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

PDF

713

Cited

2

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

[1]Naz I, Palomares-Rius J E, Saifullah, Blok V, Khan M R, Ali S. Ali S. In vitro and in planta nematicidal activity of Fumaria parviflora (Fumariaceae) against the southern root-knot nematode Meloidogyne incognita. Plant Pathology, 2012, Doi: 10.1111/j.1365-3059.2012. 02682.x.

[2]Dang Q L, Kim W K, Nguyen C M, Choi Y H, Choi G J, Jang K S, Park M S, Lim C H, Luu N H, Kim J. Nematicidal and antifungal activities of annonaceous acetogenins from Annona squamosa against various plant pathogens. Journal of Agricultural and Food Chemistry, 2011, 59: 11160-11167.

[3]Caboni P, Ntalli N G, Aissani N, Cavoski I, Angioni A. Nematicidal activity of (E,E)-2,4-decadienal and (E)-2-decenal from Ailanthus altissima against Meloidogyne javanica. Journal of Agricultural and Food Chemistry, 2012, 60: 1146-1151.

[4]Oka Y, Shuker S, Tkachi N. Nematicidal efficacy of MCW-2, a new nematicide of the fluoroalkenyl group, against the root-knot nematode Meloidogyne javanica. Pest Management Science, 2009, 65(10): 1082-1089.

[5]Mafeo T P, Mashela P W. Responses of germination in tomato, watermelon and butternut squash to a Cucumis bio-nematicide. American-Eurasian Journal of Agricultural and Environmental Science, 2009, 6(2): 215-219.

[6]Eisenhauer N, Ackermann M, Gass S, Klier M, Migunova V, Nitschke N, Ruess L, Sabais A C W, Weisser W W, Scheu S. Nematicide impacts on nematodes and feedbacks on plant productivity in a plant diversity gradient. Acta Oecologica, 2010, 36: 477-483.

[7]郑玉才, 赵兴波, 周静, 朴影, 金素钰, 贺庆华, 洪键, 李宁, 吴常信. 牦牛乳酸脱氢酶-1 两种遗传变异体的纯化及酶学特性比较. 中国农业科学, 2009, 42(3): 1047-1052.

Zheng Y C, Zhao X B, Zhou J, Piao Y, Jin S Y, He Q H, Hong J, Li N, Wu C X. Purification and enzyme property comparison of two genetic variants of yak lactate dehydrogenase-1. Scientia Agricultura Sinica, 2009, 42(3): 1047-1052. (in Chinese)

[8]Le A, Cooper C R, Gouw A M, Dinavahi R, Maitra A, Deck L M , Royer R E, Vander Jagt L V, Semenza G L, Dang C V. Inhibition of lactate dehydrogenase A induces oxidative stress and inhibits tumor progression. Proceedings of the National Academy of Sciences of the United States of America, 2010, 107(5): 2037-2042.

[9]Hsu P P, Sabatini D M. Cancer cell metabolism: Warburg and Beyond. Cell, 2008, 134(5): 703-707.

[10]Mehmeti I, Jönsson M, Fergestad E M, Mathiesen G, Nes I F, Holo H. Transcriptome, proteome, and metabolite analyses of a lactate dehydrogenase-negative mutant of Enterococcus faecalis V583. Applied and Environmental Microbiology, 2011, 77(7): 2406-2413.

[11]Sambrook J, Russell D W. Molecular Cloning: A Laboratory Manual. 3rd ed. New York: Cold Spring Harbor Laboratory Press, 2001: 99-120.

[12]Velásquez A C, Chakravarthy S, Martin G B. Virus-induced gene silencing (VIGS) in Nicotiana benthamiana and tomato. Journal of Visualized Experiments, 2009, 28: doi: 10.3791/1292.

[13]崔艳红, 贾芝琪, 李颖, 黄三文. 利用VIGS技术研究马铃薯抗晚疫病基因R3a和RB的信号传导. 园艺学报, 2009, 36(7): 997-1004.

Cui Y H, Jia Z Q, Li Y, Huang S W. Studies on signal pathways of the potato late blight resistance gene R3a, RB using VIGS. Acta Horticulturae Sinica, 2009, 36(7): 997-1004. (in Chinese)

[14]Abad P, Gouzy J, Aury J M, Castagnone-Sereno P, Danchin E G, Deleury E, Perfus-Barbeoch L, Anthouard V, Artiguenave F, Blok V C, Caillaud M C, Coutinho P M, Dasilva C, De Luca F, Deau F, Esquibet M, Flutre T, Goldstone J V, Hamamouch N, Hewezi T, Jaillon O, Jubin C, Leonetti P, Magliano M, Maier T R, Markov G V, McVeigh P, Pesole G, Poulain J, Robinson-Rechavi M, Sallet E, Ségurens B, Steinbach D, Tytgat T, Ugarte E, van Ghelder C, Veronico P, Baum T J, Blaxter M, Bleve-Zacheo T, Davis E L, Ewbank J J, Favery B, Grenier E, Henrissat B, Jones J T, Laudet V, Maule A G, Quesneville H, Rosso M N, Schiex T, Smant G, Weissenbach J, Wincker P. Genome sequence of the metazoan plant- parasitic nematode Meloidogyne incognita. Nature Biotechnology, 2008, 26(8): 909-915.

