Cloning of Heat Shock Protein Related Gene CmHSP70-5 and Expression Analysis under Heat Stress in Cucurbita moschata HU Yan-ping 1,ZHOU Yang 2,YANG Chun 3,LIAO Dao-long 1,ZHU Bai-bi 1,WANG Min 1,HUANG Wen-feng 1,ZHANG Wen 1,YUN Tian-hai 1*
(1Institute of Vegetables ,Hainan Academy of Agricultural Sciences ,Hainan Key Laboratory of Vegetable Biology ,Hainan Vegetable Breeding Engineering Technology Research Center ,Haikou 571100,Hainan ,China ;2College of Horticulture ,Hainan University ,Haikou 570228,Hainan ,China ;3Hainan Academy of Agricultural Sciences ,Haikou 571100,Hainan ,China )数字人>绥靖政策
Abstract :A heat shock protein gene ,naming CmHSP70-5,was isolated from pumpkin cDNA by polymerase chain reaction (PCR )
中国重型机械总公司method based on pumpkin genome sequence .Phylogenetic relationship of HSP70-5 and its homology were analyzed using bioinformation method .The expression quantity of CmHSP70-5 under heat stress was studied by real-time PCR .The results showed that CmHSP70-5 gene was 1 953 bp ,and its coding CmHSP70-5 protein had the closest relative with Arabidopsis At HSP70-5.C
mHSP70-5 gene had a high expression level in roots than in stems and leaves under normal conditions .Real-time PCR results indicated that under heat stress ,the expression of CmHSP70-5 gene in roots ,stems and leaves were all up-regulated and could reach the maximum quantity 3 hours after treatment ,indicating that the expression of CmHSP70-5 gene was related to heat stress in Cucurbita moschata .
Key word :Cucurbita moschata ;Heat stress ;CmHSP70-5 gene ;Gene expression
2019年7月,国际刊物《Plant Biotechnology Journal 》在线发表了华中农业大学园艺植物生物学教育部重点实验室生菜团队陈炯炯课题组题为“Characterization of four polymorphic genes controlling red leaf color in lettuce that have undergone disruptive selection since domestication ”的研究论文,报道了通过正向遗传学的方法克隆到4个调控生菜花青素合成的基因,阐明了生菜叶的遗传和分子机理,为生菜改良和育种奠定了理论基础。 花青素是一种重要的代谢产物,能帮助植物抵抗生物和非生物逆境,它的高抗氧化活性,具有极高的保健功能。生菜作为世界上最重要的蔬菜之一,其叶存在很大的变异。生菜叶由多个位点调控,导致其遗传分析变得异常复杂。在本研究中,课题组利用BSA+RNA-seq 方法对一个生菜叶分离体进行遗传分析,鉴定到多个调控位点。通过对潜在位点构建单基因分离体,并利用这些单基因分 离体对相关基因进行精细定位、成功克隆了4个调控生菜叶的基因,它们的功能得到转基因验证。这4个基因分别编码bHLH 、R2R3-MYB 、R3-MYB 和WD-40。其中bHLH 和R2R3-MYB 为正调控因子,R3-MYB 和WD-40为负调控因子。当生菜中编码bHLH 转录因子的基因发生突变时,花青素代谢途径被阻断,从而产生绿
叶片。而当另外3个基因(R2R3-MYB ,R3-MYB ,WD-40)发生突变时,会促进花青素的积累,从而产生不同颜的生菜。
Y2H 发现R2R3-MYB 和R3-MYB 均能与bHLH 结合,表明R3-MYB 能竞争R2R3-MYB 与bHLH 的结合从而抑制花青素的合成。而Y2H 发现WD-40与R2R3-MYB 之间没有互作,说明WD-40并非通过竞争LsTTG1来抑制花青素的合成。进一步序列分析发现WD-40为AtRUP1/AtRUP2的同源体,但与拟南芥的调控途径不同的是,生菜WD-40并非通过调控HY5来影响花青素的合成。最后通过对野生生菜和栽培生菜进行序列分析时发现这4个基因的功能突变均发生在驯化之后,表明生菜叶在育种过程中经历了歧化选择,即仅含有少量花青素的野生生菜通过驯化形成高花青素含量的红生菜和不含花青素的绿生菜。该研究利用BSA 结合高通量测序的方法,成功克隆调控复杂性状的多个基因,对其他作物复杂性状的遗传分析具有借鉴作用。本研究系统地阐明了生菜花青素积累的遗传和分子机理,为生菜叶的选育提供了方法指导和理论依据。
论文链接:https ://onlinelibrary.wiley/doi/abs/10.1111/pbi.13213
(华中农业大学园艺林学学院)诺顿安全特警
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生菜叶的遗传和分子机理研究取得进展
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