_??_1995 The Japan Mendel Society Cytologia 60: 353-358, 1995

Karyotypes of sect. Vicilla from Northeast China

Ruijun Li and Haoyou Wang

Department of Biology, Harbin Normal University, Harbin 150080, China

Accepted September 6, 1995

Since Kupicha (1976) have made a revision of Vicia L. in the world, sect. Vicilla included about fifteen . Three taxa among them, V. pisiformis, V. dumotorum and V. sylvatica mainly distributed in Europe, and V. sylvatica distributed to the North of Asia. The most taxa in this section belongs to Asian species, distributed in Siberia in Russia, Northeast, North China and West China in China, Mongolia, Japan and Korea, such as V. amoena, V. amurensis and V.japonica et al.. Chromosome numbers and karyotypes of Asian sect. Vicilla were reported by Huiziwara and Kodon (1963), Lee (1972), Nikiforova (1984), Liu (1986), Rudyka (1986), Luo and Wang (1989) and Sokolovskaya (1989). Cincura (1973) and Terziiski (1974) reported karyotypes of V. pisiformis and V. dumetorum distributed in Europe. The present paper reported karyotypes of three taxa of sect. Vicilla in Northeast China. It was discussed the relationships among taxa of sect. Vicilla and evolutionary significance.

Material and methods

The of the taxa used in the present work were directly collected from wild population in Northeast China (Table 1). The seeds were put on a moist filter paper, and germinated at 22•Ž. When the roots grew and elongated about 1-2cm long, the root tips were cut off and pretreated with socurated ƒÀ-dichlorobenzene solution about 4-6hr. Then they were fixed in Carnoy's solution 3-24hr.

The materials were hydrolyzed in 1N HCL at 60•Ž for 6-8min, stained in modified Carbol Fuchsin solution and squashed in 45% acetic acid. And photography of the metaphase cells were taken using OLYMPUS BH2 microscope. For karyotype analysis, the systems proposed by Levean et al. (1964) and Stebbins (1971) were followed.

Results

1. V. japonica (Figs. 1, 2): The Chromosome number of this species, 2n=24, was consistent with the finding of Huziwara and Kodon (1963). The metaphase chromosomes ranged in length from 4.93ƒÊm to 7.55ƒÊm and in arm ratio from 1.02 to 6.60. The 24 chromosomes were classed into three groups; four pairs of metacentric, five pairs of sub metacentric and three pairs of subtelocentric chromosomes. The 2nd and 4th pairs of chromosomes were SAT-chromosomes, in which satellites were on short arms of those chromosomes. Karyotype showed moderate symmetry (2A type). 2. V. amurensis (Figs. 3, 4): The somatic chromosome number was found to be 2n=12.

The somatic metaphase chromosomes varied in length from 4.95ƒÊm to 7.45ƒÊm and arm ratio from 1.23 to 3.95. These chromosomes were classed into three groups; two pairs of metacen tric, three pairs of submetacentric and one pair of subtelocentric chromosomes. Two pairs of satellites present respectively on short arm of the 2nd and 3rd pairs of chromosomes. Karyo type showed moderate symmetry (2A). 3. V. pseudo-orbus (Figs. 5, 6): The chromosome number was 2n=12, which was 354 Ruijun Li and Haoyou Wang Cytologia 60

Table 1. The chromosome numbers and origins of three Vicia species in this paper

confirmed in the previous reports (Liu 1986, Nikiforowa 1984). The metaphase chromosomes had a range of 4.09ƒÊm to 7.45ƒÊm and 1.28 to 3.69 in arm ratio. They were divided into three

groups consisting of one pair of metacentric, four pairs of submetacentric and one pair of subtelocentric chromosomes. SAT-chromosomes were the 2nd and 3rd pairs of chromosomes in complement. Karyotype showed moderate symmetry (2A type).