[15]The C. elegans Sequencing Consortium. Genome sequence of the nematode C. elegans: a platform for investigating biology. Science, 1998, 282: 2012-2018.

[16]Smits T H M, Rezzonico F, Kamber T, Goesmann A, Ishimaru C A, Stockwell V O, Frey J E, Duffy B. Genome sequence of the biocontrol agent Pantoea vagans strain C9-1. Journal of Bacteriology, 2010, 192(24): 6486-6487.

[17]Patel H K, da Silva D P, Devescovi G, Maraite H, Paszkiewicz K, Studholme D J, Venturi V. Draft genome sequence of Pseudomonas fuscovaginae, a broad-host-range pathogen of plants. Journal of Bacteriology, 2012, 194(10): 2765-2766.

[18]王宏芝, 李瑞芬, 王国英, 马荣才, 魏建华. 病毒诱导的基因沉默及其在植物功能基因组学研究中的应用. 自然科学进展, 2005, 15(1): 8-14.

Wang H Z, Li R F, Wang G Y, Ma R C, Wei J H. Application of virus-induced gene silencing in plant functional genomics. Natural Science Progress, 2005, 15(1): 8-14. (in Chinese)

[19]Spitzer B, Zvi M M B, Ovadis M, Marhevka E, Barkai O, Edelbaum O, Marton I, Masci T, Alon M, Morin S, Rogachev I, Aharoni A, Vainstein A. Reverse genetics of floral scent: Application of tobacco rattle virus-based gene silencing in petunia. Plant Physiology, 2007, 145: 1241-1250.

[20]Nagamatsu A, Masuta C, Senda M, Matsuura H, Kasai A, Hong J, Kitamura K, Abe J, Kanazawa A. Functional analysis of soybean genes involved in flavonoid biosynthesis by virus-induced gene silencing. Plant Biotechnology Journal, 2007, 5: 778-790.

[21]Di Stilio V S, Kumar R A, Oddone A M, Tolkin T R, Salles P, McCarty K. Virus-induced gene silencing as a tool for comparative functional studies in Thalictrum. PLoS ONE, 2010, 5(8): e12064. 

[22]Lu H C, Hsieh M H, Chen C E, Chen H H, Wang H I, Yeh H H. A high-throughput virus-induced gene-silencing vector for screening transcription factors in virus-induced plant defense response in orchid. Molecular Plant-Microbe Interactions, 2012, 25(6): 738-746.

[23]Cakir C, Gillespie M E, Scofield S R. Rapid determination of gene function by virus-induced gene silencing in wheat and barley. Crop Science, 2010, 50: 77-84.

[24]Markert C L, Moller F. Multiple forms of enzymes: tissue, ontogenetic, and species specific patterns. Proceedings of the National Academy of Sciences of the United States of America, 1959, 45: 753-763.

[25]王丹卉, 秦资, 韩国斌, 王世贵, 唐斌, 张帆. 异色瓢虫乳酸脱氢酶基因的克隆与序列分析. 中国生物防治学报, 2011, 27(4): 458-463.

Wang D H, Qin Z, Han G B, Wang S G, Tang B, Zhang F. Cloning and sequence analysis of lactic dehydrogenase gene from Harmonia axyridis (Coleoptera: Coccinellinae). Chinese Journal of Biological Control, 2011, 27(4): 458-463. (in Chinese)

[26]Lemire J, Mailloux R J, Appanna V D. Mitochondrial lactate dehydrogenase is involved in oxidative-energy metabolism in human astrocytoma cells (CCF-STTG1). PLoS ONE, 2008, 3(2): e1550.

[27]Mangia S, Simpson I A, Vannucci S J, Carruthers A. The in vivo neuron-to-astrocyte lactate shuttle in human brain: evidence from modeling of measured lactate levels during visual stimulation. Journal of Neurochemistry, 2009, 109(Suppl. 1): 55-62.

[28]Sultana R, Butterfield D A. Oxidatively modified, mitochondria- relevant brain proteins in subjects with Alzheimer disease and mild cognitive impairment. Journal of Bioenergetics and Biomembranes, 2009, 41: 441-446.

[29]Dubreuil G, Magliano M, Dubrana M P, Lozano J, Lecomte P, Favery B, Abad P, Rosso M N. Tobacco rattle virus mediates gene silencing in a plant parasitic root-knot nematode. Journal of Experimental Botany, 2009, 60(14): 4041-4050.

