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。     

基于AFLP标记的中国西藏近缘野生大麦遗传多样性分析

选取7对引物组合, 构建了36份西藏近缘野生大麦和4份栽培大麦的AFLP指纹图谱, 共获得清晰可辨的条带227条, 其中多态性带194条, 占85.46％, 从DNA分子水平显示出所试材料遗传多样性较为丰富。计算得各样品间的遗传距离(欧氏距离)介于2.646~10.488之间, 应用离差平方和法对供试材料的AFLP数据结果进行聚类, 建立了40份大麦材料的AFLP树状图, 聚类结果将40份材料分为5类, 进一步揭示出供试材料间遗传背景的相似性和复杂性。结合Nei’s遗传一致性分析结果, 发现近缘六棱野生大麦较之近缘二棱野生与栽培大麦的亲缘关系更近, 支持栽培大麦是从野生二棱大麦起源, 而野生六棱大麦是进化过程中的过渡类型的大麦系统发生观点。     

应用SSR标记对61个国家大麦遗传多样性的研究

使用全自动基因分析仪（ABI-3700 DNA Analyzer）,用35对SSR荧光标记引物对来自61个国家的2625份大麦种质资源的遗传多样性进行了分析,得出如下结果：（1）35对引物在2625份大麦种质资源中共检测到2063个等位变异,每个位点的等位变异数为30～79个,平均为58.94;多样性指数为1.60～3.66,平均为2.80;（2）在大麦的7条染色体中,每条染色体的等位变异数不等,染色体等位变异数从多到少排序为5、7、2、6、3、4、1,遗传多样性指数从高到低依次为2、6、7、3、5、4、1,综合这两项指标,第2、7条染色体遗传多样性较高,第1条染色体的遗传多样性最低;（3）约旦、伊朗、叙利亚和利比亚等国家大麦的遗传多样性指数最高,分别为2.58、2.53、2.50和2.38;而埃塞俄比亚、叙利亚、伊朗和约旦等国家的大麦资源则具有较多的等位变异,分别为29.03、25.57、25.11和24.60,综合分析,约旦、伊朗、叙利亚和土耳其大麦遗传多样性较高。     

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SSR标记分析中国糯大麦种质的遗传多样性

利用SSR标记对来自中国不同省市的76份糯大麦种质的遗传多样性进行分析,并以遗传相似系数为基础进行聚类分析。SSR标记分析表明,50对大麦SSR引物在76份糯大麦中共扩增出203个等位基因,属于高度多态性位点范畴。当遗传相似系数为0.70时可将76个糯大麦品种划分为5个类群,在一定程度上反映了与材料的地理来源和皮裸性。糯大麦资源平均Shan-non多样性指数显示,来自云南和西藏的糯大麦品种遗传多样性略高。SSR标记分析表明,中国糯大麦具有丰富的遗传多样性,对揭示糯大麦的起源与传播以及对资源的有效利用具有现实意义。     

应用SSR标记对61个国家大麦遗传多样性的研究

使用全自动基因分析仪(ABI-3700 DNA Analyzer),用35对SSR荧光标记引物对来自61个国家的2625份大麦种质资源的遗传多样性进行了分析,得出如下结果(1)35对引物在2625份大麦种质资源中共检测到2063个等位变异,每个位点的等位变异数为30～79个,平均为58.94;多样性指数为1.60～3.66,平均为2.80;(2)在大麦的7条染色体中,每条染色体的等位变异数不等,染色体等位变异数从多到少排序为5、7、2、6、3、4、1,遗传多样性指数从高到低依次为2、6、7、3、5、4、1,综合这两项指标,第2、7条染色体遗传多样性较高,第1条染色体的遗传多样性最低;(3)约旦、伊朗、叙利亚和利比亚等国家大麦的遗传多样性指数最高,分别为2.58、2.53、2.50和2.38;而埃塞俄比亚、叙利亚、伊朗和约旦等国家的大麦资源则具有较多的等位变异,分别为29.03、25.57、25.11和24.60,综合分析,约旦、伊朗、叙利亚和土耳其大麦遗传多样性较高.     

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.     

