基于油橄榄全基因组参与水胁...NAC基因的鉴定及表达分析_王丽娟.pdf

分子植物育种，2023 年，第 21 卷，第 3 期，第 736-746 页Molecular Plant Breeding,2023,Vol.21,No.3,736-746研究报告Research Report基于油橄榄全基因组参与水胁迫 NAC 基因的鉴定及表达分析王丽娟1,2王毅2*李贤忠1张璋1,2冯发玉1胡青2陆斌2*1 西南林业大学林学院,昆明,650224;2 云南省林业和草原科学院,云南省森林植物培育与开发利用重点实验室,国家林业局云南珍稀濒特森林植物保护和繁育重点实验室,昆明,650201*共同通信作者,dog_;摘要本研究以干旱和水淹胁迫中的 佛奥 和 TYZ-1 号 2 个油橄榄品种苗木为试材，基于油橄榄全基因组数据，鉴定出 36 个油橄榄 NAC 转录因子家族成员，利用生物信息学软件进行结构及功能预测，将油橄榄 OeNAC 蛋白与模式植物拟南芥 AtNAC 蛋白构建进化树。结果表明 OeNAC 转录因子分为两个大类和14 个亚家族，14 个亚家族中有 13 个亚族同时包含了油橄榄和拟南芥 NAC 蛋白，说明拟南芥和油橄榄的NAC 家族在进化水平上有一定的相似性。结合油橄榄水分胁迫(干旱,水淹)基因表达谱，分析出 8 个 NAC转录因子(OeNAC31,OeNAC33,OeNAC25,OeNAC30,OeNAC7,OeNAC23,OeNAC20,OeNAC1)在 佛奥和 TYZ-1 号 中均呈现高表达且对水分胁迫均有响应，推测这 8 个基因是油橄榄水分调节的主要基因。将在水分胁迫下具有表达量的 OeNAC 蛋白与已知水分胁迫相关的 NAC 转录因子构建进化树发现 8 个胁迫相关的转录因子中的 OeNAC31 与已知的水分调控因子支持率更高，表明 OeNAC31 转录因子可能是参与油橄榄水分调控的最主要基因，其在两个品种中都有不同的表达量，这可能是 佛奥 和 TYZ-1 号 抗性不同的原因。本研究通过油橄榄转录组测序，对油橄榄 NAC 转录因子进行全基因组鉴定、蛋白理化性质、系统发育、保守元件及基因表达谱分析，探究油橄榄 NAC 转录因子在 TYZ-1 号 和 佛奥 中基因表达模式，也为油橄榄 NAC 基因家族的功能验证及水胁迫反应关键基因的挖掘提供参考。关键词油橄榄;NAC 转录因子;生物信息学;基因表达谱;水分胁迫Identification and Expression Analysis of NAC Gene Involved in WaterStress Based on the Whole Genome of Olea europaeaWang Lijuan1,2Wang Yi2*Li Xianzhong1Zhang Zhang1,2Feng Fayu1Hu Qing2Lu Bin2*1 College of Forestry,Southwest Forestry University,Southwest Forestry University,Kunming,650224;2 Laboratory of Forest Plant Cultivation andUtilization,The Key Laboratory of Rare and Endangered Forest Plants of State Forestry Administration,Yunnan Academy of Forestry&Grassland Sci-ence,Kunming,650201*Co-corresponding authors,dog_;DOI:10.13271/j.mpb.021.000736AbstractIn this study,seedlings of Frantoio and TYZ-1 Olea europaea varieties were used as experimentalmaterials under drought stress and water flooding stress.Based on the whole genome data of Olea europaea,36members of NAC gene family were identified.Using bioinformatics software to predict the structure and function;Olea europaea OeNAC protein and model plant Arabidopsis AtNAC protein were used to construct the evolutionarytree.The results showed that the OeNAC transcription factors could be divided into two major groups and 14基金项目：本研究由国家重点研发计划项目(2020YFD1000703)、中央财政林草技术推广项目(云 2021 TG01 号)和云南省林业和草原科学院创新基金项目(LKYTD-2020-001)共同资助引用格式：Wang L.J.,Wang Y.,Li X.Z.,Zhang Z.,Feng F.Y.,Hu Q.,and Lu B.,2023,Identification and expression analysis of NACgene involved in water stress based on the whole genome of Olea europaea,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),21(3):736-746.