中国近缘野生大麦醇溶蛋白的遗传多态性研究

随机选取来源于中国西藏24个不同行政县的181份近缘野生大科材料,其中包括47份六棱野生大麦,134份二棱野生大麦,选用青藏高原的二棱野生大麦（W2）,欧洲的代表品种Betzes大麦及以色列的二棱野生大麦（Is)作对照。利用A-PAGE（Acidic-polyacrylamide gel electrophoresis)法进行了醇溶蛋白遗传多态性的研究。结果表明,184份供试材料共得到60种不同的电泳图谱,说明西藏近缘野生大麦醇溶蛋白遗传多态性非常丰富,其中有38种图谱为单一材料所独有,以ZYM0019和ZYM1488材料为代表的2种醇溶蛋白图谱占供试材料的29.3%,在西藏分布较为普遍,聚类分析表明地理环境相似的地区有着相似的图谱类型,图谱类型与地理生态环境具有一定的相关性。讨论了中国近缘野生大麦醇溶蛋白的多态性与地理分布的关系。以及栽培大麦的起源中心等问题。     

应用微卫星标记研究西藏野生大麦的遗传多样性

以西藏不同地区的106份野生大麦为材料,其中包括50份野生二棱大麦（HS）,27份野生瓶形大麦（HL）和29份野生六棱大麦（HA）,用Liu等（1996）发表的SSR连锁图的每个连锁群的两个臂的不同位置上选取3～5个共30个SSR标记,研究了西藏3类野生大麦的遗传多样性。结果表明,这3类野生大麦在遗传组成及等位变异频率分布上存在着明显的遗传分化。在总样本中,共检测到229个等位变异,平均每个SSR位点检测到7．6个等位变异,其中70个为这3类野生大麦间共同的等位变异,等位变异数在这3类野生大麦间有明显的差异,亚种问的遗传多样性明显高于亚种内的遗传多样性。其遗传多样性大小顺序为HS〉HL〉HA。聚类分析表明,野生二棱大麦、野生六棱大麦分别聚在不同的两类,而野生瓶形大麦中各有约50％的材料分别聚在这两类。根据本研究及前人研究结果,我们认为中国栽培大麦是从野生二棱大麦经野生瓶形大麦向野生六棱大麦进化的。该结果支持了栽培大麦起源的“野生二棱大麦单系起源论”的观点。     

利用RAPD标记分析大麦种质资源的遗传多样性

利用RAPD标记对19份西藏近缘野生大麦材料、33份我国不同省市的地方品种以及8份国外引进大麦品种共60份大麦种质资源的遗传多样性进行检测.结果表明材料间遗传差异明显.32个RAPD引物中,有25个引物(占78.13%)可扩增出清晰且具多态性的条带,另外7个引物能扩增出1～3条清晰但无多态性的条带.每个引物可扩增出1～8条多态性带,平均为3.72条.32个引物共产生119条DNA片段,其中87条具有多态性,多态性比率(PPB)为73.11%,平均多态信息量(PIC)为0.434;每个位点平均有效等位基因数(Ne)为2.304;材料间遗传相似系数GS变化范围为0.757～0.981,平均值为0.871.19份来源于西藏的近缘野生大麦材料间GS值变幅为0.818～0.969,平均为0.892;33份我国栽培大麦地方品种间的GS值变化范围为0.783～0.981,平均为0.879;8份分别来自8个国家的栽培大麦品种间的GS值变幅为0.820～0.956,平均为0.882.根据RAPD标记分析的结果,对60份大麦种质资源进行聚类分析,在平均GS值0.871水平上60份大麦材料可聚为5类,聚类结果能在一定程度上反应材料的地理分布关系,但某些相同地理来源的材料也较分散地分布在整个聚类树中.本研究从分子水平上进一步证明了我国栽培大麦丰富的遗传多样性,是世界栽培大麦的遗传多样性中心之一.     