利用SSR标记分析云南、西藏和新疆小麦的遗传多样性

用185对SSR引物对52份中国西部特有小麦的遗传多样性进行了研究分析。在31份云南小麦材料中,共检测到488个等位变异,每一个SSR引物可检测到1至9个等位变异,平均为2.64个;平均PIC值为0.2764。在15份西藏小麦材料中,共检测到472个等位变异,每个引物可扩增出1到8个等位变异,平均为2.55个;平均PIC值为0.3082。在6份新疆小麦材料中,共检测到308个等位变异,每一个SSR引物可检测1到5个等位变异,平均为1.66个;平均PIC值为0.1944。185对SSR引物在云南、西藏和新疆小麦的21条染色体、7个部分同源群和3个染色体组上检测到的等位位点的多态性存在明显差异。云南、西藏和新疆小麦均以3B染色体较高,而1D染色体最低;在7个部分同源群中,均以第三部分同源群最高,第六部分同源群最低;在A、B和D染色体组上,均以B染色体组最高,D染色体组最低,A染色体组居中。利用185对SSR引物计算了云南、西藏和新疆小麦群体内及其群体间的遗传距离(GD)和平均遗传距离,结果显示,西藏小麦和云南小麦群体内的平均遗传距离要高于新疆小麦,而云南小麦和西藏小麦间的平均遗传距离低于两者与新疆小麦的平均遗传距离。聚类分析结果也表明,云南小麦和西藏小麦的亲缘关系较近,但两者与新疆小麦的亲缘关系相对较远。     

浙江赤霉病抗性不同的大麦地方品种遗传多样性分析

采用SSR标记方法研究了43份对大麦赤霉病有不同抗性的浙江地方品种的遗传多样性。结果显示29对SSR引物在上述品种中共检测到87个等位基因,每对引物等位基因数在2~9之间,平均为3个;平均多态性信息含量（PIC）为0.3509,平均Shannon指数(I)为0.6951,平均Nei’s基因多样性指数(H)为0.4268。品种间的遗传相似系数变幅为0.082~0.986,平均值为0.467。聚类分析将参试品种分为4大类。浙江省地方品种遗传多样性较高,二棱品种的遗传变异明显高于六棱品种;抗和中抗赤霉病品种的遗传变异也较大。聚类结果与品种来源无任何关系;主成分分析结果与聚类分析结果基本一致。     

中国大麦属（Horseum）的核型研究

作者研究了我国野生二棱大麦(H. spontaneum)、栽培二棱大麦(H. distichon)、野生六棱大麦(H. agriocrithon)、野生瓶形大麦(H. lagunculiforme)、栽培六棱大麦(H. vulgare)和各种变种及过渡类型的染色体数目、核型和Giemsa N—带,结果表明,大麦种间核型差异甚微,带型则有一定区别,对大麦的进化途径进行了讨论。     

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.     

Comparison of restriction fragment length polymorphisms in wild and cultivated barley.

This study was undertaken to assess the relative level of molecular diversity between cultivated barley, Hordeum vulgare ssp. vulgare (HV), and one of its wild relatives, H. vulgare ssp. spontaneum (HS), and to identify possible restriction fragment length polymorphism (RFLP) patterns that may provide information concerning the phylogenetic relationship between these two barley groups. A total of 363 barley accessions were assayed, including 95 entries of HV collected from 36 major barley growing countries of the world and 268 entries of HS from 25 natural populations in Israel and Iran. The 26 RFLP marker loci used in the survey represent single-copy, low-copy, and repetitive DNA sequences and mark all of the chromosome arms. A randomization test, on the basis of equal sample sizes, showed that HS is more polymorphic than HV, as evaluated by the number of alleles and diversity indices. The analysis also indicated extensive RFLP differentiation between these two barley groups; highly significant differences of allele frequencies were detected at the majority of the loci. The HV sample can be subdivided according to winter or spring growth habits, and two- or six-rowed spikes. Analysis of genetic polymorphisms in these subgroups showed that levels of diversity were about equal in spring and winter groups and also in the groups with two- and six-rowed spikes. However, significant differences of allelic frequencies were detected between subgroups of the two divisions.     

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.     

A phylogenetic analysis based on nucleotide sequence of a marker linked to the brittle rachis locus indicates a diphyletic origin of barley

BACKGROUND AND AIMS: Barley (Hordeum vulgare ssp. vulgare) cultivation started between 9500 and 8400 years ago, and was a major part of ancient agriculture in the Near East. The brittle rachis is a critical trait in the domestication process. METHODS: A DNA sequence closely linked to the brittle rachis complex was amplified and resequenced in a collection of cultivated barleys, wild barleys (H. vulgare ssp. spontaneum) and weedy brittle rachis varieties (H. vulgare ssp. vulgare var. agriocrithon). The sequence was used to construct a phylogenetic tree. KEY RESULTS: The phylogeny separated the W- (btr1-carrying) from the E- (btr2-carrying) cultivars. The wild barleys had a high sequence diversity and were distributed throughout the W- and E-clades. Some of the Tibetan var. agriocrithon lines were closely related to the E-type and others to the W-type cultivated barleys, but an Israeli var. agriocrithon line has a complex origin. CONCLUSIONS: The results are consistent with a diphyletic origin of barley. The W- and E-type cultivars are assumed to have evolved from previously diverged wild barley via independent mutations at Btr1 and Btr2.     

