马铃薯StPR1基因克隆及表达特异性分析
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摘要
为了探究病程相关蛋白(Pathogenesis related protein,PR蛋白)与马铃薯抗病的相关性,检验其在马铃薯中抗软腐病、青枯病、干腐病的能力。在前期的马铃薯抗病性研究中对PRs的17个家族基因结合转录组数据进行分析及基因的筛选,最终确定PR1为试验的研究对象。选用马铃薯大西洋品种为材料,由黑龙江省农科院提供,主要进行StPR1基因的克隆,并对其进行生物信息学分析;将真菌接骨木镰孢、燕麦镰孢以及导致软腐病和青枯病等细菌为胡萝卜软腐欧文氏菌、菊欧氏菌、胡萝卜软腐欧文氏菌马铃薯黑胫亚种、茄科雷尔氏菌接种到马铃薯块茎后进行StPR1基因表达特异性分析。结果显示,StPR1基因长度为540 bp,编码179个氨基酸,蛋白质疏水性及亲水性预测PR1蛋白序列的GRAVY值在+4～-3之间,含有亲水区及疏水区,蛋白质二级结构预测其属于混合型蛋白,蛋白质三级结构预测发现其为紧密复杂的螺旋结构,具有SCP_PR-1_like保守结构域,其与辣椒PR1基因在一个进化分支上,相似性较高。qRT-PCR结果显示,StPR1基因的表达量表现为抗真菌胁迫高于细菌胁迫,并预测其抗干腐病的能力强于抗软腐病及青枯病的能力。综上所述,当马铃薯受到外界病原菌侵害时,StPR1在马铃薯的抗病过程中发挥着重要的作用。
In order to explore the correlation between pathogenesis related protein(PR protein) and potato disease resistance, the ability to resist soft rot, bacterial wilt and dry rot in potato was tested. In the previous potato disease resistance study, 17 family gene-binding transcriptome data of PRs were analyzed and the genes were screened, and PR1 was finally determined. The potato Atlantic variety was selected as the material, which was provided by Heilongjiang Academy of Agricultural Sciences. The StPR1 gene was cloned and analyzed by bioinformatics. F.sambucinum and F.avenaceum were used. And E.carotovora subsp. Carotovora Borgey(Ecc), E.chrysanthemi Burkholder. Atroseptica Dye(Ech), can cause soft rot and bacterial wilt. The specific analysis of StPR1 gene expression was carried out after inoculation of potato tuber with E. carotovora subsp. Mc Fadden et Dimock(Eca) and Ralstonia solanacearum(RS). The results showed that the StPR1 gene was 540 bp in length and encoded 179 amino acids. The hydrophobicity and hydrophilicity of the protein are predicted.The GRAV value of the PR1 protein sequence was between +4 and-3, containing hydrophilic and hydrophobic regions, and protein secondary structure. It is predicted to be a mixed protein. The tertiary structure of the protein is predicted to be a tightly complex helical structure with a SCP_PR-1_like conserved domain, which is highly similar to the pepper PR1 gene on an evolutionary branch. The results of qRT-PCR showed that the expression level of StPR1 gene was higher than that of bacterial stress in antifungal stress, and its ability to resist dry rot was better than that against soft rot and bacterial wilt. In summary, when the potato is attacked by external pathogens, StPR1 plays an important role in the disease resistance of the potato.In summary, when the potato is attacked by external pathogens, StPR1 plays an important role in the disease resistance of the potato.
