|本期目录/Table of Contents|

[1]孙家旗,唐维,刘永胜. 猕猴桃CHS基因RNA干涉载体在果实中的瞬时表达可以有效影响花青素积累[J].应用与环境生物学报,2014,20(05):929-933.[doi:10.3724/SP.J.1145.2014.03021]
 SUN Jiaqi,TANG Wei,LIU Yongsheng. Transient expression of CHS-RNAi effectively influences the accumulation of anthocyanin in fruit of kiwifruit (Actinidia chinensis)[J].Chinese Journal of Applied & Environmental Biology,2014,20(05):929-933.[doi:10.3724/SP.J.1145.2014.03021]





 Transient expression of CHS-RNAi effectively influences the accumulation of anthocyanin in fruit of kiwifruit (Actinidia chinensis)
 SUN Jiaqi TANG Wei LIU Yongsheng
1Ministry of Education Key Laboratory for Bio-resource and Eco-environment, College of Life Sciences, State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, China,
2School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei 230009, China
1四川大学生命科学学院生物资源与生态环境教育部重点实验室水力学与山区河流工程国家重点实验室 成都 6100642合肥工业大学生物技术与食品工程学院 合肥 230009
 kiwifruit CHS anthocyanin transient expression qPCR
近来猕猴桃基因组草图已经完成,然而对猕猴桃功能基因的研究还非常有限. 我们拟建立RNA干扰(RNAi)系统来研究猕猴桃的基因功能. 将红阳猕猴桃中编码查耳酮合成酶的CHS基因片段插入到载体pHB 和pTCK303中,构建35S::CHS-RNAi 和UBI::CHS-RNAi载体. 将两个重组质粒转入根癌土壤杆菌EHA105中,分别将0.7 mL农杆菌(D = 0.8)从果实底部注入花后106 d(果肉颜色开始变红)的猕猴桃果实中. 5 d后,与空载体相比,注射了CHS-RNAi载体的猕猴桃果肉颜色变浅,花青素含量、CHS的表达量显著降低. 预测这种快捷的瞬时表达系统将为猕猴桃基因功能研究提供有效手段.
Functional genomic studies on kiwifruit are very limited despite the fact that its genomic data is available. In the present study, we intended to establish an efficient system to investigate gene function by using RNA interference (RNAi) technique. A segment of cDNA putatively coding for chalcone synthase (CHS) derived from kiwifruit cv. Hongyang (Actinidia chinensis) was inserted into vectors pHB and pTCK303, to produce recombinant constructs 35S::CHS-RNAi and UBI::CHS-RNAi, respectively. The two recombinant plasmids were transformed into Agrobacterium tumefaciens EHA105 by liquid nitrogen. These two engineered A. tumefaciens strains were subsequently injected into the kiwifruits 106 days after full bloom (DAFB) when the red pigment of inner pericarp started to accumulate. About 0.7 mL engineered A. tumefaciens culture (about OD 0.8) was injected through fruit bottom into core tissue. As a result, 5 days after injection, as compared with blank control, a significant reduction of red pigment accumulation was visualized in inner pericarps injected with the engineered A. tumefaciens strains containing CHS-RNAi vectors, with remarkably decreased contents of anthocyanins detected at the same time. In addition, the CHS expression level was lower with the RNAi repression than that injected with empty vector. We anticipate that this rapid and transient expression system will be useful for studying the function of plant structural and regulatory genes in kiwifruit.


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 四川省科技厅科技支撑项目(2013NZ0014)和973计划重大项目(2011CB100401)资助 Supported by the Key Project from the Government of Sichuan Province (2013NZ0014), and the National Basic Research Program of China (973 Program) (2011CB100401)
更新日期/Last Update: 2014-10-30