Genetic diversity revealed by genomic-SSR and EST-SSR markers among common wheat, spelt and compactum

Spelt (TriticumspeltaL .) ,oncebeingthemaincropinEurope ,hasthesamegenome (AABB DD)ascommonwheat (TriticumaestivumL .) .Althoughitiscultivatedonasmallscale ,theproduc tionofspeltisclosetoorevenexceedsthatofcom monwheatinsomeregions[1] .Speltiswidelyadapt edtoavarietyofenvironmentconditions ,itsgrainscontainlargequantitiesofproteinandaresuitableforthemanufactureofsoftbread .Therefore ,speltisre ceivingincreasingattentionforplantbreeders[2 ] .ThemostrecentstudyelucidatedthattheF1generationb…     

Relationships of Aegilops tauschii revealed by DNA fingerprints: The evidence for agriculture exchange between China and the West

Genetic diversity and relationships of wild goat grass (Aegilops tauschii Cosson) from Iran and Xinjiang,west China,as well as its weedy type from the Yellow River region of Shaanxi and Henan provinces in China were analyzed by simple sequence repeat (SSR) fingerprints.A high level of genetic diversity in Aegilops tauschii accessions from Iran was observed,and the richness of genetic diversity was followed by accessions from Shaanxi,Henan,and Xinjiang.The weedy type of Aegilops tauschii showed a close genetic relationship with the wild type from different regions in Iran.The results indicated that the weedy Aegilops tauschii found in the Yellow River region was most likely introduced from lran-the diversity center of Aegilops tauschii.The weedy Aegilops tauschii populations found in the Yellow River region may be brought into the central part of China as a weed species together with common wheat (Triticum aestivum L.) from the West during various periods of time in history.This finding has provided strong evidence for the introduction of common wheat from the West into China via the Silk Road,and also demonstrated the important role of the Silk Road in the exchange of agri-culture and other relevant technologies between China and the West.     

小麦染色体2DS雌性育性位点遗传多态性分析

小麦雌性不育系XND126属于生态遗传型不育系.通过SSR分子标记分析,在2DS染色体上定位了一个雌性育性主效QTL位点.为了构建高密度遗传图谱并精细定位该主效位点,用2DS参考遗传图谱上的14对SSR标记,研究了59个育性正常的普通小麦品种与XND126的DNA多态性,筛选到不同生态型多态性较高的品种共12个,每个品种多态性标记达12~13个,这些品种可以用作杂交亲本,构建新的QTL精细定位群体.在品种组成的群体中,与主效基因位点最近的标记,表现出有较多的品种与雌性不育系具有差异.     

Relationships of Aegilops tauschii revealed by DNA fingerprints: the evidence for agriculture exchange between China and the West

Genetic diversity and relationships of wild goat grass (Aegilops tauschii Cosson) from Iran and Xinjiang, west China, as well as its weedy type from the Yellow River region of Shaanxi and Henan provinces in China were analyzed by simple sequence repeat (SSR) fingerprinting. A high level of genetic diversity in Aegilops tauschii accessions from Iran was observed, and the richness of genetic diversity was followed by accessions from Shaanxi, Henan, and Xinjiang. The weedy type of Aegilops tauschii showed a close genetic relationship with the wild type from different regions in Iran. The results indicated that the weedy Aegilops tauschii found in the Yellow River region was most likely introduced from Iran—the diversity center of Aegilops tauschii. The weedy Aegilops tauschii populations found in the Yellow River region may be brought into the central part of China as a weed species together with common wheat (Triticum aestivum L.) from the west during various periods of time in history. This finding has provided strong evidence for the introduction of common wheat from the West into China via the Silk Road, and also demonstrated the important role of the Silk Road in the exchange of agriculture and other relevant technologies between China and the West.     

