立枯丝核菌AG-1IA对水稻致病力及粗毒素活性的测定方法研究
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摘要
水稻纹枯病是严重影响水稻生产的真菌病害,水稻相对病斑高度法作为评估立枯丝核菌AG-1 IA致病力的传统方法,需要将水稻培养至分蘖末期或抽穗期,再进行活体接种,耗时耗力,且试验条件不易控制。本研究以水稻相对病斑高度法(活体接种)为对照,利用水稻离体叶鞘法和离体叶片法测定了水稻纹枯病菌致病力和粗毒素活性,比较活体接种法与离体接种法的相关性。结果表明水稻离体叶片法、水稻离体叶鞘法和水稻相对病斑法测定致病力和粗毒素活性结果均呈显著正相关,水稻离体叶片法和离体叶鞘法测定结果能够准确反映立枯丝核菌AG-1 IA的致病力和粗毒素活性,离体评估方法操作方便,试验条件易于控制。因此,水稻离体叶片法、叶鞘法可以替代活体接种法作为测定立枯丝核菌AG-1 IA致病力和粗毒素活性的方法。水稻离体叶片法、叶鞘法与活体接种法结合起来可作为测定立枯丝核菌AG-1 IA毒素活性测定的新方法。本研究的结果还表明,水稻不同组织对水稻纹枯病菌毒素敏感性不同,水稻的叶鞘组织比叶片组织对纹枯病菌敏感。由于粗毒素的致病力与活体菌株接种的致病力存在显著差异,因此纹枯病菌毒素以外的致病因子,也是不容忽视的。在定量分析立枯丝核菌AG-1 IA与水稻的互作中,需要将不同水稻的组织进行区分,才能更加精准地了解互作机制。
Rice sheath blight is the serious fungual diseases in rice.The traditional method to test the pathogenicity of Rhizoctonia solani based on the relative height of lesion on rice needs to inoculate rice at late tillering or heading stage which is time-consuming and difficult to control the conditions.In this study,compared with the traditional way,the evaluation of Rhizoctonia virulence and bioactivity of crude toxin was conducted on the way of detached rice leaf and rice leaf sheath.The results showed that,although all the three approaches of inoculation were significantly positive to evaluate the pathogenicity and crude toxin activity,the detached ways seem to be more accurate with the advantage of easy handling and condition controlling.Meanwhile,we noticed the rice sheath are more sensitivity to crude toxin of R.solani than rice leaf which might be the variation of sensitivity of the different rice tissues to the toxin.Therefore,the detached rice leaf and leaf sheath methods may take place of the traditional way of height of lesion to determine virulence of R.solani.The combination of the three approaches can be used as a new method to determine toxin activity of R.solani.The lesions difference between rice leaf and leaf sheath suggested that the susceptibility of different rice tissues to the fungus is varied.In addition,the significant differences with the severity of disease index caused by crude toxin and isolate inoculation imply that the other virulence factors should be taken into concern besides the toxin.During the quantitative analyzing the interactions of Rhizoctonia solani AG-1 IA and host rice,the different rice tissues should be taken consideration together for more accurate exploring of the interaction mechanism.
Rice sheath blight is the serious fungual diseases in rice.The traditional method to test the pathogenicity of Rhizoctonia solani based on the relative height of lesion on rice needs to inoculate rice at late tillering or heading stage which is time-consuming and difficult to control the conditions.In this study,compared with the traditional way,the evaluation of Rhizoctonia virulence and bioactivity of crude toxin was conducted on the way of detached rice leaf and rice leaf sheath.The results showed that,although all the three approaches of inoculation were significantly positive to evaluate the pathogenicity and crude toxin activity,the detached ways seem to be more accurate with the advantage of easy handling and condition controlling.Meanwhile,we noticed the rice sheath are more sensitivity to crude toxin of R.solani than rice leaf which might be the variation of sensitivity of the different rice tissues to the toxin.Therefore,the detached rice leaf and leaf sheath methods may take place of the traditional way of height of lesion to determine virulence of R.solani.The combination of the three approaches can be used as a new method to determine toxin activity of R.solani.The lesions difference between rice leaf and leaf sheath suggested that the susceptibility of different rice tissues to the fungus is varied.In addition,the significant differences with the severity of disease index caused by crude toxin and isolate inoculation imply that the other virulence factors should be taken into concern besides the toxin.During the quantitative analyzing the interactions of Rhizoctonia solani AG-1 IA and host rice,the different rice tissues should be taken consideration together for more accurate exploring of the interaction mechanism.
引文
[1]Wang Y,Li G Q.Identification of Streptomyces platensi strain F-1 and the control effect on rice sheath blight(in Chinese)[J].Journal of Shandong Agricultural University(Natural Science Edition)(山东农业大学学报(自然科学版)),2014,45(3):321-327.
[2]Yang Y Q,Yang M,Lan B fit al.Research progress in pathogenic mechanisms of rice rheath blight pathogen(in Chinese)[J].Chinese Agricultural Science Bulletin(中国农学通报),2014,30(28):245-250.