(王丽娟,王毅,李贤忠,张璋,冯发玉,胡青,陆斌,2023,基于油橄榄全基因组参与水胁迫 NAC 基因的鉴定及表达分析,分子植物育种,21(3):736-746.)subfamilies,of which 13 subfamilies contained the NAC proteins of Olea europaea and Arabidopsis thaliana,sug-gesting that the NAC families of Arabidopsis thaliana and Olea europaea had some similarities at the evolutionarylevel.By analyzing the gene expression profile of Olea europaea(under drought and water flooding conditions),itwas found that 8 NAC transcription factors(OeNAC31,OeNAC33,OeNAC25,OeNAC30,OeNAC7,OeNAC23,OeNAC20,OeNAC1)were highly expressed in both varieties and were highly responsive to water stress.It wasspeculated that these 8 genes might be the main genes of water regulation in Olea europaea.The phylogenetic treewas constructed between OeNAC protein with expression under water stress and known NAC transcription factorsrelated to water stress.It was found that OeNAC31 had the highest support rate with known water regulatoryfactors,indicating that OeNAC31 transcription factor may be the main gene involved in water regulation of olive.Moreover,there were different expression levels of OeNAC31 in both cultivars,which might be the reason for thedifferent resistance between Frantoio and TYZ-1.In this study,the whole genome of the Olea europaea NACtranscription factors was identified by transcriptome sequencing,and the physicochemical properties,phylogeny,conserved elements and gene expression profiles of the proteins were analyzed to explore the expression patternsof the Olea europaea NAC transcription factors in TYZ-1 and Frantoio.It also provides reference for functionalverification of NAC gene family in Olea europaea and mining of key genes in water stress response.KeywordsOlea europaea;NAC transcription factor;Bioinformatics;Gene expression profile;Water stress油橄榄(Olea europaea)是木犀科木樨榄属的重要油料作物，1964 年从阿尔巴尼亚引进油橄榄种苗开始栽培试验，目前中国已有 20 多个省区引种栽培油橄榄，数量达到 1 200 万株(吴文俊等,2020,中国果树,205(5):111-113)。随着中国经济的持续发展和人们健康意识的增强，最近 20 年中国橄榄油的消费呈连续快速增长的趋势，而生产发展明显滞后，国产橄榄油远远不能满足市场的需求(朱申龙等,2020)。从全国范围来看，气候条件是影响油橄榄不能持续正常生长发育和开花结实的主要原因，夏季高温多雨和土壤粘重的地区，表现出生长过速、树体高大、落叶多、枯枝多和叶幕层薄的早衰现象；冬季寒冷干燥导致生长缓慢，植株弱小，叶片枯萎、黄化，甚至脱落、死亡，因此选育抗旱、抗涝的砧木品种是解决油橄榄栽培问题瓶颈的关键。油橄榄在自然界中生长，通常会遇到气候问题带来的非生物胁迫，为了抵御非生物胁迫对其生长发育的影响，油橄榄体内会建立一系列信号传导和分子调控机制，通过相关的转录因子(Transcription factor)与顺式作用元件特异性结合来调控下游基因的表达，从而适应环境的变化(张丹和马玉花,2019;崔喜艳等,2021,私人通信)。NAC 转录因子是植物最大的转录因子家族之一。它的结构特征最初来自 NAM、ATAF1/2 和CUC2 的共有序列(陈倩,2020,私人通信)。N 端蛋白结构域是由约 150 个氨基酸组成的 DNA 结合结构域，含有保守的 A、C、D 和相对多变的 B、E 五个亚结构域，C 端转录激活域可以激活或抑制转录(Olsenet al.,2005)。大量研究也证明了 NAC 转录因子在植物生长发育及非生物胁迫中起关键的调控作用，如OsNAC52 增强转基因植物的耐旱性(Fengetal.,2010)，甘蔗(Saccharu