克里本类芽孢杆菌PS04诱导水稻抗病相关基因的表达分析

doi:10.7668/hbnxb.20190312

摘要/Abstract

摘要： 克里本类芽孢杆菌PS04是一个水稻纹枯病的高效生防菌，为研究该菌诱导水稻抗性相关基因的表达谱，用PS04菌株发酵液处理水稻24 h后于3，6，9，12，24 h取样，利用荧光定量PCR检测样品中抗性相关基因的相对表达量。结果如下：PS04菌株发酵液能够诱导8个防卫反应基因（OsPR1、OsPR2、OsPR3、OsPR4、OsPR5、OsPR10、OsPR16和NPR1）、2个水杨酸途径基因（PAL和OsNAC4）和3个茉莉酸/乙烯途径基因（AOS2、LOX和EIN2）的表达。其中NPR1和LOX基因在第3小时表达量达最高随后下降；OsPR3、OsPR4、OsPR5、OsPR16和PAL基因在第6小时表达量达最高随后下降；OsPR1和OsNAC4基因在第9小时表达量达最高随后下降；OsPR10基因在第12小时表达量达最高随后下降；OsPR2和EIN2基因在第24小时表达量达到最大值；AOS2基因的表达受抑制，在第6小时表达量达最高随后下降。结果表明，PS04菌株可以同时诱导水稻系统性获得抗性和诱导性系统抗性。

关键词: 水稻, 类芽孢杆菌, 抗病相关基因, 诱导抗病性, 生物防治

Abstract: Paenibacillus kribbensis PS04 is a high-efficiency bacterium for the biocontrol of rice sheath blight. In order to investigate the expression profiles of various disease resistance-related genes,rice leaves were harvested at 3,6,9,12,24 h after 24 h sprayed with fermentation liquid of PS04 strain. Real-time PCR analysis were performed to monitor the expression levels of rice disease resistance-related genes after treatment with PS04 strain. The fermentation liquid of PS04 strain could induce the expression of defense related genes(OsPR1, OsPR2, OsPR3, OsPR4, OsPR5, OsPR10, OsPR16 and NPR1 ),salicylic acid pathway genes(PAL and OsNAC4 ),jasmonic acid and ethylene pathways genes(AOS2, LOX and EIN2 ). The expression of NPR1 and LOX genes reached a maximum level at 3 h and then decreased. The expression of OsPR3, OsPR4, OsPR5, OsPR16 and PAL genes reached a maximum level at 6 h and then decreased. The expression of OsPR1 and OsNAC4 genes reached a maximum level at 9 h and then decreased. The expression of OsPR10 genes reached a maximum level at 12 h and then decreased. The expression of OsPR2 and EIN2 genes reached a maximum level at 24 h. The expression of AOS2 gene was inhibited,and the expression reached the highest level at 6 h and then decreased. The result of this study showed that the PS04 strain could induce both systemic acquired resistance and induced systemic resistance in rice plant.

Key words: Rice, Paenibacillus, Resistance-related genes, Inducing resistance, Biological control

中图分类号:

S432.2+3

程妍, 曲玮茵, 郑文博, 杨媚, 廖美德, 周而勋, 舒灿伟. 克里本类芽孢杆菌PS04诱导水稻抗病相关基因的表达分析[J]. 华北农学报, 2020, 35(2): 203-209. doi: 10.7668/hbnxb.20190312.

CHENG Yan, QU Weiyin, ZHENG Wenbo, YANG Mei, LIAO Meide, ZHOU Erxun, SHU Canwei. Expression Analysis of Disease Resistance-related Genes in Rice Plant Induced by Paenibacillus kribbensis PS04[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(2): 203-209. doi: 10.7668/hbnxb.20190312.

