Genetic variations in Lycoris radiata var. radiata in Japan

The genetic variations of Lycoris radiata var. radiata, a completely sterile triploid from Japan, were examined by comparing the nucleotide sequences of genomic DNA regions in 11 triploid strains sampled from Japan and four triploid strains sampled from China, and in two diploid strains of Lycoris radiata var. pumila, which is endemic to China and fertile. For this purpose, two genes were analyzed, the lectin gene in the nuclear genome and the maturase gene in the chloroplast genome. A clear genetic constancy was observed in their DNA nucleotide sequences. For both genes, completely identical nucleotide sequences were detected in the 11 Japanese and four Chinese triploid strains and also between the two Chinese diploid strains. However, some genetic variations were observed between the Japanese and Chinese triploid strains, and between the triploid and diploid strains. These results are consistent with the findings obtained from previous chromosome karyotype analyses and allozyme analyses. In addition, in our preliminary FISH analysis of the physical mapping of the rRNA gene family, the 18S-5.8S-26S rRNA and 5S rRNA loci were localized on six and four chromosomes, respectively. Regarding the 18S-5.8S-26S rRNA loci, two were associated with two SAT chromosomes. The remaining four were distinguished by having no secondary constriction. Localization of 5S rRNA loci to chromosome spreads revealed three sites on the proximal part of the long arm of three acrocentric chromosomes and one site on the distal part of the long arm of the SAT chromosome; the latter site was juxtaposed to the 18S-5.8S-26S rRNA loci. These findings indicate that L. radiata var. radiata is not a typical autotriploid. The present paper discusses the possible origin of L. radiata var. radiata from a diploid variety of L. radiata var. pumila, based on the molecular cytogenetic analysis and DNA sequence analysis.     

基于叶绿体DNA atpB-rbcL区序列探讨石蒜属种间系统发育关系

摘要：为了探讨石蒜属（Lycoris Herb.）的种间系统发育关系,对石蒜属95个材料包括15种、4变种及2个人工杂种的叶绿体 DNA atpB-rbcL间隔区进行了测序,结合花部形态和核型特征,探讨了石蒜属种间系统关系及其可能的杂交起源,结果表明：在系统发育树上亲缘关系近的材料聚在一起,其中矮小石蒜（L. radiata var. pumila）和换锦花（L. sprengeri）与2个人工杂交种（Hybrid 1、Hybrid 2）、麦秆石蒜（L. straminea）、江苏石蒜（L. houdyshelii）、短蕊石蒜（L. caldwellii）和乳白石蒜（L. albiflora）具有密切的亲缘关系。atpB-rbcL序列揭示的石蒜属种间关系与染色体核型的分类结果部分一致,主要表现在具有近端部着丝粒(A)染色体的种与具有中部(M)和端部(T)着丝粒染色体的种各成一支,与形态和染色体分类结果一致;不同之处在于具有中部、端部和近端部着丝粒染色体的种分散在两个主要分支内,进一步验证了具有中部、端部和近端部3种着丝粒类型染色体组的石蒜如麦秆石蒜、江苏石蒜、短蕊石蒜和乳白石蒜等是杂交起源的假设,结合2个人工杂交种分析,揭示了短蕊石蒜和乳白石蒜的近端部着丝粒染色体来源于换锦花;麦秆石蒜和江苏石蒜近端部着丝粒染色体来源于矮小石蒜。     

中国石蒜属种间亲缘关系ITS序列分析

本文利用核糖体DNA内转录间隔区(ITS)序列对石蒜属13个种(含变种)的亲缘关系进行分析。结果表明,各样品的ITS1长度为259~260 bp,ITS2为230 bp,分别有多个特异性信息位点。以ITS序列为依据对石蒜属植物亲缘关系进行分析,表明石蒜属13个种可分为三大类,其中类Ⅰ包括中国石蒜、地笑、安徽石蒜和长筒石蒜,核型为M+T型;类Ⅱ包括矮小石蒜、换锦花、玫瑰石蒜和红蓝石蒜,核型为ST型;类Ⅲ包括稻草石蒜、乳白石蒜、短蕊石蒜和两种人工杂交种,核型为ST+M+T。系统进化树与核型分析结果相似,第Ⅲ类可能为自然杂交种。     