In order to explore the correlation between pathogenesis related protein(PR protein) and potato disease resistance, the ability to resist soft rot, bacterial wilt and dry rot in potato was tested. In the previous potato disease resistance study, 17 family gene-binding transcriptome data of PRs were analyzed and the genes were screened, and PR1 was finally determined. The potato Atlantic variety was selected as the material, which was provided by Heilongjiang Academy of Agricultural Sciences. The StPR1 gene was cloned and analyzed by bioinformatics. F.sambucinum and F.avenaceum were used. And E.carotovora subsp. Carotovora Borgey(Ecc), E.chrysanthemi Burkholder. Atroseptica Dye(Ech), can cause soft rot and bacterial wilt. The specific analysis of StPR1 gene expression was carried out after inoculation of potato tuber with E. carotovora subsp. Mc Fadden et Dimock(Eca) and Ralstonia solanacearum(RS). The results showed that the StPR1 gene was 540 bp in length and encoded 179 amino acids. The hydrophobicity and hydrophilicity of the protein are predicted.The GRAV value of the PR1 protein sequence was between +4 and-3, containing hydrophilic and hydrophobic regions, and protein secondary structure. It is predicted to be a mixed protein. The tertiary structure of the protein is predicted to be a tightly complex helical structure with a SCP_PR-1_like conserved domain, which is highly similar to the pepper PR1 gene on an evolutionary branch. The results of qRT-PCR showed that the expression level of StPR1 gene was higher than that of bacterial stress in antifungal stress, and its ability to resist dry rot was better than that against soft rot and bacterial wilt. In summary, when the potato is attacked by external pathogens, StPR1 plays an important role in the disease resistance of the potato.In summary, when the potato is attacked by external pathogens, StPR1 plays an important role in the disease resistance of the potato.
引文
[1]孙海军.东北地区马铃薯常见病害的发生及防治[J].吉林农业,2016(15):92.doi:10.14025/j.cnki.jlny.2016.15.038.Zhang H J.Occurrence and control of common diseases of potato in Northeast China[J].Jilin Agriculture,2016(15):92.
[2]张春.马铃薯常见病害的识别与防治[J].农业科技与信息,2016(28):103,105.doi:10.15979/j.cnki.cn62-1057/s.2016.28.065.Zhang C.Identification and control of common diseases of potato[J].Agricultural Technology and Information,2016(28):103,105.
[3]金飞跃,赵振杰,任乐乐,曹克强,胡同乐.十一种杀菌剂防治马铃薯晚疫病的适宜施用时期[J].农药学学报,2016,18(2):213-218.doi:10.16801/j.issn.1008-7303.2016.0028.Jin F Y,Zhao Z J,Ren L L,Cao K Q,Hu T L.Appropriate application period of eleven fungicides for controlling potato late blight[J].Journal of Agricultural Pharmacy,2016,18(2):213-218.
[4]张贺兰,王鑫,张若晗,杨圆圆,巩秀峰,赵永秀.利用基因工程育种提高马铃薯晚疫病抗性的研究进展[J].分子植物育种,2018,16(21):7038-7041.doi:10.13271/j.mpb.016.007038.Zhang H L,Wang X,Zhang R H,Yang Y Y,Gong XF,Zhao Y X.Advances in research on improving resistance of potato late blight by genetic engineering breeding[J].Molecular Plant Breeding,2018,16(21):7038-7041.
[5]Van Loon L C,Van Strien E A.The families of pathogenesis-related proteins,their activities,and comparative analysis of PR-1 type proteins[J].Physiological and Molecular Plant Pathology,1999,55(2):85-97.doi:10.1006/pmpp.1999.0213.
[6]Breiteneder H,Radauer C.A classification of plant food allergens[J].J Allergy Clin Immunol,2004,113(5):821-830.doi:10.1016/j.jaci.2004.01.779.
[7]Liu J J,Akm E.The family 10 of plant pathogenesis-related proteins:their structure,regulation,and function in response to biotic and abiotic stresses[J].Physiological&Molecular Plant Pathology,2006,68(1):3-13.doi:10.1016/j.pmpp.2006.06.004.
[8]Kim Y J,Jang M G,Lee H J,Jang G H,Sukweenadhi J,Kwon W S,Yang D C.Functional characterization of the pathogenesis-related protein family 10 gene,Pg PR10-4,from Panax ginseng,in response to environmental stresses[J].Plant Cell Tissue&Organ Culture,2014,118(3):531-543.doi:10.1007/s11240-014-0505-5.
[9]Sels J,Mathys J,De Coninck B M A,Cammue B P A,De Bolle M F C.Plant pathogenesis-related(PR)proteins:A focus on PR peptides[J].Plant Physiology and Biochemistry,2008,46(11):941-950.doi:10.1016/j.plaphy.2008.06.011.
[10]Ga~3Mez-Ga~3Mez L,Rubio-Moraga A,Ahrazem O.Molecular cloning and characterisation of a pathogenesis-related protein Cs PR10 from Crocus sativus[J].Plant Biology,2011,13(2):297-303.doi:10.1111/j.1438-8677.2010.00359.