Genomic and genic sequence variation in synthetic hexaploid wheat （AABBDD） as compared to their parental species

In order to understand the genomic changes during the evolution of hexaploid wheat, two sets of synthetic hexaploid wheat from hybridization between maternal tetraploid wheat （AABB） and paternal diploid goat grass （DD） were used for DNA-AFLP and single strand conformation polymorphism （SSCP） analysis to determine the genomic and genic variation in the synthetic hexaploid wheat. Results indicated that more DNA sequences from paternal diploid species were eliminated in the synthetic hexaploid wheat than from maternal tetraploid wheat, suggesting that genome from parental species of lower ploidity tends to be eliminated preferentially. However, sequence variation detected by SSCP procedure was much lower than those detected by DNA-AFLP, which indicated that much less variation in the genic regions occurred in the synthetic hexaploid wheat, and sequence variations detected by DNA-AFLP could be derived mostly from non-coding regions and repetitive sequences. Our results also indicated that sequence variation in 4 genes can be detected in hybrid F1, which suggested that this type of sequence variation could be resulted from distant hybridization. It was interesting to note that 3 out of the 4 genes were mapped and clustered on the long arm of chromosome 2D, which indicated that variation in genic sequences in synthetic hexaploid wheat might not be a randomized process.     

Genomic and genic sequence variation in synthetic hexaploid wheat (AABBDD) as compared to their parental species

In order to understand the genomic changes during evolution of hexaploid wheat, two sets of synthetic hexaploid wheat from hybridization between maternal tetraploid wheat (AABB) and paternal diploid goat grass (DD) were used for DNA-AFLP and single strand conformation polymorphism (SSCP) analysis to determine the genomic and genic variation in the synthetic hexaploid wheat. Results indicated that more DNA sequences from paternal diploid species were eliminated in the synthetic hexaploid wheat than from maternal tetraploid wheat, suggesting that genome from parental species of lower ploidity tends to be eliminated preferentially. However, sequence variation detected by SSCP procedure was much lower than those detected by DNA-AFLP, which indicated that much less variation in the genic regions occurred in the synthetic hexaploid wheat, and sequence variations detected by DNA-AFLP could be derived mostly from non-coding regions and repetitive sequences. Our results also indicated that sequence variation in 4 genes can be detected in hybrid F1, which suggested that this type of sequence variation could be resulted from distant hybridization. It was interesting to note that 3 out of the 4 genes were mapped and clustered on the long arm of chromosome 2D, which indicated that variation in genic sequences in synthetic hexaploid wheat might not be a randomized process.     

Genomic and genie sequence variation in synthetic hexaploid wheat(AABBDD)as compared to their parental species

In order to understand the genomic changes during the evolution of hexaploid wheat,two sets of synthetic hexaploid wheat from hybridization between maternal tetraploid wheat (AABB) and paternal diploid goat grass(DD)were used for DNA-AFLP and single strand conformation polymorphism (SSCP) analysis to determine the genomic and genie variation in the synthetic hexaploid wheat.Results indicated that more DNA sequences from paternal diploid species wen eliminated in the synthetic hexaploid wheat than from maternal tetraploid wheat,suggesting that genome from parental species of lower ploidity tends to be eliminated preferentially.However,sequence variation detected by SSCP procedure was much lower than those detected by DNA-AFLP.which indicated that much less variation in the genie regions occurred in the synthetic hexaploid wheat.and sequence variations detected by DNA-AFLP could be derived mostly from non-coding regions and repetitive sequences.Our results also indicated that sequence variation in 4 genes can be detected in hybrid F1.which suggested that this type of sequence variation could be resulted from distant hybridization.It was interesting to note that 3 out of the 4 genes were mapped and clustered on the long alTll of chromosome 2D,which indicated that variation in genic sequences in synthetic hexaploid wheat might not be a randomized process.     

Molecular verification of the integration of Tripsacum dactyloides DNA into wheat genome through wide hybridization

RAPD and RFLP analyses of double haploid lines which derived from hybridization between hexaploid wheat (Triticum aestivum L.2n=42) and eastern gamagrass (Tripsacum dactyloides L.2n=4x=72) are reported.Two of the 340 Operon primers have been screened,which stably amplified Tripsacum dactyloides (male parent) specific bands in the double haploid lines.These results confirm the fact that Tripsacum dactyloides DNA has been integrated into wheat genome by sexual hybridization at molecular level.This idea has been further testified by RFLP analysis.Application and potentials of transferring Tripsacum dactyloides DNA into wheat genome by sexual hybridization in wheat breeding are discussed.     

