Intra-specific Polyploidy and Possible Occurrence of Some Genetic Types for Pollen Development in Cayratia japonica, Vitaceae.

7 Pages·2003·1.2 MB·English

by  Okada, Hiroshi

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ISSN13647565 Acta Phytotax. Geobot.S4(1)6:9-75(20e3)

Intra-specifiPcolyploidy PossibleOccurrence Some Genetic
and of

Types forPollenDevelopment in Cayratial'aponicVait,aceae.

HIROSHIOKADAi TSUKAYA2 MOTOHARU OKAMOT03
and
,HIROKAZU
iBotanical Gaizlen sf,i'acul toyfScien cOsea,ko City Uhivensit Kyt,itano ,Osaka 576-OO04 ,JZipan 2;?Vdetional
institutefBoarsicBiologyvCenterfoirntagrativBeioscienceM,),odat7i-Okeahzoa,ki444-8585,lapan;30saka

imseum ofNaturHailstoTyNagaiPburkH,igashi-Sumi.voshi-Oksua,ka546-O042,Jdpan

Cayratia intra-specific i.e,2,nz40 (diploid)60 (triploid).
1'aponica showed polyploidy of x=:20, and
Individua lwisth 2n=40 produced fruit san,d fbrrne d20 bivalent ast metaphase I in meiosis, but their
polle nfertili twiicleesly varied from about 30 to 900, GI.ndividual swith 2n-60 did not produce any ffuits,
formed a variable number of triva]ent s,bivalen tasnd univalents at rnctaphasc I ,and showed low
pollc nfertili tofileesss than 30%. The possibl oeccurrcnce ofsome gencti tcypcs for polle ndevelopment
indiploidsisdiscussed,

Key words: ymponica, chromosomc, mciosis, ploidy ,po]lendcx,clopmen pto,lle nfertility.
VitaceacCayiutia
CliyratiaJ'aponi c(aThunb G.ag)n. is well known as observation). At some places b,oth fuii tan-d non-
a wild climbing plant in Japan ,Attenti voebservers fruit-individ guraolws closely together ,and insects
know that this species contains both individualssimilarly visit both individual Tsh.erefore e,nvi-
with imit sand without ffui tisn nature. From a gen- ronmental conditions do not seem to be the limiting
eral view point ,individual swithout fruit s(here-facto rin frui tformatio nS.ome species-specific sit-
after non-fruit-individuals) are frequent lobyserved uation rnust affbct the reprQductive traits .In some
at locatio nunsder strong horticultu irmaplact ,such cases the cyto'genetic aspects affect reproductive
as newly established squares after land develop- trait ss,uch as ploidy level sand meiotic aspects
ment, hedges and green belt salong roads, while Cfo erxamples; Clausen et al. 1945, Cleland 1972,
individuals firuit(shereaftfiemrit-individuals)Sakamoto 1982,Noda & Kawano 1988).Infactt,he
with
grow mainly in more natural habitats ,such as the chromosome number of this species was reported as
boundaries forestbsu,sh, house inthe 2n-=40inChina(Huang 1988) 60inJapan
of and gardens et al, and
countryside. We want to know the reasen why some (Mitsuk &ur Hiayashi 1953) .Hence, it is plausible
plant sproduce fruit sb,ut others do not, On the that the differenc ebsetwee nthe frui ta-nd non-
other hand, the two morphs can be recognized by imit-individu amiigsht be due to cytological dif
leaf shape, namely, the fruit-individ furaelqsuentlyferences .To examine the hypothesi sabove, we
develo ptrifbliat eleaves i,n addition to standard, examined these two morphs cytologically. In the
pentafbli alteeaves ,while the non-fruit-individuals course of the presen sttudM additionallM we encoun-
scarcely develep trifblia tleeaves (ou urnpublished tered peculia vrariations in the fenilit iofcposllen of
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the fruit-individua lVsV.℃ attempted to clarify the exceptions. At metaphase II and anaphase II ,each
cause efthis phenomenon and discus sthe possible nucleus contained 20 chromosomes (Fi gI.C).
genetic differentia taimoonng fruit-individ uofalsTetrads were formed normally (Fig .ID),
C. J'qponiea, Microspores were almost the same size. At
division2,0
microspore cell chromosomes were
Materials and Methods observed (Fi g1E.), and two nuclei appeared te be
fbrmed norrnally. In some individual st,wo kinds of
The materials examined are liste idn thble 1. As pollen grains ,namely, normal and degenerative,
Cayratia.fupon icacna propagat evegetatively, sani- were observed (Fi g2.) .The degenerati vpeollen
ples were collected at at leas t1OO m interva wlisth- grains had a broken pollen wall, and their cyto-
in thc sarne lecalit yI,n some cases, we collected plasm was lost (Fig 2.B), Pollen fertilit iveasried
seeds, which were germinated after concentrated from 31% to 97% among the plant sexamined (Fig,
sulfuric acid treatment fbr 30 min. They were plant- 1F, [Iabl 1e),
ed at the Botanical Gardens ,Facult yof Science, Contrar yto the individua lwisth 2n=40, the
Osaka City Universit Xyl.ouche srpecimens are kept individua wlisth 2n=60 formed trivalents .bivalents
in Osaka Museum of Natural History (OSA) and and univalents, ofwhich the number varied among
Botanica Glardens ,Facult yof Science O,saka City cells, fbr example 1lm+12Ji+3 i1,0m+14u+2i (Figs-
University. 3C, D), and 9m+15if+3 [A.t metaphase II ,the po]len
Both somatic and meiotic chromosomes were mother cell contained nuclei with various chromo-
observed with the same methods as in a prcvious some numbers, for instanc ei,n some cases, both
report (Okad 1a984), Microspores wcre stained by nuclei had 30 ¢hromosomes, but in other cases,
the Feulgen' sreaction. Tb estimate the polle nfer- one consisted of33 chromosomes, while the other
tilit yp,ollen grains were stained with lacto-phenol was 27 (Fi g4.A). There were various abnormal
cotton blue solution, and darkl ystained and globose polle ngrains i,.e. w,ith a broken pollen wall, with-
grain swere counted as ferti lgerain sfrom a total out cytoplasm, or smaller or large rsized grains
sum of500 to 1,OOO grains (Fig .4B), Pollen fertiliti ewesre less than 30%
(Tabl1)e,