A Study on Karyotypes of the Genus Lycoris

The genus Lycoris (Amaryllidaceae) consists of about 20 species, all of which are confined to temperate China, Japan and Korea. Cytological investigations, including a reexamination of the karyotypes of 14 taxa, measurements of relative nuclear DNA content, and meiotic configuration observations on some specific forms and interspecific hybrids, have been carried out by the present authors in order to re-evaluate the mode of karyotype evolution and the role of hybridization in the speciation of Lycoris. These have resulted in a new theory for explaining the karyotype evolution in the genus, which will be considered elsewhere. The present paper deals with observations on karyotypes of 11 species, 1 variety and 2 artificial hybrids. Results obtained through karyotype analysis, as shown by the data in Table 1, Plates I-VI and Figs. 1-2, reveal that: (1) the karyotypes of Lycoris rosea, L. radiata var. pumila, L. sprengeri, L. haywardii, L. caldwellii, L. squamigera and L. radiata are, on the whole, consistent with those reported by the previous authors^{[1,2,3,4,5,8,10,12]};(2) the I (rodshaped) chromosomes of L. chinensis and L. longituba are all T’s (telocentric) instead of t’s (acrocentric) or t(Sat)’s; (3) the three materials of L. aurea of different sources have shown a karyotypic differentiation: one with 2n=14=8m+6T, and the others with 2n=16=6m+10T: (4) both of the karyotypes of L. straminea and L. albiflora are 2n=19=3V+6I, inconsistent with 2n=16=6V+10I for the former and with 2n=17=5V+12I for the latter as reported by Inariyama (1953), Bose and Flory (1963) and Kurita (1987). The following aspects are worthwhile discussing: 1. The types of chromosomes. Karyotype analyses reveal the existence of three major chromosome types in Lycoris: (1) m (metacentric) chromosomes: (2) t (acrocentric) chromosomes, with short arms, (3) T (telocentric) chromosomes, sometimes with dot-like terminal centromeres. To distinghish t’s from T’s is of paramount importance for solving the problem of karyotype evolution in Lycoris. Bose (1963) pointed out that in the species with 2n=22, all I chromosomes were t’s, while in species with 2n=12-16, all I chromosomes were T’s. Our results of chromosome observations are consistent with Bose’s remarks. Some authorst^[3,6] have probably mistaken the dot-like terminal centromeres of T’s of L. longituba and L. chinensis as the short arms of t’s. 2. The significance of Robertsonian change in karyotype evolution. Although chromosome numbers and karyotypes are very variable in Lycoris, as shown in Table 1, the total number of arms of a chromosome complement of any species is always multiples of 11. Hence, it seems likely that Robertsonian changes have taken part in karyotype alteration, The genus has a series of basic chromosome numbers: 6, 7, 8 and 11. But which is the most primitive one? It is uncertain whether a successive decrease in chromosome numbers as a result of Robertsonian fusion or a gradual increase in chromosome numbers brought about by fission (fragmentation) has been the essential mechanism for karyotype evolution and speciation in Lycoris. These problems are of crucial importance and will be discussed in our subsequent papers. 3. The origin of polyploids. As evident from Table 1, there are two levels of ploidy differentiation in Lycoris: (1) di ploids with 2n=22 or the equivalent of 22, (2) triploids with 2n=33 or the equivalent of 33. The most common way of origination of triploids in plants is the hybridization of diploids with Tetraploids. But tetraploids have never been found in Lycoris. Thus, it is suggested that the triploids have originated from the combination of an unreduced gamete of a diploid with a normal gamete of another diploid. 4. The role of hybridization in speciation. Results of karyotype analyses show that hybridization has taken an important part in the speciation of Lycoris. Two types of hybrids have been found: (1) 2n=19= 3V+ 16I, L. straminea, L. albiflora and the two artificial hybrids L. sprengeri×L. chinensis and L. haywardii× L. chinensis all possess this karyotype. It could be seen from the above chromosome number and karyotype that this sort of karyotype is exactly half of the total sum of 2n=22I and 2n=16= 6V+10I. It is, therefore, quite evident that taxa possessing this karyotype are all diploid hybrids of 2n=22 and 2n=16, (2) 2n=27=6V+21I, L. caldwellii and L. squamigera possess this karyotype. It is reasonable to assume, too, that they are segmental allotriploids and have arisen from the combination of an unreduced diploid gamete of 2n=16 and a normal haploid gamete of 2n=22. The origin of the hybrid karyotype 2n=17=5V+12I reported by Inari- yama (1953) is similar to that of 2n=19, except that one of the parents possesses 2n=12= 10V+2I instead of 2n=16=6V+10I. The origin of the other hybrid karyotype 2n=30=3V+ 27I reported by Bose (1963) is similar to that of 2n=27, but the diploid gamete comes from taxa possessing 2n=22 instead of 2n=16.     

