
the author draws the preliminary conclusion t h a t the inhibitory factor in the pistil of Nicotiana
alata LINK et OTTO is already present before pollination.
These questions were investigated b y means of artificial cultivation of pollen tubes on agar
in the presence of compatible a n d incompatible pulped pistils which had been previously prepared
in various ways.
This report gives the results of the last series of experiments (part A in the scheme). I n the
series of earlier experiments no signs of a n y marked inhibition were recorded.
Material and Methods. Pollen a n d pistils of the autosterile species Nicotiana alata LINK
et OTTO var. grandiflora CoMEs were used.
I. Preparation of the pistil pulp: in each case three washed and dried pistils were deprived
of their stigmas (which might contain substances stimulating pollen-tube growth) a n d nectaries
(because of high content of earotenoids). They were then quickly pulped in a m o r t a r with one
drop of distilled water. The pulp was immediately transferred to glass plates covered with
thin agar films (1.5~o agar). The required pollen was immediately scattered on the pulp a n d the
surrounding agar (for control), a n d the plates were placed in damp chambers a n d left for 16 to
18 hours a t a temperature of a b o u t 20 ~ C.
II. Three days before the experiment the pistils used for the preparations were a) pollinated
incompatibly, b) compatibly, c) left unpollinated. Pollen scattered on the pulp from these
pistils was: A. incompatible (from the same p l a n t as the pistils), B. compatible (from a plant
of a different genotype t h a n the pistils). The arrangement of the experiments is shown in the
table.

328
 AUTOSTERILITY IN NICOTIA1VA A L A T A 329

III. Evaluation of the preparations was carried out twice: in the fresh state, and following
fixation mud staining, which made it e~sier to distinguish the objects under observation from
the pulp. The evaluation was made b y comparing the lengths of the tubes with the pollen grain
diameter, according to a given scale of inhibition grades.
The preparations: fixation in 70% alcohol, following complete drying stained for 1~/2minutes
in 0.25% safranin solution in 70% alcohol; staining completed by brief immersion in 1% aqueous
aniline blue solution. The bright purple-stained pollen grains with dark blue tubes stood out
in the bluish-stained pollen tissue pulp. Preparations preserved by covering with a kopolymere
film (mcthylmetakrylate), or b y normal mounting in Canadian balsam.
Results. Considerable inhibition was exhibited in the main sector of the experiment, series A,
in its variants b (in 61.11% cases) and c (53.330/0), in ear. a) 10.50% cases of inhibition only.
Its frequency in variant b) is significantly higher t h a n in variant a): Z2 = 10.397, the probab-
ility of the occurrence of the observed difference in a homogeneous population is only about 2%;
the frequency of inhibition in variant c) is almost significantly higher than in variant a): Z~
= 7.401, with probability of occurrence of t h a t difference being in the region of 5% significance
limit. Thus inhibition of the growth of pollen tubes is exhibited in variants b) and c).

Experimental scheme
Pulp from pistils Series Pulp from pistils Series
of plant 1. A B of plant 2. B A
Pollen from Pollen from
plant plant

vat. pistll: pistil:

a pollinated with pollinated with
incomp, pollen 1 2 incomp, pollen 1 2
(from plant 1) (from plant 2)

b pollinated with pollinated with

comp. pollen 1 2 comp. pollen 1 2
(from plant 2) (from plant 1)

o unpollinated 1 2 unpollinated 1 2

BIOL. PLANT. l (4) : 328--329, 1959

I~ npo6~eMe aBTO(~TepHJIbHOCTIIy Nicotiana alata LINK et OTTO

(06paaomanHe nHrn6HRnonnoro Havana n necTnre).

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(~auwop HCCOBMeCTgMOCTn Haxo~HTCF[ B necwgKe Nicotiana alata LINX e t OTTO e a r . grandi-
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Address: Jarmila TomkovA, Prom. Biol.,Genetick:~ fistav, Biologick~ fakulta University Kaxlovy,
Vini~ng 5, Praha 2.