玉米ZmSNAC13等位变异对抗旱性的调控研究

doi:10.13304/j.nykjdb.2021.0213

摘要/Abstract

摘要：

NAC家族转录因子是植物特有的，在植物的生长发育、胁迫应答和激素调节等方面具有重要的生物学功能。前期根据干旱胁迫处理RNA-Seq筛选到NAC转录因子基因ZmSNAC13。为了探究ZmSNAC13在干旱胁迫响应中的作用，以耐旱性不同的玉米自交系B73（干旱敏感）和Qi319（耐旱）为材料，进行荧光定量PCR分析，发现干旱处理后ZmSNAC13在2个材料根中均下调表达，干旱处理12 d后在茎中均上调表达，Qi319干旱处理12 d后在叶中上调表达，而B73干旱处理8和10 d后在叶中下调表达，表明ZmSNAC13基因响应干旱胁迫。利用双荧光素酶融合报告载体分析ABA处理对ZmSNAC13基因启动子活性的影响，结果表明，ABA处理下Qi319的ZmSNAC13启动子活性显著高于B73，在Qi319的ZmSNAC13基因5’-UTR区发现9个能够提高启动子活性的高度连锁变异，并且干旱处理12 d时， Qi319叶中ZmSNAC13的表达量高于B73，因此推测干旱胁迫下ZmSNAC13基因5’-UTR区遗传等位变异通过影响启动子活性进而控制基因的表达水平，最终调控玉米对干旱胁迫的适应性。上述结果为解析玉米抗旱的遗传机制奠定了基础。

关键词: 玉米（Zea mays L.）, 干旱, ZmSNAC13, 等位变异, 基因表达

Abstract:

NAC family is a class of plant specific transcription factors， which has important biological functions in plant growth and development， stress response and hormone regulation. A NAC transcription factor gene， namedas ZmSNAC13， was selected from differentially expressed genes under drought stress by RNA-Seq. In order to clarify the role of ZmSNAC13 in response to drought stress， maize inbred lines B73 （drought sensitive） and Qi319 （drought tolerant） with different drought tolerance were used as materials， qRT-PCR analysis showed that the expression of ZmSNAC13 was down regulated in roots of 2 materials， up regulated in stems and leaves of Qi319 after 12 d of drought treatment， down regulated in leaves of B73 after 8 and 10 d of drought treatment， indicating that ZmSNAC13 responded to drought stress. The promoter activity of ZmSNAC13 under ABA treatment was analyzed by dual-luciferase fusion reporter vector. The results showed that the promoter activity of ZmSNAC13 in Qi319 was significantly higher than that in B73 under ABA treatment， furthermore， 9 highly linked variants in 5’-UTR region of ZmSNAC13 in Qi319 were found to improve the promoter activity. Moreover， the expression of ZmSNAC13 in Qi319 leaves was higher than that in B73 after 12 d of drought treatment. It was speculated that genetic allelic variations in 5’- UTR region of ZmSNAC13 affected promoter activity， and then controlled gene expression level， and ultimately regulated the adaptability of maize to drought. Above results laid foundations for understanding the genetic mechanism of drought resistance in maize.

Key words: maize （Zea mays L.）, drought, ZmSNAC13, allelic variations, gene expression

中图分类号:

S513

程名, 朱莹, 王晓楠, 罗平, 陈勇, 郝转芳, 席章营. 玉米ZmSNAC13等位变异对抗旱性的调控研究[J]. 中国农业科技导报, 2022, 24(5): 24-31.

Ming CHENG, Ying ZHU, Xiaonan WANG, Ping LUO, Yong CHEN, Zhuanfang HAO, Zhangying XI. Drought Resistance Regulated by Allelic Variations of ZmSNAC13 in Maize[J]. Journal of Agricultural Science and Technology, 2022, 24(5): 24-31.

图/表 5

表1 本研究所用引物

Table 1 Primers used in this study

引物名称 Primer name	引物序列 Primer sequence（5’-3’）	用途 Function
ZmSNAC13 qRT-PCR	F：AAGTTGGATGAGTGGGTGT	荧光定量PCR
ZmSNAC13 qRT-PCR	R：TCGTGCGTCTGGAAGCT	qRT-PCR
GAPDH	F：CATCACCACGGACTACAT	内参
GAPDH	R：GACTCCACGACATACTCA	Internal reference
ZmSNAC13 B73-P	F：ACATAGTGGAGAAACCGCTT	启动子序列扩增
ZmSNAC13 B73-P	R：ACCCCCTCCTCCTCCTCGTC	Amplification of promoter sequence
ZmSNAC13 Qi319-P	F：CACAATGTTCTCCTTTTTACTTA	启动子序列扩增
ZmSNAC13 Qi319-P	R：ACCCCCTCCTCCTCCTCCTCCT	Amplification of promoter sequence
ZmSNAC13 B73-V	F：GCAGCCCGGGGGATCC ACATAGTGGAGAAACCGCTT	载体构建
ZmSNAC13 B73-V	R：TAGAACTAGTGGATCC ACCCCCTCCTCCTCCTCGTC	Construction of vector
ZmSNAC13 Qi319-V	F：GCAGCCCGGGGGATCC CACAATGTTCTCCTTTTTACTTA	载体构建
ZmSNAC13 Qi319-V	R：TAGAACTAGTGGATCC ACCCCCTCCTCCTCCTCCTCCT	Construction of vector

图1 不同物种NAC转录因子系统进化树

Fig. 1 Phylogenetic tree of NAC transcription factors among different species

图2 干旱处理下玉米B73与Qi319 根、茎和叶中 ZmSNAC13基因的相对表达量注：*和**分别表示与处理0 d相比在P<0.05和P<0.01水平差异显著。

Fig. 2 Relative expression of ZmSNAC13 gene in roots， stems and leaves of maize B73 and Qi319 under drought stressNote：* and ** indicate significant difference compared to 0 d at P<0.05 and P<0.01 levels， respectively.

图3 ABA处理下ZmSNAC13基因的启动子活性分析注：*表示在P<0.05水平差异显著。

Fig. 3 Activity of the ZmSNAC13 promoter under ABA treatmentNote：* indicates significant difference at P<0.05 level.

图4 ZmSNAC13的9个连锁变异影响启动子活性注：*表示在P<0.05水平差异显著。

Fig. 4 9 linkage variants of ZmSNAC13 affect promoter activityNote：* indicates significant difference at P<0.05 level.

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