生物技术通报 ›› 2023, Vol. 39 ›› Issue (4): 246-258.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1045

• 研究报告 • 上一篇    下一篇

茶树SMAS基因家族的鉴定及互作分析

王艺清1,2(), 王涛1,2, 韦朝领3, 戴浩民1,2, 曹士先4, 孙威江1,2(), 曾雯1,2()   

  1. 1.福建农林大学园艺学院,福州 350002
    2.海峡两岸特色作物安全生产省部共建协同创新中心,福州 350000
    3.安徽农业大学茶与食品科技学院,合肥 230061
    4.武夷星茶业有限公司,南平 354301
  • 收稿日期:2022-08-23 出版日期:2023-04-26 发布日期:2023-05-16
  • 通讯作者: 孙威江,男,博士,教授,研究方向:茶树种质资源创新与品质化学;E-mail: swj8103@126.com
    曾雯,女,博士,讲师,研究方向:茶树分子生物学;E-mail: wenzeng_t@163.com
  • 作者简介:王艺清,女,硕士研究生,研究方向:茶树分子生物学;E-mail: 784726205@qq.com
  • 基金资助:
    茶树生态经济型品种筛选及配套栽培技术(2019YFD1001601);乌龙茶萎凋关键技术装备创新研究与示范(2021N3010)

Identification and Interaction Analysis of SMAS Gene Family in Tea Plant(Camellia sinensis

WANG Yi-qing1,2(), WANG Tao1,2, WEI Chao-ling3, DAI Hao-min1,2, CAO Shi-xian4, SUN Wei-jiang1,2(), ZENG Wen1,2()   

  1. 1. College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002
    2. Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, Fuzhou 350000
    3. School of Tea and Food Science & Technology, Anhui Agriculture University, Hefei 230061
    4. Wuyi Star Tea Industrial Company Limited, Nanping 354301
  • Received:2022-08-23 Published:2023-04-26 Online:2023-05-16

摘要:

S-腺苷甲硫氨酸合成酶(S-adenosylmethionine synthase, SAMS)是催化甲硫氨酸和ATP合成S-腺苷甲硫氨酸(SAM)的唯一酶,研究表明SAMS参与木质素的生物合成。本研究对‘黄棪’茶树(Camellia sinensis)中鉴定的SAMS基因家族,进行表达模式及蛋白互作网络分析,挖掘可能参与木质素合成的 CsSAMS候选基因。以‘黄棪’茶树基因组为参考基因组,通过生物信息学鉴定CsSAMS基因家族成员,并分析其蛋白理化性质、系统进化树、染色体定位、基因结构、蛋白结构、表达模式,通过酵母双杂交技术(Y2H)研究其蛋白互作网络,通过紫外分光光度计方法对‘黄棪’‘铁观音’‘金观音’‘福鼎大毫茶’一芽二叶部位的木质素含量进行测定。生物信息学分析结果表明,‘黄棪’茶树中共鉴定到4个CsSAMS家族成员,其编码氨基酸个数为345-519,等电点为6.12-6.47。亚细胞定位预测结果表明,CsSAMS1定位于叶绿体,CsSAMS2CsSAMS3定位于细胞质,CsSAMS4定位于细胞骨架。通过对不同茶树品种的CsSAMS表达量和木质素含量检测发现,CsSAMS2CsSAMS3CsSAMS4可能潜在调控木质素的含量,另外酵母双杂交结果表明,CsSAMS4可以与自身形成同源二聚体。本研究鉴定并分析了4个CsSAMS成员的理化性质并预测其功能,明确了不同组织部位和氮、氟处理下CsSAMS基因的表达模式,以及CsSAMS对木质素合成过程的潜在参与作用。

关键词: 茶树, SAMS, 生物信息学分析, 木质素, 表达分析, 互作蛋白

Abstract:

S-adenosylmethionine synthase(SAMS)is the only enzyme that catalyzes the synthesis of S-adenosylmethionine(SAM)from methionine and ATP. Research shows that SAMS participates in lignin biosynthesis. In this study, we analyzed the expression patterns and protein interaction networks of the SAMS gene family identified from ‘Huangdan’ tea plant(Camellia sinensis)to explore candidate genes of CsSAMS that may be involved in lignin synthesis. Then we identified the members of CsSAMS gene family by bioinformatics based on the genome of ‘Huangdan’, and further analyzed their protein physicochemical properties, phylogenetic tree, chromosome location, gene structure, protein structure and expression pattern. We also studied the protein interaction network by yeast two-hybrid technology(Y2H). Finally, we determined the lignin contents of the first bud and the two leaves of ‘Haungdan’, ‘Tieguanyin’, ‘Jinguanyin’ and ‘Fuding Dahao’ by UV spectrophotometer. The results of bioinformatics analysis showed that 4 CsSAMS gene family members were identified in ‘Huangdan’. The number of encoded amino acids was 345-519, and the isoelectric point ranged from 6.12 to 6.47. The prediction results of subcellular localization revealed that CsSAMS1 was located in the chloroplast, CsSAMS2 and CsSAMS3 were located in the cytoplasm, and CsSAMS4 was located in the cytoskeleton. Expression of CsSAMS and lignin content detection of different tea varieties indicated that CsSAMS2, CsSAMS3 and CsSAMS4 might potentially regulate lignin content. In addition, Y2H results showed that CsSAMS4 formed homodimers with itself. In this study, the physicochemical properties of four CsSAMS members are identified and analyzed, and their functions are predicted. The expression patterns of CsSAMS genes in different tissue sites, under nitrogen and fluoride treatments, as well as the potential involvement of CsSAMS in lignin synthesis process, are clarified.

Key words: Camellia sinensis, SAMS, bioinformatics analysis, lignin, expression analysis, interacting proteins