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中国油料作物学报 ›› 2019, Vol. 41 ›› Issue (3): 340-.doi: 10.7505/j.issn.1007-9084.2019.03.005

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Construction of a lhRNAi library related to rapeseed flower development mediated by rolling circle replication

  

  1. Oil Crops Research Institute of Chinese Academy of Agricultural Sciences / Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, 430062, China
  • Online:2019-06-28 Published:2019-09-02

Abstract:

Allopolyploid B. napus contains a large number of homologous and redundant genes. It is difficulty to identify novel genes related to important agronomic traits by traditional T-DNA insertional, physical and chemical mutagenesis methods. In this study, a genome-wide inflorescence lhRNAi library of B. napus was constructed by the rolling circle amplification (RCA) - mediated lhRNAi construction technology. Subsequently, the inflorescence lhRNAi library was transformed into B. napus to produce 763 T0 transgetic plants. A total of 74 transgenetic lines exhibited visible mutant phenotypes, including petal reduction in the number, stamen degeneration shrinkage and number reduction, stigma distortion and exposure, dead or closed buds and male sterility. These results demonstrated the construction method of lhRNAi library mediated by rolling circle replication and were successfully applied to rapeseed, which provided an important research platform for the study of gene functions related to rape flower inflorescence development.

Key words: Brassica napus L., rolling circle amplification, long hairpin RNAi library;flower development