利用RAPD—BSA分子标记技术筛选金针菇色泽基因研究

色泽是金针菇的重要性状之一,受一对等位基因的控制。以FL19(黄色)、8801(白色)及其后代(F1代、F2代)菌株和相应单核菌株为材料,采用分离群体分组分析(BulkedSegregantAnalysis,BSA)方法,对金针菇基因组DNA进行RAPD分析,通过对200个引物的筛选,获得了一个与白色性状基因紧密连锁的RAPD标记S3751800,并在亲本及其后代菌株进行了验证,具有较好的重复性和稳定性。可用于白色金针菇新品种的辅助选育。     

肝素、咖啡因、丙酮酸钠、BSA对延边黄牛卵母细胞体外受精效果的影响

试验为延边黄牛体外胚胎产业化生产奠定基础,进行了肝素、咖啡因、丙酮酸钠、BSA对延边黄牛体外受精效果的影响研究。结果表明,以改良BO液为基础液,添加20 μg/mL肝素时,精子顶体反应率和受精率分别为82.52%和75.34%,高于添加20 μg/mL咖啡因组(P<0.05)和添加10 μg /mL肝素与10 μg/mL咖啡因协同作用组(P>0.05);结合使用肝素(20 μg/mL),添加0.0020 g/mL丙酮酸钠时,精子顶体反应率和受精率分别为78.13%和71.57%, 高于添加0.0015 g/mL和0.0025 g/mL丙酮酸钠组(P<0.05);结合使用肝素(20 μg/mL),添加0.006 g/mL BSA时,精子顶体反应率和受精率分别为80.78%和71.44%,高于添加0.001 g/mL和0.003 g/mL BSA组(P<0.05)。因此,添加适当浓度的肝素、丙酮酸钠和BSA可显著提高延边黄牛精子体外获能效果及体外受精率。     

利用BSA法发掘玉米抗灰斑病主效QTL

以玉米高抗灰斑病自交系齐319与高感病自交系Ye478构建的RILS(重组自交系)为试材,通过两年田间表型鉴定,选取极端表型家系高抗16个,高感15个,利用SSR分子标记,并结合群体分离分析方法(BSA)筛选玉米抗灰斑病连锁标记并进行基因定位。结果表明,在玉米第1连锁群上检测到1个主效抗病基因位点(QTL),与两侧的分子标记umc2614和bnlg1803遗传图距分别为4.74 c M和3.78 c M,该抗病基因位点可解释40.9%的表型变异率,抗病基因来源于齐319,加性效应达到了-7.817 5。     

菠菜性别基因X/Y连锁标记的筛选及应用

以菠菜杂交一代品种‘冬绿1号’中的雌株与雄株杂交构建的F2分离群体为试验材料,采用SRAP-BSA法筛选与性别基因X/Y连锁的分子标记。从256对SRAP引物组合中筛选到了3个与性别基因X/Y紧密连锁SRAP标记：SRAP4.3、SRAP5.7和SRAP9.5。将SRAP5.7和SRAP9.5分别转化为SCAR标记（S5.7、S9.5）,将SRAP4.3转化为dCAPs标记（D4.3）。SCAR标记S5.7、S9.5与Y基因共分离,dCAPs标记D4.3与X/Y基因紧密连锁,遗传距离为0.3 cM。利用S5.7、S9.5和D4.3在其它7个菠菜群体中进行雌雄性别鉴定,分子标记鉴定结果与表型鉴定结果一致,因此S5.7、S9.5和D4.3可以用于分子标记辅助选择育种。     

玉米抗甘蔗花叶病毒SCAR分子标记开发

 【目的】玉米矮花叶病（由甘蔗花叶病毒引起）是中国和欧洲玉米产区的重要病害,开展分子标记辅助育种可以明显提高抗病育种效率。【方法】本文采用改进的BSA法（bulked segregant analysis,BSA）构建玉米抗感自交系DNA池,结合AFLP技术筛选多态性标记,转化后获得实用性强的SCAR标记,并借助100份自交系抗性鉴定结果对SCAR标记进行相关验证。【结果】获得了2个多态性稳定的AFLP标记P66M38-220和P55M51-240,其中将P66M38-220转化为SCAR112标记,此标记与甘蔗花叶病毒抗性高度相关。【结论】改良BSA法是一种发掘性状基因紧密连锁标记的有效方法;开发的SCAR112 标记可以用于玉米抗甘蔗花叶病毒的分子标记辅助选择。     