Discussion

1. Karyotypic features of the three taxa and their taxonomic implication The somatic chromosome numbers made in three species of the present study are agreed partly with the earlier reports (Nikiforova 1984, Nishikawa et al. 1985, Liu 1986, Rudyka 1986, Luo and Wang 1989, Sokolovskaya et al. 1989). Rudyka (1986) observed presence of B chromosomes in V. amurensis and V, pseudo-orbus. It is interested that diploid cytotypes in V. japonica with different basic chromosome number (x=6 and 7) were reported respectively by Nishikawa et al. (1985) and Sokolovskaya et al. (1989), but we didn't find any diploid cytotype of this taxon in Northeast China. Karyological investigations of Vicia amurensis and V. pseudo-orbus in present paper differed considerably from the previous results in Japan (Huziwara and Kodon 1963) and Siberia (Nikiforova 1984). The chromosome complement of V. pseudo-orbus in Japan (Huzi wara and Kodon 1963) was mainly consisted of subtelocentric chromosomes. The numbers of the subtelocentric chromosomes decreased obviously in one of Siberia (Nikiforova 1984). Nikiforova (1984) observed four pairs of subtelocentric chromosomes it V. amurensis and didnot find any SAT-chromosome. Intraspecific variation of karyotype in the two species occurred. The karyotype reported in this paper was more symmetric than ones of Japan and Siberia. In the present investigation it was shown that V. amurensis and V. pseudo-orbus were diploid and had similar karyotypes. As they all had the same karyotype symmetry (2A), comparable chromosome size and the same type of SAT-chromosomes. Two types of SAT -chromosomes were found in the complements of those species: length of satellite was euqual to one of short arm in one type, and in other type length of satellite is longer than one short arm. However, they differ in some finer details. The first type of SAT-chromosome was respectively the 2nd pair of chromosomes it V. amurensis and 3rd pair of chromosomes in V. pseudo-orbus, and 2nd type of SAT-chromosomes were respectively the 3rd pair in the former and 2nd pair of chromosomes in the latter. The karyotype of V. japonica observed in this study was different from one of Nikiforowa (1984), who did not observe any SAT-chromosomes. Thus, all the 1995 Karyotypes of Vicia sect . Vicilla from Northeast China 355

Figs. 1-6. Somatic metaphase chromosomes and karyotypes. 1, 2. Vicia japonica . 3, 4. V. amurensis. 5, 6. V. pseudo-orbus. Bar=10ƒÊm.

three taxa of Vicia, having comparative chromosome size, karyotype symmetry and chromo some type, exhibited close relationship which indicates origin from a common ancestor . 356 Ruijun Li and Haoyou Wang Cytologia 60

Table 2. Karyotypes of three Vicia species () in Northeast China

Relative length of short arm included length of satellite.

2. The origin and diversity of sect. Vicilla in Northeast China Most researchers accepted that basic chromosome number of x=7 was a primitive basic chromosome number in the genus and the other was derived from it (Rousi 1961). According to data of chromosomes reported in Vicia (Fedorov 1974, Goldblatt 1981, 1984, 1985, 1988, 1995 Karyotypes of Vicia sect. Vicilla from Northeast China 357

1991), distribution patterns of basic chromosome number in sect. Vicilla were differentiated between Europe and Asia. European taxa most had basic chromosome number of x=7, such as V. dumetorum and V. sylvatica. And taxa distributed in Asia were polytypic in basic chromosome number, having basic chromosome numbers of x=5, 6 and 7. Most of them were diploids at basic chromosome number of x=6, and intraspecific and interspecific tetraploids were ocuured. In addition, European taxa were different from Aisan taxa in chromosome complement (Cincura 1973, Terziiski 1974, Li et al. 1991, 1992, 1993, Li 1993, Li and Wang 1994). Karyotypes of the former were consisted of metacentric and submetacentric chromo somes. For example, complements of V. dumetorum and V pisiformis respectively had four and five pairs of metacentric chromosomes. Karyotype of the latter was consisted of three types of chromosomes, i.e. metacentric, submetacentric and subtelocentric chromosomes. In those karyotypes, the number of metacentric chromosomes decreased obviously, and the number of submetacentric chromosomes increased and one or two pairs of subtelocentric chromosomes were observed. The above results showed that karyotypes of European taxa were more primitive and symmetric than Asian taxa. Stebbins (1971) considered that species with more symmetric karyotype were specialized in morphological features. It was noticed that Asian taxa were polytypic in morphology. According to tendrill and stem, those taxa can be divided into two kinds: without tendrill and with eric stem, and plants with tendrill and ascending stem. The former distributed in forest, the latter distributed in various habits such as roadside, forest edge et al. Hence present authors suggested that Aisa was a centre of modern diversification of sect. Vicilla, and Europe was an origin centre of this section.

Summary

In this paper, chromosome numbers and karyotypes of three species, Vicia amurensis, V. pseudo-orbus and V. japonica, were investigated. V. amurensis and V. pseudo-orbus were diploid taxa, and V. japonica was a tetraploid taxon. The karyotype formulae were as follows: V. amurensis 2n=12=4m(2SAT)+6sm(2SAT)+2st; V. pseudo-orbus 2n=12=2m(2SAT)+ 8sm(2SAT)+2st; V. japonica 2n=24=8m(4SAT)+10sm+6st. The karyotypes of the three species belonged to '2A'. According to karyotypic features, the authors consisdered that karyotypes of the above taxa were similar. Finally, it was discussed that variation and evolution of karyotypes in Vicia sect. Vicilla.

Acknowledgment

This project supported by National Natural Sciences Foundation of China and Cai Huosi Foundation for Biological Science Development.

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