[3]Lee F N,Rush M C.Rice sheath blight:a major rice disease[J].Plant Disease,1983,67(7):829-832.
[4]Stanton M A,Rothrock C S,Stemwart J M.Response of A-genome cotton germplasm to the seedling disease pathogens,Rhizoctonia solani and Pythium ultimum[J].Genetic Resources and Crop Evolution,1994,41(1):9-12.
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[6]Zhou E X,Yang M.A rapid and simple technique for the isolation of Rhizoctonia solani from diseased plant tissues(in Chinese)[J].J.South China Agric.Univ.(华南农业大学学报),1998,19(1):123-126.
[7]Balali G R,Neate S M,Kasalkheh AM,etal.Intraspecific variation of Rhizoctonia solani AG 3 isolates recovered from potato fields in Central Iran and South Australia[J].Mycopathologia,2007,163(2):105-115.
[8]Hang W W,Wang L,Liu L M,et al.Classification and genetic diversity research progress of rice sheath blight by Rhizoctonia solani(in Chinese)[J].China Rice(中国稻米),2010,16(3):34-38.
[9]Duan C F,Yang G H,Ni Z G,et al.Occurrence of foliar rot of Chinese cabbage,mint and lettuce caused by Rhizoctonia solani AG-1IB in China(in Chinese)[J].Journal of Yunnan Agricultural University(云南农业大学学报),2008,23(3):422-425.
[10]Men L J.Separation,purification,composition analysis and inactivation of the toxin of Rhizoctonia solani(in Chinese)[D].Yangzhou:Yangzhou University(扬州:扬州大学),2009.
[11]Chen J,Tang C R,Zou Q D,et al.On pathogenicity factors in maize sheath blight(in Chinese)[J].Journal of Shenyang Agricultural University(沈阳农业大学学报),1999,30(3):189-194.
[12]Vidhyasekaran P,Ponmalar T R,Samiyappan R,et al.Host-specific toxin production by Rhizoctonia solani,the rice sheath blight pathogen[J].Phytopathology,1997,87(12):1258-1263.
[13]Tao J F.Studies on the infection process of Rhizoctonia solani in rice(in Chinese)[J].J.Sichuan Agric.Univ.(四川农业大学学报),1992,10(3):471-477.
[14]Lang B Y,Li J,Xie J fit al.Screening of Trichoderma activity inhibiting five kinds of pathogens(in Chinese)[J].Biological Disaster Science(生物灾害科学),2013,36(2):142-147.
[15]Chen X J,Pan C H,Men L J,et al.Separation,purification,and preliminary composition analysis of the toxin of Rhizoctonia solani,the pathogen of rice sheath blight(in Chinese)[J].Journal of Yangzhou University(Agricultural and Life Science Edition)(扬州大学学报:(农业与生命科学版)),2011,32(1):44-48.
[16]Zuo S M,Chen X J,Chen H Q,et al.Defense response and physiological difference of rice cultivars with different sheath blight resistance levels to the toxins produced by Rhizoctonia solani(in Chinese)[J].China J.Rice Sci.(中国水稻科学),2014,28(5):551-558.
[17]Xun J Y,Zhang H D,Zhang H fit at.Toxin produced by Rhizoctonia solani and its relationship with pathogenicity of the fungus(in Chinese)[J].Journal of Yangzhou University(Agricultural and Life Science Edition)(扬州大学学报(农业与生命科学版)),2004,25(2):61-64.
[18]Zheng A P,Lin R M,Zhang DH,et al.The evolution and pathogenic mechanisms of the rice sheath blight pathogen[J].Nat.Commun,2013,4:1424.doi:10.1038/ncomms2427.
[19]Meng Q Z,Liu Z H,Wang HY.et al.Research progress in rice sheath blight(in Chinese)[J].Journal of Shenyang Agricultural University(沈阳农业大学学报),2001,32(5):376-381.
[20]Tong Y H,Xu J Y,Pan X B fit al.A primary study on the resistant mechanism of rice plants to the infection of Rhizoctonia solani(in Chinese)[J].Jiangsu Agricultural Research(江苏农业研究),2000,21(4):45-47.
[21]Zhang G L,Yan D M,He Z H.Cytological characteristics of infection process by Rhizoctonia solani in rice(in Chinese)[J].Chinese Journal of Cell Biology(细胞生物学杂志),2010,32(3):451-455.
[22]Pan X B,Chen Z X,Xu J Y,et al.The effects of different methods of inoculation and investigation on genetic research of resistance to rice sheath blight(in Chinese)[J].Journal of Jiangsu Agricultural College(江苏农学院学报),1997,18(3):27-32.
[2]Yang Y Q,Yang M,Lan B fit al.Research progress in pathogenic mechanisms of rice rheath blight pathogen(in Chinese)[J].Chinese Agricultural Science Bulletin(中国农学通报),2014,30(28):245-250.