Different maternal genome donor to Kengyilia species inferred from chloroplast trnL-F sequences

To reveal the maternal donor of species in genus Kengyilia, the chloroplast trnL-F sequences of 14 Kengyilia species and several related diploid species were analyzed by using Maximum Parsimony (MP) and Bayesian Inference (BI) methods. The species in Kengyilia were clustered in different clades, which indicated that Agropyron (P) is the likely maternal genome donor to Kengyilia melanthera, K. mutica and K. thoroldiana, while the maternal donor to Kengyilia batalinii, K. nana, K. kokonorica, K. kaschgarica, K. hirsuta, K. alatavica, K. gobicola, K. zhaosuensis, K. rigidula, K. longiglumis and K. grandiglumis was St or Y Roegneria genome.     

安徽产石蒜两个居群的核型研究

观察了石蒜(Lycoris radinta)两个不同居群植物的染色体数目和核型,发现野生石蒜在一个植株的不同根尖细胞里,存在两种倍性的细胞,如生于宣城敬亭山的居群既有正常三倍体：2n=33=18st 15T,属于“4A”核型;还有异常二倍体：2n=20 1B=2st 18T 1B,属于“4B”核型;生于芜湖的居群核型为：2n=20 1B=lm 9T 4t 6st 1B和2n=20 1B=1M 9T 10st 1B,属于“3B”和“3C”核型。     

Molecular phylogeny of diploid Hordeum species and incongruence between chloroplast and nuclear datasets

The phylogeny of diploid Hordeum species has been studied using both chloroplast and nuclear gene sequences. However, the studies of different nuclear datasets of Hordeum species often arrived at similar conclusions, whereas the studies of different chloroplast DNA data generally resulted in inconsistent conclusions. Although the monophyly of the genus is well supported by both morphological and molecular data, the intrageneric phylogeny is still a matter of controversy. To better understand the evolutionary history of Hordeum species, two chloroplast gene loci (trnD-trnT intergenic spacer and rps16 gene) and one nuclear marker (thioreoxin-like gene (HTL)) were used to explore the phylogeny of Hordeum species. Two obviously different types of trnD-trnT sequences were observed, with an approximately 210 base pair difference between these two types: one for American species, another for Eurasian species. The trnD-trnT data generally separated the diploid Hordeum species into Eurasian and American clades, with the exception of Hordeum marinum subsp. gussoneanum. The rps16 data also grouped most American species together and suggested that Hordeum flexuosum has a different plastid type from the remaining American species. The nuclear gene HTL data clearly divided Hordeum species into two clades: the Xu+H genome clade and the Xa+I genome clade. Within clades, H genome species were well separated from the Xu species, and the I genome species were well separated from the Xa genome species. The incongruence between chloroplast and nuclear datasets was found and discussed.     

