园艺学报 ›› 2021, Vol. 48 ›› Issue (6): 1067-1078.doi: 10.16420/j.issn.0513-353x.2020-0674

• 研究论文 • 上一篇    下一篇


谷思, 刘璐, 李安然, 张伟伟, 赵帅琪, 邢宇()   

  1. 北京农学院植物科学技术学院,林木分子设计育种高精尖创新中心,北京102206
  • 收稿日期:2020-10-19 修回日期:2021-03-03 出版日期:2021-06-25 发布日期:2021-07-07
  • 通讯作者: 邢宇
  • 基金资助:

Construction of Yeast Two Hybrid Library and Screening of FvM4K1 Interacting Protein in the Fruit of Fragraria vesca

GU Si, LIU Lu, LI Anran, ZHANG Weiwei, ZHAO Shuaiqi, XING Yu()   

  1. Advanced Innovation Center for Forest Molecular Design and Breeding,College of Plant Science and Technology,Beijing University of Agriculture,Beijing 102206,China
  • Received:2020-10-19 Revised:2021-03-03 Online:2021-06-25 Published:2021-07-07
  • Contact: XING Yu


通过构建草莓不同发育时期果实的cDNA文库,筛选与促分裂原活化蛋白激酶FvM4K1互作的蛋白,探究FvM4K1参与调控果实发育的分子机理。以森林草莓(Fragraria vesca ‘Hawaii 4')为材料,提取不同发育时期果实的总RNA,建立cDNA文库,其库容为9.6 × 10 6 CFU,重组率96%,插入片段平均长度大于1 000 bp。同时构建诱饵载体pGBKT7-FvM4K1,并检测其在酵母中无自激活活性。利用共转化方法,从文库中筛选到13个与FvM4K1互作的蛋白。通过Gene Ontology(GO)通路注释表明候选互作蛋白主要集中在细胞代谢过程及具有催化活性的分子过程。候选蛋白预测功能主要集中在调控花粉发育和花粉管伸长以及开花时间等。筛选到的13个蛋白编码基因在果实发育过程中表达模式不同,其中4个蛋白预测功能与草莓果实发育密切相关。

关键词: 草莓, 森林草莓, cDNA文库, MAPK


cDNA library of strawberry fruit at different stages was constructed and the proteins interacting with strawberry mitogen activated kinase FvM4K1 were screened which revealed the molecular mechanism of FvM4K1 in regulating strawberry fruit development. The total RNA ofFragraria vesca ‘Hawaii 4'was extracted at different fruit development stages related to FvM4K1,and the strawberry cDNA library was established. Specifically,the library capacity was 9.6 × 10 6 CFU,the recombination rate was 96%,and the average length of inserted fragment was more than 1 000 bp. At the same time,the bait vector pGBKT7-FvM4K1 was constructed,and no self-activation activity was detected in yeast. Then 13 proteins interacting with FvM4K1 were screened from the library by co-transformation. The function of these candidate proteins mainly focused on floral development such as pollen development and pollen tube elongation,also affecting flowering time and so on. Meanwhile,the Gene Ontology(GO)pathway annotation of these proteins showed that they mainly participated in cell metabolism process and molecular processes with catalytic activity. Besides,the 13 proteins screened had different expression patterns during fruit development,and four of them were predicted to be closely related to strawberry fruit development.

Key words: strawberry, Fragaria ananassa, cDNA library, MAPK