小麦粘类CMS育性恢复基因的SSR分子标记与定位

利用SSR技术,对小麦粘类CMS（细胞质雄性不育）的1对近等基因系（NILs）和回交群体（BC₁'）进行分析,筛选与粘类CMS育性恢复基因相连锁的分子标记并进行恢复基因定位,以进一步探讨小麦粘类CMS育性恢复的遗传机理。结果表明：在18对位于1BS上的SSR引物中,有6对引物在近等基因系间扩增出了稳定的多态性差异;经分离群体验证表明,恢复基因与4对SSR引物的扩增位点Xwmc406、Xbarc8、Xwmc611和Xgwm273有连锁关系,遗传距离分别为2.7、2.8、21.4和26.2cM,这些标记可稳定应用于小麦粘类CMS育性恢复基因的辅助选择;研究还表明,小麦粘类雄性不育系的育性恢复主要受1BS上的1对主效恢复基因及一些微效基因共同控制;本研究标记出的恢复基因应为Rfv1;上述4个标记与该主效恢复基因Rfv1之间的位置顺序依次为Xwmc406、Rfv1、Xbarc8、Xwmc611、Xgwm273。     

棉花铃重AFLP标记的初步研究

本文以陆地棉铃重近等基因系为材料(大铃≥5.4g,小铃≤1.7g),构建大小铃两个近等基因池.利用AFLP标记技术对两基因池进行分析,64对引物组合共产生3840条稳定、清晰扩增带.五条扩增带在两基因池间呈现多态性,其中四条差异带出现在大铃基因池,编号分别为LB-1、LB-2、LB-3和LB-4;一条出现在小铃基因池,编号为SB-1.模板浓度梯度分析表明,在一定范围内,酶切模板浓度对AFLP结果影响较小.利用稍作修改的CTAB法提取的棉花DNA可用于AFLP分析.     

中国水稻主产区白叶枯病菌致病型分析及近等基因系鉴别寄主的构建

【目的】分析中国不同稻区白叶枯病菌（Xanthomonas oryzae pv. oryzae）致病型,建立近等基因系鉴别寄主,为白叶枯病菌群体结构田间实时准确监测、抗性品种应用以及抗病育种提供科学依据。【方法】利用中国鉴别寄主、IR24以及15个抗白叶枯病近等基因系等共21个鉴别寄主,采用人工剪叶接种方法,对2018—2021年采自广东、广西、海南、浙江、湖南、辽宁、云南共7个省（自治区）的954个单菌落分离菌株进行致病型测定,探明白叶枯病菌致病型种类、分布及毒性分化;基于测试菌株与15个近等基因系及IR24的抗感互作,应用主成分因子分析法,开展近等基因系与病菌互作的变量因子分析,构建白叶枯病菌致病型近等基因系鉴别寄主;基于抗病基因与测试菌株的抗感反应,分析抗病基因聚合效应。【结果】954个测试菌株在中国鉴别寄主上鉴定出11个致病型,包括SRRRR（I）、SSRRR（Ⅱ）、SSSRR（Ⅲ）、SSSSR（Ⅳ）、SSRRS（V）、SRSRR（Ⅵ）、SSSSS（Ⅸ）、SSSRS（新型1）、SRSRS（新型2）、SRSSS（新型3）以及SSRSS（新型4）,占测试菌株的比率分别为11.53％、4.82％、7.34％、6.18％、7.23％、1.05%、59.96％、1.57%、0.10%、0.10%、0.10%。Ⅸ型菌作为致病性最广的强毒菌系已上升为华南和长江中下游湖南和浙江稻区的优势致病型,西南稻区的云南以Ⅳ型菌为主,东北稻区的辽宁以I型菌为主。15个水稻抗白叶枯病近等基因系对954个菌株的抗感性分析结果表明,测试的15个近等基因系可分为5种类型,第Ⅰ类为高感基因系,包括IRBB1、IRBB2、IRBB10、IRBB11、IRBB4;第Ⅱ类为中感基因系,包括IRBB3、IRBB203、IRBB14;第Ⅲ类为中抗基因系,包括IRBB8、IRBB13;第Ⅳ类为抗病基因系,有IRBB21;第Ⅴ类为高抗基因系,包括IRBB5、IRBB7、CBB23、GDBB23;测试菌株中,出现可侵染抗病基因xa5的有42个、Xa7的有34个、Xa23的有31个。对以白叶枯病近等基因系为主的16个品种（系）与954个菌株组成的抗感互作变量数据矩阵进行因子分析,以解释总变量>85.0%为界,提取出8个主成分因子,组建了以近等基因系为主的10个品种(系)组成白叶枯病菌近等基因系鉴别寄主,按其对变量方差贡献大小,这些寄主分别为IRBB10（Xa10）、IRBB4（Xa4）、GDBB23（Xa23）、IRBB5（xa5）、IRBB7（Xa7）、IRBB21（Xa21）、IR24（Xa18）、IRBB13（xa13）、IRBB3（Xa3）、金刚30;新鉴别寄主可将954个测试菌株划分为55个致病型,对测试稻区的白叶枯病菌菌株表现出较好的鉴别力。基因聚合联合抗性分析表明,不同抗病基因聚合对病菌的抗性频率有一定的提升,不同抗病基因对测试菌株的抗性具有一定的互补性。【结论】监测稻区的白叶枯病菌系趋向多样化,毒性分化明显,强毒菌系Ⅸ型菌在部分稻区已上升为优势致病型,侵染xa5、Xa7及Xa23等广谱抗性基因的菌株有上升趋势;抗病基因聚合可拓宽品种对病原菌系的抗性谱,是培育广谱抗性品种的有效途径;近等基因系鉴别寄主的建立与应用可为白叶枯病发生流行的精准监测以及田间实时预警提供技术支撑。     

