碳代谢基因gabT调控生防菌Snea253的γ-氨基丁酸代谢途径影响杀线虫活性

于冬梅1 朱峰2 邢志富1 范海燕1 朱晓峰1 段玉玺1 王媛媛3 刘晓宇4 陈立杰1

(1.沈阳农业大学植物保护学院, 辽宁沈阳 110866)
(2.吉林省农业科学院植物保护研究所, 吉林公主岭 130033)
(3.沈阳农业大学生物科学技术学院, 辽宁沈阳 110866)
(4.沈阳农业大学理学院, 辽宁沈阳 110866)

【摘要】【背景】委内瑞拉链霉菌Snea253是本实验室前期获得的具有杀植物线虫活性的生防放线菌,通过生物信息学分析,γ-氨基丁酸转氨酶基因(gabT)是参与Snea253碳代谢的重要基因之一。【目的】明确gabT基因通过调控Snea253的γ-氨基丁酸(γ-Aminobutyric acid,GABA)代谢通路,从而影响菌株的活性。【方法】以紫外诱变所得弱毒株(Snea253-R)为材料,以南方根结线虫为靶标,在弱毒株中过表达gabT基因,通过酶联免疫法(ELISA)和高效液相色谱法(HPLC)分别检测菌株中GABA和下游代谢产物琥珀酸的含量及杀线虫活性,同时检测在不同碳源培养条件下野生型菌株gabT基因表达水平、产物含量和杀线虫活性。【结果】过表达菌株R-p IB139的gabT基因上调表达,GABA含量降低,琥珀酸含量升高,杀线虫活性提高了39%;在8种不同碳源培养条件下,gabT基因在野生株中相对表达量较高的培养基碳源是可溶性淀粉和玉米淀粉,其发酵液中GABA含量较低,发酵液中下游代谢产物增多,杀线虫活性较高。【结论】通过改变gabT基因的表达,明确GABA支路在调控Snea253代谢以提高杀线虫的过程中发挥重要作用。

【关键词】 γ-氨基丁酸转氨酶基因gabT; 委内瑞拉链霉菌; γ-氨基丁酸(GABA); 琥珀酸; 杀线虫活性;

【DOI】

【基金资助】 国家重点研发计划(2017YFD0201104) National Key Research and Development Program of China(2017YFD0201104) 国家自然科学基金(31471748) National Natural Science Foundation of China(31471748)

Download this article

    References

    [1] Abad P, Favery B, Rosso MN, et al. Root-knot nematode parasitism and host response:molecular basis of a sophisticated interaction[J]. Molecular Plant Pathology, 2003, 4(4):217-224

    [2] Zhao L, Duan YX, Bai CM, et al. Occurrence and control of vegetable root-knot nematodes under protected cultivation in Liaoning province[J]. Plant Protection, 2011, 37(1):105-109(in Chinese)

    [3] Gong B, Zhang LL, Sui SL, et al. Effects of garlic straw application on controlling tomato root-knot nematode disease and rhizospheric microecology[J]. Scientia Agricultura Sinica, 2016,49(5):933-941(in Chinese)

    [4] Stadler M, Anke H, Sterner O. Linoleic acid——the nematicidal principle of several nematophagous fungi and its production in trap-forming submerged cultures[J]. Archives of Microbiology,1993, 160(5):401-405

    [5] Shemshura ON, Bekmakhanova NE, Mazunina MN, et al.Isolation and identification of nematode-antagonistic compounds from the fungus Aspergillus candidus[J]. FEMS Microbiology Letters, 2016, 363(5):1-9

    [6] Zuckerman BM, Matheny M, Acosta N. Control of plant-parasitic nematodes by a nematicidal strain of Aspergillus niger[J]. Journal of Chemical Ecology, 1994, 20(1):33-43

    [7] Morgunov IG, Kamzolova SV, Dedyukhina EG, et al. Application of organic acids for plant protection against phytopathogens[J].Applied Microbiology and Biotechnology, 2017, 101(3):921-932

    [8] Kamzolova SV, Vinokurova NG, Shemshura ON, et al. The production of succinic acid by yeast Yarrowia lipolytica through a two-step process[J]. Applied Microbiology and Biotechnology,2014, 98(18):7959-7969

    [9] Botura MB, dos Santos JDG, da Silva GD, et al. In vitro ovicidal and larvicidal activity of Agave sisalana Perr.(Sisal)on gastrointestinal nematodes of goats[J]. Veterinary Parasitology,2013, 192(1/3):211-217

