玉米bZIP基因应答逆境胁迫的表达模式分析

doi:10.7668/hbnxb.20192475

摘要/Abstract

摘要：

为了探讨玉米bZIP基因家族成员在玉米抗逆中的作用,以玉米骨干自交系郑58为试验材料,设置200 mmol/L的NaCl溶液、20% PEG6000(Polyethylene Glycol 6000)、4 ℃低温和硝态氮或铵态氮缺乏胁迫试验,利用qPCR技术,对9个ZmbZIP基因应答各类逆境胁迫的响应表达模式进行了分析。系统进化树分析结果表明,9个ZmbZIP基因进一步细分为3个亚组。qPCR分析结果表明,8个基因在不同组织器官中表达,ZmbZIP80没有被检测到,其可能是假基因。在人为模拟盐、干旱、低温和氮胁迫条件下,8个ZmbZIP在应答不同胁迫时,呈现不同的表达模式,ZmbZIP37和ZmbZIP53受NaCl胁迫明显诱导,而ZmbZIP49和ZmbZIP79则受到显著抑制;硝态氮缺乏胁迫显著抑制叶片中ZmbZIP37和ZmbZIP53基因表达,ZmbZIP42和ZmbZIP49则受到铵态氮缺乏胁迫的明显诱导。这些结果表明,8个ZmbZIP在玉米抵御逆境胁迫时发挥着广泛的作用;9个ZmbZIP基因在玉米不同组织中和响应不同逆境胁迫时的表达模式明显不同。研究结果可为进一步研究这些基因的生物学功能提供科学数据。

关键词: 玉米自交系, ZmbZIP, 逆境胁迫, 表达模式

Abstract:

In order to study the role of ZmbZIPs in the stress tolerance of maize,using maize inbreed line Zheng 58 for exprerimental material,expression analysis of 9 ZmbZIPs were performed under 200 mmol/L NaCl,20% PEG6000,4 ℃ low temperature and nitrate or ammonium deficiency stresses.Evolutionary tree analysis indicated that 9 ZmbZIPs were divided 3 subgroup.RT-qPCR analysis showed that 8 ZmbZIPs were detected in maize tissues,whereas ZmbZIP80 were not,indicated it could be pseudogene.Under simulated salt,drought,low temperature and nitrogen stress conditions,8 ZmbZIPs exhibited different expression pattern in response to various stresses,ZmbZIP37 and ZmbZIP53 were induced while ZmbZIP49 and ZmbZIP79 were downregulated obviously under NaCl stress,ZmbZIP37 and ZmbZIP53 of leaves significantly suppressed under nitrate deficit stress while ZmbZIP42 and ZmbZIP49 were upregulated in responsive to ammonium deficient stress.The result indicated 8 ZmbZIP played widely roles against stress conditions.Expression pattern of 9 ZmbZIPs gene differed in different tissues or under various adverse stresses.The study can offer scientific data for further reveal ZmbZIP biological function.

Key words: Maize inbreed line, ZmbZIP, Adversity stress, Expression pattern

贾利强, 刘讯, 丁波. 玉米bZIP基因应答逆境胁迫的表达模式分析[J]. 华北农学报, 2022, 37(1): 9-17. doi: 10.7668/hbnxb.20192475.

JIA Liqiang, LIU Xun, DING Bo. The Expression Pattern of ZmbZIP Gene under Adversity Stresses[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(1): 9-17. doi: 10.7668/hbnxb.20192475.

