Genetic transformation of Citrus sinensis cv. Hamlin with hrpN gene from Erwinia amylovora and evaluation of the transgenic lines for resistance to citrus canker

被引：40

作者：

Barbosa-Mendes, Janaynna Magalhaes ^{[2
]}

Alves Mourao Filho, Francisco de Assis ^{[2
]}

Bergamin Filho, Armando ^{[2
]}

Harakava, Ricardo ^{[3
]}

Beer, Steven V. ^{[4
]}

Januzzi Mendes, Beatriz Madalena ^{[1
]}

机构：

[1] Univ Sao Paulo, Ctr Energia Nucl Agr, BR-13400970 Piracicaba, SP, Brazil

[2] Univ Sao Paulo, Escola Super Agr Luiz de Queiroz, BR-13418900 Piracicaba, SP, Brazil

[3] Inst Biol Sao Paulo, BR-04014002 Sao Paulo, Brazil

[4] Cornell Univ, Ithaca, NY 14853 USA

来源：

SCIENTIA HORTICULTURAE | 2009年 / 122卷 / 01期

基金：

巴西圣保罗研究基金会;

关键词：

Disease resistance; Harpin; Sweet orange; Xanthomonas axonopodis pv. citri; HYPERSENSITIVE RESPONSE; FIRE BLIGHT; DISEASE RESISTANCE; SWEET ORANGE; TOBACCO; PROTEIN; PLANTS; ACTIVATION; EXPRESSION; PROMOTER;

D O I：

10.1016/j.scienta.2009.04.001

中图分类号：

S6 [园艺];

学科分类号：

0902 ;

摘要：

Transgenic Citrus sinensis (L.) Osb. cv. Hamlin plants expressing the hrpN gene were obtained by Agrobacterium tumefaciens (Smith and Towns) Conn-mediated transformation. hrpN encodes a harpin protein, which elicits the hypersensitive response and systemic acquired resistance in plants. The gene construct consisted of gst1, a pathogen-inducible promoter, a signal peptide for protein secretion to the apoplast, the selection genes nptI1 or aacC1 and the Nos terminator. The function of gst1 in citrus was evaluated in transgenic C. sinensis cv. Valencia harboring the reporter gene uidA (gus) driven by this promoter. Histochemical analysis for gus revealed that gst1 is activated in citrus leaves by both wounding and inoculation with Xanthomonas axonopodis Starr and Garces pv. citri (Hasse) Vauterin et al. Genetic transformation was confirmed by Southern blot hybridization in eight cv. Hamlin acclimatized plants. RT-PCR confirmed hrpN gene expression in seven cv. Hamlin transgenic lines before pathogen inoculation. Some hrpN transgenic lines showed severe leaf curling and abnormal growth. Six hrpN transgenic lines were propagated and evaluated for susceptibility to X axonopodis pv. citri. RT-PCR confirmed gene expression in all six hrpN transgenic lines after pathogen inoculation. Several of the hrpN transgenic lines showed reduction in susceptibility to citrus canker as compared with non-transgenic plants. One hrpN transgenic line exhibited normal vegetative development and displayed very high resistance to the pathogen, estimated as up to 79% reduction in disease severity. This is the first report of genetic transformation of citrus using a pathogen-inducible promoter and the hrpN gene. Further evaluations of the transgenic plants under field conditions are planned. Nevertheless, the evidence to date suggests that the hrpN gene reduces the susceptibility of citrus plants to the canker disease. (C) 2009 Elsevier B.V. All rights reserved.

引用

页码：109 / 115

页数：7

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