运用随机扩增多态性DNA标记检测中国东北大豆灰斑病菌株遗传变异

 运用致病力和DNA多态性检测中国东北地区的35个大豆灰斑病菌分离物的遗传变异、根据菌株在9个品种(系)上的致病力反应可将其分为7个组。利用13个随机引物扩增供试菌株共计产生105个RAPD标记,其中78.1%具有多态性。通过聚类分析计算了各菌株间的遗传距离,并产生树状图,发现同一地区内及不同地区间的病菌表现遗传变异、致病性和DNA多态性间具有一定相关性。     

瓣化型胡萝卜雄性不育基因atp6的CAPS标记建立

根据atp6的基因序列设计特异扩增引物,以瓣化型胡萝卜核质互作雄性不育系H05A及其保持系H05B为试材,扩增获得了1条约500bp的特异扩增片段。用限制性内切酶BglII对该片段进行酶切,不育系产生1条保持系中没有的特异片段,其分子量约为60bp。建立了atp6基因的CAPS标记,该分子标记可用于胡萝卜雄性不育分子标记辅助育种。     

文心兰种质遗传多样性的SRAP分析

利用SRAP标记分析文心兰部分种质资源的遗传多样性，采用30对引物组合对33份文心兰种质进行扩增，从中筛选出19个多态性较高的引物组合，共产生277条多态性条带，平均每个引物组合产生14.6个多态性条带，相似系数在0.280～0.927。通过聚类分析发现，在值为0.37时，可将33份文心兰种质划分为4大类群。第Ⅰ类厚叶大花品种，第Ⅱ类为厚叶小花品种，第Ⅲ、第Ⅳ类均为类薄叶品种。 结果表明，文心兰种质的聚类与其遗传背景有很大的关系。     

晋大62×诱处4号杂交后代群体遗传多样性分析

选用晋大62×诱处4号杂交亲本和后代群体为试验材料,通过微卫星(SSR)标记分析,利用SPSS软件对其进行了聚类分析,探讨了亲本及后代群体间的遗传特性和亲缘关系,研究了大豆群体的遗传多样性,为大豆杂交育种亲本的选配、杂交后代的筛选、种质创新及大豆遗传多样性研究提供了理论和实践依据。结果表明,以亲本的基因组DNA为模板,选用40对不同的引物进行电泳共筛选出23对位于多条染色体上的差异引物,其多态性比例为55.0%。通过聚类分析所有材料可聚类为以下几类:母本晋大62(CK1)和后代6~13号材料聚为第一大类,父本诱处4号(CK2)和后代25~31号材料聚为第二大类,而1~5、14~20聚为第一小类,21~24、32~35聚为第二小类,这两小类材料与亲本的亲缘关系较远。     

花生DNA分子标记的研究工花生AFI——Ⅰ分析技术体系的建立与作图杂交亲本的筛选

由于缺乏传统遗传标记，花生遗传连锁的报道极为罕见，迄今未发表经典的遗传图谱。发展稳定可靠的AFLP标记，将为基因分型应用于新品种保护、良种保纯、杂种鉴定和种质资源研究提供强有力的技术支撑。研究在建立适合花生AFLP分析的DNA提取流程的基础上，建立了花生AFLP分析的技术方案。AFLP分析结果表明，6个花生杂交亲本间存在一定差异。四对引物共计扩增出307条长度不同的条带。其中多态性条带为160条，占总条带数的52．1％。本研究筛选获得了多态性程度高的杂交亲本，为进一步构建分子连锁图谱创造了条件。     

黄瓜叶面积遗传及QTL定位分析

选用2个叶面积大的黄瓜品种（D9320、D2005）和2个叶面积小的黄瓜品种（D0455、D0401）配制杂交组合，筛选叶面积差异较大的亲本组合D0401×D9320构建F2分离群体，采用加性－显性（AD）模型进行遗传分析。结果表明：黄瓜叶面积是由多基因控制的数量性状，以显性效应为主，存在一定的加性效应，狭义遗传力和广义遗传力较高。采用SSR分子标记构建遗传连锁图谱，利用复合区间法对黄瓜叶面积进行QTL定位。结果表明：CSWACC03、CSWCT05BZ和CSCTTT15a位于同一个连锁群上，连锁群总长度为68.3 cM；检测出1个与黄瓜叶面积有关的QTL，与最近标记的距离为3.6 cM，且变异贡献率为8.65 %。     

