荔枝漆酶(LAC)基因家族鉴定及表达分析  

Identification and Expression Analysis of Laccase(LA C)Genes in Litchi(Litchi chinensis Sonn.)

作　　者：李郑华 姜成东[1] 王鹏[3] 李芳 王树军[2] 刘耀婷 李焕苓[2] 王果[2] 王家保[2,3] Li Zhenghua;Jiang Chengdong;Wang Peng;Li Fang;Wang Shujun;Liu Yaoting;Li Huanling;Wang Guo;Wang Jiabao(College of Horticulture,Hainan University,Haikou,570228;Danzhou Scientific Observing and Experimental Station of Agro-Environment(Ministry of Agriculture and Rural Affairs),Environment and Plant Protection Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101;Tropical Crops Genetic Resources Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101)

机构地区：[1]海南大学园艺学院,海口570228 [2]中国热带农业科学院环境与植物保护研究所,农业农村部儋州农业环境科学观测实验站,海口571101 [3]中国热带农业科学院热带作物品种资源研究所,海口571101

出　　处：《分子植物育种》2023年第7期2174-2187,共14页Molecular Plant Breeding

基　　金：国家重点研发计划项目(No.2019YFD1000900);国家现代农业产业技术体系专项(No.CARS-32-03);海南省自然科学基金项目(No.319QN270)共同资助。

摘　　要：探索荔枝漆酶(LAC)基因家族的数量、结构和进化关系,可以为解析漆酶基因功能提供理论依据,为探索荔枝果皮褐变机制提供基础。本研究利用生物信息学分析手段,对LAC基因家族成员数量、保守结构域、家族成员的亚细胞定位及染色体位置、系统发育和顺式作用元件,以及基因家族成员在不同组织器官和采后贮藏果皮中的表达情况进行了分析。结果表明,荔枝基因组中共鉴定出61个LAC家族成员,编码的氨基酸残基长度在127~958 aa,分子量在14.07~103.47 Da,等电点在4.42~10.34;亚细胞定位分析表明,有19个LAC家族成员定位在叶绿体中,17个LAC家族成员定位在液泡中,其他成员分别定位在内质网、细胞外基质等。61个荔枝LAC家族成员分别定位在10条染色体上,其中第13号和8号染色体上LAC基因数量最多,且呈簇状分布于染色体一端。系统发育树将荔枝LAC家族分为5个亚组,第Ⅴ亚组最大,有30个家族成员,第Ⅳ亚组最小,仅有8个家族成员。荔枝LAC家族的结构域组织相对保守,具有典型的Cu-oxide漆酶结构域。荔枝LAC基因启动子序列分析发现了大量的顺式作用元件,可能与漆酶家族功能多样性相关。10个荔枝组织器官的转录组测序分析表明,除Lch08g07950-mRNA-1基因外,其余61个基因在荔枝各组织中表达量均较低。实时荧光定量RT-PCR结果与转录组分析结果基本一致,Lch08g07950-mRNA-1在所有组织中都有较高的表达水平,并且其在采后荔枝果皮中的表达水平与褐变发生具有相关性。Elucidation of the function and evolutionary relationship of the laccase(LA C)gene family in litchi can provide a basis for interpreting the browning mechanism of litchi pericarps and a theoretical basis for analyzing the function of the laccase genes.The number of LAC gene family members,conserved structure,subcellular and chromosomal locations,phylogeny and cis-acting elements,gene family members'expression in different tissues and pericarp of postharvest fruit were analyzed by bioinformatics analysis in this study.The results showed that a total of 61 litchi LAC family members were identified in the litchi genome.The length of the encoded amino acid residues were 127~958 aa,the molecular weights were 14.07~103.47 Da,and the isoelectric points were between 4.42~10.34.Subcellular location analysis revealed that 19 members of the litchi LAC family were located in the chloroplast,17 members were located in the vacuole,and the other members were located in the endoplasmic reticulum,extracellular matrix,and others,respectively.Sixty-one litchi LAC genes were distributed in ten chromosomes,among which the most were in chromosomes 13 and 8,distributed in clusters at one end of the chromosomes.Phylogenetic analysis divided the litchi LAC genes into five subgroups,among which subgroup V was the biggest(30 members),and subgroup IV was the smallest(8 members).The domain organization of the litchi LAC family was relatively conservative,harboring a typical Cu-oxide Laccase domain.Sequence analysis of the litchi LAC gene promoters revealed many cis-acting elements,which may be related to the functional diversity of the litchi LAC family.Transcriptomic sequencing analysis of 10 litchi tissues showed that except for the Lch08g07950-mRNA-1 gene,the expression levels of the other 60 genes in all litchi tissues were lower.Results of real-time quantitative RT-PCR were consistent with those of transcriptomic analysis.Lch08g07950-mRNA-1 showed relatively high expression in all tissues,and its expression level in pericarp of postharv

关 键 词：荔枝漆酶基因 基因家族 生物信息学分析 基因表达 荔枝果皮褐变 

分 类 号：S667.1[农业科学—果树学]