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| Piriformospora indica Inducing Tobacco Resistance to Black Shank and Its Preliminary Mechanism |
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Abstract Piriformospora indica, isolated from Indian Thar desert, s identified as a plant root endophytic fungus which plays an important role in inducing resistance against biotic and abiotic stresses. Recently, P. indica has been regarded as a fungal model to study the interaction between endophytic fungus and plants. In order to study the influence of P.indica on black shank resistance in Nicotiana tobacum, we analyzed the differences of the incidence, antioxidant enzyme activities, including phenylalnine ammonialyase (PAL), peroxidase (POD) and polyphenol oxidasei(PPO), and the expression levels of disease resistance-related genes in P.indica-colonized and non-colonized plants after Phytophthora parasitica var. nicotianae inoculation. We also studied whether P.indica influenced the growth of P. parasitica var. nicotianae by antibiosis assay. The results showed that: (1)The disease severity of P. indica-colonized N. tobacum was significantly lower than that of non-colonized ones; (2) After P. parasitica var. nicotianae inoculation from 1 to 10 days, the activities of PAL, POD and PPO in P. indica-colonized tobacco were significantly higher than those in non-colonized plants. P. indica-colonized plants showed 3.99-fold greater PAL activity in comparison with controls after P.parasitica var. nicotianae inoculation for 1 day. Meanwhile, compared with controls, POD and PPO activity in P. indica-colonized plants were elevated by 1.5- and 2.26-fold after infestation with P. parasitica var. nicotianae for 2 and 4 days, respectively; (3) Real-time quantitative PCR (qRT-PCR) results showed that the expression levels of disease resistance-related genes (PR1b, P450-2, hrs203J, GST, NmIMSP and P450-1) were up-regulated in P. indica-colonized N. tobacum after P. parasitica var. nicotianae inoculation at 6 to 96 h, respectively. The expression levels of PR1b, P450-2, hrs203J were up-regulated by 3.8, 15.7 and 3.1 folds in P. indica-colonized N. tobacum compared with controls after P. parasitica var. nicotianae inoculation for 12 h, respectively. The NmIMSP expression reached a level of 2.6-fold higher in P. indica-colonized plants in contrast to the controls after P. parasitica var. nicotianae inoculation for 48 h. The P450-1 was 1.5 folds of the control ones after P. parasitica var. nicotianae inoculation for 72 h. (4) The antibiosis assay showed that P. indica didn’t inhibit P. parasitica var. nicotianae growth. Thus, the resistance induced by P.indica against P.parasitica var. nicotianae in N.tobacum was not dependent on the antibiotic secretion, but on the up-regulation of resistance-related genes and improved antioxidant enzyme activity in N. tobacum. This study reveals the role of P. indica conferring N. tobacum disease resistance against P. parasitica var. nicotianae and correlative mechanism, and provides a new way for the biological control of black shank.
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Received: 28 October 2014
Published: 01 March 2015
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| Chacon O, Hernandez I, Portieles R, Lopez Y, Pujol M, Orlando B H, 2009. Identification of defense-related genes in tobacco responding to black shank disease. Plant Science, 177:175~180Chen J X and Wang X F, eds., 2002. Plant physiology experiment guide, South China University of Technology Press, Guangzhou, China, pp.119~124 (陈建勋, 王晓峰, 主编, 2002. 植物生理学实验指导, 华南理工大学出版社, 中国, 广州, pp.119~124)Chen Y Y, Lou B G, Gao Q K, Lin F C, 2013. Preliminary Study on Mechanisms of Drought Resistance in Brassica napus L. Conferred by Piriformospora indica. Journal of Agricural Biotechnology, 21(3): 272~281(陈佑源, 楼兵干, 高其康, 林福呈, 2013. 印度梨形孢诱导油菜抗旱性机理的初步研究. 农业生物技术学报, 21(3): 272~281)Dolatabadi H K, Goltapeh E M, Jaimand K, Rohani N, Varma A, 2011. Effects of Piriformospora indica and Sebacina vermifera on growth and yield of essential oil in fennel(Foeniculum vulgare) under greenhouse conditions. J Basic Microbiol, 51:33~39Dolatabadi H K, Goltapeh E M, Mohammadi N, Rabiey M, Rohani N, Varma A , 2012. Biocontrol potential of root endophytic fungi and Trichoderma species against Fusarium wilt of lentil under in vitro and greenhouse conditions. J Agr Sci Technol, 14:407~420Gosal S K, Karlupia A, Gosal S S, Chhibba I M, Varma A, 2010. Biotization with Piriformospora indica and Pseudomonas fluorescens improves survival rate, nutrient acquisition, field performance and saponin content of micropropagated Chlorophytum sp. Indian J Biotechnol, 9:289~297Heitz T, Geoffroy P, Stintzi A, Fritig B, Legrandf M,1993. cDNA Cloning and Gene Expression Analysis of the Microbial Proteinase Inhibitor of Tobacco. The Journal of Biological Chemistry, 268:16987~16992Hernandez I, Chacon O, Rodriguez R, Portieles R, Lopez Y, Pujol M, Orlando B H, 2009. Black shank resistant tobacco by silencing of glutathione S-transferase. Biochemical and Biophysical Research Communications, 387:300~304Hui F Q, Peng B, Lou B G, Lin F C, Nie C C, Liu J,2014. Piriformospora indica Improves Salt Tolerance in Nicotiana tobacum by Promoting the Synthesis of Osmolyte and Inducing the Expression of Stress Resistance Genes. Journal of Agricural Biotechnology,22(2):168~176(惠非琼,彭兵,楼兵干,林福呈,聂长春,刘剑,2014.