贵州野生大麦麦芽品质性状的SSR标记分析

采用6对啤酒大麦的麦芽浸提和糖化力紧密连锁引物对103份采自贵州的野生大麦材料进行SSR标记。结果表明,贵州野生大麦麦芽品质性状存在丰富的变异,6对SSR引物共检测出38个等位变异,每个位点平均6.33个等位变异,其中GMS001位点对贵州野生大麦基因组DNA变异检测最有效。UPGMA聚类图显示,该6对与麦芽品质紧密连锁的SSR引物对区分野生大麦在贵州不同的资源产地和棱性是有效的,表现为遵义地区野生大麦遗传多样性丰富,而来自贵州凯里地区的野生大麦资源遗传多样性狭窄。麦芽品质性状标记结果表明贵州野生六棱大麦较四棱大麦的遗传差异更显著,表明进行贵州啤酒大麦人工育种的亲本应在亲缘关系较远的六棱大麦之间选择。     

青藏高原近缘野生大麦5S rRNA基因染色体原位杂交定位

采用原位杂交技术,以５Ｓ ｒＲＮＡ基因为探针,对产于青藏高原的４份近缘野生大麦和栽培大麦,即：二棱野生大麦Ｈｏｒｄｅｕｍ ｖｕｌｇａｒｅ Ｌ．ｓｓｐ．ｓｐｏｎｔａｎｅｕｍ（Ｋｏｃｈ）Ｈｓｕｅ,六棱野生大麦Ｈ．ｖｕｌｇａｒｅ Ｌ．ｓｓｐ．ａｇｒｉｏｃｒｉｔｈｏｎ（Ａｂｅｒｇ） Ｈｓｕｅ,六棱瓶形野生大麦Ｈ．ｖｕｌｇａｒｅ Ｌ．ｓｓｐ．ａｇｒｉｏｃｒｉｔｈｏｎ ｖａｒ．ｌａｇｕｎｃｕｌｉｆｏｒｍ（Ｂａｋｈｔ） Ｈｓｕｅ,栽培大麦Ｈ．ｖｕｌｇａｒｅ．Ｌ．进行了研究,将杂交结果进行观察与统计,并建立起５Ｓ ｒＲＮＡ基因定位的模式图。结果表明５Ｓ ｒＲＮＡ基因在染色体上的位点呈现动态变化,由二棱野生大麦、六棱瓶形野生大麦到六棱野生大麦、栽培大麦、位点数目有递增的趋势,而且位置也发生了某些改变。探讨了５Ｓ ｒＲＮＡ基因进化     

基于SSR标记分析小豆及其近缘植物的遗传关系

本研究利用87对SSR引物分析了80份栽培小豆(Vigna angularis)、22份野生小豆(V.angularis var.nipponensis)以及10份豇豆属(共7个种)近缘植物,旨在比较豇豆属不同种的遗传多样性,并分析种间的遗传关系.结果显示87对SSR引物在112份小豆及其近缘植物资源中共检测到667个等位变异.其中有75个、71个和82个SSR位点分别在栽培小豆、野生小豆和近缘植物中表现为多态.随机抽样分析发现,平均每SSR位点检测到的等位变异数目为近缘植物>野生小豆>栽培小豆,与多态信息含量(PIC)值一致,说明近缘植物及野生小豆中蕴含着丰富的遗传变异,是栽培小豆育种的重要基因来源.聚类分析显示,栽培小豆、野生小豆和近缘植物间的遗传分化比较明显,分别聚为三大类,其中栽培小豆的遗传背景与其生态环境相对应;近缘植物又可以分为三个亚类,亚类间的遗传距离与其亲缘关系相对应.本研究结果也说明利用SSR标记辅助豇豆属的种间分类是可行的.     

