Jellyfish Green Fluorescent Protein (GFP) as a Reporter for Fusarium gramminearum Development on Wheat

doi:10.1016/S2095-3119(14)60875-3

Journal of Integrative Agriculture ›› 2014, Vol. 13 ›› Issue (10): 2177-2183.DOI: 10.1016/S2095-3119(14)60875-3

Jellyfish Green Fluorescent Protein (GFP) as a Reporter for Fusarium gramminearum Development on Wheat

QI Jun-xian, LIU Tai-guo, XU Ying, CHEN Huai-gu, GAO Li, LIU Bo , CHEN Wan-quan

1、State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences,
Beijing 100193, P.R.China
2、Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, P.R.China

收稿日期:2013-05-15 出版日期:2014-10-01 发布日期:2014-10-10
通讯作者: LIU Tai-guo, E-mail: tgliu@ippcaas.cn; CHEN Wan-quan, E-mail: wqchen@ippcaas.cn
作者简介:QI Jun-xian, E-mail: qijunxian521@126.com
基金资助:
The work was supported in part by the Special Fund for Agro-scientific Research in the Public Interest (201303016) and the China Agriculture Research System (CARS-03) from the Ministry of Agriculture of China; in part by the Project of International Scientific and Technical Cooperation (2013DFG31930), the National Key Technologies Research and Development Program of China (2012BAD19B04), the Breeding and Cultivation of Novel GM Varieties (2013ZX08002001), and 863 Program (2012AA101501) from the Ministry of Science and Technology of China.

Jellyfish Green Fluorescent Protein (GFP) as a Reporter for Fusarium gramminearum Development on Wheat

QI Jun-xian, LIU Tai-guo, XU Ying, CHEN Huai-gu, GAO Li, LIU Bo , CHEN Wan-quan

1、State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences,
Beijing 100193, P.R.China
2、Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, P.R.China

Received:2013-05-15 Online:2014-10-01 Published:2014-10-10
Contact: LIU Tai-guo, E-mail: tgliu@ippcaas.cn; CHEN Wan-quan, E-mail: wqchen@ippcaas.cn
About author:QI Jun-xian, E-mail: qijunxian521@126.com
Supported by:
The work was supported in part by the Special Fund for Agro-scientific Research in the Public Interest (201303016) and the China Agriculture Research System (CARS-03) from the Ministry of Agriculture of China; in part by the Project of International Scientific and Technical Cooperation (2013DFG31930), the National Key Technologies Research and Development Program of China (2012BAD19B04), the Breeding and Cultivation of Novel GM Varieties (2013ZX08002001), and 863 Program (2012AA101501) from the Ministry of Science and Technology of China.

摘要/Abstract

摘要： The plasmid pGPDGFP under the control of pgpdA promotor was used together with vector pAN7-1 containing the hygromycin resistance cassette to co-transform protoplasts of HG1, Fusarium graminearum from Hubei Province, China. Twelve out of 14 hygromycin-resistant transformants showed green signal under the UV light and contained one or several copies of gfp, as indicated by Southern analysis of genomic DNA digested with different restriction enzymes and hybridized to the gfp probe. A single gfp copy transformant (HG1C5) was selected for further evaluation of 80 Chinese wheat cultivars or advanced lines. The results showed different resistance type to F. graminearum were observed. GFP signals observed in the rachis and adjacent spikes of 70 Chinese wheat lines such as Chuanchongzu 104 indicated both type I (host resistance to the initial infection by the fungus) and type II (resistance to the spread of FHB symptoms within an infected spike) were not observed. While other 10 lines showed type II resistance to F. graminearum with GFP signals only in inoculated spikelets. Development of the mycelium can be intuitively observed and the resistance of wheat to F. graminearum can be identified at 7 days post inoculation (dpi) in this way. The results showed no differences were evaluated between the transformed HG1C5 and the non-transgene artificial inoculation by SAS paired chi-square test and McNemar’s test (P=0.0625).

关键词: Fusarium head blight (FHB) , green fluorescent protein (GFP) , resistance evaluation

Abstract: The plasmid pGPDGFP under the control of pgpdA promotor was used together with vector pAN7-1 containing the hygromycin resistance cassette to co-transform protoplasts of HG1, Fusarium graminearum from Hubei Province, China. Twelve out of 14 hygromycin-resistant transformants showed green signal under the UV light and contained one or several copies of gfp, as indicated by Southern analysis of genomic DNA digested with different restriction enzymes and hybridized to the gfp probe. A single gfp copy transformant (HG1C5) was selected for further evaluation of 80 Chinese wheat cultivars or advanced lines. The results showed different resistance type to F. graminearum were observed. GFP signals observed in the rachis and adjacent spikes of 70 Chinese wheat lines such as Chuanchongzu 104 indicated both type I (host resistance to the initial infection by the fungus) and type II (resistance to the spread of FHB symptoms within an infected spike) were not observed. While other 10 lines showed type II resistance to F. graminearum with GFP signals only in inoculated spikelets. Development of the mycelium can be intuitively observed and the resistance of wheat to F. graminearum can be identified at 7 days post inoculation (dpi) in this way. The results showed no differences were evaluated between the transformed HG1C5 and the non-transgene artificial inoculation by SAS paired chi-square test and McNemar’s test (P=0.0625).

Key words: Fusarium head blight (FHB) , green fluorescent protein (GFP) , resistance evaluation

QI Jun-xian, LIU Tai-guo, XU Ying, CHEN Huai-gu, GAO Li, LIU Bo , CHEN Wan-quan. Jellyfish Green Fluorescent Protein (GFP) as a Reporter for Fusarium gramminearum Development on Wheat[J]. Journal of Integrative Agriculture, 2014, 13(10): 2177-2183.

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Jellyfish Green Fluorescent Protein (GFP) as a Reporter for Fusarium gramminearum Development on Wheat

Jellyfish Green Fluorescent Protein (GFP) as a Reporter for Fusarium gramminearum Development on Wheat

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