Isozyme evidence for the allotriploid origin of Lycoris flavescens (Amaryllidaceae)

 Variation in isozyme patterns from ten populations of the Korean endemic Lycoris species was used to test the hypothesis that L. flavescens originated from natural hybridization between diploid L. chinensis and L. sanguinea var. koreana. Lycoris sanguinea var. koreana shows fixed heterozygosity at four of nine loci assayed, suggesting that this species is an allotetraploid instead of a diploid. Electrophoretic data suggest that Lycoris flavescens is an allotriploid species derived from the hybridization between diploid L. chinensis and tetraploid L. sanguinea var. koreana. The patterns of allelic distribution in populations of L. flavescens suggest multiple origins of the allotriploid. Within the L. flavescens complex, our isozyme data support the recognition of two taxa, L. flavescens and a recently recognized species, L. uydoensis. Received August 28, 2000 Accepted December 27, 2000     

Phylogenetic relationships and the maternal donor of Roegneria (Triticeae: Poaceae) based on three nuclear DNA sequences (ITS,Acc1, and Pgk1) and one chloroplast region (trnL-F)

Roegneria is a polyploid perennial genus in the tribe Triticeae. Some species of Roegneria are morphologically similar to genus Elymus and have been classified in Elymus. To investigate the delimitation and phylogenetic relationships of Roegneria, nuclear (ITS, Acc1, and Pgk1) and chloroplast (trnL–trnF) DNA regions were sequenced for 38 allopolyploid species and 32 diploid species of Triticeae. Phylogenetic analyses of nuclear DNA revealed that all Roegneria species were included in the St and Y genome clades, and that the Y genome was closely related to the V and Xp genomes. The chloroplast DNA dataset showed that Roegneria species were grouped with Pseudoroegneria species. The Pseudoroegneria species from the Middle East (P. libanotica and P. tauri) and Central Asia (P. strigosa) were more closely related to Roegneria species. The results suggested that: (i) the species containing the St and Y genomes should be segregated from Elymus and treated as a distinct genus, Roegneria, based on the genomic constitution; (ii) P. libanotica, P. tauri, and/or P. strigosa potentially served as the maternal donor of the St genome in Roegneria; (iii) The Y genome of Roegneria originated from a diploid Y genome species, and the V and Xp genomes may have contributed to Y genome formation; (iv) among Roegneria species of previously uncertain genomic constitution, R. seriotina was tetraploid and possessed the StY genomes, E. calcicolus was hexaploid with the StYH genomic constitution and should be classified in Campeiostachys, R. glaucifolia possessed the StStY genomes, and R. tschimganica had the genomic constitution St₁St₂Y.     

Phylogenetic Analysis of Leymus (Poaceae: Triticeae) Inferred from Nuclear rDNA ITS Sequences

To investigate the phylogenetic relationships of polyploid Leymus (Poaceae: Triticeae), sequences of the nuclear rDNA internal transcribed spacer region (ITS) were analyzed for 34 Leymus accessions representing 25 species, together with three Psathyrostachys species (Ns genome), two Pseudoroegneria (St genome) species, Lophopyrum elongatum (E(e) genome), and Thinopyrum bessarabicum (E(b) genome). The phylogenetic analyses (maximum likelihood and Bayesian inference) supported two major clades, one including 21 Leymus species and three Psathyrostachys species, the other with nine Leymus species and four diploid species. The ITS RNA secondary structure of the Leymus species was compared with that of their putative diploid donor. It is suggested that (1) the species from the same areas or neighboring geographic regions are closely related to each other; (2) L. coreanus, L. duthiei, L. duthiei var. longearistatus, and L. komarovii are closely related to other Leymus species, and it is reasonable to transfer these species from the genus Hystrix to Leymus; (3) the ITS sequences of Leymus are evolutionarily distinct; (4) the different Leymus species and different distribution of a species derived their Ns genome from different Psathyrostachys species; and (5) there is a close relationship among Leymus, Pseudoroegneria, Lophopyrum, and Thinopyrum, but it is difficult to presume that the St, E(e), and E(b) genome may be the Xm genome donor of the Leymus species.     