玉米高赖氨酸突变奥帕克-2和奥帕克-16近等基因系的营养品质分析

为了评价高赖氨酸玉米近等基因系的品质,以便为其遗传与应用研究奠定基础,用近红外分析仪分析了玉米高赖氨酸突变奥帕克-2(o2)和奥帕克-16(o16)近等基因系BC5F3种子的营养品质。结果表明:普通玉米自交系QCL2179和QCL2180籽粒的赖氨酸、油分、蛋白质和淀粉平均含量分别为0.33%、6.26%、13.51%和65.94%。其o2近等基因系籽粒的赖氨酸、油分、蛋白质和淀粉平均含量分别提高了22%、15%、16%和1.7%。其o16近等基因系不透明和带修饰基因胚乳籽粒的赖氨酸、油分、蛋白质和淀粉平均含量分别提高了21%、18%、-16%和1.5%。其o16近等基因系半不透明(透明度较低)胚乳籽粒的赖氨酸、油分、蛋白质和淀粉平均含量分别提高了3%、-3.5%、-5%和1.5%。可见,o2近等基因系、o16不透明和带修饰基因胚乳近等基因系可作为o2和o16两基因遗传研究和应用的重要材料。     

抗白叶枯病水稻近等基因系的选育

以沈农15为轮回亲本(RP),IRBB60为Xa-21基因供体(DP),经3代回交,2代自交,在分子标记辅助选择(molecular marker assisted selection,MAS)条件下,构建了携有抗白叶枯病基因Xa-21的近等基因系。近等基因系农艺性状与沈农15相似,抗C2(河北菌株)、C5(广东菌株)等白叶枯小种。本研究所运用的分子育种程序既能消除外源遗传背景,又能使水稻品种的农艺性状得到有效改良。     

Dough properties and bread-making quality-related characteristics of Yumechikara near-isogenic wheat lines carrying different Glu-B3 alleles

We investigated the relationships of three allelic variations in Glu-B3 (ab, g, and h) with dough properties and bread-making quality-related characteristics using near-isogenic lines (NILs) of ‘Yumechikara’ that commonly carry Glu-A1a, Glu-B1b, Glu-D1d, Glu-A3f, Glu-B3ab and Glu-D3a. Measurement of peak time (PT) in a 2-g mixograph indicated that Glu-B3g was the most effective for a strong dough property, followed by Glu-B3ab, with Glu-B3h being the least effective. The results of measurement of mixing time during bread-making were similar to those for PTs, i.e., the lines carrying Glu-B3g showed the longest mixing time, followed by those of Glu-B3ab, and those of Glu-B3h showed the shortest mixing time. Since two parameters of bread-making quality, loaf volume (LV) and specific loaf volume (SLV), were affected by flour protein contents in all groups of the Glu-B3 genotype, we compared the effects of the three Glu-B3 alleles on those parameters using analysis of covariance (ANCOVA) to remove the effect of protein content. The results indicated that the Glu-B3h group showed the largest SLV, followed by the Glu-B3ab group, and the Glu-B3g group showed the smallest SLV. These results suggest that the introduction of Glu-B3h into ‘Yumechikara’ makes it possible to breed varieties with good bread-making quality-related characteristics.     

