摘要
光敏色素互作因子(Phytochrome-interacting factors,PIFs)属于bHLH家族成员,在植物光信号传导、光形态建成和抗逆性等方面发挥重要作用.本研究基于已发表的多年生黑麦草转录组数据,克隆了逆境响应差异表达PIF基因LpPIL15-like,并初步分析其基因特征、转录模式及耐逆功能.LpPIL15-like基因具有5个外显子,编码区全长为1671 bp,编码 556个氨基酸,与硬直黑麦草LrPIL15-like基因亲缘关系最近.LpPIL15-like蛋白具有典型的bHLH结构域及APB和APA功能域,定位于细胞核.LpPIL15-like启动子中含有ABRE、CGTCA-motif和G-box等激素、逆境和光响应顺式作用元件.定量分析表明,LpPIL15-like在伸展叶中表达量最高,在茎、根和叶鞘中表达量较低.此外,LpPIL15-like基因的表达受昼夜节律调控,在光照条件下的表达量显著高于黑暗条件.PEG、NaCl、CdCl2、高温胁迫以及6-BA、ABA等激素处理0.5、2、6、12、24 h显著增强了叶片中LpPIL15-like基因表达,而逆境胁迫和激素处理0.5、2、6、12 h均显著抑制了LpPIL15-like基因在根中的表达.异源过量表达LpPIL15-like基因显著提高了酵母的耐盐、高温和干旱的能力.综上,LpPIL15-like的转录受昼夜节律、非生物逆境和植物激素调控,可作为黑麦草耐逆遗传改良的候选基因.
Abstract
Phytochrome interacting factors(PIFs)belong to the bHLH family and play crucial roles in plant light signal transduction,photomorphogenesis and stress resistance.Based on the published transcrip-tome data of perennial ryegrass,we cloned a stress-responsive differentially expressed PIF gene named LpPIL15-like,and preliminarily analyzed its gene characteristics,transcription patterns and stress resis-tance function.LpPIL15-like gene has 5 exons,a total coding length of 1671 bp,and encoding 556 amino acids,which is the closest genetic relationship with the LrPIL15-like gene of Lolium rigidum.LpPIL15-like protein has a typical bHLH domains and APB and APA functional domains,and is located in the nucleus.The promoter of LpPIL15-like contains hormones,stress and light response cis-regulatory ele-ments such as ABRE,CGTCA motif and G-box.Quantitative analysis showed that the expression of LpPIL15-like was the highest in extended leaves,and the expression was lower in stem,root and leaf sheaths.Furthermore,the expression of LpPIL15-like gene was regulated by circadian rhythm,and the expression of LpPIL15-like gene was significantly higher under light than under dark conditions.PEG,NaCl,CdCl2,and high temperature stress and 6-BA,ABA and other hormones treatment at 0.5,2,6,12,and 24 h significantly enhanced the expression of LpPIL15-like in leaves,while stress and hormone treatments at 0.5,2,6,and 12 h significantly inhibited the expression of LpPIL15-like gene in roots.Het-erologous overexpression of LpPIL15-like significantly improved the salt,high temperature and drought tol-erance of yeast.In conclusion,the transcription of LpPIL15-like is regulated by circadian rhythm,abiotic stress and plant hormone,and can be used as a candidate gene for genetic improvement of stress tolerance in ryegrass.
基金项目
中央高校基本科研业务费(XUEKEN2022020)
国家自然科学基金(32371778)
中国博士后基金(2023M731733)
NETL
NSTL
维普
万方数据