[11]Uknes S.Regulation of pathogenesis-related protein-1a gene expression in tobacco[J].The Plant Cell Online,1993,5(2):159-169.doi:10.1105/tpc.5.2.159.
[12]Yun H K,Yi S Y,Yu S H,Choi D.Cloning of a pathogenesis-related protein-1 gene from Nicotians glutinosa L.and its Salicylic acid-independent induction by copper andβ-aminobutyric acid[J].Journal of Plant Physiology,1999,154(3):327-333.doi:10.1016/s0176-1617(99)80175-8.
[13]张玉,张增林,蒋彩虹,常爱霞,杨爱国,罗成刚,王绍美,王元英.烟草病程相关蛋白Nt PR10基因克隆与表达分析[J].中国烟草科学,2017,38(3):1-7.doi:10.13496/j.issn.1007-5119.2017.03.001.Zhang Y,Zhang Z L,Jiang C H,Chang A X,Yang AG,Luo C G,Wang S M,Wang Y Y,Cloning and expression analysis of tobacco pathogenesis related protein Nt PR10 gene[J].Chinese Tobacco Science,2017,38(3):1-7.
[14]Chinnasri B,Borsics T,Christopher D A,Sipes B S.Induction of pathogenesis-related gene 1(PR-1)by acibenzolar-s-methyl application in pineapple and its effect on reniform nematodes(Rotylenchulus reniformis)[J].Agriculture&Natural Resources,2016,50(5):368-373.doi:10.1016/j.anres.2016.04.008.
[15]Sandhu D,Tasma I M,Frasch R,Bhattacharyya M K.Systemic acquired resistance in soybean is regulated by two proteins,Orthologous to Arabidopsis NPR1[J].Bmc Plant Biology,2009,9(1):105.doi:10.1186/1471-2229-9-105.
[16]Fu Z Q,Dong X.Systemic acquired resistance:turning local infection into global defense[J].Annual Review of Plant Biology,2013,64(1):839-863.doi:10.1146/annurev-arplant-042811-105606.
[17]杨德翠,张玉喜,郑国生.牡丹病程相关蛋白1基因的克隆及表达分析[J].园艺学报,2013,40(8):1583-1590.doi:10.16420/j.issn.0513-353x.2013.08.031.Yang D C,Zhang Y X,Zheng G S.Cloning and expression analysis of the disease related protein 1 gene of peony[J].Journal of Horticulture,2013,40(8):1583-1590.
[18]张青,赵景梅,黄东益,肖鑫辉,夏薇,许云,黄小龙,吴文嫱.大薯病程相关蛋白1(PR1)基因及其启动子序列的克隆与分析[J].分子植物育种,2018(7):2078-2084.doi:10.13271/j.mpb.016.002078.Zhang Q,Zhao J M,Huang D Y,Xiao X H,Xia W,Xu Y,Huang X L,Wu W Q.Cloning and analysis of the sequence of the large potato disease path related protein 1(PR1)gene and its promoter[J].Molecular Plant Breeding,2018(7):2078-2084.
[19]王乐,杨柳,郭志鸿,张玉宝,王亚军,杨果,谢忠奎.东方百合(Lilium)杂交品种索邦病程相关蛋白(PR1)基因的克隆与原核表达[J].中国沙漠,2018,38(3):584-591.doi:10.7522/j.issn.1000-694X.2017.00031.Wang L,Yang L,Guo Z H,Zhang Y B,Wang Y J,Yang G,Xie Z K.Cloning and prokaryotic expression of the disease related protein(PR1)gene of the lily(Lilium)hybrid[J].Chinese Desert,2018,38(03):584-591.
[20]马立功,张匀华,孟庆林,石凤梅,刘佳,李易初,王志英.向日葵病程相关蛋白Ha PR1基因的克隆与功能[J].作物学报,2015,41(12):1819-1827.doi:10.3724/SP.J.1006.2015.01819.Ma L G,Zhang Y H,Meng Q L,Shi F M,Liu J,Li YC,Wang Z Y.Cloning and function of Ha PR1 gene associated with sunflower disease path[J].Crop Journal,2015,41(12):1819-1827.
