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园艺学报 ›› 2020, Vol. 47 ›› Issue (2): 220-232.doi: 10.16420/j.issn.0513-353x. 2019-0555

• 研究论文 • 上一篇    下一篇

三倍体枇杷花期调控基因 EjSPL5 的克隆、亚细胞定位及表达分析

孙伟雄,石 敏,薛宝贵,夏 燕,郭启高,王淑明,吴 頔,景丹龙*,梁国鲁   

  1. 南方山地园艺学教育部重点实验室,西南大学南方山地作物逆境生物学国家级培育基地,西南大学农业科学研究院,园艺园林学院,重庆 400715
  • 出版日期:2020-02-25 发布日期:2020-02-25
  • 基金资助:
    国家重点研发计划项目(2019YFD1000905);中央高校基本科研业务费专项资金项目(XDJK2019AA001,XDJK2020B058);重庆市科技局项目(cstc2018jscx-mszdX0054,cstc2017jcyjAX0433);重庆市高校创新研究群体项目(CXQT19005)

Cloning,Subcellular Localization and Expression Analysis of the EjSPL5 Gene in Triploid Loquat

SUN Weixiong,SHI Min,XUE Baogui,XIA Yan,GUO Qigao,WANG Shuming,WU Di,JING Danlong*,and LIANG Guolu   

  1. Key Laboratory of Horticulture Science for Southern Mountains Regions of Ministry of Education,National Cultivation Base of Crop Adversity Biology for Southern Mountains Regions of Southwest University,Academy of Agricultural Sciences,College of Horticulture and Landscape Architecture,Southwest University,Chongqing 400715,China
  • Online:2020-02-25 Published:2020-02-25

摘要: 利用同源克隆和 cDNA 末端快速扩增(RACE)技术从三倍体枇杷‘Q27’中分离得到 1 个调控花期的 SQUMOSA PROMOTER BINDING PROTEIN LIKE(SPL)转录因子家族基因 EjSPL5。该基因 cDNA 序列全长为 778 bp,其中 5′非翻译区(UTR)序列为 24 bp,3′UTR 序列为 214 bp,包含 1 个长为 549 bp 的开放阅读框(ORF),编码 182 个氨基酸。蛋白序列比对分析表明,EjSPL5 和苹果(Md)、拟南芥(At)及金鱼草(Am)的 SPL 蛋白序列具有较高的相似性,均具有保守的 SBP 结构域和核定位信号(NLS)。分子系统进化分析表明,EjSPL5 与苹果 MdSPL5 具有较高的同源性,聚为同一分支。EjSPL5的亚细胞定位发现,其行使功能区域定位在细胞核。实时荧光定量 PCR 分析表明,EjSPL5 在枇杷萼片和幼果中的表达量较高;二倍体和三倍体枇杷的早、晚花品种中 EjSPL5 表达量存在显著差异,其表达主要集中在花发育的前 6 个时期,在花蕾露白期和盛花期的表达量较低。三倍体枇杷 EjSPL5 表达量在花芽形态分化期最高,二倍体枇杷在花序侧生支轴快速伸长期的表达量最高。早花枇杷品种的 EjSPL5 相对表达量高于晚花品种.

关键词: 枇杷, 三倍体, EjSPL5, 亚细胞定位, 基因表达

Abstract: In this study,SQUMOSA PROMOTER BINDING PROTEIN LIKE5(EjSPL5)was isolated by homologous cloning and rapid amplification of cDNA ends(RACE)techniques from the triploid loquat‘Q27’. Full length cDNA sequence of EjSPL5 was 778 base pairs(bp),including a 24 bp 5′UTR and a 214 bp 3′UTR. Open reading frame(ORF)of EjSPL5 was 549 bp and encodes 182 amino acids. Sequence alignment analysis showed that EjSPL5 is highly similar to SPL proteins of Malus × domestica(Md),Arabidopsis thaliana(At)and Antirrhinum majus(Am),and has a conserved SBP domain and nuclear localization signal(NLS). There were high homology of amino acid sequences in EjSPL5 and MdSPL5 of Malus × domestica. Subcellular localization of EjSPL5 was detected to be localized to nucleus. Real-time quantitative PCR analysis suggested that EjSPL5 was highly expressed in sepal and young fruit. There was a significant difference in the expression levels of EjSPL5 in the early and late flowering cultivars of diploid and triploid loquat. The expression of EjSPL5 was mainly concentrated in the first six stages of flower development,while the expression level of EjSPL5 was lower in the stages of white corollas and full bloom. However,the expression level of EjSPL5 was the highest in the morphological differentiation of flower bud in triploid loquat and the rapid elongation of lateral inflorescence branches in diploid loquat. Furthermore,the expression level of EjSPL5 in early-flowering cultivars was higher than that in lateflowering ones.

Key words: loquat, triploid, EjSPL5, subcellular localization, gene expression

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