比较在前期获得的明显抑制致病疫霉菌丝生长的6株细菌的抑菌性能及其防病潜力,为开发生防制剂防治马铃薯晚疫病提供依据.采用对峙培养法和打孔法测定6株细菌活体、菌液和无菌体发酵液对病菌菌丝生长的抑制作用,利用块茎切片法评价拮抗菌菌液对晚疫病的离体防治效果.结果表明,6株菌的菌液对菌丝生长的抑制作用均明显优于活体和无菌体发酵液,抑菌带宽度均在27 mm以上,且均可引起菌丝体畸变,其中以WL2菌株的抑菌作用最强,活体、菌液和发酵液的抑菌带宽均在26 mm以上,致畸作用也最强;此外,6株菌的菌液在离体块茎切片上的防病效果也以WL2菌液处理最好,病情指数最低(13.7),相对保护率最高(80.5%).这表明菌株WL2比其他5个菌株在防治马铃薯晚疫病方面有更大应用潜力.
The six bacterial strains including WL1,WL2,WR1,WR2,WK1 and M15,obtained in previously experiments,had the obvious inhibition to Phytophthora infestans mycelia growth.In order to clarify the biocontrol potential,the inhibition property and the disease prevention effects on in vitro tissue of six strains were compared in this experiment.The dual-culture method and punch method were adopted to determine the inhibition of living bacteria,bacterial liquid,as well as fermented liquid on P.infestans mycelia growth.The potato tuber slices method was used to evaluate the disease prevention effect of bacterial liquid.The results showed that the inhibition effects of six bacterial liquid to P.infestans growth were significantly greater than that of living bacteria and fermented liquid,and the inhibitory zones were above 27 mm,and all of them caused mycelial malformation.In which,bacterial liquid of WL2 strain displayed the strongest inhibitory effect,the inhibitory zones were above 26 mm,and caused the most serious malformation.In- addition,WL2 bacterial liquid acquired the best disease prevention on in vitro tissue in comparison with other 5 strains,the disease index was only 13.7,and the relative protection ratio was up to 80.5%.These results indicate that WL2 strain has more potential in control potato late blight in the future.
[1] 蒋晨晨.马铃薯主粮化的供求调适研究[D].泰安:山东农业大学,2016. JIANG C C.Research on supply and demand adjustment measures of the potatoes staple food[D].Taian:Shandong Agricultural University,2016.
[2] 卢肖平.马铃薯主粮化战略的意义、瓶颈与政策建议[J].华中农业大学学报(社会科学版),2015(3):1-7.DOI:10.13300/j.cnki.hnwkxb.2015.03.001. LU X P.Strategy of potato as staple food:significance,bottlenecks and policy suggestions[J].Journal of Huazhong Agricultural University(Social Sciences Edition),2015(3):1-7.DOI:10.13300/j.cnki.hnwkxb.2015.03.001.
[3] JAHAN S N,SMAN A K,CORCORAN P,et al.Plant-mediated gene silencing restricts growth of the potato late blight pathogen Phytophthora infestans[J].Journal of Experimental Botany,2015,66(9):2785-2794.DOI:10.1093/jxb/erv094.
[4] 陈晓梅.马铃薯致病疫霉生理小种多样性及其空间分布[D].福州:福建农林大学,2014. CHEN X M.Race diversity and its spatial distribution in the potato late blight pathoen Phytophthora infestans[D].Fuzhou:Fujian Agriculture & Forestry University,2014.
[5] 杨胜先,龙国,张绍荣,等.马铃薯晚疫病菌生理小种研究进展[J].江苏农业科学,2015,43(9):9-13.DOI:10.15889/j.issn.1002-1302.2015.09.003.
[6] 吴艳清,蒋继志,梁廷银.致病疫霉拮抗芽孢杆菌的筛选及脂肽类物质分离的研究[J].安徽农业科学, 2012,40(14):8111-8112.DOI:10.13989/j.cnki.0517-6611.2012.14.103. WU Y Q,JIANG J Z,LIANG T Y.Screening of antagonistic Bacillus spp.strains against Phytophthora infestans(Mont.)de bary and isolation of lipopeptide substances[J].Journal of Anhui Agri Sci,2012,40(14):8111-8112.DOI:10.13989/j.cnki.0517-6611.2012.14.103.
[7] 黄英菊,蒋继志,冯丽娜,等.致病疫霉颉颃细菌研究进展[J].中国植保导刊,2013,33(8):18-22.DOI:10.3969/j.issn.1672-6820.2013.08.007. HUANG Y J,JIANG J Z,FENG L N,et al.Research progress on antagonistic bacteria against Phytophthora infestans[J].China Plant Protection,2013,33(8):18-22.DOI:10.3969/j.issn.1672-6820.2013.08.007.
[8] CLERMONT N,LERAT S,BEAULIEU C.Genome shuffling enhances biocontrol abilities of Streptomyces strains against two potato pathogens[J].Journal of Applied Microbiology,2011,111(3):671-682.DOI:10.1111/j.1365-2672.2011.05078.x.