[21]秦朝,王斐,刘祥,王向东,眭顺照,李名扬.蜡梅CpPR-4基因的克隆及植物表达[J].西南师范大学学报(自然科学版),2012,37(12):98-101.doi:10.13718/j.cnki.xsxb.2012.12.012.Qin C,Wang F,Liu X,Wang X D,Sui S Z,Li M Y.Cloning and plant expression of Cp PR-4 gene from Chimonanthus praecox[J].Journal of Southwest China Normal University(Natural Science Edition),2012,37(12):98-101.
[22]肖栋,韦艳萍,李英,侯喜林.不结球白菜病程相关蛋白基因Bc PR5的克隆及表达分析[J].南京农业大学学报,2018,41(4):640-646.doi:10.7685/jnau.201708031.Xiao D,Wei Y P,Li Y,Hou X L.Cloning and expression analysis of a disease-related protein gene Bc PR5 in non-heading Chinese cabbage[J].Journal of Nanjing Agricultural University,2018,41(4):640-646.
[23]李雪姣,范伟,牛东东,关明俐,缪刘杨,史佳楠,窦世娟,魏健,刘丽娟,李莉云,刘国振.水稻病程相关PR1家族蛋白质在叶片生长及与白叶枯病菌互作反应中的表达[J].植物学报,2014,49(2):127-138.doi:10.3724/SP.J.1259.2014.00127.Li X J,Fan W,Niu D D,Guan M L,Mu L Y,Shi JN,Dou S J,Wei J,Liu L J,Li L Y,Liu G Z.Expression of PR1 family proteins related to rice pathogenesis in leaf growth and interaction with Xanthomonas oryzae[J].Plant Journal,2014,49(2):127-138.
[24]Gao L,Wang S,Li X Y,Wei X J,Zhang Y J,Wang H Y,Liu D Q.Expression and functional analysis of a Pathogenesis-related protein 1 gene,Tc Lr19PR1,involved in wheat resistance against leaf rust fungus[J].Plant Molecular Biology Reporter,2015,33(4):797-805.doi:10.1007/s11105-014-0790-5.
[25]Darwiche R,El Atab O,Baroni R M,Teixeira P J P L,Mondego J M C,Pereira G A G,Schneiter R.Plant pathogenesis related proteins of the cacao fungal pathogen Moniliophthora perniciosa differ in their lipid-binding specificities[J].Journal of Biological Chemistry,2017,292(50):20558-20569.doi:10.1074/jbc.m117.811398.
[26]栗小英,刘景坤,张艳俊,王海燕,刘大群.信号分子与叶锈菌诱导下小麦病程相关蛋白1基因的表达分析[J].江苏农业科学,2015,43(9):28-31.doi:10.15889/j.issn.1002-1302.2015.09.007.Li X Y,Liu J K,Zhang Y J,Wang H Y,Liu D Q.Expression analysis of protein 1 gene related to wheat pathogenesis induced by signal molecule and leaf rust[J].Jiangsu Agricultural Science,2015,43(9):28-31.
[2]张春.马铃薯常见病害的识别与防治[J].农业科技与信息,2016(28):103,105.doi:10.15979/j.cnki.cn62-1057/s.2016.28.065.Zhang C.Identification and control of common diseases of potato[J].Agricultural Technology and Information,2016(28):103,105.
[3]金飞跃,赵振杰,任乐乐,曹克强,胡同乐.十一种杀菌剂防治马铃薯晚疫病的适宜施用时期[J].农药学学报,2016,18(2):213-218.doi:10.16801/j.issn.1008-7303.2016.0028.Jin F Y,Zhao Z J,Ren L L,Cao K Q,Hu T L.Appropriate application period of eleven fungicides for controlling potato late blight[J].Journal of Agricultural Pharmacy,2016,18(2):213-218.
[4]张贺兰,王鑫,张若晗,杨圆圆,巩秀峰,赵永秀.利用基因工程育种提高马铃薯晚疫病抗性的研究进展[J].分子植物育种,2018,16(21):7038-7041.doi:10.13271/j.mpb.016.007038.Zhang H L,Wang X,Zhang R H,Yang Y Y,Gong XF,Zhao Y X.Advances in research on improving resistance of potato late blight by genetic engineering breeding[J].Molecular Plant Breeding,2018,16(21):7038-7041.
[5]Van Loon L C,Van Strien E A.The families of pathogenesis-related proteins,their activities,and comparative analysis of PR-1 type proteins[J].Physiological and Molecular Plant Pathology,1999,55(2):85-97.doi:10.1006/pmpp.1999.0213.
