首页|Development of a rapid and efficient system for CR genes identification based on hairy root transformation in Brassicaceae

Development of a rapid and efficient system for CR genes identification based on hairy root transformation in Brassicaceae

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Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection.Breeding of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this disease.The traditional way of R gene functional validation requires stable transformation that is both time-and labor-consuming.In this study,a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was developed.The transformation positive rate was over 80％in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable transformation.The system was applicable to different B.napus varieties and other cruciferous crops including Brassica rapa and Brassica oleracea.In particular,two known CR genes,CRA3.7.1 and CRA8.2.4 were used respectively,as example to show that the system works well for CR gene study combined with subsequent P.brassicae infection in B.napus.Most importantly,it works both in over-expression that led to disease resistance,as well as in RNAi which led to disease susceptible phenotype.Therefore,this system can be used in batch-wise identification of CR genes,and also offered the possibility of manipulating key genes within the P.brassicae genome that could improve our knowledge on host-pathogen interaction.

BrassicaceaeAgrobacterium rhizogenesHairy root transformationClubrootGene function

Wenlin Yu、Lu Yang、Yuanyuan Xiang、Rongde Li、Xueqing Zhou、Longcai Gan、Xianyu Xiang、Yunyun Zhang、Lei Yuan、Yanqing Luo、Genze Li、Youning Wang、Yinhua Chen、Peng Chen、Chunyu Zhang

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National Key Laboratory of Crop Genetic Improvement and College of Plant Science and Technology,Huazhong Agricultural University,Wuhan,Hubei 430070,China

National Agro-Tech Extension and Service Center,Beijing 100125,China

Industrial Crops Research Institute,Yunnan Academy of Agricultural Sciences,Kunming,Yunnan 650221,China

College of Agronomy,Northwest A & F University,Yangling,Shaanxi 712100,China

National Key Laboratory for Tropical Crop Breeding,Sanya Nanfan Research Institute,Hainan University,Sanya,Hainan 572025,China

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Wuhan Science and Technology Major Project on Key techniques of biological breeding and Breeding of new varietiesspecial project for sustainable development agenda of innovation demonstration zoneNational Key R & D Program of China

2022021302024851202204AC100001-A042022YFD1200400

2024

10.1016/j.hpj.2024.05.002

园艺学报(英文版)

园艺学报(英文版)

CSTPCD
ISSN：
年,卷(期)：2024.10(4)
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