结缕草ZjNAC基因的克隆与表达分析

doi:10.11686/cyxb2015453

草业学报 ›› 2016, Vol. 25 ›› Issue (4): 239-245.DOI: 10.11686/cyxb2015453

结缕草ZjNAC基因的克隆与表达分析

张胤冰¹, 孙鑫博², 樊波¹, 韩烈保¹, 张雪¹, 袁建波¹, 许立新^{1*, *}

1.北京林业大学草坪研究所,北京 100083;
2.河北省作物生长调控重点实验室,河北农业大学,河北保定 071001

收稿日期:2015-09-23 出版日期:2016-04-20 发布日期:2016-04-20
作者简介:张胤冰(1990-),女,黑龙江哈尔滨人,在读硕士。E-mail:13020019042@163.com
基金资助:
中央高校基本科研业务费专项资金(BLX2012034)和国家高新技术研究发展计划项目(2013AA102607)资助

Cloning and expression of ZjNAC from Zoysia japonica

ZHANG Yin-Bing¹, SUN Xin-Bo², FAN Bo¹, HAN Lie-Bao¹, ZHANG Xue¹, YUAN Jian-Bo¹, XU Li-Xin^{1, *}

1.Institute of Turfgrass Science, Beijing Forestry University, Beijing 100083, China;
2.Key laboratory of crop growth regulation of Hebei Province, Agricultural University of Hebei, Baoding 071001, China

Received:2015-09-23 Online:2016-04-20 Published:2016-04-20

摘要/Abstract

摘要： NAC类家族的基因是植物特有的转录因子,并且它在植物的生长发育中发挥着重要的作用。本研究以结缕草为研究材料,对结缕草转录组数据库利用local blast的办法克隆出1个NAC类转录因子。为深入探讨和研究NAC类转录因子在植物逆境生理上的作用奠定基础。通过与水稻的OsNACs家族成员进行进化树分析,结合蛋白分析,QRT-PCR等的方式探索其生物学功能。实验结果表明,该基因长度为1496 bp,包含1个完整开放阅读框(ORF),长度为1332 bp,编码449个氨基酸,该蛋白含有1个NAM结构域。与水稻的NAC类转录因子家族的系统进化树对比发现,它与OsNAC036近缘性最高,因此将该基因命名为ZjNAC036。QRT-PCR结果显示,该基因在干旱情况下表达量呈上升趋势,并且干旱处理8 h的时候,差异倍数最大,达到了50倍左右,而在盐胁迫、低温胁迫下没有规律性变化。这表明ZjNAC036在干旱逆境生理上可能有重要的作用,而在高盐及寒冷的逆境生理方面可能作用不大。

Abstract: Proteins in the NAC family (including NAM, ATAF1/2, and CUC2 proteins) are plant-specific transcription factors that play critical roles in plant development. To explore and exploit the excellent resistance-gene resources of zoysia (Zoysia japonica), a gene encoding a NAC transcription factor (ZjNAC) was cloned by blasting the Zoysia transcriptome database. The biological function of the cloned ZjNAC gene was explored by a phylogenetic analysis with OsNACs of rice (Oryza sativa) combined with protein and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analyses. The bioinformatics analysis showed the cloned ZjNAC gene was 1496 bp long, and contained a 1332 bp open reading frame (ORF) encoding 449 amino acids with a NAM domain. In a phylogenetic analysis, the cloned ZjNAC gene showed the highest homology to OsNAC036 from rice, and so the cloned gene was designated as ZjNAC036. The qRT-PCR analysis showed that ZjNAC transcript levels increased under drought stress, reaching peak levels (approximately 50-fold that in the control) after 8 h of drought stress. The transcript levels of ZjNAC did not show obvious variations under salt stress or cold stress. Together, these results show that the protein encoded by ZjNAC036 may play an important role in the drought stress response, rather than the salt or cold stress responses.

张胤冰, 孙鑫博, 樊波, 韩烈保, 张雪, 袁建波, 许立新. 结缕草ZjNAC基因的克隆与表达分析[J]. 草业学报, 2016, 25(4): 239-245.

ZHANG Yin-Bing, SUN Xin-Bo, FAN Bo, HAN Lie-Bao, ZHANG Xue, YUAN Jian-Bo, XU Li-Xin. Cloning and expression of ZjNAC from Zoysia japonica[J]. Acta Prataculturae Sinica, 2016, 25(4): 239-245.

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结缕草ZjNAC基因的克隆与表达分析

Cloning and expression of ZjNAC from Zoysia japonica

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