水稻直链淀粉含量的分子遗传研究

为选育高产、优质、直链淀粉含量中等的杂交稻组合，用23个非糯水稻纯系品种（系）及其所配22个组合的F1代，研究了微卫星标记484／485的PCR扩增产物（CT）n多态性，484／W2R的PCR扩增产物被AccⅠ酶切多态性，二者与直链淀粉含量的相关性，亲本与F1直链淀粉含量的相关性，结果表明，供试纯系材料有3种多态片段，即（CT）11，（CT）18和（CT）20，其中（CT）11或（CT）18的品种数多；484／W2R扩增产物能或不能被AccⅠ酶切的品种数比较接近；484／485扩增产物（CT）n多态性和484／W2R扩增产物被ACCⅠ酶切的多态性与直链淀粉含量极显著相关；杂种F1直链淀粉含量与双亲平均值极显著正相关，直链淀粉含量偏高的籼稻品种与直链淀粉含量偏低的粳稻品种杂交，杂种F1直链淀粉含量中等的组合数多。     

冬小麦种质“矮孟牛”姊妹系遗传差异

采用田间试验鉴定和基因组分子标记分析相结合的方法,对冬小麦种质“矮孟牛” 7个类型的性状和基因组遗传差异比较分析。结果表明,7个类型在调查的株高、穗长、穗粒数、粒长、粒宽、单株穗数等16个性状方面存在差异,其中矮孟牛V型的主要产量构成因素等性状比较协调,总体优于其他6个类型。在检测的2 210对SSR、EST-SSR和STS标记中,有656对标记可在矮孟牛三亲本中显示多态性,将其用于矮孟牛7个类型全基因组遗传差异分析,并利用GGT2.0绘制7个类型的遗传差异图谱。在矮孟牛V型的1A、1B、2D、3A、4D、7A染色体上检测到8个特异位点,利用矮丰3号/牛朱特和孟县201/牛朱特构建的2个F₂分离群体,经IciMappingV3.0单标记QTL作图分析,发现矮孟牛V型部分特异位点附近存在与产量构成因素等重要性状相关的QTL。矮孟牛V型的基因组特异位点可能是它作为骨干亲本区别于其他姊妹系的重要基因组特征。     

Genetic diversity in mung bean germplasm revealed by RAPD markers

Knowledge of the genetic relationships among landraces is useful to gene bank managers because it permits a better organization of the crop's gene pool management, more efficient sampling of the available germplasm resources and better access to useful genetic variation for breeders. Genetic diversity of 19 landraces of the cultivated mung bean, Vigna radiate, and three weedy and wild relatives including Vigna mungo, Vigna luteola and Vigna radiate var. sublobata, was investigated at the DNA level with the random amplified polymorphic DNA (RAPD) procedure. Sixty random decamer primers were employed in amplification reactions; 28 of these were informative and yielded 246 bands, of which 229 were polymorphic with a mean of 8.2 bands per primer. A genetic distance matrix based on Nei and Li coefficient was converted to a dendrogram and a two-dimensional plot using multidimensional scaling (MDS). The accessions studied were separated into three main clusters, which included V. radiate landraces, V. mungo and V. luteola, respectively. The variation of this cluster supports the view that the genetic distance of V. mungo and V. luteola varies considerably from the accession VO2955 (V. radiata). The multidimensional scaling plot confirmed that V. mungo, V. luteola and most of the accessions of V. radiata formed distinct clusters with no overlap, and two mung bean accessions (PI177493 and VO4134–1 from Turkey and India, respectively) were genetically distant from other V. radiata landraces. V. radiata and V. mungo are positioned in separate botanical species and V. radiata var. sublobata is classified within other V. radiata landraces. Based on the limited range of accessions tested, the approach holds promise for the classification of mung bean germplasm, identification of mung bean landraces and applications of molecular markers to mung bean breeding.     

