园艺学报 ›› 2020, Vol. 47 ›› Issue (6): 1087-1097.doi: 10.16420/j.issn.0513-353x.2019-0738

• 研究论文 • 上一篇    下一篇


田绍泽1,刘思禹1,2,王 坤1,岳远征1,3,马方芳1,杨赵楠1,殷 倩1,胡惠蓉1,*   

  1. 1华中农业大学园艺林学学院,农业农村部华中都市农业重点实验室,园艺植物生物学教育部重点实验室,武汉430070;2武汉琴台绿化工程有限责任公司,武汉 430070;3南京林业大学风景园林学院,南京 210037
  • 出版日期:2020-06-25 发布日期:2020-06-25
  • 基金资助:

Cloning and Functional Analysis of PhTs2 Gene in Petunia hybrida

TIAN Shaoze1,LIU Siyu1,2,WANG Kun1,YUE Yuanzheng1,3,MA Fangfang1,YANG Zhaonan1,YIN Qian1,and HU Huirong1,*   

  1. 1College of Horticulture and Forestry Sciences,Huazhong Agricultural University,Key Laboratory of Urban Agriculture in Central China,Ministry of Agriculture and Rural Affairs,Key Laboratory of Horticultural Plant Biology,Ministry of Education,Wuhan 430070,China;2Wuhan Qintai Greening Engineering Co.,Ltd.,Wuhan 430070,China;3College of Landscape Architecture,Nanjing Forestry University,Nanjing 210037,China
  • Online:2020-06-25 Published:2020-06-25

摘要: 从矮牵牛‘幻想’中克隆了花发育相关基因PhTs2,该基因CDS长855 bp,编码284个氨基酸,具有保守的adh-short结构域。进化分析表明,PhTs2蛋白与同科植物烟草和番茄的同源蛋白亲缘关系较近。实时荧光定量PCR分析发现该基因在矮牵牛花药发育的早期特异性表达。为分析其功能,构建PhTs2超量表达载体,转化了烟草,并对转基因烟草进行了鉴定:表型观测发现其花药形态异常,花粉数量减少;花粉活力和萌发率极显著降低;扫描电镜观测发现其花粉粒畸形,外壁纹饰不规则;半薄切片显示其绒毡层发育过程紊乱。该研究为培育雄性不育系提供了潜在的基因资源。

关键词: 矮牵牛, 育性, 花药, 绒毡层, 雄性不育, 花粉

Abstract: PhTs2,a gene related to flower development,was cloned from Petunia hybrida‘Fantasy Red’. The CDS of this gene is 855 bp long and encodes 284 amino acids with a conserved adh-short domain. The phylogenetic analysis showed that the PhTs2 protein was closely related to the homologous proteins in tobacco and tomato. Quantitative real-time PCR analysis indicated that the gene was specifically expressed in the early developmental stage of anther in petunia. To analyze its function,an overexpression vector of PhTs2 was constructed and transformed into tobacco,then the transgenic tobaccos were identified:Phenotypic observation of transgenic tobaccos showed abnormal anther morphology and reduced pollen count;pollen vigor and germination rate were extremely reduced;the malformed pollen grains with irregular outer wall texture was observed by scanning electron microscopy;the semi-thin section showed the disorder of the tapetum development and the abnormal morphology of pollen grains. This study provides potential genetic resources for the cultivation of male sterile lines.

Key words: Petunia hybrida, fertility;anther, tapetum, male sterility, pollen