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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (12): 1819-1827.doi: 10.3724/SP.J.1006.2015.01819

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Function Analysis of Pathogenesis Related Protein Gene HaPR1 from Sunflower (Helianthus annuus)

MA Li-Gong1,2,ZHANG Yun-Hua2,*,MENG Qing-Lin2,SHI Feng-Mei2,LIU Jia2,LI Yi-Chu2,WANG Zhi-Ying1,*   

  1. 1 College of Forestry, Northeast Forestry University, Harbin 150040, China; 2 Institute of Plant Protection, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China
  • Received:2015-04-20 Revised:2015-07-20 Online:2015-12-12 Published:2015-08-11
  • Contact: 王志英, E-mail: zyw0451@sohu.com, Tel: 13069873855; 张匀华, E-mail: yhzhang9603@126.com, Tel: 13603639603 E-mail:maligong0@163.com
  • Supported by:

    This research was supported by the Modern Agro-industry Technology Research System (CARS-16) and the Agricultural Science and Technology Innovation Program of Heilongjiang Province (QN015).

Abstract:

Pathogenesis-related proteins are commonly used as markers of plant defense responses. The full-length cDNA of pathogenesis-related protein 1 (PR1) named HaPR1 in Helianthus annuus was cloned based on the transcriptome of H.annuus induced by Sclerotinia sclerotiorum, and its expression model and function were analyzed in this study. Sequence analysis showed that the cDNA of HaPR1 (GenBank No. KR071874) contained a 489 bp ORF encoding a protein of 162 amino acids residues with the molecular mass of 17.52 kD and theoretical pI of 8.19, HaPR1 possessed six conserved cysteine and four conserved allergen V5/Tpx-1 related domain. The HaPR1 was highly homologous with PR1 in other species. Real-time PCR analysis showed that expression level of HaPR1 was the highest in leaf, and was significantly induced by drought, salt stress, oxalic acid, S. sclerotiorum and its metabolites. Then the HaPR1 gene was transformed into tobacco by Agrobacterium tumefaciens to further verify its function. The results showed that the expression of HaPR1 improved the resistance of transgenic lines, and significantly increased SOD, POD, and CAT activities and reduced the content of MDA. It suggested that HaPR1 has a function of resistance to S. sclerotiorum.

Key words: Helianthus annuus, Pathogenesis-related protein 1, Gene clone, Function analysis

[1]谢纯政, 刘海燕, 李玲, 梁炫强. 植物病程相关蛋白PR10研究进展. 分子植物育种, 2008, 6: 949–953



   Xie C Z, Liu H Y, Li L, Liang X Q. Advances on class 10 pathogenesis-related proteins. Mol Plant Breed, 2008, 6: 949–953 (in Chinese with English abstract)



[2]Loon L C, Strien E A. The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol Mol Plant Pathol, 1999, 55: 85–97



[3]Van Loon L C, Rep M, Pieterse C M J. Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopathol, 2006, 44: 135–162



[4]牛吉山, 刘瑞, 郑磊. 小麦PR-1、PR-2、PR-5基因的白粉菌和水杨酸诱导表达分析及白粉病抗性研究. 麦类作物学报, 2007, 27: 1132–1137



    Niu J S, Liu R, Zheng L. Expression analysis of wheat PR-1, PR-2, PR-5 activated by Bgt and SA, and powdery mildew resistance. J Triticeae Crops, 2007, 27: 1132–1137 (in Chinese with English abstract)



[5]Xie X Z, Xue Y J, Zhou J J, Zhang B, Chang H, Takano M. Phytochromes regulate SA and JA signaling pathways in rice and are required for developmentally controlled resistance to Magnaporthe grisea. Mol Plant, 2011, 4: 688–696



[6]Nandi A, Kachroo P, Fukushige H, Hildebrand D F, Klessig D F, ShahJ. Ethylene and jasmonic acid signaling affect the NPR1-independent expression of defense genes without impacting resistance to Pseudomonas syringae and Peronospora parasitica in the Arabidopsis ssi1 mutant. Mol Plant Microbe Interact, 2003, 16: 588–599



[7]Flors V, Ton J, van Doorn R, Jakab G, García-Agustín P, Mauch-Mani B. Interplay between JA, SA and ABA signalling during basal and induced resistance against Pseudomonas syringae and Alternaria brassicicola. Plant J, 2008, 54: 81–92



[8]Alexander D, Goodman R M, GutRella M, Glascock C, Weymann K, Friedrich L, Maddox D, Ahlgoy P, Luntz T, Ward E. Increased tolerance to two oomycete pathogens in transgenic tobacco expressing pathogenesis-related protein 1a. Proc Natl Acad Sci USA, 1993, 90: 7327–7331



[9]NiKi T, Mitsuhara I, Seo S. Antagonistic effect of Salicylic 330 acid and jasmonic acid on the expression of pathogeneis-relation (PR) protein genes in wounded mature tobacco leaves. Plant Cell Physiol, 1998, 39: 500–507



[10]Sarowar S, Kim Y J, Kim E N. Overexpression of a pepper basic pathogenesis-related protein1 gene in tobacco plants enhances resistance to heavy metal and pathogen stresses. Plant Cell Rep, 2005, 24: 216–224