野生大麦与栽培大麦醇溶蛋白遗传多样性比较

利用A-PAGE(acid-polyacrylamide gel electrophoresis)法对采自以色列的野生大麦的一个野生自然群体的15个系和来自世界不同国家的14份栽培大麦品种醇溶蛋白的遗传多样性进行了分析.结果表明:在所有的29份供试材料中,共发现52条相对迁移率不同的谱带.52条谱带的出现频率为3.44%～93.1%,多样性指数为0.066～0.368;以中国春醇溶蛋白为标准,ω区大麦醇溶蛋白的谱带数最多,其次是β区;野生大麦Shannon多样性指数依次为β区>ω区>α区>γ区,而栽培大麦Shannon多样性指数依次为ω区β>区>γ区>a区;野生大麦自然群体和栽培大麦品种间的遗传相似系数变幅相当,且聚类分析结果显示,野生大麦自然群体和来自全球不同区域栽培大麦品种间的醇溶蛋白遗传多样性同样丰富.以上结果说明,野生大麦中保存了较栽培大麦更为丰富的基因资源,今后栽培大麦的品质改良应该重视野生大麦资源的合理利用.     

应用RAPD指纹探讨黄麻属种间遗传多样性及其亲缘关系

采用RAPD标记构建了黄麻属(Corchorus)植物10个种27份材料的指纹图谱,从119个随机引物中筛选出清晰且多态性高的25个引物,共扩增出329条：DNA片段,分子量在0．3～3．0kb之间,其中294条谱带具有遗传多态性,多态比率(PPB)为89．36％,平均每个引物扩增出13．16条带。用Biol D^ 数据分析软件,计算Nei氏相似性系数,建立了UPCMA聚类图。结果表明：(1)供试黄麻属15份野生种和12份栽培种具有丰富的遗传多样性,遗传相似系数在0．49～0．98;(2)在聚类分析中,当L1聚值水平D=0．785时,可将两个栽培种及其近缘野生种(C．capsulris和C．olititorius)与原始黄麻野生种划分为3个不同类群。反映出栽培种及其近缘野生种与原始野生种间有明显的遗传差异;(3)当L2取值水平D=0．850时,可将供试27份材料划分为10个以物种为单元的亚类群或个类,①即假黄麻C．aestuans(3份),②三齿种C．tridens,③梭状种C．fascicularis,④假长果种C．psendo-olitorius,⑤假圆果种C．pseudo-capsularis,⑥三室种C．tilacularis,⑦甜麻(新种未定名),⑧圆果种C．capsularis(9份),⑨长果种C．olitorius(7份),⑩荨麻叶种C．uriticifolius,结果与10个种的经典分类相吻合,揭示了种间的遗传差异性。其中圆果种C．cap—sularis与长果种C．olitorius两个种亲缘关系较近,与荨麻叶种C．uriticifolius种间亲缘关系较远;(4)非洲荨麻叶种C．uriticifolius和中国的甜麻(新种)、假黄麻C．aestuans3个种与其他种间的遗传差异较大,处在分子聚类树较基础的地位,为较原始的黄麻野生种;(5)非洲的三室种21C为三室种一个生态型亚种;采集于中国的黄麻野生种河南南阳的粘粘菜、福建漳浦的麻菜,云南开远的猪菜为3个不同生态类型的假黄麻;海南野生圆果为圆果黄麻栽培种的近缘野生种;漳浦野生长果、河南野生长果、马里野生长果为长果栽培种的近缘野生种,种内不同材料间遗传相似性较高,亲缘关系较密切;(6)在两个栽培种中,品种基因型相似性系数圆果栽培种高于长果栽培种。     

福建栽培大豆品种RAPD标记多样性分析

利用RAPD分子标记技术对福建18份栽培大豆品种遗传多样性进行研究.结果表明,12个引物共扩增出91条带,其中多态条带65条,多态性程度为71.43%.D=0.62时,聚类分析结果可以将供试材料分为3个大类群,即菜用大豆品种、杂交育成品种和地方品种、有半野生血缘的品种各聚成1类,揭示了福建省栽培大豆品种间的亲缘关系,同时还反映出品种间关系与地理起源有一定的相关.     