应用流式细胞术测定17种中国野生蔷薇核DNA含量

以17种中国野生蔷薇为试材,采用改良的LB 01裂解液,以4种不同的标准植物——大豆（Glycine max Merr.‘Polanka’）、绿豆（Vigna radiata（L.） Wilczek）、番茄（Lycopersicon esculentum Miller）和玉米（Zea mays L.）为外标,以二倍体材料丽江蔷薇（Rosa×lichiangensis Yü et Ku）为内部参照,利用流式细胞术对其核DNA含量及染色体倍性进行检测,并采用常规染色体压片法验证倍性准确性。本研究首次检测了3个二倍体种——商城蔷薇（Rosa shangchengensis T.C.Ku）、广东蔷薇（Rosa kwangtungensis Yü et Tsai）和无刺刺梨（Rosa roxburghii f.inermis S.D.Shi）,1个三倍体种——伞房蔷薇（Rosa corymbulosa Rolfe）和1个四倍体种——弯刺蔷薇（Rosa beggeriana Schrenk）的核DNA含量及基因组大小。结果表明,流式细胞术检测结果与常规染色体压片法结果一致,可对中国野生蔷薇的倍性研究进行补充。本研究结果可丰富中国蔷薇属植物的细胞遗传学背景资料并为繁育新品种提供理论依据。     

石蒜属植物分支系统学分析

基于37个形态学、解剖学、孢粉学和细胞学性状及解剖学性状之外的28个形态学、孢粉学和细胞学性状,分别对石蒜属进行分支系统学分析,试图建立石蒜属种间的系统发育关系。利用PAUP^*软件分别构建了最大简约树(MP),所得树的拓扑结构是一致的。同时,基于解剖学9个性状,对石蒜、换锦花、忽地笑、江苏石蒜、长筒石蒜、乳白石蒜、夏水仙、红兰石蒜、安徽石蒜、短蕊石蒜、中国石蒜11个种进行系统发育树构建,其结果也是支持上述系统发育树的。系统发育树结构结果表明,石蒜属16种明显聚为两大类:石蒜、玫瑰石蒜、稻草石蒜和江苏石蒜;广西石蒜、红兰石蒜、换锦花、香石蒜、夏水仙、长筒石蒜、安徽石蒜、中国石蒜、忽地笑、乳白石蒜、短蕊石蒜和陕西石蒜。除换锦花、红兰石蒜及江苏石蒜系统发育位置不同之外,大类群的划分与RAPD指纹图谱基本一致。类群一均属于石蒜亚属(Lycoris亚属)。类群二又可以聚为两小类:广西石蒜、红兰石蒜、换锦花、香石蒜、夏水仙归为一类;长筒石蒜、安徽石蒜、中国石蒜、忽地笑、乳白石蒜、短蕊石蒜和陕西石蒜归为一类。前一子类群除广西石蒜外,都属于整齐花亚属(Symman thus亚属)。后一子类群除长筒石蒜与安徽石蒜外,均属于石蒜亚属(Lycoris亚属)。因此,花冠整齐与否是一个重要的分类特征,但作为石蒜属植物亚属的划分依据,没有得到本研究支持。而在本文中,雄蕊与花被片的位置关系可以作为大分类群划分依据,能否依此来对石蒜属植物亚属进行划分,仍需探讨。另外研究表明叶微形态特征在研究种间亲缘关系时,具有一定的作用。而在种间亲缘关系鉴定时,出叶期不应成为重要的依据。同时研究还表明中国石蒜与忽地笑具有非常近的亲缘关系,与形态学研究一致。     

Molecular evidence for an Asian origin and a unique westward migration of species in the genus Castanea via Europe to North America

The genus Castanea (Fagaceae) is widely distributed in the deciduous forests of the Northern Hemisphere. The striking similarity between the floras of eastern Asia and those of eastern North America and the difference in chestnut blight resistance among species has been of interest to botanists for a century. To infer the biogeographical history of the genus, the phylogeny of Castanea was estimated using DNA sequence data from different regions of the chloroplast genome. Sequencing results support the genus Castanea as a monophyletic group with Castanea crenata as basal. The three Chinese species form a strongly supported sister clade to the North American and European clade. A unique westward expansion of extant Castanea species is hypothesized with Castanea originating in eastern Asia, an initial diversification within Asia during the Eocene followed by intercontinental dispersion and divergence between the Chinese and the European/North American species during the middle Eocene and a split between the European and the North American species in the late Eocene. The differentiation within North America and China might have occurred in early or late Miocene. The North America species are supported as a clade with C. pumila var. ozarkensis, the Ozark chinkapin, as the basal lineage, sister to the group comprising C. pumila var. pumila, the Allegheny chinkapin, and Castanea dentata, the American chestnut. Morphological evolution of one nut per bur in the genus may have occurred independently on two continents.     