Molecular verification of the integration ofTripsacum dactyloides DNA into wheat genome through wide hybridization

RAPD and RFLP analyses of double haploid lines which derived from hybridization between hexaploid wheat (Triticum aestivum L. 2n=42) and eastern gamagrass (Tripsacum dactyloides L. 2n=4x=72) are reported. Two of the 340 Operon primers have been screened, which stably amplified 处留情 (male parent) specific bands in the double haploid lines. These results confirm the fact that Tripsacum dactyloides DNA has been integrated into wheat genome by sexual hybridization at molecular level. This idea has been further testified by RFLP analysis. Application and potentials of transferring Tripsacum dactyloides DNA into wheat genome by sexual hybridization in wheat breeding are discussed.     

基于EST-SSR标记鉴定猴耳环自由授粉的全同胞子代

【目的】猴耳环是重要的药用树种,具有较好的工业应用价值。利用EST-SSR标记对猴耳环自然群体1株母树的自由授粉子代进行父本鉴定,从而确定全同胞子代,为基于全同胞家系的后续研究提供材料基础。【方法】以自然群体中母树ELS31自由授粉产生的1 489株子代及该群体内挂果的38株候选父本为材料,利用15个EST-SSR标记检测子代多样性和标记多态性,基于最大似然法鉴定各子代的父本,母本父本均相同的子代即构成全同胞家系,并估算群体内花粉传播距离,检验各父本对应的全同胞家系的多样性,通过卡方检验判断标记是否合乎预期的孟德尔分离比。【结果】15个EST-SSR标记的引物(对)共扩增出89个等位片段。子代群体期望杂合度(H_e)为0.525,表明群体多样性为中等水平;基于自然群体18株无亲缘关系的单株估算的标记平均多态性信息量(PIC)为0.736,表明标记多态性高。在1 489株自由授粉的子代中,确定了857株子代(57.6%)的34株父本。子代数最多的10个父本产生的子代数为26~184株,其他24个父本仅共产生子代139株。未发现自交子代,表明猴耳环是异交物种,自交的可能性极低。对子代数量20株以上的10个父本对应的全同胞家系观测杂合度(H_o)为0.502~0.693,H_e为0.417~0.544,各家系均是H_o大于H_e,表明存在一定程度的杂合子过剩。花粉传播的范围为10.0~559.1 m,平均119.2 m,但主要传播距离在150 m以内。716株子代(83.5%)的父本(10株)与母树距离在150 m以内。距离最远的父本ELS01 (559.1 m)和ELS02 (552.2 m)分别仅产生了9和12株子代。15个标记在子代20株以上的部分或全部全同胞家系中均有不同程度的偏分离,平均每家系的偏分离标记数为8.6个;偏分离最严重的是标记ARCeSSR141,在父本ELS30的全同胞家系中卡方(χ²)值为164.55。【结论】基于15个EST-SSR标记鉴定了猴耳环自然群体1株母本的1 489株自由授粉子代的父本,获得了子代20株以上的10个父本的全同胞家系。这为后续遗传测定、遗传图谱构建和数量性状位点定位等研究提供了材料基础。     

Development, chromosome location and genetic mapping of EST-SSR markers in wheat

A number of 151695 wheat expression sequence tags （ESTs） that originated from GenBank/dbEST from July 14, 2003 to August 24, 2004 were used to search for simple sequence repeats （SSRs） with motif 2-5 bp, and 2038 simple sequence repeats （EST-SSRs）, which accounted for 1.34% of EST database, were identified. Based on these SSR sequences, 249 EST-SSR primer pairs and 166 amplified clear bands in various wheat cultivars were designed. These EST-SSR markers can be used as new molecular markers in wheat and related species. Using Chinese Spring nulli-tetrasomic lines, 93 EST-SSR primer pairs and 193 EST-SSR loci were located on 19 wheat chromosomes except for 4A and 4B. Forty-three loci were mapped on 11 chromosomes of the genetic framework map previously constructed using recombinant inbred lines.     

