Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (10): 1882-1891.doi: 10.3864/j.issn.0578-1752.2015.10.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Functional Analysis of COI1 Genes in Oilseed Rape （Brassica napus L.）

WANG Wen-jing, YANG Xiao-chuan, DING Yong-qiang, YIN Guo-ying, MA Hao-ran, ZHANG Jie, SHI Xiao-yu, ZHANG Ding-yu, LI Jia-na, ZHANG Hong-bo   

  1. College of Agronomy and Biotechnology, Southwest University/Chongqing Engineering Research Center for Rapeseed, Chongqing 400716
  • Received:2014-12-04 Online:2015-05-16 Published:2015-05-16

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Abstract: 【Objective】 CORONATINE INSENSITIVE 1 (COI1) is a critical component of jasmonate (JA) receptor complex. The objective of this study is to investigate the spatial expression pattern and regulatory roles of COI1 gene in oilseed rape(Brassica napus), a staple oil crop in the world. 【Method】 The COI1 genes in oilseed rape(B. napus) and its parental species B. rapa and B. oleracea were analyzed based on the genomic data. The spatial transcription pattern of COI1 in oilseed rape was analyzed by RT-PCR with specific primers according to the conserved region of COI1 gene homologs. Then, a cDNA fragment of the conserved region was cloned into vector pTRV2 of the tobacco rattle virus-based VIGS (virus-induced gene silencing) system. And, VIGS techniques were applied to silence COI1 genes in oilseed rape. The rape plants shown to have COI1 genes silenced were used to investigate male fertility and aphid resistance. 【Result】 Analysis of the genomic data of B. rapa, B. oleracea, and B. napus revealed that the genomes of B. rapa and B. oleracea contained 7 highly homologous COI1 genes, which could be classified into 4 subgroups, i.e., COI1a, COI1b.1, COI1b.2, and COI1c, while 8 COI1 genes were presented in the genome of B. napus. Transcriptional assay revealed that the expression of COI1 in oilseed rape is tissue specific. A 505bp fragment of the conserved region of COI1 was introduced into the VIGS vector pTRV2 to develop COI1-silenced plants via VIGS techniques. Twenty-five plants with the transcripts of COI1 down-regulated by over 70% were identified by transcriptional analyses, from which the ten plants with the lowest expression level of COI1 were selected to investigate male fertility and aphid resistance. The results showed that the fertility of COI1-silenced plants was extremely impaired, with no seeds in their siliques. Furthermore, the filaments of COI1-silenced plants were shorter than those of control plants, and over 80% pollens of the COI1-silenced plants were irregular in shape. Aphid resistance assay showed that aphids grew and reproduced much faster on the COI1-silenced plants than on the control plants cultured under the same condition. 【Conclusion】 This study revealed that the expression of COI1 gene in oilseed rape is tissue specific. VIGS-induced silencing of COI1 greatly impaired the male fertility and decreased the aphid resistance of oilseed rape. This work has provided an important start point to dissect the mechanism of JA signaling in oilseed rape.

Key words: Brassica napus L., COI1 gene, jasmonate, male sterility, aphid resistance

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