印度梨形孢通过促进渗透调节物质的合成和诱导抗逆相关基因的表达提高烟草耐盐性.农业生物技术学报, 22(2):168~176)Kozlowska M, Fryder K, Wolko B, 2001. Peroxidase involvement in the defense response of red raspberry to Didymella applanata(Niessl/Sacc). Acta Physiologiae Plantarum, 23:303~310Kumar M, Yadav V, Tuteja N, Johri A K, 2009. Antioxidant enzyme activities in maize plants colonized with Piriformospora indica. Microbiology, 155:780~790Lee Y C, Johnson J M, Chien C T, Sun C, Cai D G, Lou B G, Oelmuller R, Yeh K W, 2011.Growth promotion of Chinese cabbage and Arabidopsis by Piriformospora indica is not stimulated by mycelium-synthesized auxin. Molecular Plant-Microbe Interactions, 24(4):421~431Lou B G, Sun C, Cai D G,2007. Piriformospora indica with multiple funcations and its application prospects. Acta Phytophylacica Sinica, 34(6):653~656(楼兵干, 孙超, 蔡大广, 2007. 印度梨形孢的多种功能及其应用前景.植物保护学报, 34(6):653~656)Molitor A, Zajic D, M. Voll L, Kühnemann J P, Samans B, Kogel K H, Waller F, 2011. Barley Leaf Transcriptome and Metabolite Analysis Reveals New Aspects of Compatibility and Piriformospora indica–Mediated Systemic Induced Resistance to Powdery Mildew . Molecular Plant-Microbe Interactions, 24(12):1427~1439Pfitzner.M U, Goodman.M H, 1987. Isolation and characterization of cDNA clones encoding pathogenesis-related proteins from tobacco mosaic virus infected tobacco plants. Nucleic Acids Research, 15:4449~4465Pontler D, Godiard L, Marco Y, Roby D,1994. hsr203J, a tobacco gene whose activation is rapid, highly localized and specific for incompatible plant/pathogen interactions. The Plant Journal, 5(4) :507~521Shang Z Q, 2007. Recent research advance on pathogen and disease occurrence of tobacco black shank and its integrated control. Review of China Agricultural Science and Technology, 9(2):73~76(尚志强,2007. 烟草黑胫病病原、发生规律及综合防治研究进展. 中国农业科技导报,9(2):73~76)Silva Y, Portieles R, Pujol M, Terauchi R, Matsumura H, Serrano M, Orlando B H,2013. Expression of a microbial serine proteinase inhibitor gene enhances the tobacco defense against oomycete pathogens. Physiological and Molecular Plant Pathology,84:99~106Su Z G, 2011. Construction and Analysis of Tobacco SSH Library Induced by Race 0 of Phytophthora parasitica var.Thesis for M.S.,Tobacco Research Institute of CAAS Chinese Academy of Agricultural Sciences,Supervisor:Wang Y Y,pp.39~40(苏振刚, 2011. 烟草黑胫病0号生理小种诱导的抗性SSH文库构建及其基因分析.硕士学位论文，中国农业科学院烟草研究所，导师:王元英, pp.39~40)Sun C, Johnson J, Cai D G, Sherameti I, Oelmüller R and Lou B G, 2010. Piriformospora indica confers drought tolerance in Chinese cabbage leaves by stimulating antioxidant enzymes, the expression of drought-related genes and the plastid-localized CAS protein. Journal of Plant Physiology, 167(12): 1009~1017Takemoto D, Hayashi M, Doke N, Nishimura M, Kawakita K,1999. Molecular Cloning of a Defense-Response-Related Cytochrome P450 Gene from Tobacco. Plant Cell Physiol. 40(12):1232~1242Wang H C, Chen X J, Cai L T, Cao Y, Lu N, Xia H Q, Wang M S, Shang S H, 2013. Race distribution and distribution of sensitivities to mefenoxam among isolates of Phytophthora parasitica var. nicotianae in Guizhou province of China. Crop Protection, 52:136~140Wang H C, Li W H, Feng Y G, Shi J X,2011. History and current status of chemical control of tobacco black shank. Acta Tabacaria Sinica,17(5):96~102（汪汉成,李文红,冯勇刚,石俊雄,2011. 烟草黑胫病化学防治的历史与现状.中国烟草学报，17(5):96~102）Wang X K, eds.,2005.Principles and Techniques of Plant Physiological Biochemical Experiment (2nd Edition), Higher Education Press, Beijing, China, pp.224~225(王学奎，主编，2005.植物生理生化原理与技术，高等教育出版社，中国，北京，pp.224~225)Zarea M J, Hajinia S, Karimi N, Goltapeh E M, Rejali F, Varma A, 2012. Effect of Piriformospora indica and Azospirillum strains from saline or non-saline soil on mitigation of the effects of NaCl. Soil Biol Biochem, 45:139~146Zeng Y S, Wang Z Z, 1999. Role of Phenylalanine ammonia-lyase in plant disease resistance. Jour nal of Zhongkai Agrotechnical College, 12(3):56~65(曾永三,王振中, 1999.苯丙氨酸解氨酶在植物抗病反应中的作用.仲恺农业技术学院学报, 12(3):56~65) Zhu Z Y, Zhang Y M, XU T, 2004. Induced resistance of tobacco to Phytophthora nicotianae by synthetic oligosaccharides. Acta Phytopathologica Sinica, 34(3):231~236(朱振元, 张勇民, 徐同, 2004. 化学合成寡糖诱导烟草抗黑胫病的初步研究.植物病理学报, 34(3):231~236) |
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