红麻HcAOC1基因克隆及响应茉莉酸甲酯的表达模式分析  

Cloning and Expression Pattern Analysis of Kenaf HcA OC1 Gene in Res- ponse to Methyl Jasmonate

作　　者：李宏伟 尤婧祎 周步进[1,2] 李敏 刘一丁[1] 康浩东[1] 王颐宁 廖小芳 周瑞阳 Li Hongwei;You Jingyi;Zhou Bujin I;Li Min;Liu Yiding;Kang Haodong;Wang Yining;Liao Xiaofang;Zhou Ruiyang(Key Laboratory of Plant Genetics and Breeding,College of Agriculture,Guangxi University,Nanning,530004;Institute of Cash Crops,Guangxi Academyof Agricultural Sciences,Nanning,530007)

机构地区：[1]广西大学农学院,植物遗传育种重点实验室,南宁530004 [2]广西农业科学院经济作物研究所,南宁530007

出　　处：《分子植物育种》2023年第1期148-156,共9页Molecular Plant Breeding

基　　金：广西自然科学基金项目(2018JJB130045,2019JJA130200)资助

摘　　要：茉莉酸对植物的生长发育、花药开裂具有重要调控作用。丙二烯氧化物环化酶基因(AOC)是茉莉酸合成途径关键基因,在植物生长发育、育性调控过程也发挥关键作用。为了探究红麻茉莉酸合成途径中关键基因(HcAOC1)的调控机理,本研究以红麻细胞质雄性不育系UG93A为材料。利用同源克隆的方法,以UG93A的g DNA和cDNA为模板,成功克隆获得HcAOC1基因全长。其CDS序列为780 bp,编码259个氨基酸残基,蛋白质分子量为28.4 kD,等电点为9.28。HcAOC1系统发育进化树分析结果表明,红麻与棉花、蓖麻、榴莲的亲缘关系较近,与小果野芭蕉、萝卜的亲缘关系较远(GenBank登录号:MW520098)。此外,在花期采用不同浓度的外源茉莉酸甲酯(MeJA)处理红麻。在处理的第0天、5天、10天和15天后取样成熟期花药,利用RT-qPCR分析不同处理时间下HcAOC1的表达差异,发现HcAOC1表达量随着处理时间延长存在先增高后降低的趋势,即0.5 mmol/L (T1)、1 mmol/L (T2)、2 mmol/L (T3)处理在第10天HcAOC1的表达量达最高值,均极显著高于对照组,其在T1浓度处理下HcAOC1表达量最高;在第15天时,三种处理Hc AOC1表达量均明显下降且趋于对照。研究结果表明,Me JA处理能诱导红麻UG93A中Hc AOC1的表达,但在不同浓度的MeJA诱导下,HcAOC1的表达有显著差异,特别在低浓度MeJA诱导下,HcAOC1基因表达量上升较明显。综上所述,HcAOC1的表达受Me JA的诱导变化,暗示MeJA诱导可在茉莉酸合成途径发挥调控作用,为分析红麻UG93A雄性不育机理提供参考依据。Jasmonic acid plays an important role in the regulation of plant growth, development and anther dehis cence. Allene oxide cyclase gene(AOC) is a key gene in the jasmonic acid synthesis pathway and also plays a key role in the regulation of plant growth, development and fertility. In order to explore the regulation mechanism of the key gene HcAOC1 in the Kenaf jasmonic acid synthesis pathway. Kenaf cytoplasmic male sterile line UG93A was used as material in this study. Using g DNA and c DNA of UG93A as templates, the full length of HcAOC1 gene was successfully cloned by homology cloning strategy, and homologous cloning methods have been used. The CDS sequence is 780 bp, encoding 259 amino acid residues, molecular weight of the protein is 28.4 kD, and the isoelectric point is 9.28. The phylogenetic tree analysis of HcAOC1 showed that Kenaf is closely related to cotton,castor bean and durian, but is far related to Musa acuminata and radish(GenBank accession number MW520098).In addition, different concentrations of exogenous methyl jasmonate(MeJA) were used to treat kenaf at flowering stage. Mature anthers were sampled at 0, 5, 10, and 15 days after treatment, the expression of HcAOC1 in different treatment times was analyzed by RT-qPCR. It was found that the expression of HcAOC1 increased at first and then decreased with the extension of treatment time, that is, the expression of HcAOC1 in 0.5 mmol/L(T1), 1 mmol/L(T2)and 2 mmol/L(T3) treatment reached the highest value on the 10th day, which was significantly higher than that in the control group, and the expression of HcAOC1 was the highest at T1 concentration. On the 15th day, the expres sion of HcAOC1 in all three treatments decreased significantly and tended to be the control group. The results showed that MeJA treatment could induce the expression of HcAOC1 in Kenaf UG93A, but the expression of HcAOC1 is significantly different under the induction of different concentrations of MeJA, especially under the induction of low concentrations of MeJA, the expression o

关 键 词：红麻 HcAOC1 茉莉酸甲酯 差异表达分析 

分 类 号：S563.5[农业科学—作物学]