橡胶树肌动蛋白解聚因子的表达和功能鉴定  

Expression analysis and functional characterization of an actin depolymerizing factor in Hevea brasiliensis

作　　者：邓治[1] 刘向红[1,2] 李德军[1] 

机构地区：[1]中国热带农业科学院橡胶研究所侬业部橡胶树生物学与遗传资源利用重点实验室,海南儋州571737 [2]海南大学农学院,海南海口570228

出　　处：《湖南农业大学学报（自然科学版）》2016年第2期129-135,共7页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(31270651;31200514)

摘　　要：为探寻肌动蛋白解聚因子(actin depolymerizing factor,ADF)在调控肌动蛋白解聚和聚合平衡过程中发挥的作用,从橡胶树中获得了Hb ADF基因组序列,经测定,该序列长2 258 bp,含有2个内含子和3个外显子,在裂殖酵母中过表达Hb ADF,获得相对分子质量约38 000的融合蛋白。对裂殖酵母细胞形态进行观察,发现诱导表达Hb ADF的酵母细胞长度显著增加,双核和多核比例达67%。实时荧光定量PCR结果表明,Hb ADF的表达受3%KI处理调控,在橡胶树不同死皮阶段和不同排胶时间段的表达存在变化,说明Hb ADF可能参与了橡胶树排胶和死皮的过程。Actin depolymerizing factor（ADF） plays an important role in regulating actin assembly and disassembly for cells. In this study, the genomic sequence of Hb ADF, a 2 258 bp-length with two introns and three exons, was obtained from Hevea brasiliensis with PCR and sequencing methods. The recombinant protein with relative molecular mass about 38 000 was successfully induced by overexpressing Hb ADF in fission yeast. The cell morphology and the proportion of binucleate or coenocytic cells were further analyzed in the yeast overexpressing Hb ADF. The results showed that the length of yeast cells from overexpressing Hb ADF was markedly increased. In addition, it was observed that about 67% of the cells overexpressing Hb ADF were binucleate or coenocytic after induction. Real-time PCR analysis indicated that Hb ADF was regulated by 3% KI treatment and its expression varied with tapping panel dryness（TPD） stages and latex flow times. These results suggested that Hb ADF might play important roles in regulating TPD occurrence and latex flow in Hevea brasiliensis.

关 键 词：橡胶树 肌动蛋白解聚因子 表达分析 功能鉴定 排胶 细胞形态 

分 类 号：S794.1[农业科学—林木遗传育种]