[6]Breiteneder H,Radauer C.A classification of plant food allergens[J].J Allergy Clin Immunol,2004,113(5):821-830.doi:10.1016/j.jaci.2004.01.779.
[7]Liu J J,Akm E.The family 10 of plant pathogenesis-related proteins:their structure,regulation,and function in response to biotic and abiotic stresses[J].Physiological&Molecular Plant Pathology,2006,68(1):3-13.doi:10.1016/j.pmpp.2006.06.004.
[8]Kim Y J,Jang M G,Lee H J,Jang G H,Sukweenadhi J,Kwon W S,Yang D C.Functional characterization of the pathogenesis-related protein family 10 gene,Pg PR10-4,from Panax ginseng,in response to environmental stresses[J].Plant Cell Tissue&Organ Culture,2014,118(3):531-543.doi:10.1007/s11240-014-0505-5.
[9]Sels J,Mathys J,De Coninck B M A,Cammue B P A,De Bolle M F C.Plant pathogenesis-related(PR)proteins:A focus on PR peptides[J].Plant Physiology and Biochemistry,2008,46(11):941-950.doi:10.1016/j.plaphy.2008.06.011.
[10]Ga~3Mez-Ga~3Mez L,Rubio-Moraga A,Ahrazem O.Molecular cloning and characterisation of a pathogenesis-related protein Cs PR10 from Crocus sativus[J].Plant Biology,2011,13(2):297-303.doi:10.1111/j.1438-8677.2010.00359.
[11]Uknes S.Regulation of pathogenesis-related protein-1a gene expression in tobacco[J].The Plant Cell Online,1993,5(2):159-169.doi:10.1105/tpc.5.2.159.
[12]Yun H K,Yi S Y,Yu S H,Choi D.Cloning of a pathogenesis-related protein-1 gene from Nicotians glutinosa L.and its Salicylic acid-independent induction by copper andβ-aminobutyric acid[J].Journal of Plant Physiology,1999,154(3):327-333.doi:10.1016/s0176-1617(99)80175-8.
[13]张玉,张增林,蒋彩虹,常爱霞,杨爱国,罗成刚,王绍美,王元英.烟草病程相关蛋白Nt PR10基因克隆与表达分析[J].中国烟草科学,2017,38(3):1-7.doi:10.13496/j.issn.1007-5119.2017.03.001.Zhang Y,Zhang Z L,Jiang C H,Chang A X,Yang AG,Luo C G,Wang S M,Wang Y Y,Cloning and expression analysis of tobacco pathogenesis related protein Nt PR10 gene[J].Chinese Tobacco Science,2017,38(3):1-7.
[14]Chinnasri B,Borsics T,Christopher D A,Sipes B S.Induction of pathogenesis-related gene 1(PR-1)by acibenzolar-s-methyl application in pineapple and its effect on reniform nematodes(Rotylenchulus reniformis)[J].Agriculture&Natural Resources,2016,50(5):368-373.doi:10.1016/j.anres.2016.04.008.
[15]Sandhu D,Tasma I M,Frasch R,Bhattacharyya M K.Systemic acquired resistance in soybean is regulated by two proteins,Orthologous to Arabidopsis NPR1[J].Bmc Plant Biology,2009,9(1):105.doi:10.1186/1471-2229-9-105.
[16]Fu Z Q,Dong X.Systemic acquired resistance:turning local infection into global defense[J].Annual Review of Plant Biology,2013,64(1):839-863.doi:10.1146/annurev-arplant-042811-105606.
[17]杨德翠,张玉喜,郑国生.牡丹病程相关蛋白1基因的克隆及表达分析[J].园艺学报,2013,40(8):1583-1590.doi:10.16420/j.issn.0513-353x.2013.08.031.Yang D C,Zhang Y X,Zheng G S.Cloning and expression analysis of the disease related protein 1 gene of peony[J].Journal of Horticulture,2013,40(8):1583-1590.
[18]张青,赵景梅,黄东益,肖鑫辉,夏薇,许云,黄小龙,吴文嫱.大薯病程相关蛋白1(PR1)基因及其启动子序列的克隆与分析[J].分子植物育种,2018(7):2078-2084.doi:10.13271/j.mpb.016.002078.Zhang Q,Zhao J M,Huang D Y,Xiao X H,Xia W,Xu Y,Huang X L,Wu W Q.Cloning and analysis of the sequence of the large potato disease path related protein 1(PR1)gene and its promoter[J].Molecular Plant Breeding,2018(7):2078-2084.