Polymorphic microsatellite loci for the genetic analysis of Lycoris radiata (Amaryllidaceae) and cross-amplification in other congeneric species

Lycoris radiata is a perennial herb that has been used in traditional Chinese medicine for a long time and has two main medicinal components in its bulb, lycorine and galanthamine. However, the original microsatellite loci have not been developed for any species of Lycoris. Total genomic DNA was extracted from fresh bulbs using a modified CTAB protocol. We isolated 10 microsatellite loci from 21 L. radiata individuals of a natural population from Yellow Mountain in Anhui Province, China. The number of alleles ranged from two to nine. The observed and expected heterozygosities ranged from 0.238 to 0.952 and from 0.455 to 0.784, respectively. One locus significantly deviated from Hardy-Weinberg equilibrium and no significant linkage disequilibrium was found between pairs of loci. Cross-species amplification of these microsatellite loci was characterized in additional five species (L. sprengeri, L. anhuiensis, L. albiflora, L. longituba, and L. chinensis) of Lycoris. The results suggest that these microsatellite markers would contribute to the population genetic studies of L. radiata and other related species.     

A hypothesis on the origin of genetic heterozygosity in diploids and triploids in Japanese Cayratia japonica species complex (Vitaceae)

We previously reported the occurrence of triploid strains in Japanese populations of Cayratia japonica (Thunb.) Gagnep. Interestingly, the triploid and most diploid strains had variably reduced pollen fertility. Two questions emerged from this earlier work: (1) How do triploids arise, and are they allotriploids or autotriploids? and (2) Why is there low pollen fertility in some diploid plants? We used a molecular genetic approach to determine the phylogenetic origins of triploids in C. japonica and the closely related species Cayratia tenuifolia (Wight & Arn.) Gagnep. In our analysis, we compared the sequences of the nuclear single-copy genes LEAFY and ASYMMETRIC LEAVES1. As a result, most triploids and diploids were heterozygous for the loci examined; the triploid genome shared an allele with the diploid genome, but other alleles differed between the ploidies. Therefore, Japanese populations of C. japonica and C. tenuifolia almost certainly arose from repeated hybridization events among genetically differentiated strains. Using our sequence data, we discuss possible scenarios accounting for the occurrence of triploids in the two species of Cayratia.     

Ascertaining maternal and paternal lineage within Musa by chloroplast and mitochondrial DNA RFLP analyses.

In banana, the maternal transmission of chloroplast DNA and paternal transmission of the mitochondrial DNA provides an exceptional opportunity for studying the maternal and paternal lineage of clones. In the present study, RFLP combined with hybridization of heterologous mitochondrial and chloroplastic probes have been used to characterize 71 wild accessions and 131 diploid and 103 triploid cultivated clones. In additon to Musa acuminata and Musa balbisiana, other species from the four Musa sections were studied to investigate their contribution to the origin of cultivated bananas. These molecular analyses enable the classification of the Musa complex to be discussed. Results ascertain relationships among and between the wild accessions and the mono- and interspecific diploid and triploid bananas, particularly for the acuminata genome. Parthenocarpic varieties are shown to be linked to M. acuminata banksii and M. acuminata errans, thus suggesting that the first center of domestication was in the Philippines - New Guinea area.     