[30]Valentine T A, Randall E, Wypijewski K, Chapman S, Jones J, Oparka K J. Delivery of macromolecules to plant parasitic nematodes using a tobacco rattle virus vector. Plant Biotechnology Journal, 2007, 5: 827-834.
[1] ZHANG Qi, CHEN ErHu, SUN DeHong, TANG PeiAn. Relationship Between Glutathione S-Transferase Genes CfGSTe1 and CfGSTd1 and Ethyl Formate Tolerance in Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2026, 59(5): 1008-1019.
[2] CHEN ErHu, TANG JingJie, HU ShunJie, TANG PeiAn. The Roles of Heat Shock Protein Genes CfHsp70-1 and CfHsp70-2 in Enhancing the High-Temperature Tolerance after Heat Acclimation in Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2025, 58(5): 918-928.
[3] XIAO ZhuoDan, QIAO JiaZheng, GAO YuLan, SHANG ZhangYin, LIU Huai, WANG Jia. Silencing of Cytochrome P450 Genes CYP6CY53 and CYP302A1 in Aphis craccivora Enhances the Sensitivity to Flonicamid [J]. Scientia Agricultura Sinica, 2025, 58(18): 3664-3675.
[4] CHEH ErHu, YUAN GuoQing, CHEN Yan, CHEN MengQiu, SUN ShengYuan, TANG PeiAn. Mitochondrial Protein-Coding Genes Nad5, Nad6 and Atp6 are Involved in Phosphine Resistance of Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2024, 57(9): 1722-1733.
[5] LUO LiDan, CHEN JiaMing, AN Qi, LIU Lei, SUN QinZhe, LIU Huan, WANG SenShan, SONG LiWen. Effects of Extreme High Temperature on Trehalose Content and Trehalose Transporter Gene in Tetranychus truncatus [J]. Scientia Agricultura Sinica, 2024, 57(6): 1091-1101.
[6] ZHAO YiYan, GUO HongFang, LIU WeiMin, ZHAO XiaoMing, ZHANG JianZhen. Effects of Apolipophorin on Ovarian Development and Lipid Deposition in Locusta migratoria [J]. Scientia Agricultura Sinica, 2024, 57(4): 711-720.
[7] YUAN GuoQing, CHEN ErHu, TANG PeiAn. The Mechanisms of Mitochondrial Protein-Coding Genes ND6 and ATP6 in Regulating Cold Tolerance of Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2024, 57(22): 4483-4494.
[8] LI ChuXin, SONG ChenHu, ZHOU JinHuan, LI JiaXin, WANG XinLiang, TIAN XuBin, SONG Zhen. Research on Prevention and Control Technology of Citrus Yellow Vein Clearing Virus Based on VIGS [J]. Scientia Agricultura Sinica, 2024, 57(22): 4473-4482.
[9] WANG Ni, SHI ZheYi, YOU YuanZheng, ZHANG Chao, ZHOU WenWu, ZHOU Ying, ZHU ZengRong. Effects of miRNA on Gene Expression of Sphingolipids Metabolism and Small RNA Analysis of Silencing NlSPT1 and NlSMase4 in Nilaparvata lugens [J]. Scientia Agricultura Sinica, 2024, 57(20): 4022-4034.
[10] CHEH ErHu, SHEN DanRong, DU WenWei, MENG HongJie, TANG PeiAn. Cuticle Protein Genes are Involved in Phosphine Resistance of Cryptolestes ferrugineus [J]. Scientia Agricultura Sinica, 2023, 56(9): 1696-1707.
[11] SHAO HongYang, MENG Xiang, ZHANG Tao, CHEN Min. Analysis of Cytochrome P450 Genes in Response to Quercetin and Function of CYP6ZB2 in Hyphantria cunea [J]. Scientia Agricultura Sinica, 2023, 56(7): 1322-1332.
[12] GUAN RuoBing,LI HaiChao,MIAO XueXia. Commercialization Status and Existing Problems of RNA Biopesticides [J]. Scientia Agricultura Sinica, 2022, 55(15): 2949-2960.
[13] YIN Fei,LI ZhenYu,SAMINA Shabbir,LIN QingSheng. Expression and Function Analysis of Cytochrome P450 Genes in Plutella xylostella with Different Chlorantraniliprole Resistance [J]. Scientia Agricultura Sinica, 2022, 55(13): 2562-2571.
[14] WU Wei,XU HuiLi,WANG ZhengLiang,YU XiaoPing. Cloning and Function Analysis of a Serine Protease Inhibitor Gene Nlserpin2 in Nilaparvata lugens [J]. Scientia Agricultura Sinica, 2022, 55(12): 2338-2346.
[15] CHEN ErHu,MENG HongJie,CHEN Yan,TANG PeiAn. Cuticle Protein Genes TcCP14.6 and TcLCPA3A are Involved in Phosphine Resistance of Tribolium castaneum [J]. Scientia Agricultura Sinica, 2022, 55(11): 2150-2160.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd