龙眼微管蛋白家族成员全基因组鉴定及其在龙眼体胚发生早期的表达模式  

Genome-wide identification and expression pattern of tubulin gene family during the early somatic embryogenesis of longan

作　　者：路保顺 张舒婷[1] 赵桦 张梓浩[1] 赖钟雄[1] 林玉玲[1] LU Baoshun;ZHANG Shuting;ZHAO Hua;ZHANG Zihao;LAI Zhongxiong;LIN Yuling(Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学园艺植物生物工程研究所,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2022年第3期329-338,共10页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金项目(31672127);福建省自然科学基金项目(2020J01543);福建省高原学科建设经费项目(102/71201801101);福建农林大学科技创新专项(CXZX2019033S、CXZX2018078).

摘　　要：为研究微管蛋白(tubulin)家族成员在龙眼体胚发生早期的表达模式,基于龙眼基因组数据库,采用生物信息学方法对龙眼tubulin家族蛋白理化性质、系统进化关系、基因扩张、进化选择模式、染色体分布、基因结构、蛋白保守基序和启动子顺式作用元件进行分析.基于龙眼体胚发生早期转录组数据库,分析tubulin家族成员在体胚发生早期的表达模式,并采用实时荧光定量PCR(qRT-PCR)技术进行验证.结果表明:龙眼tubulin家族共有15个成员,其中,α-tubulin类型成员6个,β-tubulin类型成员8个,γ-tubulin类型成员1个(TUBG2),所有成员均包含tubulin和tubulin_C两个保守结构域.同一类型tubulin家族成员间的亲缘关系较近,不同类型间的亲缘关系较远.龙眼tubulin家族15个成员分布在10条染色体上,氨基酸数目为197~473个,分子质量为21.88~53.25 ku,等电点为4.44~5.67,为酸性蛋白.龙眼tubulin家族基因外显子数目为3~11个,其编码的蛋白具有高度保守的基序.龙眼tubulin家族基因启动子区域存在光、激素、细胞周期、分生组织和抗逆境胁迫等响应元件.对tubulin家族基因在龙眼体胚发生早期的表达模式进行分析发现,龙眼tubulin家族大多数成员在体胚发生早期呈下调表达趋势,在胚性愈伤组织阶段的表达量高于球形胚阶段.以上结果表明,龙眼tubulin家族成员高度保守,且在胚性愈伤组织中大量累积,可能在龙眼胚性愈伤组织的胚性维持中发挥重要作用.To illuminate the expression pattern of tubulin gene family during the early somatic embryogenesis of longan(Dimocarpus longan Lour.), physicochemical properties and phylogenetic relationship of tubulin family, gene expansion, evolutionary selection model, chromosomal distribution, gene structure, protein conserved motif and promoter cis-acting elements were analyzed by bioinformatic methods based on longan genome datasets. The expression pattern of tubulin gene family were further analyzed based on transcriptome datasets, and confirmed by quantitative real-time polymerase chain reaction(qRT-PCR). The results showed that longan tubulin family contained 15 members(6 α-tubulin, 8 β-tubulin and 1 γ-tubulin) with both conserved domains of tubulin and tubulin_C. The phylogenetic tree analysis showed closer genetic relationship within single tubulin type and farther relationship between tubulin types. The 15 tubulin gene family members were distributed on 10 chromosomes with the number of amino acids ranging from 197 to 473 aa, molecular weight ranging from 21.88 to 53.25 ku, and isoelectric point ranging from 4.44 to 5.67. The number of exon in tubulin gene family ranged from 3 to 11, and the encoded proteins had highly conserved motifs. The promoter region of tubulin gene family contained responsive elements which regulated activities in response to light, hormone, cell cycle, meristem and adversity. The expression pattern analysis revealed that most of the tubulin gene family members were down-regulated during the early somatic embryogenesis of longan, and the expression levels were higher in embryonic callus than that in globular embryo. In conclusion, longan tubulin gene family were highly conserved, and greatly expressed in embryonic callus, indicating their important roles in the embryonic maintenance of embryonic callus.

关 键 词：龙眼 TUBULIN 体细胞胚胎发生 表达模式 

分 类 号：S667.2[农业科学—果树学]