Recovery and Biological Properties of Nitrate Non-utilizing Mutants of Rice Blast, Magnaporthe grisea

Eleven nitrate non-utilizing (nit) mutants were recovered from six isolates of Magnaporthe grisea cultured on MM media amended with 60 g/L potassium chlorate, with a frequency of 1.42 %. Some biological properties, such as growth rate, growth biomass, cultural characters, conidial production, sexual reproduction ability, and pathogenicity were compared between nit mutants and their parent isolates. Results showed that all the nit mutants were resistant to chlorate. Some important biological properties such as the growth rate on YPSA, conidial production ability on TPSA, pathogenicity, had no significant differences between nit mutants and their parent isolates. Mating type didn‘t change, but perithecia production ability of fertile isolates changed significantly as compared with that of their parent isolates. Therefore, the nit can be used as a genetic marker to study the genetics such as pathogenicity, fungicide resistance in Magnaporthe grisea.     

Bulked Segregant Analysis to Detect QTL Related to Heat Tolerance in Rice （Oryza sativa L.） Using SSR Markers

The study was undertaken to assess the genetic effect of quantitative trait loci （QTLs） conferring heat tolerance at flowering stage in rice. A population consisting of 279 F2 individuals from the cross between 996, a heat tolerant cultivar and 4628, a heat-sensitive cultivar, was analyzed for their segregation pattern of the difference of seed set rate under optimal temperature condition and high temperature condition. The difference of seed set rate under optimal temperature condition and high temperature condition showed normal distribution, indicating the polygenic control over the trait. To identify main effect of QTL for heat tolerance, the parents were surveyed with 200 primer pairs of simple sequence repeats （SSR）. The parental survey revealed 30% polymorphism between parents. In order to detect the main QTL association with heat tolerance, a strategy of combining the DNA pooling from selected segregants and genotyping was adopted. The association of putative markers identified based on DNA pooling from selected segregants was established by single marker analysis （SMA）. The results of SMA revealed that SSR markers, RM3735 on chromosome 4 and RM3586 on chromosome 3 showed significant association with heat tolerance respectively, accounted for 17 and 3% of the total variation respectively. The heat tolerance during flowering stage in rice was controlled by multiple gene. The SSR markers, RM3735 on chromosome 4 and RM3586 on chromosome 3 showed significant association with heat tolerance respectively, accounted for 17 and 3% of the total variation respectively. The two genetic loci, especially for RM3735 on chromosome 4, can be used in marker-assistant-selected method in heat tolerance breeding in rice.     

利用RIL群体和自然群体检测与花生含油量相关的SSR标记

以远杂9102×中花5号杂交后代衍生的重组近交系F₈代家系为材料，在含油量测试的基础上，选用10份低油材料(平均含油量52.91%)、12份高油材料(平均含油量58.85%)以及亲本进行SSR引物筛选，通过631对SSR引物扩增，筛选出来源于7对引物的13个有显著差异的片段可以有效区分低油材料和高油材料。以这7对差异引物在F₈ RIL群体中扩增，对20份低油家系材料(含油量<55%)和45份高油家系材料(含油量>56%)进行统计分析，获得1个与花生含油量相关的分子标记2A5-250/240，其中，标记2A5-250为低油材料(含油量<55%)所拥有，相符率为95.0%，标记2A5-240为高油材料(含油量>56%)所拥有，相符率为88.9%。用SSR标记2A5-250/240检测11份高油(平均含油量为55.93%)栽培种花生和11份低油(平均含油量为48.41%)栽培种花生，结果表明，标记2A5-240与高油栽培种花生的符合率为63.6%，2A5-250与低油栽培种花生的符合率为90.9%。在19份高油(平均含油量为58.60%)野生花生中，10份野生花生能检测到标记2A5-240。综合分析RIL群体和自然群体的研究结果表明，标记2A5-250/240可用于花生含油量分子标记辅助选择。