A Preliminary Study on Cytology of Chinese Pteris

Cytological and biosystematic studies on the genus Pteris have made a great contribution to the theory of speciation and evolution in ferns. Sino-Japanese area is one of the speciation centers of this genus. But only a few Chinese species have been known cytologically. As a preliminary study, ten species of the genus Pteris and two species of the related genera Pteridium and Histiopteris were observed. All the materials were collected from southwest and south China. Their young sori were fixed in ethanol and glacial acetic acid (3:1), in the field. The preparations were made with acetocarmine squash method. The vouchers are deposited in PE. The results of observation are summarized in Table 1. Of Pteris, four species are agamosporous, eight are polyploid, six are actually the members of species complexes. Only two species are purely sexual diploids. Pteris cretica L. var. nervosa (Thunb.) Ching et S. H. Wu has 58 autobivalents at meiosis of spore mother cell, and usually produces 32 diplospores per sporangium. It is obviously an agamosporous diploid. Apart from 32-spored sporangium, some 4-, 8-, 16-, 64-, 13-, 34-, 36-spored, completely abortive or partly abortive sporangia were also found in the fixed material of a single individual. For explaining these unusual types of sporangia at least nine sporogenesis routes can be inferred. It may not be impossible that besides mainly producing functional diplospores, P. cretica var. nervosa also gives a few functional giant spores with a doubled or multiplied somatic chromosome number, which means that polyploids may be simply derived from the spores produced by their diploid parent. The spores in a sporangium are usually tetrahedral, but a few bilateral ones are also found in some sporangia. Sometimes, the bilateral spores are even more than the tetrahedral ones in a sporangium. Pteris vittata L. from the type locality shows 58 bivalents at diakinesis of meiosis. Without doubt, it is a sexual tetraploid, which was also found in south Guangdong and south Yunnan. However, Its natural ancestral diploid has been widely found in the subtropical regions of China, such as Sichuan, Guizhou, Yunnan and Hubei provinces. A sterile triploid with the chromosome number of n=201Ⅰ+26Ⅱ+5Ⅲ and a sterile tetraploid hybrid with the chromosome number of n=9Ⅰ+45Ⅱ+3Ⅲ+21Ⅴ were found in south Yunnan and south Guangdong respec tively. It is evident that P. vittata L. sensu lato is a species complex which includes several cy totypes. Its hexaploid form was reported from south India by Abraham et al. The distribu tion pattern of different cytotypes in P. vittata complex may indicate that the tropics is more favourable to formation and surviving of polyploid than the subtropics. However, it is most possible that Chinese subtropics is the origin place of the ancestral member in the P. vittata complex. In gross morphology, the tetraploid and triploid forms are only bigger and stronger than the diploid form. But the diploid can also grow rather big in cultivation. They can har dly be distinguished from each other. Therefore, they are not given formal names here. The author fully agree with Prof. T. Reichstein when he said in 1983 that it was hoped that a new nomenclature could be adopted for the cytotypes of species complex in future. The special chromosome number of Pteris deltodon Bak .was counted in this work for the first time. It has 55 bivalents at meiosis of SMC. This number is the only exception in the genus Pteris, and shows that aneuploidy may have taken place in this genus. However, the pos sibility can not be ruled out that P. deltodon is an allotetraploid came from two diploid species respectively with the chromosome number 26 and 29. Pteris gallinopes described by Prof. Ching in i983 is an agamosporous tetraploid with 116 autobivalents at meiosis, different from the related triploid species P. dactylina Hook. and P. henryi Christ. P. ensiformis Burm^。 P. muftifida Poir. and P. semipinnata L. are sexual tetraploids with n=58. P. excelsa Gaud. and P. oshimensis Hieron. var. paraemeiensis Ching are agamosporous triploids with n = 87 autobivalents. P. wallichiana Agardh is a sexual diploid with n=29. The high frequency of polyploids and agamospory among these random sampled Chinese species futher confirms the conclusion that both polyploidy and apomixis have played an important role in speciation and evolution of the genus Pteris. Pteridium aquilimum (L.) Kunh var. latiusculum (Desv.) Underw. ex Heller and Histiopteris incisa (Thunb.) J. Sm. have n=52 and n=96 respectively. They are both sexual tetraploids. Their chromosome numbers show again that the genera Pteridium and Histiopteris are distinquished from the genus Pteris by different basic numbers. This project is supported by the National Natural Science Foundation of China.     