Cryptic species and population genetic structure of Plasmopara viticola in São Paulo State,Brazil

Downy mildew (Plasmopara viticola) is one of the most important diseases in grape-growing areas worldwide, including Brazil. To examine pathogen population biology and structure, P. viticola was sampled during the 2015/16 growing season from 516 lesions on nine grape cultivars in 11 locations in subtropical areas of São Paulo State, Brazil. For identification of cryptic species, a subsample of 130 isolates was subjected to cleaved amplified polymorphic sequence (CAPS) analysis, and for 91 of these isolates the ITS1 region was sequenced. These analyses suggest that the population of P. viticola in São Paulo State consists of a single cryptic species, P. viticola clade aestivalis. Seven microsatellite markers were used to determine the genetic structure of all 516 P. viticola isolates, identifying 23 alleles and 55 multilocus genotypes (MLGs). Among these MLGs, 34.5% were clonal and represented 93% of the isolates sampled. Four dominant genotypes were present in at least five different locations, corresponding to 65.7% of the isolates sampled. Genotypic diversity (Ĝ = 0.21–0.89) and clonal fraction (0.58–0.96) varied among locations (populations). Most populations showed significant deviation from Hardy–Weinberg expectations; in addition, excess of heterozygosity was verified for many loci. However, principal coordinate analysis revealed no clusters among locations and no significant isolation by distance was found, suggesting high levels of migration. The results indicate that downy mildew epidemics result from multiple clonal infections caused by a few genotypes of P. viticola, and reproduction of P. viticola in São Paulo State is predominantly asexual.     

正交设计优化草地早熟禾SRAP-PCR反应体系及引物筛选

采用L9(34)正交试验设计方法,对草地早熟禾(Poa pratensis)基因组DNA SRAP PCR反应体系中的Taq DNA聚合酶、Mg2+、引物及dNTP四因素的用量进行优化,并比较不同模板DNA用量对扩增的影响,建立草地早熟禾SRAP PCR最佳反应体系,同时,利用该体系对SRAP引物进行筛选。结果表明,草地早熟禾SRAP PCR最佳反应体系为Taq DNA聚合酶1.0 U、Mg2+ 1.75 mmol·L-1、引物0.25 μmol·L-1、dNTP 220 μmol·L-1、40 ng模板DNA、2 μL 10×PCR buffer,总体积20 μL。运用该体系从100对SRAP引物中筛选出43对引物能够产生清晰稳定的扩增条带且多态性丰富。优化体系的建立及引物的筛选可为今后利用SRAP标记技术对草地早熟禾进行遗传多样性分析、图谱构建、种质资源鉴定奠定技术基础。     

Genetic mapping of the barley Rrs14 scald resistance gene with RFLP, isozyme and seed storage protein markers

The scald susceptible barley cultivar ‘Clipper’ and a third‐backcross (BC₃) line homozygous for the Rrs14 scald resistance gene that originally came from Hordeum vulgare ssp. spontaneum were grown in replicated field trials. The level of resistance that Rrs14 confers against field populations of the pathogen Rhynchosporium secalis, the causal agent of scald disease, was evaluated. The Rrs14 BC₃ line exhibited 80% and 88% less leaf damage than ‘Clipper’ in 1995 and 1996, respectively. Given this effectiveness of Rrs14, research was undertaken to identify a linked marker locus suitable for indirect selection of Rrs14. Based on linkage to a set of previously mapped loci, Rrs14 was positioned to barley chromosome 1H between the seed storage protein (hordein) loci Hor1 and Hor2, approximately 1.8 cM from the latter locus. The Hor2 locus is thus an ideal codominant molecular marker for Rrs14. The tight linkage between Rrs14 and Hor2 and the availability of alternative biochemical and molecular techniques for scoring Hor2 genotypes, permits simple indirect selection of Rrs14 in barley scald resistance breeding programmes.     