黄瓜白粉病抗性基因定位及候选基因分析

【目的】通过对黄瓜白粉病抗性基因的精细定位,明确抗性基因的候选区段,为进一步克隆基因及解析基因功能奠定基础。【方法】本研究选用高抗白粉病资源(自交系‘74’)和感白粉病资源(自交系‘80’)配制杂交组合,构建F_1、F_2群体,利用单囊壳白粉菌鉴定亲本及群体白粉病抗性并进行遗传分析;采用BSA-seq的方法对黄瓜白粉病抗性基因进行初步定位,在此基础上在候选区段开发分子标记,进一步精细定位白粉病抗性基因。【结果】分离群体单株表型鉴定结果表明,自交系74的抗性是由不完全隐性基因控制的。BSA-seq技术初步将抗性基因定位于5号染色体15—25 Mb位置,命名为PM 74。最终,利用分子标记将抗性基因定位于SSR15321和SSR07531之间,遗传距离为3.06 c M,物理距离为238 444 bp,可解释41.95%的表型变异。生物信息学分析表明在候选区段内包含有17个候选基因,其中Cucsa.275630为TIR-NBS-LRR类基因。【结论】本研究将黄瓜白粉病抗性基因精细定位到约238 kb的候选区段内,区段内包含有一个TIR-NBS-LRR基因,该基因将是今后研究的重点候选基因。     

四倍体马铃薯块茎蛋白含量分子标记的开发与验证

块茎蛋白含量是马铃薯的重要品质性状之一, 相关分子标记的开发研究较少。本研究以马铃薯高蛋白品种大西洋、低蛋白品种定薯1号及包含173份子代的蛋白含量分离群体为材料, 通过高通量简化基因组测序技术与集群分离分析(BSA)相结合, 开发了SCAR8-107标记, 并以此标记对分离群体的74份高蛋白、42份低蛋白子代及54个四倍体马铃薯品种验证表明, SCAR8-107标记在后代分离群体和四倍体马铃薯品种中的检测结果与表型蛋白含量的对应度分别达到了90.51%和72.97%, 经过Pearson’s双侧相关性分析, 相关性均达到了极显著水平。SCAR8-107的开发对于四倍体马铃薯块茎蛋白含量标记辅助选择与加速马铃薯品质育种具有重要意义。     

梨抗黑星病AFLP标记筛选

 Pear scab caused by Venturia nashicola is one of the most destructive diseases of pears. Molecular markers linked to scab resistance gene is expected to be useful for improving pear. In this study, the F₁ population derived from the cross of ‘Huangguan’ and ‘Yali’ was analyzed genetically. The resistance of pear to scab was proved to be controlled by a single gene in a dominant manner. Bulked segregant analysis (BSA) was conducted to screen 64 fluorescent AFLP primer pairs. A marker designated as D3-365 was found to be linked to the resistant locus. Selective genotype linkage analysis showed that the genetic distance between the marker and the resistant locus was 14.9 cM.     

玉米抗矮花叶病基因SSR分子标记定位研究

通过矮花叶病抗性鉴定筛选出玉米高抗自交系黄野四-3和高感自交系8112.遗传分析显示矮花叶病抗性表现为显性遗传.利用SSR分子标记,结合群体分离分析方法(BSA),对抗矮花叶病基因进行定位,筛选获得了与玉米抗矮花叶病基因位点连锁的SSR标记,并将该基因定位于第3连锁群,与两侧的Bnlg1035和Umc2266分子标记遗传距离分别为8.02 cM和3.04 cM.这一研究结果为快速筛选抗矮花叶病玉米新种质,培育抗性新品种提供了理论依据,为抗矮花叶病基因的分子标记辅助育种和抗病基因克隆奠定了基础.     

Resistance to the Whitefly Aleurotrachelus socialis (Hemiptera: Aleyrodidae) and SSR Marker Identification in Advanced Populations of the Hybrid Manihot esculenta subsp. Manihot flabellifolia

Genes resistant to Aleurotrachelus socialis were transferred to the F₁ from the interspecific hybrid wild species of Manihot flabellifolia to M. esculenta and two advanced generations of backcrosses (BC₁ and BC₂). We characterized the resistance of A. socialis transferred to BC₂ parents (CW67-160, CW67-130, CW67-44), MTAI-8 (BC₁), resistant (CMB9B-73) and susceptible (CMB9B-104) genotypes from contrasting pools, and resistant (MEcu-72) and susceptible (CMC-40) genotypes. Whitefly demography and biology were evaluated. SSR molecular markers associated with a phenotypic response of plant resistance were detected in segregating populations (BC₂). Results showed that although female survival time was similar on all hosts, the lowest averages of longevity, fecundity and oviposition rate were observed in the resistant control MEcu-72, only being significantly similar to the parent CW67-130. When the BC₁ and BC₂ populations were compared, it was found that A. socialis fecundity was eight times lower on CMB9B-73 progeny than on CW67-130, expressing the highest levels of resistance to the whitefly. Ten genotypes of CMB9A and CMB9B family had the best segregation. A total of 486 microsatellite primers were evaluated using bulked segregant analysis (BSA), 11 showed polymorphism between the contrasting pools and only one showed significant differences between resistant and susceptible individuals. In conclusion, fecundity was the parameter that impacted most on the intrinsic rate of A. socialis population growth.