六棱大麦种质表型遗传多样性评价

为探究六棱大麦种质资源的遗传多样性,提高六棱大麦育种组合选配的针对性和育种效率。以来自国内外的89份六棱大麦种质为材料,考察了3个不同环境下参试材料的株高、穗下节间长、穗长、主穗粒数、单株穗数、千粒重、单株粒重、单株生物重等8个性状,综合参试材料3个不同环境的表现,通过变异系数及遗传多样性指数分析参试材料的遗传多样性,通过主成分分析、聚类分析等方法对参试六棱大麦种质进行了综合评价。结果表明:(1)六棱大麦的穗长、单株粒重2个性状变异较丰富,株型性状与穗粒数变异系数较小,六棱大麦育种的增益效应主要体现在穗长和粒重的适度增加;(2)六棱皮大麦与六棱裸大麦仅在千粒重性状上差异显著,六棱皮大麦各性状多样性指数普遍高于六棱裸大麦;(3)参试六棱大麦种质分为5类,其中第4类基本均为中国地方种质,分布广泛,但遗传距离较近;其他4类为国内外选育的种质,各具特性;(4)利用主成分二维排序分析筛选到以高产为基础,分别兼具矮秆、长穗且分蘖能力强、大粒及高生物重的4类优异种质;基于主成分构建了六棱大麦种质资源的综合评价方程。     

RAPD Analysis and the Probable Evolutionary Route of Wild Relatives of Barley from China

he genetic relationships among 12 wild relatives and cultivar of barley ( Hordeum vulgare L.) as well as 1 perennial wild barley grass (H. brevisubulatum (Trin.) Link) from China were investigated by RAPD analysis. 36 out of 63 arbitrary primers produced 285 distinctive bands in total, 219 of which were polymorphic. Clearly resolved bands were treated as independent characters and scored for their presence or absence in a binary data matrix. Simple matching coefficients and Nei's similarity coefficients were calculated respectively. Dendrograms were generated by using the PHYLIP 3.5c software. The results revealed that the cultivated barley and their wild relatives from China were clustered into one group, among which, the two-rowed wild relatives of barley ( H. vulgare L. ssp. spontaneum (Koch) Hsü) and the six-rowed wild forms (H. vulgare L. ssp. agriocrithon (Aberg) Hsü) were respectively clustered into different subgroups. It was considered that wild relatives of barley from China were subspecies of H.vulgare. And it was proposed that the cultivated barley was originally evolved from the two-rowed wild barley. The retrogressive two-rowed wild barley and the bottle-shaped wild forms (H. vulgare L. ssp. agriocrithon var. lagunculiforme Bakht Hsü) were the intermediate types in the evolutionary route from the two-rowed wild barley to the six-rowed wild forms and eventually evolved to the cultivated barley.     

On the origin of six-rowed barley with brittle rachis, agriocrithon [Hordeum vulgare ssp. vulgare f. agriocrithon (Aberg) Bowd.], based on a DNA marker closely linked to the vrs1 (six-row gene) locus

The origin of six-rowed cultivated barley has been revealed to be more complex since the discovery of agriocrithon, a six-rowed barley with brittle rachis. The present study investigates whether such six-rowed brittle barley is wild or hybrid in nature, by analyzing genetic diversity at the cMWG699 marker locus, which is closely linked to the vrs1 (six-row gene) locus. DNA sequence analysis for 42 accessions showed only three types in six-rowed brittle barleys; in contrast, nine sequence types were found in ten wild barleys, ssp. spontaneum, in our previous study. Nucleotide diversities for the six-rowed brittle barley were 2.8–4.5 times lower than that for the ssp. spontaneum at this marker locus. The three sequence types found in the six-rowed brittle barley also appeared in the six-rowed cultivated barley. A cross-allelism test confirmed that the six-rowed character of the six-rowed brittle barley was controlled by the vrs1 locus. The nucleotide diversity and genealogy demonstrated that f. agriocrithon does not have the same level of diversity as found in wild barley, ssp. spontaneum. Consequently, f. agriocrithon does not appear to represent genuinely wild populations, but more probably originated from hybridization between ssp. spontaneum and six-rowed cultivated barley.     