[11]Li Z T, Dhekney S A, Gray D J. PR-1 gene family of grapevine: a uniquely duplicated PR-1 gene from a Vitis interspecific hybrid confers high level resistance to bacterial disease in transgenic tobacco. Plant Cell Rep, 2011, 30: 1–11



[12]Bonasera J M, Kim J F, Beer S V. PR genes of apple: identification and expression in response to elicitors and inoculation with Erwinia amylovora. BMC Plant Biol, 2006, 6: 23



[13]Seo P J, Lee A K, Xiang F N, Park C M. Molecular and functional profiling of Arabidopsis pathogenesis-related genes: insights into their roles in salt response of seed germination. Plant Cell Physiol, 2008, 49, 334–344



[14]Stintzi A, Heitz T, Kauffmann S, Legrand M, Fritig B. Identification of a basic pathogenesis-related, thaumatin-like protein of virus-infected tobacco as osmotin. Physiol Mol Plant Pathol, 1991, 38: 137–146



[15]Agrawal G K, Jwa N S, Rakwal R. A novel rice (Oryza sativa L.) acidic PR1 gene highly responsive to cut, phytohormones, and protein phosphatase inhibitors. Biochem Biophys Res Commun, 2000, 274: 157–165



[16]Agrawal G K, Rakwal R, Jwa N S. Rice (Oryza sativa L.) OsPR1b gene is phytohormonally regulated in close interaction with light signals. Biochem Biophys Res Commun, 2000, 278: 290–298



[17]Agrawal G K, Rakwal R, Jwa N S, Agrawal V P. Signaling molecules and blast pathogen attack activates rice OsPR1a and OsPR1b genes: a model illustrating components participating during defence/stress response. Plant Physiol Biochem, 2001, 39: 1095–1103



[18]Livak K, Schmittgen, T. Analysis of relative gene expression data using real-time quantitative PCR and the 2-delta delta CT method. Methods, 2001, 25: 402–408



[19]邹琦. 植物生理生化实验指导. 北京: 农业出版社, 1991



    Zou Q. Plant Physiological and Biochemical Experiments. Beijing: Agriculture Press, 1991 (in Chinese)



[20]朱广廉, 钟文海, 张爱琴. 植物生理实验. 北京: 北京大学出版社, 1991



    Zhu G L, Zhong W H, Zhang A Q. Plant Physiology Experiment. Beijing: Peking University Press, 1991 (in Chinese)



[21]汤章城, 王国强, 史益敏. 现代植物生理学实验指南. 北京: 科学出版社, 1999



    Tang Z C, Wang G Q, Shi Y M. Modern Experiment Guide for Plant Physiology. Beijing: Science Press, 1999 (in Chinese)



[22]陈建勋, 王晓峰. 植物生理学实验指导. 广州: 华南理工大学出版社, 2002. pp 31–32, 117–120



    Chen J X, Wang X F. Experiment Manual for Plant Physiology. Guangzhou: South China University of Technology Press, 2002. pp 31–32, 117–120 (in Chinese)



[23]Kim S T, Yu S, Kang Y H. The rice pathogen-related protein 10 (Jiospr10) is induced by abiotic and biotic stresses and exhibits ribonuclease activity. Plant Cell Rep, 2008, 27: 593–603



[24]张计育, 渠慎春, 薛华柏, 高志红, 郭忠仁, 章镇. 湖北海棠病程相关蛋白MhPR8基因的克隆与表达. 中国农业科学, 2012, 45: 1568–1575



    Zhang J Y, Qu S C, Xue H B, Gao Z H, Guo Z R, Zhang Z. Isolation and expression of pathogenesis-related protein gene MhPR8 from Malus hupehensis. Sci Agric Sin, 2012, 45: 1568–1575 (in Chinese with English abstract)



[25]栗小英, 高琳, 张艳俊, 王海燕, 刘大群. 叶锈菌及信号分子诱导小麦TcLr35中β-1,3-葡聚糖酶基因的表达分析. 中国农业科学, 2014, 47: 2774–2783



    Li X Y, Gao L, Zhang Y J, Wang H Y, Liu D Q. Expression and analysis of β-1,3-glucanase gene in wheat TcLr35 induced by wheat leaf rust pathogen and signal molecule. Sci Agri Sin, 2014, 47: 2774–2783 (in Chinese with English abstract)



[26]侯丽霞, 高超, 车永梅, 赵方贵, 刘新. 葡萄病程相关蛋白1基因的克隆和表达分析. 植物生理学报, 2012, 48: 57–62



    Hou L X, Gao C, Che Y M. Gene cloning and expression analysis of pathogenesis-related protein 1 in Vitis vinifera L. Plant Physiol J, 2012, 48: 57–62 (in Chinese with English abstract)



[27]王艳, 陈西, 周莲洁, 杨中敏. 费尔干猪毛菜病程相关蛋白基因SfPR-1的表达规律和植物表达载体构建. 生物技术通报, 2014, 1: 117–124



    Wang Y, Chen X, Zhou L J, Yang Z M. Expression profiles of pathogen-related protein gene (SfPR-1) from Salsola ferganica and construction of plant expression vectors. Biotechnol Bull, 2014, 1: 117–124 (in Chinese with English abstract)

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