http://www.hbnxb.net/CN/Y2020/V35/I2/203

参考文献

[1] 俞寅达,孙婳珺,夏志辉. 水稻纹枯病生物防控研究进展[J]. 分子植物育种,2019, 17(2):600-605. doi:10.13271/j.mpb.017.000600.
Yu Y D, Sun H J, Xia Z H. Progress on biological control of rice sheath blight[J]. Molecular Plant Breeding,2019, 17(2):600-605.
[2] 李勇. 水稻纹枯病防治效果试验报告[J]. 现代农业,2015(4):8-9. doi:10.3969/j.issn.1008-0708.2015.04.005.
Li Y. Experimental report on control effect of rice sheath blight[J]. Modern Agriculture,2015(4):8-9.
[3] 李涛,路雪君,廖晓兰,欧晓明. 水稻纹枯病的发生及其防治策略[J]. 江西农业学报,2010, 22(9):91-93. doi:10.19386/j.cnki.jxnyxb.2010.09.027.
Li T, Lu X J, Liao X L, Ou X M. Occurrence and control strategy of rice sheath blight[J]. Acta Agriculturae Jiangxi,2010, 22(9):91-93.
[4] 张云欢,郑文博,赵美,王陈骄子,周而勋,舒灿伟. 水稻纹枯病菌谷胱甘肽S-转移酶基因的生物信息学及表达分析[J]. 华北农学报,2018, 33(1):45-51. doi:10.7668/hbnxb.2018.01.008.
Zhang Y H, Zheng W B, Zhao M, Wang C J Z, Zhou E X, Shu C W. Bioinformatics and expression analysis of glutathione S-transferase gene in Rhizoctonia solani AG1-IA[J]. Acta Agriculturae Boreali-Sinica,2018, 33(1):45-51.
[5] 杨迎青,杨媚,兰波,周而勋,李湘民. 水稻纹枯病菌致病机理的研究进展[J]. 中国农学通报,2014, 30(28):245-250. doi:10.11924/j.issn.1000-6850.2014-1538.
Yang Y Q, Yang M, Lan B, Zhou E X, Li X M. Research progress in pathogenic mechanisms of rice sheath blight pathogen[J]. Chinese Agricultural Science Bulletin,2014, 30(28):245-250.
[6] 蔡新忠,郑重. 植物系统性获得抗病性的产生机理和途径[J]. 植物保护学报,1999, 26(1):83-90. doi:10.13802/j.cnki.zwbhxb.1999.01.017.
Cai X Z, Zheng Z. Mechanisms and pathways of plant systemic acquired resistance[J]. Acta Phytophylacica Sinica,1999, 26(1):83-90.
[7] Lucas W J, Groover A, Lichtenberger R, Furuta K, Yadav S R, Helariutta Y, He X Q, Fukuda H, Kang J L, Brady S M, Patrick J W, Sperry J, Yoshida A, López-Millán A F, Grusak M A, Kachroo P. The plant vascular system:evolution, development and functions[J]. Journal of Integrative Plant Biology,2013, 55(4):294-388. doi:10.1111/jipb.12041.
[8] 白薇. 水稻系统获得抗性相关基因的分子研究[D]. 呼和浩特:内蒙古农业大学, 2010.
Bai W. Molecular study of disease resistance in rice[D]. Huhhot:Inner Mongolia Agricultural University, 2010.
[9] 陈英,谭碧玥,黄敏仁. 植物天然免疫系统研究进展[J]. 南京林业大学学报(自然科学版),2012, 36(1):129-136. doi:10.3969/j.issn.1000-2006.2012.01.027.
Chen Y, Tan B Y, Huang M R. Recent advances in plant immune system[J]. Journal of Nanjing Forestry University(Natural Science Edition),2012, 36(1):129-136.
[10] 刘晓光,高克祥,康振生,何邦令. 生防菌诱导植物系统抗性及其生化和细胞学机制[J]. 应用生态学报,2007, 18(8):1861-1868. doi:10.13287/j.1001-9332.2007.0304.
Liu X G, Gao K X, Kang Z S, He B L. Systemic resistance induced by biocontrol agents in plants and its biochemical and cytological mechanisms[J]. Chinese Journal of Applied Ecology,2007, 18(8):1861-1868.
[11] 陈英,黄敏仁,诸葛强,王明庥. 植物抗病信号传导途径及其相互作用[J]. 南京林业大学学报(自然科学版),2002, 26(3):85-90. doi:10.3969/j.issn.1000-2006.2002.03.022.
Chen Y, Huang M R, Zhuge Q, Wang M X. Signaling pathways and interplay in plant disease resistance[J]. Journal of Nanjing Forestry University (Natural Science Edition),2002, 26(3):85-90.
[12] Feys B J, Parker J E. Interplay of signaling pathways in plant disease resistance[J]. Trends in Genetics,2000, 16(10):449-455. doi:10.1016/S0168-9525(00)02107-7.
[13] 陈志谊,许志刚,陆凡,刘永锋. 拮抗细菌B-916对水稻植株的抗性诱导作用[J]. 西南农业学报,2001, 14(2):44-48. doi:10.16213/j.cnki.scjas.2001.02.012.
Chen Z Y, Xu Z G, Lu F, Liu Y F. The inducing resistance effect of antagonistic bacterium B-916 on rice plant[J]. Southwest China Journal of Agricultural Sciences,2001,14(2):44-48.
[14] 李德全,陈志谊,聂亚锋. 生防菌Bs-916及高效突变菌株抗菌物质及其对水稻抗性诱导作用的研究[J]. 植物病理学报,2008, 38(2):192-198. doi:10.13926/j.cnki.apps.2008.02.008.
Li D Q, Chen Z Y, Nie Y F. Antifungal substances producted by a high-yielding mutant of Bs-916 and their effects inducting-resistance on rice plant[J]. Acta Phytophologica Sinica,2008,38(2):192-198.