Identification of allele specific AFLP markers linked with bacterial wilt [Ralstonia solanacearum (Smith) Yabuuchi et al.] resistance in hot peppers (Capsicum annuum L.)

Even though the bacterial wilt is identified as the most destructive disease in hot peppers world-wide, robust molecular markers that facilitate marker assisted selection are absent till date. Kerala Agricultural University (India) has released two hot peppers named Ujwala and Anugraha which show high level field resistance to this pathogen. The variety Anugraha was developed through backcross breeding between a high yielding but highly susceptible variety Pusa Jwala with the highly resistant Ujwala, using Pusa Jwala as a recurrent parent. Thus, Pusa Jwala and Anugraha are near isogenic lines (NILs) differing for the resistance to bacterial wilt only and the resistance is governed by a homozygous recessive (rr) gene action. The F₁s of Anugraha × Pusa Jwala were selfed to generate the segregating F₂ population. The F₂ population has been field screened, 10 highly susceptible and 10 most resistant plants were identified and DNA from these plants were bulked separately. Bulked segregant analysis with AFLP primer combination EcoACT + MseCAC was done using the DNA from donor parent Ujwala, susceptible parent Pusa Jwala, resistant parent Anugraha, bulked susceptible F₂ and bulked resistant F₂ plants. On resolution using capillary electrophoresis system in genetic analyzer, the AFLP products have yielded three polymorphic bands (103, 117, and 161 bp) which were linked with the resistant recessive allele and three polymorphic bands (183, 296, 319 bp) linked with the dominant susceptible allele of the bacterial wilt resistance gene. The results were confirmed through co-segregation analysis in most resistant and susceptible plants of F₂ segregating population.     

利用分子标记辅助选择优质蛋白玉米近等基因系

为了扩增优质蛋白玉米育种新种质,用普通玉米种质转育创制优质蛋白玉米(QPM)近等基因系;利用SSR标记phi057进行辅助选择,以优质蛋白玉米种质为o2基因供体,转导、构建来自不同遗传背景普通玉米种质为受体的QPM近等基因系。结果表明：利用共显性SSR标记引物phi057在o2基因供体自交系与普通玉米受体自交系间表现多态性,能区分O2O2、O2o2和o2o2等3种基因型,来自不同遗传背景种质所构建获得的QPM近等基因系赖氨酸含量有不同程度提高,赖氨酸含量达到0.36%~0.42%,不同遗传背景受体自交系赖氨酸增加幅度9.1%~64.0%。普通玉米种质导入供体o2基因后能够提高种质赖氨酸含量。     

大豆叶柄角的QTL定位分析

叶柄角是大豆株型的重要构成因素,影响大豆冠层结构、光合作用效率以及最终产量。解析大豆叶柄角的遗传基础对提升大豆产量具有重要意义。本研究以2个叶柄角具有显著差异的亲本BLA和SLA以及它们衍生的RIL群体为材料,构建高密度的遗传图谱,对大豆不同部位的叶柄角进行QTL分析,并利用近等基因系验证部分QTL。遗传分析结果显示,叶柄角呈连续正态分布,符合数量性状遗传特征。利用GBS技术构建了包含859个Bin标记的大豆高密度遗传图谱,总遗传长度为2326.9cM,标记间平均距离为2.763cM;共检测到14个调控叶柄角的QTL,LOD值在2.58~4.80之间,可解释遗传变异范围在6.9%~12.4%之间,其中5个QTL定位在第12染色体上且成簇存在;构建的qLA12和qLA18的近等基因系表型结果显示,叶柄角在同一对近等基因家系间差异显著,表明qLA12和qLA18是2个可信的QTL。本研究为进一步克隆调控叶柄角功能基因奠定了基础,为大豆理想株型育种提供了遗传材料。