Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (16): 3384-3391.doi: 10.3864/j.issn.0578-1752.2013.16.009

• PLANT PROTECTION • Previous Articles     Next Articles

Molecular Clone and Its RNA Interference Effect Analysis of Lactate Dehydrogenase Gene in Meloidogyne incognita

MEI Mei1,2, HUANG Yong-hong2,3, MAO Zhen-chuan2, LIU Zhi-min1,4, XIE Bing-yan2   

  1. 1.College of Horticulture and Landscape, Hunan Agriculture University, Changsha 410128
    2.Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081
    3.Fruit Research Institute, Guangdong Academy of Agricultural Science/Key Laboratory of Biology and Genetic Resource Utilization of Fruit Tree in South Subtropics, Ministry of Agriculture, Guangzhou 510640
    4.Hunan First Normal University, Changsha 410205
  • Received:2013-02-01 Online:2013-08-15 Published:2013-05-10

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Abstract: 【Objective】The objective of this study is to clone and analyze the lactate dehydrogenase gene in Meloidogyne incognita (mildh), and to determine the inhibitory effect of mildh silencing on the diseases caused by Meloidogyne spp..【Method】In the preliminary studies, the number of RNA interference (RNAi) phenotype genes of nematodes in their genomes were predicted by bioinformatics methods. In this study, using special primers designed based on the predicted nematode ldh, the ldh in M. incognita (mildh) were cloned which was transferred into tomato plantlets by virus-induced gene silence (VIGS) technique after analyzing the sequence characters to investigate the influence of the mildh silencing on the amounts of root knot on the tomato plantlet inoculated with M. incognita.【Result】The cloned mildh contained a complete coding sequence that coded 315 amino acids. The identity between the cloned mildh and the predicted mildh was as high as 94.9% in nucleotide sequence and 95.5% in the deduced amino acid sequence. The silencing vector pTV-mildhi, mediated by Tobacco rattle virus (TRV), was constructed. The tomato plantlets were inoculated with M. incognita after treated by pTV-mildhi. Sixty days later, statistical analysis showed that the amounts of root knot on tomato plantlets of which the mildh was silenced decreased by 48.6% compared to that of the empty vector control, by 48.4% compared to that of the water control. 【Conclusion】The study revealed the mildh silencing had a significant inhibitory effect on root-knot nematode disease caused by M. incognita. It also showed that mildh genes may be involved in the pathogenesis of nematode, which provided a theoretical basis to the research of nematode pathology and to the control of nematode disease.

Key words: Meloidogyne spp. , lactate dehydrogenase , virus-induced gene silence , RNA interference

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