【作者】 任羽；

【导师】 张银东； 王得元；

【作者基本信息】 华南热带农业大学 ， 作物遗传育种， 2005， 硕士

【摘要】 辣椒杂种优势育种是目前辣椒育种中的主要育种技术。辣椒自交系间的遗传关系对杂种优势育种的成败具有重要意义。针对华南地区绿皮尖椒的育种目标,本研究利用新型分子标记——SRAP标记对31个辣椒自交系和4个杂交种的遗传关系进行研究,并和传统的基于基因型值的遗传关系评价方法进行了比较。主要取得如下结果: 对辣椒SRAP反应体系的程序、模板、Mg²⁺等参数进行优化研究,建立了辣椒SRAP体系:辣椒的PCR程序为94℃预变性5min,94℃变性1min,35℃复性1min,72℃延伸1min,5循环,94℃变性1min,50℃复性1min,72℃延伸1min,35循环,最后72℃延伸10min;25μl反应体系中,模板量为15ng,Mg²⁺为2.0mmol/L。新建立的辣椒SRAP体系重复性好、稳定性强。 采用30个引物组合对35个样品进行扩增,其中27个引物组合可扩增出DNA条带。27个引物组合共扩增出310个多态性条带,每个组合的多态性条带数为3～28条不等,平均每个引物组合产生11.5个多态性条带。可见SRAP标记是进行辣椒自交系遗传关系研究的一种有效的工具。 利用5种相似系数对31个自交系的310个SRAP标记基于UPGMA法进行研究,认为Yule相似系数计算方法是辣椒基于分子标记分类的较为合适的方法;基于Yule相似系数,在相似系数GS为0.55处,可将辣椒的两个变种分开,即Capsicum annuum var. grossum变种群和Capsicum annuum var. longum变种群;在相似系数GS为0.7处,将31个自交系分为5类。第一类:7号和9号;第二类:26、27号;第三类:23、25、22、21、20、19、18、17、14、6、11、24、10、15、13、8、16、12、5、4号;第四类是29、28号;第五类是31、30、2、3、1号。 利用5种遗传距离对31个自交系的10个农艺性状的基因型值基于UPGMA法进行研究,认为Steuclid遗传距离计算方法是较为合适的方法;基于Steuclid遗传距离,在遗传距离GD为6.0处,可将辣椒的两个变种分开,即Capsicum annuum var. grossum变种群和Capsicum annuum var. longum变种群;在遗传距离GD为4.6处,可将31个自交系分为5类。第一类:7号,灯笼椒变种;第二类:9号和5号,植株较高大、晚熟、感病;第三类:25号,中抗、矮杆、早熟;第四类:6、12、10、19、14、11、29、31、30、24、16、20、18、17、21、15、13、23、22、8、4号,这一类在植株更多还原

【Abstract】 Heterosis breeding is one of the main techniques in hot pepper (Capsicum annuum L) breeding. The genetic relationship between inbred lines of hot pepper is very important in heterosis breeding. According to the breeding goal of the green hot pepper in south China region, the genetic relationship among thirty-one inbred lines and four hybrid were studied by a new molecular marker, SRAP (Sequence-related amplified polymorphism), and a comparison between the analysis based on SRAP markers and that based on the classic genotype values was also carried out. The main results were as follows:The SRAP PCR reaction was established by optimizing the parameters including protocol, concentration of DNA template and Mg2+. The SRAP PCR reaction system was initially denaturing at 94°C for 5 min, The first five cycles are run at 94°C, 1 min, 35°C, 1min, and 72°C,1 min ,for denaturing, annealing and extension, respectively. Then the annealing temperature is raised to 50°C for another 35 cycles. In this system of SRAP PCR, the optimum concentration of DNA template and Mg2+were 15ng/25μl, 2.0mmol/L, respectively. The new established SRAP PCR system of hot pepper was fully reproducible and good stability.Twenty-seven out of thirty primer combinations amplified 310 polymorphic bands in thirty-five hot pepper materials. 3-28 polymorphic bands could be amplified from each primer combination, with an average being 11.5 bands. This suggested SRAP was an effective marker in hot pepper genetics field.310 SRAP markers of thirty-one hot pepper inbred lines were studied by UPGMA (Unweighted pair group method arithmetic average) and five similar coefficient calculation methods. The results showed that Yule coefficient was a preferably method in hot pepper classification based on 310 SRAP markers. In the cluster analysis based on Yule coefficient, C. annuum var. longum and C. annuum var. grossum were divided at GS0.55; At GS0.70, thirty-one pepper inbred lines were classified into five groups: group I including No. 7and No. 9; group II including No. 26 and No. 27; group III including No. 23, 25, 22, 21, 20, 19,18, 17, 14, 6, 11, 24, 10, 15, 13, 8, 16,12, 5 and 4; group IVincluding No. 29更多还原