Massive production of all-female diploids and triploids in the crucian carp

In many species of aquaculture importance, all-female and sterile populations possess superior productivity due to faster growth and a relatively homogenous size of individuals. However, the production of all-female and sterile fish in a large scale for aquaculture is a challenge in practice, because treatments necessary for gynogenesis induction usually cause massive embryonic and larval mortality, and the number of induced gynogens is too small for their direct use in aquaculture. Here we report the massive production of all-female triploid crucian carp by combining artificial gynogenesis, sex reversal and diploid-tetraploid hybridization. Previously, we have obtained an allotetraploid carp population (4n = 200) by hybridization between red crucian carp (Carassius auratus red var; ♀) and common carp (Cyprinus carpio; ♂). We induced all-female diploid gynogens of the Japanese crucian carp (Carassius cuvieri; 2n = 100). We also generated male diploid gynogens of the same species treated gynogenetic fry with 17-α-methyltestosterone, leading to the production of sex-revered gynogenetic males. Finally, these males were used to cross with the female diploid Japanese crucian carp gynogens and the allotetraploid females, resulting in the production of fertile all-female diploid Japanese crucian carp (2n=100) and sterile all-female triploid hybrids (3n = 150), respectively. Therefore, diploid crucian carp gynogenetic females and sex-reversed male together with an allotetraploid line provide an opportunity to produce all-female triploid populations in a large scale to meet demands in aquaculture industry.     

Phylogenetic relationships in Leymus (Triticeae; Poaceae): Evidence from chloroplast trnH-psbA and mitochondrial coxII intron sequences

Leymus Hochst. is a polyploid genus with a diverse array of morphology, cytology, ecology, and distribution in Triticeae. To investigate the phylogenetic relationships and maternal genome donor of polyploid Leymus, the chloroplast trnH-psbA region and mitochondrial coxII intron sequences of 33 Leymus taxa were analyzed with those of 36 diploid perennial species representing 19 basic genomes in Triticeae. The results showed that reticulate evolution occurred in Leymus species, with the cytoplasmic lineage of Leymus contributed by different progenitors. Interspecific relationships of Leymus were also elucidated on the basis of orthologous comparison. Our data suggested that: (i) due to incomplete lineage sorting and/or difference in the pattern of chloroplast and mitochondrial inheritance, the genealogical conflict between the two genealogical patterns suggest the contribution of Psathyrostachys Nevski, Agropyron J. Gaertn, Eremopyrum (Ledeb.) Jaub. & Spach, Pseudoroegneria (Nevski) Á. Löve, Thinopyrum Á. Löve, and Lophopyrum (Host) Á. Löve to the cytoplasmic lineage of Leymus; (ii) there is a close relationship among Leymus species from the same area or neighboring geographic regions; (iii) L. coreanus (Honda) K. B. Jensen & R. R.-C. Wang, L. duthiei (Stapf) Y. H. Zhou & H. Q. Zhang ex C. Yen, J. L. Yang & B. R. Baum, L. duthiei var. longearistatus (Hack.) Y. H. Zhou & H. Q. Zhang ex C. Yen, J. L. Yang & B. R. Baum, and L. komarovii(Roshev.) C. Yen, J. L. Yang & B. R. Baum are closely related to other Leymus species, and it is reasonable to transfer these species from the genus Hystrix Moench to Leymus; (iv) Leymus species from North America are closely related to L. coreanus from the Russian Far East and L. komarovii from northeast China but are evolutionarily distinct from Leymus species from Central Asia and the Qinghai–Tibet Plateau. The occurrence of multiple origin and introgression could account for the rich diversity and ecological adaptation of Leymus species.     

Genetic diversity and the population structure at two ploidy levels of Lycoris radiata as revealed by SCoT analysis

Ploidy differences can occur between and within species. To decipher the genetic structure of a species with different ploidy levels, we investigated an area of China where diploid and triploid individuals of Lycoris radiata co-occur. Twenty-six populations of L. radiata were sampled across its natural range, and SCoT markers were used to assess the genetic diversity and population structure. Chromosome counts revealed that 10 populations were diploid and 16 were triploid, each population showed a single cytotype. SCoT analysis revealed high genetic diversity at the species level (PPB = 93.1%; H = 0.258; I = 0.396). However, compared to the triploid strains of Lycoris, the diploid strains had a relatively higher genetic diversity. One possible interpretation for these results is that the triploid strains only propagated using bulb division, while the diploid strains reproduced using sexual propagation and bulb division. Our investigation of the genetic structure, based on UPGMA and PCoA cluster analyses, revealed that populations of L. radiata are divided into three genetic groups. In addition, our results indicate that Yangtze River can affect population diffusion, which played an important role in the genetic differentiation of L. radiata. Lastly, our results suggest that the triploid sterile species of L. radiata might possess two origins.