title: Jasmonate-Sensitivity-Assisted Screening and Characterization of Nicotine Synthetic Mutants from Activation-Tagged Population of Tobacco (Nicotiana tabacumL.).
abstract: Nicotine is a secondary metabolite that is important to the defense system and commercial quality of tobacco (Nicotiana tabacumL.). Jasmonate and its derivatives (JAs) are phytohormone regulators of nicotine formation; however, the underlying molecular mechanism of this process remains largely unclear. Owing to the amphitetraploid origin ofN. tabacum, research on screening and identification of nicotine-synthetic mutants is relatively scarce. Here, we describe a method based on JA-sensitivity for screening nicotine mutants from an activation-tagged population of tobacco. In this approach, the mutants were first screened for abnormal JA responses in seed germination and root elongation, and then the levels of nicotine synthesis and expression of nicotine synthetic genes in the mutants with altered JA-response were measured to determine the nicotine-synthetic mutants. We successfully obtained five mutants that maintained stable nicotine contents and JA responses for three generations. This method is simple, effective and low-cost, and the finding of transcriptional changes of nicotine synthetic genes in the mutants shows potentials for identifying novel regulators involved in JA-regulated nicotine biosynthesis.
authors.affiliation: College of Agronomy and Biotechnology, Southwest University Chongqing, China.
authors.forename: Guoying
authors.initials: G
authors.lastname: Yin
authors.affiliation: Tobacco Research Institute, Chinese Academy of Agricultural Sciences Qingdao, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Agronomy and Biotechnology, Southwest University Chongqing, China.
authors.forename: Haixia
authors.initials: H
authors.lastname: Niu
authors.affiliation: College of Agronomy and Biotechnology, Southwest University Chongqing, China.
authors.forename: Yongqiang
authors.initials: Y
authors.lastname: Ding
authors.affiliation: College of Agronomy and Biotechnology, Southwest University Chongqing, China.
authors.forename: Dingyu
authors.initials: D
authors.lastname: Zhang
authors.affiliation: College of Agronomy and Biotechnology, Southwest University Chongqing, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Tobacco Research Institute, Chinese Academy of Agricultural Sciences Qingdao, China.
authors.forename: Guanshan
authors.initials: G
authors.lastname: Liu
authors.affiliation: College of Agronomy and Biotechnology, Southwest University Chongqing, China.
authors.forename: Sangen
authors.initials: S
authors.lastname: Wang
authors.affiliation: Tobacco Research Institute, Chinese Academy of Agricultural Sciences Qingdao, China.
authors.forename: Hongbo
authors.initials: H
authors.lastname: Zhang
keywords: activation-tagged population; jasmonate-sensitivity; mutant; nicotine; tobacco