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园艺学报 ›› 2019, Vol. 46 ›› Issue (8): 1458-1472.doi: 10.16420/j.issn.0513-353x.2018-1020

• 研究论文 • 上一篇    下一篇

山梨B-box基因PuBBX24表达特性及其在童期调控中的功能分析

王日红,宋敏燕,王 然,杨英杰*   

  1. 青岛农业大学园艺学院,青岛市园艺植物遗传改良与育种重点实验室,山东青岛 266109
  • 出版日期:2019-08-25 发布日期:2019-08-25
  • 基金资助:

    国家现代农业产业技术体系建设专项资金项目(CARS-28-07);国家自然科学基金项目(31401848,31572109);山东省重点研发计划项目(2017CXGC0209);青岛农业大学人才基金项目(631323)

Expression Pattern and Function in Juvenile Regulation of B-box Gene PuBBX24 in Pyrus ussuriensis

WANG Rihong,SONG Minyan,WANG Ran,and YANG Yingjie*   

  1. College of Horticulture,Qingdao Agricultural University;Qingdao Key Lab of Genetic Improvement and Breeding of Horticultural Plants,Qingdao,Shandong 266109,China
  • Online:2019-08-25 Published:2019-08-25

摘要:

为了解PuBBX24基因在梨生长发育过程中的功能,以山梨(Pyrus ussuriensis Maxim.)实生苗为试材,分离PuBBX24及其启动子序列,qRT-PCR分析PuBBX24在山梨根、茎、花、叶片以及果实中的表达特性,对启动子顺式作用元件进行预测,获得PuBBX24启动子驱动GUS基因表达的转基因拟南芥并进行不同组织、不同生长发育阶段,以及ABA、非生物胁迫和不同光处理下PuBBX24基因启动子活性分析,同时在拟南芥中异源表达PuBBX24并验证其功能。结果表明,PuBBX24在山梨根、茎、叶、花以及果实中均有表达,但在茎中表达量最高,叶片表达量最低,且在由童期向成年期发育的叶片中表达水平逐渐降低。PuBBX24上游启动子序列中包含一系列响应激素、光、胁迫等顺式作用元件。而PuBBX24启动子驱动的GUS基因主要在转基因拟南芥叶片、下胚轴以及萼片中表达,且在叶片中的表达水平随童期向成年期的发育而逐渐降低,在根中表达不均匀,在柱头和花柄中表达水平较低,在角果中未检测到GUS活性。PuBBX24启动子显著响应ABA、光、低温、渗透以及盐胁迫处理,且PuBBX24的过量表达影响了转基因拟南芥植株童期—成年期阶段转变进程。这些结果表明,山梨PuBBX24可能参与ABA、光以及胁迫响应,同时在童期—成年期阶段转变过程中起负调控作用。

关键词: 山梨, PuBBX24, 表达特性, 启动子, 童期

Abstract:

To understand the function of PuBBX24 in pear growth and development process,the gene of PuBBX24 and its promoter sequence were cloned from Pyrus ussuriensis seedling for molecular biological analysis. The spatial expression pattern of PuBBX24 was analyzed in roots,stems,leaves,flowers,and fruits. The cis-elements of PuBBX24 promoter were predicted and subsequently transferred into Arabidopsis with GUS gene for spatial-temporal,as well as under conditions of ABA,different light and other abiotic stresses analysis of its activity. The function of PuBBX24 was also heterologously analyzed in Arabidopsis. The results showed that PuBBX24 had different expression patterns in roots,stems,leaves,flowers,and fruits,with the highest expression level in stems and lowest expression level in leaves. The PuBBX24 expression level decreased gradually in the leaves from juvenile to adult period. A series of cis-elements of PuBBX24 promoter were found with annotations of hormone-responsive,light-responsive and stress-responsive elements. The expression of GUS in transgenic Arabidopsis was mainly detected in leaves,hypocotyls,and sepals,with no uniform levels in roots,lower level in stigma and pedicel. In addition,GUS showed a gradually decreased expression pattern in the rosette leaves from juvenile to adult period and undetectable GUS activity was found in silique. PuBBX24 promoter showed obvious response to stresses of ABA,light,low temperature,osmosis and salt treatments,and over-expression of PuBBX24 in Arabidopsis affected the juvenile-adult transition. These results indicated that PuBBX24 might be involved in responses to ABA,light,and other abiotic stresses and play an important role in regulating the juvenile-adult transition process.

Key words: Pyrus ussuriensis, PuBBX24, expression pattern, promoter, juvenile period

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