Production of Transgenic Anliucheng Sweet Orange (Citrus sinensis Osbeck) with Xa21 Gene for Potential Canker Resistance

doi:10.1016/S2095-3119(13)60675-9

Journal of Integrative Agriculture ›› 2014, Vol. 13 ›› Issue (11): 2370-2377.DOI: 10.1016/S2095-3119(13)60675-9

Production of Transgenic Anliucheng Sweet Orange (Citrus sinensis Osbeck) with Xa21 Gene for Potential Canker Resistance

LI Ding-li, XIAO Xuan, GUO Wen-wu

1、Key Laboratory of Horticultural Plant Biology, Ministry of Education/College of Horticulture & Forestry Sciences, Huazhong Agricultural
University, Wuhan 430070, P.R.China
2、College of Life and Environmental Sciences, Gannan Normal University, Ganzhou 341000, P.R.China

收稿日期:2013-10-28 出版日期:2014-11-06 发布日期:2014-11-14
通讯作者: GUO Wen-wu, Tel: 86-27-87281543, Fax: 86-27-87280622,E-mail: guoww@mail.hzau.edu.cn
作者简介:LI Ding-li, E-mail: qauldl@163.com; XIAO Xuan, E-mail: xiaoxuanxx@163.com;* These authors contributed equally to this study.
基金资助:
This research was financially supported by the National High- Tech R&D Program of China (863, 2011AA100205), and the National Natural Science Foundation of China (31125024). The authors thank Prof. Zhu Lihuang (the Institute of Genetics & Developmental Biology, Chinese Academy of Sciences, Beijing) for providing the plasmid pCXK1301.

Production of Transgenic Anliucheng Sweet Orange (Citrus sinensis Osbeck) with Xa21 Gene for Potential Canker Resistance

LI Ding-li, XIAO Xuan, GUO Wen-wu

1、Key Laboratory of Horticultural Plant Biology, Ministry of Education/College of Horticulture & Forestry Sciences, Huazhong Agricultural
University, Wuhan 430070, P.R.China
2、College of Life and Environmental Sciences, Gannan Normal University, Ganzhou 341000, P.R.China

Received:2013-10-28 Online:2014-11-06 Published:2014-11-14
Contact: GUO Wen-wu, Tel: 86-27-87281543, Fax: 86-27-87280622,E-mail: guoww@mail.hzau.edu.cn
About author:LI Ding-li, E-mail: qauldl@163.com; XIAO Xuan, E-mail: xiaoxuanxx@163.com;* These authors contributed equally to this study.
Supported by:
This research was financially supported by the National High- Tech R&D Program of China (863, 2011AA100205), and the National Natural Science Foundation of China (31125024). The authors thank Prof. Zhu Lihuang (the Institute of Genetics & Developmental Biology, Chinese Academy of Sciences, Beijing) for providing the plasmid pCXK1301.

摘要/Abstract

摘要： Citrus canker, an epidemic quarantine disease caused by Xanthomonas axonopodis pv. citri, has brought a great damage in citrus production worldwide. Herein, a rice PRR (pattern recognition receptor) gene Xa21 together with GUS reporter gene and hygromycin phosphotransferase gene (HPT) was introduced into Anliucheng sweet orange (Citrus sinensis Osbeck) via Agrobacterium-mediated transformation of embryogenic callus. The transgenic calluses were screened on MT basal medium containing hygromycin (HYG) and detected by histochemical GUS staining. The transgenic plantlets were recovered through somatic embryogenesis pathway. The regenerated plantlets were accustomed to and maintained in the greenhouse. The transgene integration of recovered plantlets was identified by PCR and Southern blot hybridization. It showed that all the transgenic plantlets tested had undergone single copy integration, the expression of Xa21 in eight different transgenic lines detected by qRT-PCR can be divided into three grades, high for T5 and T6, middle for T4 and low for the rest. The tolerance to citrus canker disease of the three recovered transgenic lines T2, T4 and T6 was assessed by in vitro pin-puncture inoculation. The results showed that all the three transgenic lines conferred improved resistance to citrus canker bacterium infection and the T4 transgenic line displayed the highest resistance. The mechanism and feasibility of rice Xa21 in triggering innate immunity in citrus was briefly discussed.

关键词: citrus canker , Xa21 , sweet orange , pattern recognition receptors (PRRs)

Abstract: Citrus canker, an epidemic quarantine disease caused by Xanthomonas axonopodis pv. citri, has brought a great damage in citrus production worldwide. Herein, a rice PRR (pattern recognition receptor) gene Xa21 together with GUS reporter gene and hygromycin phosphotransferase gene (HPT) was introduced into Anliucheng sweet orange (Citrus sinensis Osbeck) via Agrobacterium-mediated transformation of embryogenic callus. The transgenic calluses were screened on MT basal medium containing hygromycin (HYG) and detected by histochemical GUS staining. The transgenic plantlets were recovered through somatic embryogenesis pathway. The regenerated plantlets were accustomed to and maintained in the greenhouse. The transgene integration of recovered plantlets was identified by PCR and Southern blot hybridization. It showed that all the transgenic plantlets tested had undergone single copy integration, the expression of Xa21 in eight different transgenic lines detected by qRT-PCR can be divided into three grades, high for T5 and T6, middle for T4 and low for the rest. The tolerance to citrus canker disease of the three recovered transgenic lines T2, T4 and T6 was assessed by in vitro pin-puncture inoculation. The results showed that all the three transgenic lines conferred improved resistance to citrus canker bacterium infection and the T4 transgenic line displayed the highest resistance. The mechanism and feasibility of rice Xa21 in triggering innate immunity in citrus was briefly discussed.

Key words: citrus canker , Xa21 , sweet orange , pattern recognition receptors (PRRs)

LI Ding-li, XIAO Xuan, GUO Wen-wu. Production of Transgenic Anliucheng Sweet Orange (Citrus sinensis Osbeck) with Xa21 Gene for Potential Canker Resistance[J]. Journal of Integrative Agriculture, 2014, 13(11): 2370-2377.

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Production of Transgenic Anliucheng Sweet Orange (Citrus sinensis Osbeck) with Xa21 Gene for Potential Canker Resistance

Production of Transgenic Anliucheng Sweet Orange (Citrus sinensis Osbeck) with Xa21 Gene for Potential Canker Resistance

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