小麦高效转基因受体品系CB037的抗条锈性分析

Genetic transformation is used for efficient gene cloning, gene editing and gene engineering, etc. Obtaining recipient lines amenable to transformation and with pure genetic background is critical for high efficiency transformations. For recent years, the wheat inbreed line CB037 had been widely used as a recipient for transgenes and obtain its high transformation potential. Despite having stable agronomic traits, the CB037 is genetically heterogeneous for resistance to wheat stripe rust (Puccinia striiformis f. sp. tritici, Pst). In this study, the Pst-resistant line CB037-PstR and Pst-susceptible line CB037-PstS were isolated, and their F₂ population was created. Genetic analysis showed that the CB037-PstR carried a single dominant resistance gene. The identified resistance gene was mapped on the short arm of chromosome 1B using BSR-seq and molecular marker analysis. GISH results further revealed that CB037-PstR is a 1BL/1RS translocation line and likely carried Yr9. This study segregated genetic heterogeneity in CB037 for stripe rust resistance and isolated its Pst-susceptible and resistant lines. These results will facilitate trait-oriented use of CB037 in genetic engineering of wheat.     

基于BSR Seq分析筛选Y1499条锈病抗性相关基因

条锈病是危害小麦生产安全的重要病害之一,而携带不同抗病基因小麦品种的利用与合理布局是降低病害威胁最经济可靠的途径。为挖掘和丰富小麦条锈病抗性相关基因资源,本研究以筛选到的抗条锈病硬粒小麦种质Y1499和感病材料Gaza为亲本构建遗传群体,在室内对其进行苗期抗性鉴定和统计分析;利用衍生后代中选择的F_3∶4群体构建混池,进行RNA测序,对测序数据进行相关分析。结果表明,Y1499对CYR32的抗性由两个主效基因互补遗传控制。利用BSR-Seq关联分析在鉴定到3个小麦材料Y1499中条锈病抗性基因候选区间。GO富集结果显示,F_3∶4抗感池间差异表达基因涉及多个过氧化物酶等抗氧化反应相关基因。KEGG富集结果表明,显著差异表达基因主要富集于谷胱甘肽代谢和苯丙烷类生物合成等代谢通路。     

基于BSR-Seq对甘蔗脱叶性的研究和生物信息学分析

甘蔗的叶片是制糖过程中杂质的主要来源,育种过程中倾向于选育易脱叶的品种。为了研究甘蔗脱叶性相关的主效、稳定基因,本研究采用BSR-Seq技术,从‘ROC25’ב云蔗89-7’的F1后代中选取极端难脱叶品系和极端易脱叶品系的+6叶位叶鞘离区,分别构建混池,并和亲本一起进行RNA转录组测序分析,共得到60.46 Gb的有效片段,4个样品的Q30值均在93%以上,GC含量都大于51%,与参考基因组R570比对,比对率都超过41%。4个样品共检测出4085个差异基因,候选区域定位于9号染色体上的4个位置,总长1.40 Mb,注释到基因86个,亲本间注释到的非同义突变15个,挖掘出2个主效差异基因（Sh09_g020620,Sh09_g020080）,Sh09_g020080在亲本的表达差异和子代中是同向的,而Sh09_g020620在2个亲本和2个子代混池之间的差异是异向的,表明可能仅Sh09_g020080对易脱叶性有所贡献。选取BSR亚群体中极端易脱叶品系40-159和极端难脱叶品系5-94的+1~+7叶进行荧光定量分析,发现Sh09_g020080基因在单一品系的表达和子代混池的+6叶的表达是同向的。取另一个‘B35-9’בCP08-1553’的杂交后代个体中的极端易脱叶品系16-226和极端难脱叶品系16-224的+1~+7叶位的叶鞘离区进行荧光定量分析,结果显示Sh09_g020080在2个群体中+4、+5、+6叶位的表达都为易脱叶的品系高于难脱叶品系。注释显示Sh09_g020080具有σ因子活性,参与红光、远红光、蓝光反应。根据理化性质和蛋白质结构,推测Sh09_g020080编码蛋白具有4个保守区域,且具有积累叶绿素的作用。该研究为甘蔗脱叶相关主效候选基因的挖掘提出了新的见解。     

利用BSR-Seq定位小麦品种郑麦103抗条锈病基因YrZM103

郑麦103是一个高抗条锈病的小麦新品种,为明确其携带的抗病基因,用郑麦103与感条锈病品种农大399杂交构建分离群体,用条锈菌CYR32、CYR33和CRY34(V26)混合菌系进行田间接种和成株期抗性鉴定,对214个F2:3家系的条锈病抗性进行遗传分析,初步确定郑麦103的抗条锈性由单个主效基因控制,定名为Yr ZM103。通过BSR-Seq技术开发了6个与Yr ZM103紧密连锁的分子标记,将Yr ZM103定位于染色体臂7BL分子标记ZM215和ZM221之间,遗传距离分别为11.8 c M和6.9 c M。利用7BL染色体上与其他已知抗条锈病基因紧密连锁的分子标记进行比较作图,发现Yr ZM103是不同于7BL末端其他抗条锈病基因的新基因。     

SlGH9-15 regulates tomato fruit cracking with hormonal and abiotic stress responsiveness cis-elements

Fruit cracking occurs easily during the late period of fruit development when plants encounter an unsuitable environment, dramatically affecting fruit production and marketing. This study conducted the bulked segregant RNA-Seq (BSR) to identify the key regulatory gene of fruit cracking in tomatoes. BSR-Seq analysis illustrated that two regions associated with irregularly cracking were located on chromosomes 9 and 11, containing 127 candidate genes. Further, through differentially expression analysis and qRT-PCR in cracking-susceptible and cracking-resistant genotypes, the candidate gene SlGH9-15 (Solyc09g010210) with significantly differential expression levels was screened. Bioinformatics analysis of the GH9 gene family revealed that 20 SlGH9 genes were divided into three groups. The phylogenetic analysis showed that SlGH9-15 was closely related to cell wall construction-associated genes AtGH9B1, AtGH9B6, OsGH9B1, and OsGH9B3. The cis-acting elements analysis revealed that SlGH9-15 was activated by various hormones (ethylene and ABA) and abiotic stresses. The expression pattern indicated that 13 SlGH9 genes, especially SlGH9-15, were highly expressed in the cracking-susceptible genotype. Its expression level gradually increased during fruit development and achieved maximum value at the red ripe stage. Additionally, the cracking-susceptible tomato showed higher cellulase activity and lower cellulose content than the cracking-resistant tomato, particularly at the red ripe stage. This study identified SlGH9-15 as a key gene associated with fruit cracking in tomatoes for the first time and gives new insights for understanding the molecular mechanism and complex regulatory network of fruit cracking.