摘要
[目的]探究BRL3基因在麻疯树花发育中的功能。[方法]利用RACE PCR技术成功从麻疯树花蕾中获得了JcBRL3基因的c DNA全长序列;利用原核表达体系对JcBRL3基因进行诱导表达,利用LC-MS/MS对其表达产物进行质谱鉴定,利用生物信息学方法对其蛋白质结构和基本理化性质进行分析;利用实时荧光定量PCR方法分析了Jc BRL3基因在麻疯树雌雄花发育的9个关键时期的相对表达量;利用叶盘法将JcBRL3基因转化至烟草中超表达,分析超表达JcBRL3对烟草花的形态结构影响。[结果]JcBRL3基因的开放阅读框长3618bp,编码1205个氨基酸。原核表达产物质谱鉴定表明:该基因编码1个JcBRL3蛋白(A0A067KCE7)。蛋白质结构分析表明:JcBRL3含有多个富含亮氨酸重复序列,是跨膜蛋白,并且具有保守重复序列LxxLxLxxN/CxL。荧光定量PCR分析发现:在雌花中JcBRL3的表达于单核胚囊期达到最高水平,在雄花中Jc BRL3的表达于花粉粒成熟期达到最高水平且其显著高于其它任何时期。转基因烟草的花形态结构分析表明:转JcBRL3基因烟草柱头位置低于花药,花粉粒畸形率较低。[结论]JcBRL3蛋白为富亮氨酸重复序列类受体激酶蛋白,是一个膜蛋白,JcBRL3基因可能参与了麻疯树雌花单核胚囊期和雄花花粉粒成熟的发育并促进了花丝伸长。
[Objective]To explore the function of BRL3 gene in flower development of Jatropha curcas.[Method]The full-length cDNA sequence of JcBRL3 gene was obtained by RACE PCR.The prokaryotic expression system was used to induce the expression of JcBRL3 gene.The expression products were identified by mass spectrometry using LC-MS/MS.The structure and basic physicochemical properties of the protein were analyzed by bioinformatics.The relative expression levels of JcBRL3 gene during the key stages of flower development of Jatropha curcas were analyzed by qRT-PCR.The overexpression JcBRL3 gene was transformed into tobacco by leaf plate method to analyze the influence of overexpression of JcBRL3 gene on the morphology and structure of tobacco flowers.[Result]The length of the open reading frame of JcBRL3 gene was 3618 bp and encoded 1205 amino acids.The results of mass spectrometry showed that the expression of this gene encoded a JcBRL3 protein.Protein structure analysis showed that JcBRL3 protein was a transmembrane protein with multiple leucine-rich repeats and conserved repeats:LxxLxLxxN/CxL.qRT-PCR analysis showed that the expression of JcBRL3 in female flowers reached the highest level at mononuclear embryo sac stage,and in male flowers reached the highest level at pollen grain maturation stage,which was significantly higher than at any other stage.The morphological structure analysis of transgenic tobacco flower showed that the stigma position of transgenic tobacco was lower than that of anther,and the malformation rate of pollen grains was lower.[Conclusion]JcBRL3 protein is a LRR-RLK,which is a membrane protein.JcBRL3 gene may participate in the development process of female flower mononuclear embryo sac stage and pollen grain maturation of male flower and promote filament elongation in Jatropha curcas.
作者
陈雨倩
雷世康
徐刚
CHEN Yu-qian;LEI Shi-kang;XU Gang(Institute for Forest Resources and Environment of Guizhou/Key Laboratory of Forest Cultivation in Plateau Mountain of Guizhou Province/College of Forestry,Guizhou University,Guiyang 550025,Guizhou,China;School of Life Sciences,Southwest University,Chongqing 400715,China)
出处
《林业科学研究》
CSCD
北大核心
2021年第6期46-55,共10页
Forest Research
基金
国家科学自然基金(31760198)
贵州省生态学一流学科(GNYL[2017]007)。
关键词
麻疯树
花发育
JcBRL3
功能分析
Jatropha curcas
flower development
JcBRL3
functional analysis