A DNA marker closely linked to the <Emphasis Type="Italic">vrs1</Emphasis> locus (row-type gene) indicates multiple origins of six-rowed cultivated barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

The origin of six-rowed cultivated barley was studied using a DNA marker cMWG699 closely linked to the vrs1 locus. Restriction patterns of the PCR-amplified product of the cMWG699 locus were examined in 280 cultivated (Hordeum vulgare ssp. vulgare) and 183 wild (H. vulgare ssp. spontaneum) barleys. Nucleotide sequences of the PCR products were also examined in selected accessions. Six-rowed cultivated barleys were divided into two distinct groups, types I and II. Type I six-rowed cultivated barley was distributed widely while type II six-rowed cultivated barley was found only in the Mediterranean region. The type I sequence was also found in a wild barley accession from Turkmenistan whereas the type II sequence was also found in a two-rowed cultivated barley from North Africa and a wild barley from Morocco. These results suggested that the six-rowed type I and II barleys were derived from two-rowed type I and II barleys, respectively, by independent mutations at the vrs1 locus. Received: 3 November 2000 / Accepted: 17 April 2001     

Genetic Diversity Analysis of Tibetan Wild Barley Using SSR Markers

One hundred and six accessions of wild barley collected from Tibet, China, including 50 entries of the two-rowed wild barley Hordeum vulgare ssp. spontaneum (HS), 29 entries of the six-rowed wild barley Hordeum vulgare ssp. agriocrithon (HA), and 27 entries of the six-rowed wild barley Hordeum vulgare ssp. agriocrithon var. lagunculiforme (HL), were analyzed using 30 SSR markers selected from the seven barley linkage groups for studying genetic diversity and evolutionary relationship of the three subspecies of Tibetan wild barley to cultivated barley in China. Over the 30 genetic loci that were studied, 229 alleles were identified among the 106 accessions, of which 70 were common alleles. H. vulgare ssp. spontaneum possesses about thrice more private alleles (2.83 alleles/locus) than HS (0.93 alleles/locus), whereas almost no private alleles were detected in HL. The genetic diversity among-subspecies is much higher than that within-subspecies. Generally, the genetic diversity among the three subspecies is of the order HS > HL > HA. Phylogenetic analysis of the 106 accessions showed that all the accessions of HS and HA was clustered in their own groups, whereas the 27 accessions of HL were separated into two groups (14 entries with group HS and the rest with group HA). This indicated that HL was an intermediate form between HS and HA. Based on this study and previous works, we suggested that Chinese cultivated barley might evolve from HS via HL to HA.     

A comparative analysis of genetic polymorphism in wild and cultivated barley from Tibet using isozyme and ribosomal DNA markers.

This study was conducted to address some of the issues concerning the possible significance of Tibet in the origin and evolution of cultivated barley. A total of 1757 barley accessions from Tibet, including 1496 entries of Hordeum vulgare ssp. vulgare (HV), 229 entries of the six-rowed wild barley H. vulgare ssp. agriocrithon (HA), and 32 entries of the two-rowed wild barley H. vulgare ssp. spontaneum (HS), were assayed for allozymes at four esterase loci. A subsample of 491 accessions was surveyed for spacer-length polymorphism at two ribosomal DNA loci. Genetic variation is extensive in these barley groups, and the amount of genetic diversity in cultivated barley of this region is comparable with that of cultivated barley worldwide. The level of genetic variation of HA is significantly lower than the other two barley groups, and there is also substantial heterogeneity in the level of polymorphism among different agrigeographical subregions. However, little genetic differentiation was detected among the three barley groups (HV, HA, and HS), as well as among different agrigeographical subregions. Comparison of the results from this and previous studies indicated a strong differentiation between Oriental and Occidental barley, thus favoring the hypothesis of a diphyletic origin of cultivated barley.     

