蝴蝶兰PhPIF4基因的克隆与响应GA_(3)表达分析

Cloning and Expression Analysis in Response to GA_(3) of PhPIF4 in Phalaenopsis

蝴蝶兰是我国销售量最大的年宵盆花花卉,为了在蝴蝶兰生产栽培中对花期精准调控,挖掘成花关键基因,本文研究了生长调节剂GA_(3)和6-BA对大辣椒蝴蝶兰品种的成花影响,并对转录组数据中挖掘的PhPIF4基因进行克隆、生物信息学分析及对生长调节剂GA_(3)的响应表达分析。结果表明:PhPIF4具有bHLH_SF super family(cl00081)同源异型盒家族的典型保守结构域,大辣椒PhPIF4与小兰屿蝴蝶兰亲缘关系较近,其次为建兰、墨兰和石斛兰。PhPIF4基因编码蛋白定位于非叶绿体的细胞器中。GA_(3)处理增加了花梗长度,对花梗长度影响最大的处理组为200 mg/L GA_(3),GA_(3)可使花期提前9~11 d。6-BA可显著增加蝴蝶兰花量,花量最多的处理组为300 mg/L 6-BA,但对花期和花梗长度无显著作用。二者混合使用时,对蝴蝶兰开花时间和花期的影响介于二者之间。PhPIF4与蝴蝶兰花发育过程紧密相关,随着花发育进程其表达量逐渐加倍。喷施生长调节剂GA_(3)后,PhPIF4基因在花芽、叶和根中的表达量均显著增加,且在现蕾期增加量最大,表明PhPIF4基因的表达可能受GA_(3)影响与调控,参与了赤霉素成花途径。本研究为揭示PhPIF4在赤霉素调控生长发育的调控网络中的作用奠定基础。

Phalaenopsis is the most popular Spring Festival pot flower in China.In order to accurately regulate the flowering stage and excavate the key flowering gene in the production and cultivation of Phalaenopsis,this paper stud-ied the effects of growth regulators GA_(3)and 6-BA on the flowering of Phalaenopsis Big Chilli,and the PhPIF4 gene selected from transcriptome data was cloned,and its bioinformatics analysis and response to growth regulator GA_(3)were studied.The results showed that PhPIF4 had a typical conserved domain of the bHLH_SF super family(cl00081)ho-mobox family.PhPIF4 of Big Chilli is closely related to Phalaenopsis equestris,followed by Cymbidium ensifolium,C,sinense and Dendrobium catenatum.The protein encoded by PhPIF4 was localized in non-chloroplast organelles.GA_(3)treatment increased stalk length and the maximum stalk length of the treatment group was 200 mg/L GA_(3).GA_(3)could bring forward flowering 9-11 d.6-BA significantly increased the number of flowers,and the treatment group with the most flowering amount was 300mg/L 6-BA,but 6-BA had no significant effect on flowering time and stalk length.When the two growth regulators were mixed to spray,the effect on flowering time and flowering time of Phalaenopsis was between the two.PhPIF4 was closely related to the development of flowering,and its expression gradually increased with the development of flowering.After spraying GA_(3),the expression of PhPIF4 in flower buds,leaves and roots increased,and the increase was the largest in flower bud stage,suggesting that the expression of PhPIF4 might be affected and regulated by GA_(3),and PhPIF4 might participate in the gibberellin flowering pathway.In this paper,the effects of GA_(3)and 6-BA on reproductive growth of Phalaenopsis were studied,and the expression of PhPIF4 in response to GA was investigated,which would lay a foundation for revealing the role of PhPIF4 in gibberellin regulation of growth and development regulatory network.