Results
Discussion
In this study, we observed two d{ffere nchtromo-
some numbers, 2n=40 and 60, within the species In this study it became clear that the basi cchromo-
distribu tien dJapan (Fig IsA., 3A, B). Chromosome some number of Cayratia,iaponica is 20 judging
sizes varied in lengt hfrom about 2 to 1,2 pm at the from the meiotic aspccts, that is ,all ofthe individ-
somatic metaphase. All the individua wlitsh 2n=40 uals observed with 2n=40 fbrmed 20 bivalents,
produced imit sand good seeds, while those with while thos ewith 2n=60 forrne ad variable number of
2n=60 did not produced any fruit s(Tab l1e) or trivalents ,bivale natnds univalents, All ofthe fruit-
exceptionally produced a few fruit s(dat naot individual hsad 2n-40, namely, diploi dwh,ile all of
shown). Similarl yal,l the individua lwisth 2n-40 the non-fruit-individuals possessed 2n:=60 ,i,e.,
were frequent olbyserved to develo ptrifbli alteeavcs,triploi d,Probably ,diploi dindividua lcasn propa-
while all those with 2n=60 did not (da tnoat shown). gate from both seeds and cuttings of the subter-
In meiosis, the individua wlitsh 2n40 fbrmed ranean stem. On the other hand, triploi idndividuals
20 bivalen tast metaphase l (Fi gI.B) without any may propagate only from cuttings of the subter-
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TABLE 1. Localitie sso,matic chromosome numbers, ehromosome pairin gat rneiotic metuphase l, frui ftbrrnatio annd polle fnertili-
tiesofCqy'ratia.faponica.
locality ehromosome imitfbrm-atploolnl+e4n)f+e+n+i+l+i+vto+uychcer%)
number2n
n
Nanise,Machida.Tekyo 4040404040404040404e404040 Kitamura s.n. (OSA 153352)
Shizen-kansatsu-no-mori,Ybkohama,KanagawaPrefi Kitarnura s.n. (0SA 153353)
Myodaiji-che-4,Okazaki,AichiPref. 638456853297T8s1u4k2a9y6a31106U5l

Tlashiro-cho,Higashi-ku,Nagoya,AichiPref Tsukaya103
Kaishe-no-mor iA,ichi Prefi 20]} Tsukaya107
Okazaki Castle M,yodaiji-ch oO,kazaki ,Aichi Prer. Tsukaya 106
Ohsumi-ga-oka, Kyotanabe, KyotoPref Okada5632

Kisaichi,Katano,OsakaPref, 2ell?)20]j:J Okada 5637
Bet.Gard.Kisaich,Katano,OsakaPrer, Okada560S

Hamadera, SakaL Osaka Pref. Yamamoto s.n. (OSA I496I7)
Mukos,awa, Nishinomiya, HyogoPvef, Okada 5623

Mikushi-machi, Totteri ,Tottor Piref. 201ib ++ Okada5631
Campus of Tottori Unix,. 1- , Koyama, Tottori, Okada 5629
Tbttor iPref,
Campus ef Totter iUniv.-2, Koyama, Tottori, 40 + 76 Okada5606
Tbtto rPiref.
Motobu-cho ,Okinawa lsl .O,kinawa Pref. 40 + 85 Okada5638
Mt. Azvma, Scndai,MiyagiPreil 606060606U6060 4) YbkoyamaOI09?O