AFLP analysis of genetic polymorphism and evolutionary relationships among cultivated and wild Nicotiana species.

Amplified fragment length polymorphism (AFLP) analysis was used to determine the degree of intra- and inter-specific genetic variation in the genus Nicotiana. Forty-six lines of cultivated tobacco (Nicotiana tabacum L.) and seven wild Nicotiana species, including N. sylvestris, N. tomentosiformis, N. otophora, N. glutinosa, N. suaveolens, N. rustica, and N. longiflora, were analyzed, using at least eight different oligonucleotide primer combinations capable of detecting a minimum of 50 polymorphic bands per primer pair. The amount of genetic polymorphism present among cultivated tobacco lines (N. tabacum) was limited, as evidenced by the high degree of similarity in the AFLP profiles of cultivars collected worldwide. Six major clusters were found within cultivated tobacco that were primarily based upon geographic origin and manufacturing quality traits. A greater amount of genetic polymorphism exists among wild species of Nicotiana than among cultivated forms. Pairwise comparisons of the AFLP profiles of wild and cultivated Nicotiana species show that polymorphic bands present in N. tabacum can be found in at least one of three proposed wild progenitor species (i.e., N. sylvestris, N. tomentosiformis, and N. otophora). This observation provides additional support for these species contributing to the origin of N. tabacum.     

双孢蘑菇遗传多样性分析

应用AFLP指纹技术对双孢蘑菇的20个野生菌株和5个栽培品种的遗传多样性进行了研究。AFLP指纹揭示出20个野生异核体菌株所固的基因型。5个商业品种表现出比较一致的AFLP指纹,但也显示出它们之间的一些差别。由单孢分离获得的同核体菌株携带着部分异核体菌株的AFLP指纹;由同一子实体分离得到的大部分单孢菌株是异核体菌株,它们具有与其亲本一致的AFLP指纹。UPGMA分析揭示出2个与地理分布（美洲、欧洲）和相对应的组。研究结果表明：（1）在野生菌株之间存在着明显的遗传差异;（2）大多数单孢分离的菌株具有与母本一致的遗传物质;（3）野生菌株间的遗传变异大于栽培品种间的变异;（4）在双孢蘑菇的遗传多样性分析中,AFLP技术是非常有效的工具。     

The genetic diversity of the Vigna angularis complex in Asia.

A selected set of accessions of components of the azuki bean (Vigna angularis) complex comprising 123 cultivated accessions and 23 wild or weedy accessions from Bhutan, China (including Taiwan), India, Japan, Korea, and Nepal was analyzed using amplified fragment length polymorphism (AFLP) methodology. Using 12 AFLP primer pairs, 580 unambiguous bands were generated, 313 (53.9%) of which were polymorphic among azuki bean accessions. All 580 bands were used to assess phenotypic (band) and genetic (nucleotide) diversity among the 146 azuki bean accessions. The results indicate five major groups of azuki bean germplasm primarily associated with geographic origin of accessions and their status: wild, weedy, or cultivated. These five groups are (i) Himalayan wild, (ii) Nepal-Bhutan cultivated, (iii) Chinese wild, (iv) Taiwan wild - Bhutan cultivated, and (v) northeast Asian accessions. Within the northeast Asian accessions, three subgroups are present. These consist of (v1) Japanese complex - Korean cultivated, (v2) Japanese cultivated, and (v3) Chinese cultivated accessions. The results suggest domestication of azuki bean occurred at least twice, once in the Himalayan region of southern Asia and once in northeast Asia. The remarkable diversity of azuki bean germplasm in the Himalayan region compared with other regions suggests this is a rich source of germplasm for plant breeding. The results suggest there are important gaps in the germplasm collections of azuki bean and its close relatives from various parts of Asia and that specific collecting missions for Vigna germplasm related to azuki bean in the highlands of subtropical Asia are needed.