Polymorphic chloroplast simple sequence repeat primers for systematic and population studies in the genus Hordeum

In this study we report the development of primers to amplify polymorphic chloroplast simple sequence repeats in the genus Hordeum , which includes cultivated barley ( H. vulgare ssp. vulgare ) and its wild progenitor H. vulgare ssp. spontaneum . Polymorphic products were amplified in a wide range of Hordeum spp. and intraspecific variation was detected in both cultivated and wild barley. A decrease in cytoplasmic diversity was observed between sspp. spontaneum and vulgare as well as between ssp. vulgare landraces and cultivars, which is characteristic of domestication processes in many crop species. We also observed possible evidence for reticulate evolution of H. brachyantherum polyploids, with apparent multiple cytoplasmic introgressions during successive polyploidization events.     

Characterization of genetic diversity in core collection accessions of wild barley,Hordeum vulgare ssp. spontaneum

Genetic variability in the 143 core accessions of wild barley, Hordeum vulgare ssp. spontaneum, was assessed by allozyme analysis. A total of 34 alleles were detected at ten isozyme loci. All loci were polymorphic except Pgd-1, which was monomorphic. Est-2 and Est-4 were the most diverse loci, with genetic diversity values of 0.747 and 0.686, respectively. The comparison of the results with those of previous studies indicates that all alleles occurring in cultivated and wild barley are observed in this set of the wild Barley Core Collection. Only one allele (Pgd-1 Tj) was absent. It is noteworthy that one new allele at the Ndh-2 locus and another new allele at Aco-2 locus were first detected in the present study. Nine of the 34 alleles were rare and detected only in one to four accessions. The genetic similarities among the 143 accessions ranged from 0.18 to 1.00. Data analysis based on clustering and principal coordinate analysis showed that a high level of genetic variability exists in this set of core accessions, and indicated that some duplication probably exists in this set core based on the present study.     

Genetic variation of <Emphasis Type="Italic">Bmy1</Emphasis> alleles in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) investigated by CAPS analysis

The enzyme beta-amylase is one of the most important hydrolytic enzymes in the grain of malting barley and is encoded by the gene Bmy1. To learn more about its structure and function, a total of 657 barley accessions including 541 Hordeum vulgare ssp. vulgare (HV), and 116 H. vulgare ssp. spontaneum (HS) were selected for the cleaved amplified polymorphic sequence (CAPS) analysis. These materials, covering all the 16 kinds of beta-amylase phenotypes screened from more than 8,500 accessions of the world barley germplasm, were classified into 13 CAPS types in the present study. A combined assay of phenotypes and CAPS types revealed extensive genetic variation at the Bmy1 locus, and in total 23 Bmy1 allele types were identified. The newly identified alleles (A-I-11, A-II-6, A-II-7, A-II-10, B-I-3, B-I-12 and B-I-13) provided us with a novel resource for barley breeding and Bmy1 study. In HV barley, six out of seven major allele types (C-II-1, B-II-2, B-Ia-3, A-II-5, A-II-6, and A-II-7) were shared with HS barley; the B-I-8 allele, which was predominant in north European cultivated barley, was found to be unique. Remarkably, very low Bmy1 genetic variation was detected in Tibetan barleys, which puts the validity of the hypothesis that Tibet is one of the original centers of cultivated barley into question.     

Chloroplast DNA Diversity in Populations of Wild and Cultivated Barley

Chloroplast DNA (cpDNA) diversity was found within and among populations (245 accessions total) of wild barley, Hordeum vulgare L. ssp. spontaneum Koch from Israel and Iran. Three polymorphic restriction sites (HindIII, EcoRI, BclI) which define three distinct cpDNA lineages were detected. One lineage is common to populations in the Hule Valley and Kinneret of northern Israel, and in Iran. The second lineage is found predominantly in the Lower Jordan Valley and Negev. The distribution of the third lineage is scattered but widespread throughout Israel. Sixty two accessions of cultivated barleys, H. vulgare L., were found, with two exceptions, to belong to just one cpDNA lineage of wild barley, indicating that the cpDNA of cultivated barley is less variable than its wild ancestor. These results demonstrate the need for assessing intraspecific cpDNA variability prior to choosing single accessions for phylogenetic constructions at the species level and higher.     

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.