Carnpus ofUniv. of Tokyo, Hongo, Tbkyo 33+27--30+3oi} 1692e25162S Tsukaya 10S
Myedaiji-che-1,Okazaki,AichiPreL Tsukayalel

Myodaiji-cho-3,Okttzaki,AichiPref, IIIs+IIs-LIs]) T s uk av"aT]0s4ukaya
Myodalji-cho-2,Okazaki,AichiPref. IIIs+IIs+Is'ii 102

Bot,Gard,KisaichK,atano,OsakaPref 1]ls+ITs+Isi) Okada5636

Bot. Gard, Nagai ,Osaka, Osaka Pref. Okamoto s.n. (OSA 153508)
i)rnetaphasetl
:) metaphase I
]] jlls :trivalents, I[s :bivalents ,Is :univalents
4] +: produce fruit s-, :does not produce fruits
ranean stem when ornamental plants are trans- mally advanced to meiosis, but microspore degen-
planted in human horticultu racatlivities. Exccp- cration occurred during the tetrad stage and
tionally, triploid individua lpsroduced few imits, microspore cell divisio nI.n this study, we cannot
which might haye occurred due to the rather high specify what geneti cor physiologic aplhenQmena
fertili tiy,,e, ,about 9-2g%, affect microspore degeneration,
Although all of the diploid psroduced imits, Okada ( 1984) observed that an artificial diploid
they showed a wide range of variation in pollen hybri dbetween Ranuncttlu ssilerijblius and R. chi-
fertili tfyr,om 31 to 97% (Tab l1e) .Low pollen nensis exhibited almost nomial mciosis, but had a
fertili tdiied snot arise from abnormalities in mcio- very low polle nfertil itTyh.is contradiction was
sis, that is ,all of the individual swith 2n-40 nor- explained on the assumption that mu]tiple genes
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FIG. 4. Cytogenet cihacractcristics ofthe non-ftuit-individua]s of Cayixitiajapo mAc:a 3,3
chromosomes m one of the nuclei at metaphase II ,B, polle onfan individu awilth 2n= 60.
Bar in A, 4 ,um .m B, 1Oe pm,
concemcd with polle ndevelopment wcre distributedyariation (Gra n1t956) .Ifth evarious pollen fertili-
on differe nchtromosomes. If the gene combina- ties observed in diploi dCayratia ]'aponi caarose
tion was differenti abteetdween both species, tetrads from an incompatib lceombination of genes con-
of the Fi hybrid would contain a recombined gene cemed with polle ndevetopment a,s is the assump-
combination. Such recombined genes acted incom- tion in the case of thc inter-spec ihfyibcri dbetween
patibl yin pollen development ,thus reducing the Ranttnculu ssirelijblius andR. chinensis, C, J'opon-
pollen fertil iItn yf.act P,rociss iet al, (200 1sh)owed ica must be composed of multiple geneti ctypes
examples of multiple genes concerned with pollen that affect the reproductive isolati boentween types.
development in Arahidopsi tshaliana, and Fishman It is necessary to analyze whether there are any
& Willi s(2001 r)eported on the effect of recom- geneti ctypes within the species, and whether mul-
bined genes on the infertil oiftthye next generation tipl egcnes concerned with pollen developmen tin
in Mimultts. this species have differentiat ewidthin the species.
It is generall ayccepted that onc of the very
important steps in speciation is the establishrnent of References
an isQlati omnechanism between newly derived
populations (fo rexample; Stebbins 1950, Grant Claugen, J. ,D, D, Keck & W. M. Hiesey. 1945.
1981) .We have varieus examples showing repro- Experimental studics on the nature of species. II.
ductiv eiselati obny mutation of the chromosomal Plant cvolution through arnphiploidy and autoploidy
with examples from the Madiinae .Carnegie Inst.
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sterility <fb erxainple; Okada 1981) ,or by genetic EN'olution ,Academic Press ,Londen & New Ybrk,
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Columbia Univ .Press, New Ybrk. Pellti e&r S. Bonhomme, 2001, Five gametophytic
Huang, S,-F Y.:,-Q .Wang, Z.-Y LChen & X.-H, Shi ,1988. mutations afTecting polle ndevelopmcnt and pollen
Plant chromosome count$ (4) S.ubtrop. Forest Sci. tube growth in Arabidqpsis thatiana. Genetics
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Kromosomo 17-19:633-638. Agxopyron tsukushiense in the tribe Triticeae,
Neda, S. & S. Kawano. 1988. The biology ofAtlium Gramineae. Bot. Mag. [[bkyo 95:375-383.
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Recelved Nobember 14, 2002; aecqpted Mai=h 20, 2003
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