    [10] Dhakal R, Bajpai VK, Baek KH. Production of GABA(γ-Aminobutyric acid)by microorganisms:a review[J]. Brazilian Journal of Microbiology, 2012, 43(4):1230-1241

    [11] Shelp BJ, Mullen RT, Waller JC. Compartmentation of GABA metabolism raises intriguing questions[J]. Trends in Plant Science, 2012, 17(2):57-59

    [12] Shelp BJ, Bozzo GG, Trobacher CP, et al. Hypothesis/review:contribution of putrescine to 4-aminobutyrate(GABA)production in response to abiotic stress[J]. Plant Science, 2012, 193-194:130-135

    [13] Scott EM, Jakoby WB. Solubleγ-aminobutyric-glutamic transaminase from Pseudomonas fluorescens[J]. Journal of Biological Chemistry, 1959, 234(4):932-936

    [14] Shelp BJ, Bown AW, McLean MD. Metabolism and functions ofγ-aminobutyric acid[J]. Trends in Plant Science, 1999, 4(11):446-452

    [15] Zhou L, Shen BB, Bai SY, et al. RNA interference of OsGABA-T1gene expression induced GABA accumulation in rice grain[J].Acta Agronomica Sinica, 2015, 41(9):1305-1312(in Chinese)

    [16] Simpson JP, Clark SM, Portt A, et al.γ-Aminobutyrate transaminase limits the catabolism ofγ-aminobutyrate in cold-stressed Arabidopsis plants:insights from an overexpression mutant[J]. Botany, 2010, 88(5):522-527

    [17] Renault H. Fiat lux!:phylogeny and bioinformatics shed light on GABA functions in plants[J]. Plant Signaling&Behavior, 2013,8(6):e24274

    [18] Renault H, El Amrani A, Berger A, et al.γ-Aminobutyric acid transaminase deficiency impairs central carbon metabolism and leads to cell wall defects during salt stress in Arabidopsis roots[J].Plant, Cell&Environment, 2013, 36(5):1009-1018

    [19] Renault H, Roussel V, El Amrani A, et al. The Arabidopsis pop2-1mutant reveals the involvement of GABA transaminase in salt stress tolerance[J]. BMC Plant Biology, 2010, 10:20

    [20] Chen LJ, Chen JS, Zheng YN, et al. Identification of actinomycetes strain Snea253 and its activity against soybean cyst nematode[J]. Chinese Journal of Biological Control, 2009, 25(1):66-69(in Chinese)

    [21] Tian CL, Zhu F, Chen JS, et al. Variance analysis of Streptomyces venezuelae Snea253 mutants against Meloidogyne incognita[J].Journal of Nuclear Agricultural Sciences, 2014, 28(9):1541-1548(in Chinese)

    [22] Tian J, Bryk R, Itoh M, et al. Variant tricarboxylic acid cycle in Mycobacterium tuberculosis:identification ofα-ketoglutarate decarboxylase[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(30):10670-10675

    [23] Bao H, Chen XY, Lv SL, et al. Virus-induced gene silencing reveals control of reactive oxygen species accumulation and salt tolerance in tomato byγ-aminobutyric acid metabolic pathway[J].Plant, Cell&Environment, 2015, 38(3):600-613

    [24] Xiong W, Brune D, Vermaas WFJ. Theγ-aminobutyric acid shunt contributes to closing the tricarboxylic acid cycle in Synechocystis sp. PCC 6803[J]. Molecular Microbiology, 2014, 93(4):786-796

    [25] Feehily C, Karatzas KAG. Role of glutamate metabolism in bacterial responses towards acid and other stresses[J]. Journal of Applied Microbiology, 2013, 114(1):11-24

    [26] Feehily C, O’Byrne CP, Karatzas KAG. Functionalγ-aminobutyrate shunt in Listeria monocytogenes:role in acid tolerance and succinate biosynthesis[J]. Applied and Environmental Microbiology, 2013, 79(1):74-80

    [27] Metzner M, Germer J, Hengge R. Multiple stress signal integration in the regulation of the complexσS-dependent csiD-ygaF-gabDTP operon in Escherichia coli[J]. Molecular Microbiology, 2004, 51(3):799-811

    [28] Chevrot R, Rosen R, Haudecoeur E, et al. GABA controls the level of quorum-sensing signal in Agrobacterium tumefaciens[J].Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(19):7460-7464

This Article

ISSN:0253-2654

CN: 11-1996/Q

Vol 46, No. 12, Pages 3257-3266

December 2019

Downloads:0

Share
Article Outline

摘要

  • 1 材料与方法
  • 2 结果与分析
  • 3 讨论与结论
  • 参考文献