[3]Lee F N,Rush M C.Rice sheath blight:a major rice disease[J].Plant Disease,1983,67(7):829-832.
[4]Stanton M A,Rothrock C S,Stemwart J M.Response of A-genome cotton germplasm to the seedling disease pathogens,Rhizoctonia solani and Pythium ultimum[J].Genetic Resources and Crop Evolution,1994,41(1):9-12.
[5]Chen S Q,Wu B,Qin M,et al.A study on the anostomasis group of Rhizoctonia solani K(u|")hn in Sichuan Province(in Chinese)[J].Southwest China Journal of Agricultural Sciences(西南农业学报),1998,12(1):56-60.
[6]Zhou E X,Yang M.A rapid and simple technique for the isolation of Rhizoctonia solani from diseased plant tissues(in Chinese)[J].J.South China Agric.Univ.(华南农业大学学报),1998,19(1):123-126.
[7]Balali G R,Neate S M,Kasalkheh AM,etal.Intraspecific variation of Rhizoctonia solani AG 3 isolates recovered from potato fields in Central Iran and South Australia[J].Mycopathologia,2007,163(2):105-115.
[8]Hang W W,Wang L,Liu L M,et al.Classification and genetic diversity research progress of rice sheath blight by Rhizoctonia solani(in Chinese)[J].China Rice(中国稻米),2010,16(3):34-38.
[9]Duan C F,Yang G H,Ni Z G,et al.Occurrence of foliar rot of Chinese cabbage,mint and lettuce caused by Rhizoctonia solani AG-1IB in China(in Chinese)[J].Journal of Yunnan Agricultural University(云南农业大学学报),2008,23(3):422-425.
[10]Men L J.Separation,purification,composition analysis and inactivation of the toxin of Rhizoctonia solani(in Chinese)[D].Yangzhou:Yangzhou University(扬州:扬州大学),2009.
[11]Chen J,Tang C R,Zou Q D,et al.On pathogenicity factors in maize sheath blight(in Chinese)[J].Journal of Shenyang Agricultural University(沈阳农业大学学报),1999,30(3):189-194.
[12]Vidhyasekaran P,Ponmalar T R,Samiyappan R,et al.Host-specific toxin production by Rhizoctonia solani,the rice sheath blight pathogen[J].Phytopathology,1997,87(12):1258-1263.
[13]Tao J F.Studies on the infection process of Rhizoctonia solani in rice(in Chinese)[J].J.Sichuan Agric.Univ.(四川农业大学学报),1992,10(3):471-477.
[14]Lang B Y,Li J,Xie J fit al.Screening of Trichoderma activity inhibiting five kinds of pathogens(in Chinese)[J].Biological Disaster Science(生物灾害科学),2013,36(2):142-147.
[15]Chen X J,Pan C H,Men L J,et al.Separation,purification,and preliminary composition analysis of the toxin of Rhizoctonia solani,the pathogen of rice sheath blight(in Chinese)[J].Journal of Yangzhou University(Agricultural and Life Science Edition)(扬州大学学报:(农业与生命科学版)),2011,32(1):44-48.
[16]Zuo S M,Chen X J,Chen H Q,et al.Defense response and physiological difference of rice cultivars with different sheath blight resistance levels to the toxins produced by Rhizoctonia solani(in Chinese)[J].China J.Rice Sci.(中国水稻科学),2014,28(5):551-558.
[17]Xun J Y,Zhang H D,Zhang H fit at.Toxin produced by Rhizoctonia solani and its relationship with pathogenicity of the fungus(in Chinese)[J].Journal of Yangzhou University(Agricultural and Life Science Edition)(扬州大学学报(农业与生命科学版)),2004,25(2):61-64.
[18]Zheng A P,Lin R M,Zhang DH,et al.The evolution and pathogenic mechanisms of the rice sheath blight pathogen[J].Nat.Commun,2013,4:1424.doi:10.1038/ncomms2427.
[19]Meng Q Z,Liu Z H,Wang HY.et al.Research progress in rice sheath blight(in Chinese)[J].Journal of Shenyang Agricultural University(沈阳农业大学学报),2001,32(5):376-381.
[20]Tong Y H,Xu J Y,Pan X B fit al.A primary study on the resistant mechanism of rice plants to the infection of Rhizoctonia solani(in Chinese)[J].Jiangsu Agricultural Research(江苏农业研究),2000,21(4):45-47.
[21]Zhang G L,Yan D M,He Z H.Cytological characteristics of infection process by Rhizoctonia solani in rice(in Chinese)[J].Chinese Journal of Cell Biology(细胞生物学杂志),2010,32(3):451-455.
[22]Pan X B,Chen Z X,Xu J Y,et al.The effects of different methods of inoculation and investigation on genetic research of resistance to rice sheath blight(in Chinese)[J].Journal of Jiangsu Agricultural College(江苏农学院学报),1997,18(3):27-32.