‘潘那利’番茄碱性螺旋环螺旋转录因子基因的克隆与表达分析  

Cloning and Prokaryotic Expression Analysis of a Transcription Factor Gene in Solanum pennellii

作　　者：胡佳蕙 王柏柯[1] 李宁[1] 余庆辉[1] 王娟[1] HU Jiahui;WANG Baike;LI Ning;YU Qinghui;WANG Juan(Institute of Horticultural Crops,Xinjiang Academy of Agricultural Science,Urumqi 830091,China;College of Horticulture,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆农业科学院园艺作物研究所,乌鲁木齐830091 [2]新疆农业大学园艺学院,乌鲁木齐830052

出　　处：《西北植物学报》2022年第4期541-548,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：中国博士后科学基金面上资助(2019M663862);天山青年计划(2019Q048)。

摘　　要：碱性螺旋环螺旋(basic/Helix-Loop-Helix,bHLH)转录因子是植物最大的转录因子家族之一,其广泛参与植物逆境胁迫响应。该研究从野生‘潘那利’番茄(Solanum pennellii Correll)中成功克隆出bHLH转录因子基因SpbHLH89(Sol Genomics登录号Sopen04g001150),采用qRT-PCR分析其在干旱胁迫下的表达模式,并利用异源表达初步分析其对非生物胁迫的响应。结果表明:(1)SpbHLH89编码区包含684 bp,编码227个氨基酸,具有典型的碱性螺旋环螺旋区,主要定位于细胞核中;进化树结果显示,SpbHLH89转录因子高度保守,与拟绒毛烟草NtbHLH94(Nicotiana tomentosiformis)存在高度相似性。(2)qRT-PCR结果显示,SpbHLH89在‘潘那利’番茄的茎、叶和花中均有表达,其表达量受干旱胁迫诱导。(3)SDS-PAGE与Western bloting结果显示,pET-30a-SpbHLH89重组蛋白大小约为31 kD。(4)在盐胁迫(400 mmol/L NaCl)和干旱胁迫(600 mmol/L甘露醇)条件下,异源表达重组蛋白的E.coli BL21(DE3)重组菌生长速度提高,说明异源表达SpbHLH89转录因子基因可提高细菌对非生物胁迫的耐受性。Basic helix-loop-helix(bHLH)transcription factors are one of the largest transcription factor families in plants.It is extensively involved in stress response of plants.In the present study,we cloned a bHLH transcription factor gene-SpbHLH89(Sol Genomics number Sopen04g001150)from Solanum pennellii.Meanwhile,we also analyzed its expression patterns under drought stress by qRT-PCR and verified its biological function by prokaryotic expression.The results showed that:(1)an open reading frame(ORF)of SpbHLH89 was obtained from S.pennellii,which is 684 bp long and encodes 227 amino acids with typical basic-helix-basic domain and mainly located on the nucleus.Phylogenetic tree analysis showed that SpbHLH89 was highly conserved and shared a high degree of sequence similarity with NtbHLLH094 from Nicotiana tomentosiformis.(2)qRT-PCR analysis showed that the pattern of SpbHLH89 was highly expressed in flowers and also induced by drought stress.(3)SDS-PAGE and Western blotting analysis showed that the molecular weight of pET-30a-SpbHLH89 was approximately 31 kD.(4)Heteroexpression of the pET-30a-SpbHLH89 could improve the growth of the recombinant strain of E.coli BL21(DE3)under salt(400 mmol/L NaCl)and drought(600 mmol/L mannitol)stress.Taken together,the heterologous expression of SpbHLH89 transcription factor could improve the tolerance of recombinant bacteria to abiotic stress.

关 键 词：‘潘那利’番茄 SpbHLH89 亚细胞定位 表达模式 原核表达 

分 类 号：Q785[生物学—分子生物学] Q786