http://www.hbnxb.net/CN/Y2022/V37/I1/9

图/表 7

表1 ZmbZIP基因的实时荧光定量引物

Tab.1 qRT-PCR primers for expression on analysis of ZmbZIPs gene

基因名称 Gene name	基因号 Gene locus	上游引物(5'—3') Forward	下游引物(5'—3') Reverse
ZmbZIP37	AC233853.1_FGT002	GTCCGACCACAACAGCATCAACGA	TCCTCTTGACGCGCTTAGGATC
ZmbZIP42	GRMZM2G024851	GCATTTAAGATCCCTTTGTTCCAGC	CAGCTTTATTCACCGTGGGATGAG
ZmbZIP49	GRMZM2G027976	CTGTCTCCCCGTGTGGCAT	GCCTCTCGATCTCCTTCTTCAACG
ZmbZIP53	GRMZM2G120167	AGAGTGGAGAGTCTAGCACAGGAA	TGCACATCAAGGTCCCTGCTGTTAC
ZmbZIP56	GRMZM2G118870	AGGGTCACTTCCTGTTATGACCTG	CGCTTCTTGGGGAGACAGAGGAG
ZmbZIP65	GRMZM2G052102	TGCATCTCCACCTACACTGCGTTC	CTGACTGGGAACTTGCGGGATG
ZmbZIP79	GRMZM2G158313	ACAGCCACCTCAAGCAACGAATC	TGCTGCTGGTACACATGGGGTAG
ZmbZIP121	GRMZM2G170079	GGTCGCGGGTGAGGAAGCTG	TACGCTGAGCAAGAATGCAGTCGTAG

图1 ZmbZIP基因的生物信息学分析
A. ZmbZIP基因的染色体定位;B.ZmbZIP的基因结构分析;C.保守的ZmbZIP结构域多重序列比对;D.ZmbZIP蛋白的进化树分析。

Fig.1 The bioinformatic analysis of ZmbZIPs gene
A.Chromosomal distribution of ZmbZIPs;B.Gene structure analysis of ZmbZIPs;C.Multiple sequence alignment of conserved ZmbZIPs domain;D.The phylogenetic tree analysis of ZmbZIPs.

表2 玉米ZmbZIP蛋白的理化性质分析

Tab.2 The physical and chemical properties analyses of ZmbZIPs in maize

基因号 Gene ID	基因名称 Gene name	大小/aa Size	分子质量/ku MW	等电点 PI	染色体 Chromosome	可变剪切 Splice variants
AC233853.1_FGT002	ZmbZIP37	288	10.86	6.59	Chr3:3077422—3079293	2
GRMZM2G024851	ZmbZIP42	348	39.14	9.24	Chr3:139605736—139609752	3
GRMZM2G027976	ZmbZIP49	180	20.04	10.29	Chr3:199110559—199113757	2
GRMZM2G120167	ZmbZIP53	349	38.56	10.04	Chr4:17281947—17287115	1
GRMZM2G118870	ZmbZIP56	322	35.19	5.97	Chr4:114048204—114058719	5
GRMZM2G052102	ZmbZIP65	258	27.34	6.39	Chr4:231582689—231584787	1
GRMZM2G158313	ZmbZIP79	252	26.98	5.96	Chr5:147356832—147358697	1
GRMZM2G092609	ZmbZIP80	320	34.67	6.59	Chr5:174493404—174497912	6
GRMZM2G170079	ZmbZIP121	280	29.57	6.86	Chr8:179164455—179168169	1

图2 ZmbZIP基因在玉米不同器官根、茎、叶、雄花和嫩穗中的表达
不同小写字母代表差异性显著(P<0.05)。图3-5同。

Fig.2 Expression of ZmbZIPs in different tissues including root,stem,leaf,tassel and ear tissue in maize
Different small letters indicates significant difference among treatments at 0.05 level.The same as Fig.3-5.

图3 ZmbZIP应答不同逆境胁迫时的表达模式分析

Fig.3 The expression pattern of ZmbZIPs under 200 mmol/L NaCl,20% PEG6000 and 4 ℃ stress

图4 玉米叶片中的ZmbZIP应答不同氮形态缺乏胁迫的响应表达模式

Fig.4 The expression pattern of ZmbZIPs of leaves in response to nitrogen stress in maize

图5 玉米根系中的ZmbZIP在不同逆境胁迫时的表达模式分析

Fig.5 The expression pattern of ZmbZIPs of roots in response to nitrogen stress in maize

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