巴西橡胶树HbNAM基因克隆和表达分析

doi:10.3969/j.issn.1000-2006.2016.01.010

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南京林业大学学报（自然科学版） ›› 2016, Vol. 40 ›› Issue (01): 59-64.doi: 10.3969/j.issn.1000-2006.2016.01.010

巴西橡胶树HbNAM基因克隆和表达分析

康桂娟,黎瑜,曾日中^*

中国热带农业科学院橡胶研究所,农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

出版日期:2016-02-18 发布日期:2016-02-18
基金资助:
收稿日期:2015-03-20 修回日期:2015-09-22
基金项目:国家自然科学基金项目(31201143); 海南省自然科学基金项目(313061); 中国热带农业科学院橡胶研究所基本科研业务费重点项目(1630022011023)
第一作者:康桂娟(kanguijuan2010@126.com); 黎瑜(13976781768@163.com)。*通信作者:曾日中(hnzrz@aliyun.com),研究员。
引文格式:康桂娟,黎瑜,曾日中. 巴西橡胶树HbNAM基因克隆和表达分析[J]. 南京林业大学学报(自然科学版),2016,40(1):59-64.

Clone and expression analysis of HbNAM from Hevea brasiliensis Muell. Arg.

KANG Guijuan, LI Yu, ZENG Rizhong^*

Key Laboratory for Rubber Biology and Genetic Resource Utilization of Ministry of Agriculture,Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences(CATAS), Danzhou 571737, China

Online:2016-02-18 Published:2016-02-18

摘要/Abstract

摘要： 为了研究巴西橡胶树中植物特有NAC转录因子的功能,笔者通过RT-PCR和实时荧光定量PCR技术对HbNAM的开放阅读框(ORF)进行了克隆和表达分析,并通过酵母转化实验进行转录激活活性分析。结果显示,HbNAM基因ORF为1 077 bp,编码358个氨基酸,在第12—167位氨基酸之间含有一个典型的NAC结构域,属于NAC转录因子家族中的NAM亚族。酵母试验表明,HbNAM在高度变异C端具有转录激活活性,具备转录因子的基本特征。实时荧光定量PCR分析发现,HbNAM在叶片、雄花、雌花、胶乳和树皮中均表达,其中以光合组织叶片和生殖组织雄花中的表达量最高,并且胶乳中该基因的表达受割胶、植物激素或植物生长调节剂茉莉酸(JA)和乙烯利(ET)以及低温胁迫影响,推测HbNAM可能与植物的生长发育和胁迫应答过程有关。

Abstract: In order to identify the function of plant-specific NAC transcription factors in Hevea brasiliensis, the whole open reading frame(ORF)sequence of HbNAM was cloned and analysed by using reverse transcription polymerase chain reaction(RT-PCR)as well as real-time PCR, and the transactivation activity analysis was carried out in yeast cells. The results indicated that HbNAM occupied an ORF of 1 077 bp, encoding a protein of 358 amino acids belonging to NAM subgroup with a conserved NAC with domain from amino acid 12 to 167. Transactivation assay in yeast cells showed that HbNAM protein possessed transcriptional activation activity in the highly divergent C terminal region. The realtime PCR indicated that HbNAM could express in leaves, male flowers,female flowers, latex as well as bark,and the hightest expression of levels of HbNAM was in leaves and male flowers among these detected tissues. The expression of HbNAM in latex was affected by tapping,plant hormone jasmonate(JA), ethylene(ET)and cold stress treatment,which suggested that HbNAM may have relationship with plant growth, development and stress responses in rubber tree.

中图分类号:

S794.1
Q812

康桂娟,黎瑜,曾日中. 巴西橡胶树HbNAM基因克隆和表达分析[J]. 南京林业大学学报（自然科学版）, 2016, 40(01): 59-64.

KANG Guijuan, LI Yu, ZENG Rizhong. Clone and expression analysis of HbNAM from Hevea brasiliensis Muell. Arg.[J].Journal of Nanjing Forestry University (Natural Science Edition), 2016, 40(01): 59-64.DOI: 10.3969/j.issn.1000-2006.2016.01.010.

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巴西橡胶树HbNAM基因克隆和表达分析

Clone and expression analysis of HbNAM from Hevea brasiliensis Muell. Arg.

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