[19]王乐,杨柳,郭志鸿,张玉宝,王亚军,杨果,谢忠奎.东方百合(Lilium)杂交品种索邦病程相关蛋白(PR1)基因的克隆与原核表达[J].中国沙漠,2018,38(3):584-591.doi:10.7522/j.issn.1000-694X.2017.00031.Wang L,Yang L,Guo Z H,Zhang Y B,Wang Y J,Yang G,Xie Z K.Cloning and prokaryotic expression of the disease related protein(PR1)gene of the lily(Lilium)hybrid[J].Chinese Desert,2018,38(03):584-591.
[20]马立功,张匀华,孟庆林,石凤梅,刘佳,李易初,王志英.向日葵病程相关蛋白Ha PR1基因的克隆与功能[J].作物学报,2015,41(12):1819-1827.doi:10.3724/SP.J.1006.2015.01819.Ma L G,Zhang Y H,Meng Q L,Shi F M,Liu J,Li YC,Wang Z Y.Cloning and function of Ha PR1 gene associated with sunflower disease path[J].Crop Journal,2015,41(12):1819-1827.
[21]秦朝,王斐,刘祥,王向东,眭顺照,李名扬.蜡梅CpPR-4基因的克隆及植物表达[J].西南师范大学学报(自然科学版),2012,37(12):98-101.doi:10.13718/j.cnki.xsxb.2012.12.012.Qin C,Wang F,Liu X,Wang X D,Sui S Z,Li M Y.Cloning and plant expression of Cp PR-4 gene from Chimonanthus praecox[J].Journal of Southwest China Normal University(Natural Science Edition),2012,37(12):98-101.
[22]肖栋,韦艳萍,李英,侯喜林.不结球白菜病程相关蛋白基因Bc PR5的克隆及表达分析[J].南京农业大学学报,2018,41(4):640-646.doi:10.7685/jnau.201708031.Xiao D,Wei Y P,Li Y,Hou X L.Cloning and expression analysis of a disease-related protein gene Bc PR5 in non-heading Chinese cabbage[J].Journal of Nanjing Agricultural University,2018,41(4):640-646.
[23]李雪姣,范伟,牛东东,关明俐,缪刘杨,史佳楠,窦世娟,魏健,刘丽娟,李莉云,刘国振.水稻病程相关PR1家族蛋白质在叶片生长及与白叶枯病菌互作反应中的表达[J].植物学报,2014,49(2):127-138.doi:10.3724/SP.J.1259.2014.00127.Li X J,Fan W,Niu D D,Guan M L,Mu L Y,Shi JN,Dou S J,Wei J,Liu L J,Li L Y,Liu G Z.Expression of PR1 family proteins related to rice pathogenesis in leaf growth and interaction with Xanthomonas oryzae[J].Plant Journal,2014,49(2):127-138.
[24]Gao L,Wang S,Li X Y,Wei X J,Zhang Y J,Wang H Y,Liu D Q.Expression and functional analysis of a Pathogenesis-related protein 1 gene,Tc Lr19PR1,involved in wheat resistance against leaf rust fungus[J].Plant Molecular Biology Reporter,2015,33(4):797-805.doi:10.1007/s11105-014-0790-5.
[25]Darwiche R,El Atab O,Baroni R M,Teixeira P J P L,Mondego J M C,Pereira G A G,Schneiter R.Plant pathogenesis related proteins of the cacao fungal pathogen Moniliophthora perniciosa differ in their lipid-binding specificities[J].Journal of Biological Chemistry,2017,292(50):20558-20569.doi:10.1074/jbc.m117.811398.
[26]栗小英,刘景坤,张艳俊,王海燕,刘大群.信号分子与叶锈菌诱导下小麦病程相关蛋白1基因的表达分析[J].江苏农业科学,2015,43(9):28-31.doi:10.15889/j.issn.1002-1302.2015.09.007.Li X Y,Liu J K,Zhang Y J,Wang H Y,Liu D Q.Expression analysis of protein 1 gene related to wheat pathogenesis induced by signal molecule and leaf rust[J].Jiangsu Agricultural Science,2015,43(9):28-31.