南京椴群体遗传多样性和遗传结构分析

【目的】 南京椴(Tilia miqueliana)为江苏省重要的乡土树种,野外资源稀缺。南京椴野外群体遗传多样性和遗传结构的探索,可为资源保护、品种选育及遗传改良提供依据。【方法】 以南京椴5个天然群体[江苏宝华山(P1)、牛首山(P2)、安徽皇藏峪(P3)、安徽蜀山(P4)和浙江天台山(P5)]93个体为实验材料,选用15对多态性EST-SSR引物,进行遗传多样性及群体遗传结构分析。【结果】 ①用15对引物共检测等位基因数(A)总和为96,平均值为6.4,四倍体基因型(G)和四倍体特异基因型(G_i)总和分别为441和251,特异基因型比率(R₁)和种质鉴别率(R₂)均值分别为45.73%和17.99%。②在5个群体中,每个位点等位基因数(A_loc)和四倍体基因型丰富度均值(G_loc)分别为5.50±2.43和9.41±4.29;平均观测杂合度(H_o)和平均期望杂合度(H_e)为0.61±1.43和0.62±0.14。参考各群体G_loc和H_e值,确定遗传多样性较高的群体为P1和P3。③群体间遗传分化较小,遗传分化系数(G_st)仅为0.030;AMOVA分子变异分析显示,群体多样性水平变异来自于群体内(96%)。④聚类和遗传结构Structure分析显示,5个群体可划分为2组(组1包括P1、P2和P5;组2包括P3和P4)。Mental检验结果表明遗传距离与地理距离之间无显著相关。【结论】 南京椴群体均具有丰富的遗传多样性,其中宝华山群体和皇藏峪群体多样性较高,群体扩张可能是以这两个种群为中心,经人类活动迁移至其他区域。但南京椴群体间未形成明显分化,主要是由于植株寿命长,群体缺乏自然更新,加之群体间存在人为种子传播。因此,本研究提出通过建立隔离区,明确优先保护群体、加大植株异交,并采用人工繁育及种质回迁的方式保护南京椴野外群体。     

牛角藓 Cratoneuron filicinum（Hedw.）Spruce多样性的 SRAP 标记研究

利用SRAP分子标记方法对不同地区的13份牛角藓 Cratoneuron f ilicinum（Hedw ．） Spruce样本进行遗传多样性分析．结果表明,稳定且重复性高的15对引物共扩增出162条清晰的条带,其中161条具有多态性,平均多态性位点比率为99．49％,说明牛角藓的遗传变异丰富．运用PopGen-32软件分析发现,平均等位基因数 Na＝1．9938,有效等位基因数 Ne＝1．6248,Nei＇s基因多样性指数 H＝0．3606,Shannon＇s信息指数 I＝0.5366,遗传分化系数Gst为0．5736,说明牛角藓的遗传变异主要在居群之间．通过NTSYS2．1软件对14份样本的亲缘关系用U PG-M A方法进行聚类,样本的遗传距离与地理位置没有相关性．     

尖叶拟船叶藓与异猫尾藓的同工酶比较

采用聚丙烯酰胺凝胶电泳方法,对尖叶拟船叶藓和异猫尾藓的过氧化物酶（POD）、酯酶（EST）和超氧化物歧化酶（SOD）等3种同工酶进行了分析研究,结果表明：3种同工酶不仅清楚地显示出尖叶拟船叶藓和异猫尾藓同属于船叶藓科所具有的共同特征酶带,而且酶谱还显示出了种间及各居群间存在的差异.2个种在3种同工酶上表现出较高的相似性,说明拟船叶藓属和猫尾藓属之间有较近的亲缘关系.在物种水平上,异猫尾藓的遗传多样性高于尖叶拟船叶藓,在种内居群间,尖叶拟船叶藓的遗传多样性较高,且受环境因素的影响较大.     

基于EST-SSR分子标记的广西海伦兜兰遗传多样性研究

海伦兜兰（Paphiopedilum helenae）具有极高的观赏价值,为国家一级重点保护野生植物。本研究利用EST-SSR分子标记技术对广西海伦兜兰的5个居群（71个样品）的遗传多样性和遗传结构进行分析,旨在为该种质资源的有效保护与利用提供参考依据。结果表明：海伦兜兰具有中等遗传多样性,多态位点百分比（PPB）为90%-100%,期望杂合度（H_e）平均值为0.451,香农信息指数（I）平均值为0.801。海伦兜兰绝大多数（93%）遗传变异存在于居群内,仅有7%遗传变异存在于居群间,居群内的遗传变异大于居群间的变异,基因流（N_m）平均值为4.916。非加权组平均法（UPGMA）、主坐标分析（PCoA）和Structure分析结果表明,聚类并未严格按照地理位置划分,广西邦亮长臂猿国家级自然保护区靖西市（JX）和龙州县下冻镇春秀村（LZ）居群的遗传多样性较高,应作为海伦兜兰重点保护单元进行保护。     

