玉米苯丙氨酸解氨酶(PAL)家族基因分析及ZmPAL5的功能研究  

Analysis of Maize Phenylalanine Ammonia-Lyase(PAL)Family Genes and Functional Study of ZmPAL5

作　　者：曹丽茹[1] 叶飞宇 库丽霞[2] 马晨晨 庞芸芸 梁小菡 张新[1] 鲁晓民[1] CAO LiRu;YE FeiYu;KU LiXia;MA ChenChen;PANG YunYun;LIANG XiaoHan;ZHANG Xin;LU XiaoMin(Grain Crop Research Institute,Henan Academy of Agricultural Sciences/The Shennong Laboratory,Zhengzhou 450002;College of Agronomy,Henan Agricultural University,Zhengzhou 450046)

机构地区：[1]河南省农业科学院粮食作物研究所/神农种业实验室,郑州450002 [2]河南农业大学农学院,郑州450046

出　　处：《中国农业科学》2024年第12期2265-2281,共17页Scientia Agricultura Sinica

基　　金：河南省农业科学院杰出青年科技基金(2022JQ02);神农种业实验室项目(SN01-2022-02);国家自然科学基金青年基金(32201708);河南省现代农业产业技术体系建设专项资金(HARS-02-G1)。

摘　　要：【目的】挖掘玉米抗旱关键基因、揭示其抗旱分子机制,为培育抗旱玉米新品种提供基因资源和理论指导。【方法】采用转录组数据结合加权基因共表达网络(weighted gene co-expression network,WGCNA)与抗旱性生理生化指标的筛选方法,鉴定与抗旱和复水相关的ZmPAL基因。利用生物信息学方法对编码PAL的基因进行全基因组分析。运用实时荧光定量PCR(quantitative real-time polymerase chain reaction,qRT-PCR)检测ZmPAL基因在干旱处理条件下的表达情况,以及ZmPAL5在不同自交系间的表达特性和在不同组织中的表达模式。最后,利用遗传转化分析ZmPAL5在玉米中的抗旱功能,并借助CRISPR/Cas9技术对PAL5同源基因进行缺失型拟南芥突变体的抗旱性分析。【结果】鉴定了19个玉米ZmPAL基因,其中6个基因聚集在第5染色体上,其编码蛋白多为亲水性酸性蛋白,且PAL家族基因的进化相对保守。ZmPAL基因启动子区域含有大量与激素和非生物应激响应相关的顺式作用元件。确定了6个核心基因,其中4个基因在干旱处理后显著上调表达。尤其是ZmPAL5在干旱胁迫后表达量增加了8.57倍。在干旱胁迫和复水处理条件下,发现抗旱自交系郑8713中ZmPAL5的表达水平显著高于旱敏感自交系B73。同时,ZmPAL5是一个组成型表达基因,在幼茎中表现出高水平的表达。过表达ZmPAL5玉米在干旱胁迫下生长良好,其相对含水量、木质素、叶绿素、可溶性蛋白、脯氨酸含量,以及超氧化物歧化酶、过氧化物酶、过氧化氢酶和抗坏血酸过氧化物酶的活性分别是野生型的1.52、1.49、1.47、1.43、1.44、1.41、1.53、1.41和1.35倍,但丙二醛含量是野生型的0.65倍。PAL5缺失型拟南芥突变体对干旱敏感。在干旱胁迫下,其生理生化指标与过表达ZmPAL5玉米的指标变化趋势相反。【结论】筛选出6个响应干旱胁迫的核心基因(ZmPAL3、ZmPAL5、ZmPAL6、ZmPAL8、ZmPAL11和Zm【Objective】Mining the key drought-resistant genes of maize,revealing its drought-resistant molecular mechanism,and providing genetic resources and theoretical guidance for the cultivation of new drought-resistant maize varieties.【Method】Transcriptome data combined with weighted gene co-expression network(WGCNA)and screening methods for physiological and biochemical indicators of drought resistance were used to identify ZmPAL genes associated with drought resistance and rewatering.Genome-wide analysis of the genes encoding PAL was performed using bioinformatics methods.Quantitative real-time fluorescence PCR(qRT-PCR)was used to detect the expression of ZmPAL genes under drought treatment conditions,as well as the expression characteristics of ZmPAL5 among different inbred lines and the expression patterns in different tissues.Finally,genetic transformation was used to analyze the drought resistance function of ZmPAL5 in maize,and the deletion-type Arabidopsis mutant was analyzed for drought resistance with the help of CRISPR/Cas9 technology for the PAL5 homologous gene.【Result】Nineteen maize ZmPAL genes were identified,six of which were clustered on chromosome 5 and encoded proteins that were mostly hydrophilic acidic proteins and relatively evolutionarily conserved in the PAL family of genes.The promoter region of ZmPAL genes contained a large number of cis-acting elements associated with hormonal and abiotic stress responses.Six core genes were identified,four of which were significantly up-regulated for expression after drought treatment.In particular,ZmPAL5 showed an 8.57-fold increase in expression after drought stress.The expression level of ZmPAL5 was found to be significantly higher in the drought-resistant inbred line Zheng 8713 than in the drought-sensitive inbred line B73 under both drought stress and rewatering treatments.Meanwhile,ZmPAL5,a constitutively expressed gene,showed a high level of expression in young stems.Overexpressed ZmPAL5 maize grew well under drought stress,and its relativ

关 键 词：玉米 苯丙氨酸解氨酶 ZmPAL5 抗旱性 生理生化 

分 类 号：S513[农业科学—作物学]