Overexpression of a Broccoli Defensin Gene BoDFN Enhances Downy Mildew Resistance

doi:10.1016/S1671-2927(00)8639

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (7): 1137-1144.DOI: 10.1016/S1671-2927(00)8639

Overexpression of a Broccoli Defensin Gene BoDFN Enhances Downy Mildew Resistance

JIANG Ming, HE Cai-ming, MIAO Li-xiang, ZHANG Yu-chao

1.College of Life Science, Taizhou University, Linhai 317000, P.R.China
2.Institute of Horticulture, Zhejiang Academy of Agricultural Science, Hangzhou 310021, P.R.China

收稿日期:2011-07-01 出版日期:2012-07-01 发布日期:2012-07-27
通讯作者: JIANG Ming, Tel: +86-576-85305853, Mobile: 15057232729, E-mail: jiangming1973@139.com
基金资助:
This study was supported by the Zhejiang Provincial Natural Science Foundation of China (Y3080081) and the Taizhou Science and Technology Project, China (08XH02).

Overexpression of a Broccoli Defensin Gene BoDFN Enhances Downy Mildew Resistance

JIANG Ming, HE Cai-ming, MIAO Li-xiang, ZHANG Yu-chao

1.College of Life Science, Taizhou University, Linhai 317000, P.R.China
2.Institute of Horticulture, Zhejiang Academy of Agricultural Science, Hangzhou 310021, P.R.China

Received:2011-07-01 Online:2012-07-01 Published:2012-07-27
Contact: JIANG Ming, Tel: +86-576-85305853, Mobile: 15057232729, E-mail: jiangming1973@139.com
Supported by:
This study was supported by the Zhejiang Provincial Natural Science Foundation of China (Y3080081) and the Taizhou Science and Technology Project, China (08XH02).

摘要/Abstract

摘要： Plant defensins are small, basic cysteine-rich peptides that play important roles in disease resistance. A gene, designated BoDFN, was isolated from Brassica oleracea var. italica. Gene sequence has been submitted to NCBI with an accession no. of HQ436486. Complete coding sequence of BoDFN is 243 bp in length encoding 80 amino acids. Sequence comparison results showed that BoDFN shared high homology with those of crucifer plants and there were only few DNA base differences. RT-PCR results indicated an increase of gene expression in Hyaloperonospora parasitica infected leaves and revealed a significant increase at 24 and 36 h of treatment. A recombinant plasmid, named pBI121-BoDFN, was constructed and introduced into Agrobacterium tumefacien LBA4404. PCR screening for 65 regenerated plantlets, 17 positive plantlets were obtained, PCR screening results revealed that 17 out of 65 regenerated plantlets were positive. Disease resistant identification results indicated that all positive plants showed an increase in resistance to H. parasitica.

关键词: broccoli, downy mildew, defensin, overexpression

Abstract: Plant defensins are small, basic cysteine-rich peptides that play important roles in disease resistance. A gene, designated BoDFN, was isolated from Brassica oleracea var. italica. Gene sequence has been submitted to NCBI with an accession no. of HQ436486. Complete coding sequence of BoDFN is 243 bp in length encoding 80 amino acids. Sequence comparison results showed that BoDFN shared high homology with those of crucifer plants and there were only few DNA base differences. RT-PCR results indicated an increase of gene expression in Hyaloperonospora parasitica infected leaves and revealed a significant increase at 24 and 36 h of treatment. A recombinant plasmid, named pBI121-BoDFN, was constructed and introduced into Agrobacterium tumefacien LBA4404. PCR screening for 65 regenerated plantlets, 17 positive plantlets were obtained, PCR screening results revealed that 17 out of 65 regenerated plantlets were positive. Disease resistant identification results indicated that all positive plants showed an increase in resistance to H. parasitica.

Key words: broccoli, downy mildew, defensin, overexpression

JIANG Ming, HE Cai-ming, MIAO Li-xiang, ZHANG Yu-chao. Overexpression of a Broccoli Defensin Gene BoDFN Enhances Downy Mildew Resistance[J]. Journal of Integrative Agriculture, 2012, 12(7): 1137-1144.

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Overexpression of a Broccoli Defensin Gene BoDFN Enhances Downy Mildew Resistance

Overexpression of a Broccoli Defensin Gene BoDFN Enhances Downy Mildew Resistance

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