黄淮麦区部分小麦新品种(系)醇溶蛋白遗传多样性分析

采用酸性聚丙烯酰胺凝胶电泳(A-PAGE)技术分析了黄淮麦区106个小麦(Triticum aestivum L.)新品种(系)间醇溶蛋白带型的遗传差异,结果表明:106份材料间均存在带型差异,每种材料可分离10-24条谱带,106份材料共分离出63条差异带;45份材料具有1B/1R易位系标记性位点Gli-B11,占供试材料的42.5%;106份材料间的遗传距离为0.028 7～0.974 4,平均值为0.570 3.基于遗传距离的聚类分析表明,供试材料在遗传距离0.646水平上聚为5类.上述结果表明供试的黄淮麦区小麦新品种(系)具有广泛的醇溶蛋白遗传多样性.     

AFLP fingerprinting of Chinese epidemic strains of Puccinia striiformis f. sp. tritici

Amplified fragment length polymorphism (AFLP) was used to fingerprint the epidemic strains CY25, CY27, CY28, CY29, CY30, CY31, Hy3, Hy7, Sy13 and a mutant strain WV-4 of P. striiformis f. sp. tritici, the pathogen of wheat stripe rust. The results showed that (i) genetic diversity existed in the pathogen populations, and based on it a dendrogram of these strains was constructed by unweighted pair-group mean average to demonstrate the relationships of the tested strains; (ii) no significant correlation between virulence of the pathogens and the polymorphism of DNA fingerprints was found; ( iii ) AFLP fingerprints showed higher polymorphism than that of the virulence variation; (iv) several new pathotypes identified might evolve independently of the reference strains identified before.     

杉木地理种源的EST-SSR分子标记变异研究

研究采用EST-SSR分子标记对全国不同杉木种源区的42个代表性种源进行了地理种源分子标记的遗传多样性研究。实验结果表明,杉木种源水平上存在较高的遗传多态性。实验使用15个EST-SSR引物扩增出17个位点,共有52个多态性等位基因,每个位点平均具3.058 8个等位基因,平均每个座位的PIC为0.296。相对其他研究,该研究利用SSR标记在每位点检测到更多的遗传变异信息。以遗传距离10作为阀值时,42个杉木种源聚类分析可知,除了福建永春碧卿(编号2)和广西浦北(编号16)种源自成一类外,其他40个种源按从南到北、从东到西的地理分布被聚为四大类群,与以往的杉木种子区划研究结果有较高的吻合度。同时,研究也检测到SSR分子标记的等位基因频率在四大类群间存在着较大的变异,而且这些标记大多位于与植物抗逆性相关的功能基因的编码区域内。因此,笔者推测杉木的地理变异可能与这些基因的等位变异有关。     

春小麦重组自交系及多种SSR标记构建遗传图谱

 以春小麦重组自交系(RIL)宁春4 号×宁春27 号为作图群体,利用SSR 标记构建小麦遗传连锁图谱。结果表明,用1001对SSR 引物选出亲本间表现多态性的引物307 对,多态性频率为30.7%。利用307 对多态性引物对RIL 群体进行分析,共检测到266 个多态性标记位点。通过χ²检测(P<0.05),有147 个SSR 标记表现为偏分离,偏分离率为55.3%,129 个偏向母本宁春4号,其偏分离位点主要分布在B 和D 基因组上。用Mapmaker 3.0 和Mapdraw 2.1 软件将266 个SSR 位点绘制在小麦遗传连锁图上,该图谱覆盖小麦基因组全长2187.79 cM,标记间的平均遗传距离为8.22 cM。     

Assessment of genetic diversity in the forest musk deer （Moschus berezovskii） using microsatellite and AFLP markers

Microsatellite genotyping and amplified fragment length polymorphism (AFLP) techniques are often utilized in studies of conservation genetics of endangered animals. To select a more effective marker system for conserving the endangered forest musk deer, we used microsatellite and AFLP markers to estimate levels of genetic diversity of two populations, the pure mother Jinfengshan (JFS) group and the offspring Baisha (BS) group with introduction of new blood. It was expected that JFS would pos- sess significa...