利用优化的SSR分子标记技术检测杂交水稻万优481纯度

[目的]通过降低试验药剂、减少PCR循环时间、提高琼脂糖利用指数等,建立经济、快速、低污染的杂交水稻纯度鉴定体系。[方法]将杂交水稻万优481、亲本万3A和万恢481在光照培养箱内温度发芽(30℃,长到5 cm左右),用CTAB法提取DNA;按优化的PCR体系和电泳方法,用亲本万3A和万恢481的DNA从24对引物中筛选出多态性好的引物P18,用引物P18对杂交水稻万优481样本进行纯度鉴定。[结果]利用优化的SSR分子标记技术准确鉴定出万优481的纯度为98.5%,与田间鉴定结果相符,凝胶电泳成像带谱清晰,同时,与常用方法相比,酶等试验药品使用量减少20%~25%,试验所用时间较常用方法减少了50%以上。[结论]利用优化的SSR分子标记技术体系鉴定杂交水稻种子纯度,经济、快速且鉴定的结果清晰、可靠,同时大大减少琼脂糖等试验残渣的产生,降低了对环境污染,值得推广、应用。     

AFLP, ISSR and RAPD markers reveal high levels of genetic diversity among Lupinus spp.

The Lupinus genus includes a number of important crop species. The use of defined nucleotide sequences for the analysis of genetic diversity among these species has revealed modest levels of diversity. The aim of this study was to access AFLP, ISSR and RAPD markers to evaluate the genetic diversity among L. albus, L. angustifolius, L. cosentinii, L. hispanicus, L. luteus, L. mutabilis, L. pilosus and L. polyphyllus. Unexpectedly, low levels of genetic similarity were found (ranging from 0.205 to 0.432), regardless of the type of molecular marker used. Nevertheless, these techniques consistently showed a greater genetic similarity between L. pilosus and L. cosentinii, L. mutabilis and L. polyphyllus and among L. luteus, L. hispanicus and L. angustifolius, clearly separating the Old World from the New World species. Such low genetic similarity among Lupinus spp. is most unlikely to be due to differences in coding sequences but could be the result of a long diverging process concerning non‐coding regions, which would represent a very important proportion of these genomes.     

Molecular markers and ex situ conservation of the European elms (Ulmus spp.)

The enormous losses suffered by the European elms during recent Dutch elm disease outbreaks led to concern over the conservation of elm genetic resources, and the subsequent establishment of a series of ex situ collections. However, as ex situ collections are inevitably finite in size, some consideration needs to be given to selecting which samples to include in them. To contribute towards this process for European ex situ elm collections we have undertaken genetic studies on a Europe-wide sample of 535 individuals. A major aim has been to use genetic markers to clarify the identification of samples to ensure that the ex situ collections contain a representative spread of taxonomic diversity. This is important given the paucity of mature elms in the landscape due to Dutch elm disease. The lack of mature material (critical for identification) compounds identification problems in what was already a taxonomically difficult group. Our data (derived from random amplified polymorphic DNA and inter-simple sequence repeats) have provided a useful supplement to morphology in undertaking such sample identifications. The molecular data served to highlight mis-identified samples and led to extensive revisions of sample identities within individual countries. Our results were less useful in detecting regional intra-specific genetic structure, and do not provide sufficient information for prioritising within-species sample selections.     

大白菜和紫菜薹自交系染色体组DNA的RAPD分析

用８个随机引物（１０ｂｐ）对２个紫菜薹自交系的３个单株和４个大白菜自交系的４个单株的染色体组ＤＮＡ进行ＰＣＲ扩增，共有４０条带分离清晰、明亮，其中引条带在７个单株中表现出差异，可作为遗传标记（ＲＡＰＤ标记）。４个大白菜自交系之间的平均差异条带数为１２．３，２个紫菜薹自交系之间为９．５，大白菜与紫菜薹之间平均有１５．５条带的差异。用７个引物在９３Ｗ０５－７（紫菜薹）和９３Ｗ５８－５（大白菜）之间检测出对个ＲＡＰＤ标记。从同一自交系的不同单株抽提的染色体组ＤＮＡ扩增出较一致的ＤＮＡ谱带，而不同自交系间扩增出的ＤＮＡ谱带差异很大，这表明此方法也可以象ＲＦＬＰ一样，用于大白菜和紫菜薹的分子标记。