小黑杨PnsGA20ox1基因的克隆及功能分析

张力杰; 张凯旋; 魏志刚

doi:10.3969/j.issn.1000-2006.2013.06.003

小黑杨PnsGA20ox1基因的克隆及功能分析

张力杰,张凯旋,魏志刚

南京林业大学学报（自然科学版） ›› 2013, Vol. 37 ›› Issue (06) : 11-16.

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南京林业大学学报（自然科学版） ›› 2013, Vol. 37 ›› Issue (06) : 11-16. DOI: 10.3969/j.issn.1000-2006.2013.06.003

研究论文

小黑杨PnsGA20ox1基因的克隆及功能分析

张力杰,张凯旋,魏志刚^*

作者信息 +

Cloning and function analysis of PnsGA20ox1 gene in Populus simonii ×P. nigra

ZHANG Lijie,ZHANG Kaixuan,WEI Zhigang^*

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文章历史 +

摘要

根据与小黑杨(Populus simonii ×P. nigra)亲缘关系较近的几个物种的赤霉素氧化酶基因(GA20ox)保守区设计兼并引物,克隆出小黑杨GA20ox基因片段; 利用RACE技术得到其全长为1 678 bp 的cDNA序列,进行蛋白质预测,发现其编码385个氨基酸,分子质量为44.04 ku,理论等电点为5.97,核酸序列与毛果杨GA20ox1同源性高达99%,将其命名为PnsGA20ox1。荧光定量PCR分析发现,GA20ox1基因在小黑杨各个组织中均表达,其中在成熟木质部中表达量最高,在节间组织中表达量最低。转PnsGA20ox1基因烟草分析表明,过表达PnsGA20ox1基因的烟草综纤维素含量和纤维长度比对照组均有不同程度的提高,而木质素含量与对照无明显差异。

Abstract

The gene fragment of GA20ox was cloned from Populus simonii&#215;P. nigra by using degenerate primers based on the conservative of GA20ox gene homologous genes. The full-length of cDNA was isolated by using RACE.The cDNA was 1 678 bp and encoded a protein of 385 amino acids with Mr= 44. 04 ku and pI=5.97.The nucleotides shared 99% identity with the GA20ox1 gene of Populus trichocarpa.It was named PnsGA20ox1. Quantitative real-time PCR analysis showed the PnsGA20ox1 gene expressed in all organs of Populus simonii&#215;P.nigra, and the expression level of PnsGA20ox1 was the highest in mature xylem but the least in internode organizations.The transgenic tobacco showed that the holocellulose content and fiber length in overexpression of the PnsGA20ox1 gene of tobacco improved comparised with the control,while there was no significant difference in lignin content comparised with the control.

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张力杰,张凯旋,魏志刚. 小黑杨PnsGA20ox1基因的克隆及功能分析[J]. 南京林业大学学报（自然科学版）. 2013, 37(06): 11-16 https://doi.org/10.3969/j.issn.1000-2006.2013.06.003

ZHANG Lijie,ZHANG Kaixuan,WEI Zhigang. Cloning and function analysis of PnsGA20ox1 gene in Populus simonii ×P. nigra[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2013, 37(06): 11-16 https://doi.org/10.3969/j.issn.1000-2006.2013.06.003

中图分类号： Q786

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基金

收稿日期:2012-12-16 修回日期:2013-04-12
基金项目:国家自然科学基金项目(31170562); 中央高校基本科研业务费专项资金项目(DL12CA09,DL12EA03)
第一作者:张力杰,硕士生。*通信作者:魏志刚,副教授。E-mail: zhigangwei1973@163.com。
引文格式:张力杰,张凯旋,魏志刚. 小黑杨PnsGA20ox1基因的克隆及功能分析[J]. 南京林业大学学报:自然科学版,2013,37(6):11-16.