割手密GRAS基因家族鉴定及表达分析  

Identification and Expression Analysis of GRAS Gene Family in Saccharum spontaneum

作　　者：黄翠霖 林萍萍[2] 赵新旺 张木清[1] HUANG Cui-Lin;LIN Ping-Ping;ZHAO Xin-Wang;ZHANG Mu-Qing(Guangxi Key Laboratory of Sugarcane Biology/State Key Laboratory of Unitization and Conservation for Subtropical Agri-Biological Resources,Guangxi University,Nanning 530005,China;National Sugarcane Engineering Technology Research Center,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]广西大学广西甘蔗生物学重点实验室/亚热带农业生物资源保护与利用国家重点实验室,南宁530005 [2]福建农林大学国家甘蔗工程技术研究中心,福州350002

出　　处：《农业生物技术学报》2023年第8期1578-1594,共17页Journal of Agricultural Biotechnology

基　　金：国家糖料产业技术体系甘蔗抗病品种选育岗位专家项目(CARS170109)。

摘　　要：甘蔗(Saccharum officinarum)是重要的糖料和生物能源作物,其产量受到各种生物胁迫和非生物胁迫的影响。割手密(S.spontaneum)是甘蔗栽培育种过程中的重要亲本之一。GRAS(GAI-RGA-SCR)转录因子在植物光形态建成、种子萌发、根系生长及防御应答等过程中起重要调控作用。本研究利用GRAS结构域的隐马尔科夫模型(Hidden Markov Model,HMM)配置文件,从割手密基因组中鉴定出173个GRAS基因家族成员,分布于割手密30条染色体。利用最大似然法进行系统发育分析,结果显示,173个割手密GRAS家族基因可分为8个亚家族;基因结构分析显示,GRAS家族基因在C端含有至少1个保守的GRAS结构域,61.2%的GRAS家族基因存在至少1个内含子;GRAS家族基因上游2000 bp序列启动子顺式作用元件分析显示,所有GRAS启动子区域存在应答生物和非生物胁迫的顺式作用元件,表明其可能涉及多种逆境胁迫的调控过程。转录组数据分析显示,割手密GRAS家族成员在茎和叶的不同发育时期存在较大的表达差异,qRT-PCR验证9个基因在成熟茎叶组织的表达,其结果与转录组数据一致,推测GRAS家族成员在割手密生长发育调控中发挥不同作用;胁迫分析表明,SsPAT1.8-1和SsDELLA6-2受甘蔗梢腐病病原菌胁迫诱导表达;SsPAT1.8-2、SsPAT1.9、SsPAT1.10、SsDELLA2、SsSCR2、SsDELLA1-2、SsSCL3.1-2和SsHAM9受甘蔗花叶病毒(Sugarcane mosaic virus,ScMV)胁迫诱导表达;SsSCL3.2-1受干旱胁迫诱导表达。以ScMV侵染抗病品种’B48’,q RT-PCR检测结果显示,SsSCR2在+1叶中上调表达,SsHAM9在+1和-3叶中均上调表达,SsLISCL11-1在-3叶中下调表达。上述结果提示,GRAS家族基因可能在甘蔗对不同胁迫的响应中发挥调节功能。本研究为进一步解析甘蔗GRAS家族基因的功能提供基础资料,为甘蔗抗逆育种提供潜在的基因资源。Sugarcane(Saccharum officinarum)is an important sugar and biofuel crop,and its yield is affected by various biotic and abiotic stresses.S.spontaneum is one of the important parents in sugarcane cultivation and breeding.GRAS(GAI-RGA-SCR)transcription factors play important roles in the regulation of plant photomorphogenesis,seed germination,root growth and defense responses.In this study,173 members of the GRAS gene family were identified from the genome of Saccharum spontaneum using the Hidden Markov Model(HMM)configuration file of the GRAS domain,which were distributed on 30 chromosomes of S.spontaneum.Phylogenetic analysis by maximum likelihood method showed that 173 GRAS genes of S.spontaneum could be further divided into 8 subfamilies.Motif and gene structure analysis showed that 173 GRAS genes contained at least one conserved GRAS domain at the C-terminal,and 61.2% of GRAS genes contained at least one intron.The analysis of cis-acting elements in the 2000 bp sequence promoter upstream of 173 GRAS genes found that all GRAS promoter region had cis-acting elements in response to biotic and abiotic stresses,indicating that they might be involved in the regulation process of a variety of stress.The spatiotemporal transcriptional expression analysis found that the members of the GRAS family from S.spontaneum significantly expressed at different stages of stem and leaf development.qRT-PCR verified the expression of 9 genes in mature stem and leaf tissues,and the results were consistent with transcriptome data,suggesting that their family members played different roles in the growth and development regulation of S.spontaneum.The expression of SsPAT1.8-1 and SsDELLA6-2 were induced by the pathogen of sugarcane pokkah boeng disease;SsPAT1.8-2,SsPAT1.9,SsPAT1.10,SsDELLA2,SsSCR2,SsDELLA1-2,SsSCL3.1-2 and SsHAM9 were induced by Sugarcane mosaic virus(ScMV);SsSCL3.2-1 was induced by drought stress.qRT-PCR results showed that after ScMV infection,SsSCR2 was up-regulated in leaf+1 of resistant variety'B48',SsHAM9 was up-reg

关 键 词：甘蔗 GRAS(GAI-RGA-SCR)基因家族 表达分析 逆境胁迫 

分 类 号：S566.1[农业科学—作物学]