[9] 蒋继志,郭俊亭,李丽艳.致病疫霉拮抗真菌研究进展[J].河北大学学报(自然科学版),2012,32(1):105-112.DOI:10.3969/j.issn.1000-1565.2012.01.021. JIANG J Z,GUO J T,LI L Y.Advances of antagonistic fungi against Phytophthora infestans[J].Journal of Hebei University(Naturnal Science Edition),2012,32(1):105-112.DOI:10.3969/j.issn.1000-1565.2012.01.021.
[10] ANOUK G,MOUT D V,DENISE B,et al.The anti-phytophthora effect of selected potato-associated Pseudomonas strains:from the laboratory to the field[J].Frontiers in Microbiology,2015,6:1309.DOI:10.3389/fmicb.2015.01309.
[11] 李颖,金鑫,蒋继志,等.致病疫霉拮抗细菌筛选及J-28菌株鉴定[J].中国植保导刊,2016,36(1):10-15,19.DOI:10.3969/j.issn.1672-6820.2016.01.002. LI Y,JIN X,JIANG J Z,et al.Screening of antagonistic bacteria against Phytophthora infestans and identification of J-28 strain[J].China Plant Protection,2016,36(1):10-15,19.DOI:10.3969/j.issn.1672-6820.2016.01.002.
[12] 黄英菊,蒋继志,冯丽娜,等.几种拮抗菌对致病疫霉抑制作用的比较研究[J].河北农业大学学报,2014,37(4):81-85.DOI:10.13320/j.cnki.jauh 2014.0093. HUANG Y J,JIANG J Z,FENG L N,et al.Inhibition comparison of several antagonists against Phytophthora infestans[J].Journal of Agricultural University of Hebei,2014,37(4):81-85.DOI:10.13320/j.cnki.jauh 2014.0093.
[13] 郭会婧,蒋继志,李向彬,等.致病疫霉拮抗菌筛选及复合发酵液的抑菌作用研究[J].河北农业大学学报,2008,31(4):87-90.DOI:10.3969/j.issn.1000-1573.2008.04.018. GUO H J,JIANG J Z,LI X B,et al.Screening of antagonistic strains and inhibition of combination fermentation against Phytophthora infestans[J].Journal of Agricultural University of Hebei,2008,31(4):87-90.DOI:10.3969/j.issn.1000-1573.2008.04.018.
[14] 蒋继志,梁廷银,王怀远,等.致病疫霉拮抗菌M15假单胞菌的筛选及离体防病作用[J].河北农业大学学报,2013,36(3):72-76. JIANG J Z,LIANG T Y,WANG H Y,et al.Screening of antagonistic Pseudomonas fluorescens against Phytophthora infestans and disease control in vitro[J].Journal of Agricultural University of Hebei,2013,36(3):72-76.
[15] 冯丽娜,蒋继志,赵偲,等.内生真菌DC-11的分离筛选及对致病疫霉的抑制作用[J].河北农业大学学报,2015,38(2):76-81.DOI:10.13320/j.cnki jauh 2015.0038. FENG L N,JIANG J Z,ZHAO C,et al.Screening of endophytic fungi DC-11 strain and its inhibition to Phytophthora infestans[J].Journal of Agricultural University of Hebei,2015,38(2):76-81.DOI:10.13320/j.cnki jauh 2015.0038.
[16] 郭尧,李丽,蒋继志,等.致病疫霉拮抗放线菌Sy11发酵条件的优化[J].河北农业大学学报,2014,37(2):71-75.DOI:10.13320/j.cnki.jauh.2014.0040. GUO Y,LI L,JIANG J Z,et al.Optimization of fermentation conditions of Streptomyces albus Sy11 against Phytophthora infestans[J].Journal of Agricultural University of Hebei,2014,37(2):71-75.DOI:10.13320/j.cnki.jauh.2014.0040.
[17] VELIVELLI S L S,PAUL D V,PETER K,et al.Biological control agents:From field to market,problems,and challenges[J].Trends in Biotechnology,2014,32(10):493-496.DOI:10.1016/j.tibtech.2014.07.002.
[18] HUNZIKER L,BONISCH D,GROENHAGEN U,et al.Pseudomonas strains naturally associated with potato plants produce volatiles with high potential for inhibition of Phytophthora infestans[J].Applied and Environmental Microbiology,2015,81(3):821-830.DOI:10.1128/AEM.02999-14.
[19] 杨立宾,宋瑞清,邓勋,等.致病疫霉生防木霉菌株筛选鉴定及生物学特性[J].东北林业大学学报, 2013,41(7):118-122.DOI:10.13759/j.cnki.dlxb.2013.07.015. YANG L B,SONG R Q,DENG X,et al.Screening,identification and biological characteristics of Trichoderma strain on biological control of Phytophthora infestans[J].Journal of Northeast Forestry University, 2013,41(7):118-122.DOI:10.13759/j.cnki.dlxb.2013.07.015.
