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Plant function and evolutionary biology
RESEARCH ARTICLE

Selectable marker-free co-expression of Nicotiana rustica CN and Nicotiana tabacum HAK1 genes improves resistance to tobacco mosaic virus in tobacco

Li-Jun Qin A B , Dan Zhao A , Yi Zhang B and De-Gang Zhao A C
+ Author Affiliations
- Author Affiliations

A The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education), Institute of Agro-Bioengineering and College of Life Sciences, Guizhou University, Guiyang 550025, Guizhou Province, People’s Republic of China.

B The State Key Laboratory Breeding Base of Green Pesticide and Agricultural Biological Engineering, Guizhou University, Guiyang, 550025, Guizhou Province, People’s Republic of China.

C Corresponding author. Email: dgzhao@gzu.edu.cn

Functional Plant Biology 42(8) 802-815 https://doi.org/10.1071/FP14356
Submitted: 17 December 2014  Accepted: 29 April 2015   Published: 4 June 2015

Abstract

The viral disease caused by tobacco mosaic virus (TMV) is the most prevalent viral disease in many tobacco production areas. A breeding strategy based on resistance genes is an effective method for improving TMV resistance in tobacco. Also, the physiological status of plants is also critical to disease resistance improvement. Potassium ion is one of the most abundant inorganic nutrients in plant cells, and mediates plant responses to abiotic and biotic stresses. Improving K+ content in soil by fertilising can enhance diseases resistance of crops. However, the K+ absorption in plants depends mostly on K+ transporters located in cytoplasmic membrane. Therefore, the encoding genes for K+ transporters are putative candidates to target for improving tobacco mosaic virus resistance. In this work, the synergistic effect of a N-like resistance gene CN and a tobacco putative potassium transporter gene HAK1 was studied. The results showed that TMV-resistance in CN-HAK1-containing tobaccos was significantly enhanced though a of strengthening leaf thickness and reduction in the size of necrotic spots compared with only CN-containing plants, indicating the improvement of potassium nutrition in plant cells could increase the tobacco resistance to TMV by reducing the spread of the virus. Quantitative real-time polymerase chain reaction (qRT–PCR) analysis for TMV-CP expression in the inoculated leaf of the transgenic and wild-type plants also supported the conclusion. Further, the results of defence-related determination including antioxidative enzymes (AOEs) activity, salicylic acid (SA) content and the expression of resistance-related genes demonstrated CN with HAK1 synergistically enhanced TMV-resistance in transgenic tobaccos. Additionally, the HAK1- overexpression significantly improved the photosynthesis and K+-enriching ability in trans-CN-HAK1 tobaccos, compared with other counterparts. Finally, this work provides a method for screening new varieties of marker-free and safe transgenic antiviral tobacco.

Additional keywords: TMV resistance, potassium-enrichment, co-transformation, tobaccos.


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