[15] 戴秀华. 解淀粉芽胞杆菌Lx-11防治水稻细菌性条斑病的促生、控病机理研究[D]. 南京:南京农业大学, 2015.
Dai X H. Study on the mechanism of Bacillus amyloliquefaciens Lx-11 against Xanthomonas oryzae pv. oryzicola and promote the growth of rice[D]. Nanjing:Nanjing Agricultural University,2015.
[16] Rais A, Jabeen Z, Shair F, Hafeez F Y, Hassan M N. Bacillus spp., a bio-control agent enhances the activity of antioxidant defense enzymes in rice against Pyricularia oryzae[J]. PLoS One,2017, 12(11):e187412. doi:10.1371/journal.pone.0187412.
[17] Viswanathan R, Samiyappan R. Induction of systemic resistance by plant growth promoting rhizobacteria against red rot disease in sugarcane[J]. Sugar Tech,1999, 1(3):67-76. doi:10.1007/bf02945166.
[18] Sharma C K, Vishnoi V K, Dubey R C, Maheshwari D K. A twin rhizospheric bacterial consortium induces systemic resistance to a phytopathogen Macrophomina phaseolina in mung bean[J]. Rhizosphere,2018, 5:71-75. doi:10.1016/j.rhisph.2018.01.001.
[19] Droby S, Vinokur V, Weiss B, Cohen L,Daus A, Goldschmidt E E, Porat R. Induction of resistance to Penicillium digitatum in grapefruit by the yeast biocontrol agent Candida oleophila[J]. Phytopathology,2002, 92(4):393-399. doi:10.1094/PHYTO.2002.92.4.393.
[20] Kim J S, Lee J, Lee C H, Woo S Y, Kang H, Seo S G, Kim S H. Activation of pathogenesis-related genes by the rhizobacterium, Bacillus sp. JS, which induces systemic resistance in tobacco plants[J]. The Plant Pathology Journal,2015, 31(2):195-201. doi:10.5423/PPJ.NT.11.2014.0122.
[21] Martínez-Medina A, Van Wees S C M, Pieterse C M J. Airborne signals from Trichoderma fungi stimulate iron uptake responses in roots resulting in priming of jasmonic acid-dependent defences in shoots of Arabidopsis thaliana and Solanum lycopersicum[J]. Plant, Cell and Environment,2017,40(11):2691-2705. doi:10.1111/pce.13016.
[22] 梁小文,王根豪,陈晓燕,廖美德,范志金,周立峰,李肖宇,马忠岩,曾繁富,肖筱成,熊建生,曾升华,吴俊杰,钱凤,朱湘盛. 一株克里本类芽孢杆菌及其应用[P]. CN201310643992.5. 2014-04-30.
Liang X W, Wang G H, Chen X Y, Liao M D, Fan Z J, Zhou L F, Li X Y, Ma Z Y, Zeng F F, Xiao X C, Xiong J S, Zeng S H, Wu J J, Qian F, Zhu X S. A strain of Paenibacillus kribbensis and its application[P]. CN201310643992.5. 2014-04-30.
[23] Guo T, Liao M D. Suppression of Rhizoctonia solani and induction of host plant resistance by Paenibacillus kribbensis PS04 towards controlling of rice sheath blight[J]. Biocontrol Science and Technology,2014, 24(1):116-121. doi:10.1080/09583157.2013.844224.
[24] 郭田,王刘庆,廖美德. PS04菌株对水稻纹枯病的防效及对水稻2种防御性酶活性的诱导[J]. 西北农林科技大学学报(自然科学版),2013, 41(6):98-102. doi:10.13207/j.cnki.jnwafu.2013.06.016.
Guo T, Wang L Q, Liao M D. Biocontrol of PS04 against rice sheath blight and its induced activities to two rice defense enzymes[J]. Journal of Northwest A&F University (Nat Sci Ed),2013, 41(6):98-102.
[25] 牛吉山,刘靖,倪永静,尹钧. 茉莉酸对 PR-1、PR-2、PR-5和Ta-JA2 基因表达以及小麦白粉病抗性的诱导[J]. 植物病理学报,2011, 41(3):270-277. doi:10.13926/j.cnki.apps.2011.03.018.
Niu J S, Liu J, Ni Y J, Yin J. Introduction of PR-1, PR-2, PR-5, Ta-JA2 and wheat powdery mildew resistance in response to MeJA treatment[J]. Acta Phytophologica Sinica,2011, 41(3):270-277.
[26] 王帅. 芽孢杆菌及其脂肽类化合物防治植物病害和促进植物生长的研究[D]. 南京:南京农业大学, 2009.
Wang S. Plant growth promotion and control of plant disease with Bacillus spp. and lipopeptide[D]. Nanjing:Nanjing Agricultural University, 2009.
[27] 张维娜. 普城沙雷菌A21-4对辣椒诱导抗性机理的初步研究[D]. 郑州:河南农业大学, 2013.
Zhang W N. Preliminary study on mechanism of induced systemic resistance by Serratia plymuthica A21-4[D]. Zhengzhou:Henan Agricultural University, 2009.
[28] 张荣胜. 水稻细菌性条斑病菌遗传多样性研究与生防菌剂研发[D]. 南京:南京农业大学, 2012.
Zhang R S. Genetic diversity of Xanthomonas oryzae pv. oryzicola strains and research and development of biocontrol agents[D]. Nanjing:Nanjing Agricultural University, 2012.

选择文件类型/文献管理软件名称

选择包含的内容