多粘类芽孢杆菌HK18-8对辣椒炭疽病菌的抑制作用及其定殖能力
|
摘要
从健康的化香(Platycarya strobilacea Sieb. et Zucc.)根际土壤中分离获得生防菌HK18-8,为探究其对辣椒炭疽病的生防潜力,通过对峙培养和孢子萌发试验发现,HK18-8显著抑制短尖刺盘孢(Collectotrichum acutatum)的菌丝生长和分生孢子萌发,其抑菌带达到10mm,孢子萌发抑制率为70.15%;进行离体辣椒果实接种试验表明,HK18-8菌悬液对辣椒炭疽病的防治效果达到88.58%,其预防效果高于治疗效果,HK18-8的发酵液和菌悬液的防病效果显著;采用平板稀释法测定其在辣椒果实的定殖能力,处理9 d时仍能保持10~6 cfu·g~(-1)以上;根据形态特征、生理生化特性及16S rRNA序列同源性,HK18-8菌株鉴定为多粘类芽孢杆菌(Paenibacillus polymyxa)。
A promising biocontrol agent HK18-8,a strain with strong effect on pepper anthracnose,was selected from the rhizosphere of healthy dyetree plant. Its antifungal effect and colonization ability were detected by in vitro experiments. The results showed that,HK18-8 was able to inhibit the mycelial growth and conidia germination of Colletotrichum acutatum significantly in the in vitro experiment. The inhibition zone of the mycelial growth reached to 10 mm,and the inhibition rate of conidia germination was 70.15%. HK18-8 performed the successful potential control against pepper anthracnose(the control efficiency was 88.58%),and the preventive effect was higher than its therapeutic effect. In addition,HK18-8 was readily able to colonize on fruit surface of pepper,persisting with a relative population density of more than 10~6 cfu · g~(-1). Based on the morphological,physiological characteristics and 16 S rRNA sequence analysis,the strain HK18-8 was identified as Paenibacillus polymyxa.
A promising biocontrol agent HK18-8,a strain with strong effect on pepper anthracnose,was selected from the rhizosphere of healthy dyetree plant. Its antifungal effect and colonization ability were detected by in vitro experiments. The results showed that,HK18-8 was able to inhibit the mycelial growth and conidia germination of Colletotrichum acutatum significantly in the in vitro experiment. The inhibition zone of the mycelial growth reached to 10 mm,and the inhibition rate of conidia germination was 70.15%. HK18-8 performed the successful potential control against pepper anthracnose(the control efficiency was 88.58%),and the preventive effect was higher than its therapeutic effect. In addition,HK18-8 was readily able to colonize on fruit surface of pepper,persisting with a relative population density of more than 10~6 cfu · g~(-1). Based on the morphological,physiological characteristics and 16 S rRNA sequence analysis,the strain HK18-8 was identified as Paenibacillus polymyxa.
引文
Brian W,Naeem N,Alexander E W,Jacqueline M D,Yuan Z C.2016.Isolation,identification and characterization of Paenibacillus polymyxa CR1with potentials for biopesticide,biofertilization,biomass degradation and biofuel production.BMC Microbiology,16(1):244-251.
Buchanan R E,Gibbons N E.1994.Bergey’s manual of systematic bacteriology 9thed.Baltimore:Williams&Wilkins Company.
Hao T,Chen S.2017.Colonization of wheat,maize and cucumber by Paenibacillus polymyxa WLY78.PLoS ONE,12(1):e0169980.
He Hong,Shen Zhao-chang,Qiu Si-xin,Cai Xue-qing,Guan Xiong,Hu Fang-ping.2004.Fermentation conditions of endophytic antagonistic strain BS-2 of Bacillus subtilis.Chinese Journal of Biological Control,20(1):38-41.(in Chinese)何红,沈兆昌,邱思鑫,蔡学清,关雄,胡方平.2004.内生拮抗枯草芽孢杆菌BS-2菌株的发酵条件.中国生物防治,20(1):38-41.
Jiang Gui-fang,Song Li.2014.Advances on biologically controling pepper anthracnose.Hubei Agricultural Sciences,53(11):2481-2485.(in Chinese)蒋桂芳,宋力.2014.辣椒炭疽病生物防治技术的研究与展望.湖北农业科学,53(11):2481-2485.
Kloepper J W.1992.A review of issues related to messunring colonization of plant roots by bacteria.Canadian Journal of Microbiology,38(6):667-672.
Mao Ai-jun,Hu Qia,Geng San-sheng.2004.Studies on the inoculation technique for resistant varieties of pepper to anthracnose and the application.Acta Agriculturae Boreali-Sinica,(2):87-91.(in Chinese)毛爱军,胡洽,耿三省.2004.辣椒炭疽病抗病性鉴定技术及利用.华北农学报,(2):87-91.
Qin Gang,Wu Qing-li.2012.The intial screen of biological control and study of antimicrobial activity of capsicum anthracnose.Northern Horticulture,(6):124-128.(in Chinese)秦刚,吴庆丽.2012.辣椒炭疽病生防菌筛选及抗菌活性初步研究.北方园艺,(6):124-128.
Ren X L,Zhang N,Cao M H,Wu K,Shen Q L,Huang Q W.2012.Biological control of tobacco black shank and colonization of tobacco roots by Paenibacillus polymyxa strain C5.Biology and Fertility of Soil,48:618-620.
Ryu C M,Kim J W,Choi O H,Kim S H,Park C S.2006.Improvement of biological control capacity of Paenibacillus polymyxa E681 by seed pelleting on sesame.Biological Control,39(3):282-289.
Song Yong-yan,Li Ping,Zheng Ai-ping,Li Shu-jin,Tan Fu-rong.2006.Taxonomy of biocontrolling bacterial strain LM-3 and study on its antifungal protein.Journal of Sichuan University,43(5):1110-1115.(in Chinese)宋永燕,李平,郑爱萍,李姝晋,谭芙蓉.2006.生防细菌LM-3的鉴定及其抗菌蛋白的研究.四川大学学报,43(5):1110-1115.
Than P P,Haryudian P,Sitthisack P,Paul W J,Kevin D H.2008.Chili anthracnose disease caused by Colletotrichum species.Journal of Zhejiang University Science B,9(10):764-778.
Tian Jian,Zhou Hong-li,Yi You-jin,Zhou Jin-wei,Bo Lian-yang,Hu Xie-xin,Luo Cheng-yin,Xia Bo.2016.Screening of an antagonistic endophytic bacterium against Colletotrichum capsici and its fresh-keeping effects on postharvest Capsicum.Modern Food Science and Technology,32(5):151-160.(in Chinese)田建,周红丽,易有金,周金伟,柏连阳,胡谢馨,罗程印,夏菠.2016.辣椒炭疽病菌拮抗内生菌筛选及其对辣椒采后保鲜效果的研究.现代食品科技,32(5):151-160.
Wang Liu-qing,Wang Qiu-ying,Liao Mei-de.2013.The progress of biological properties and mechanisms of Paenibacillus polymyxa.Chinese Agricultural Science Bulletin,29(11):158-163.(in Chinese)王刘庆,王秋影,廖美德.2013.多粘类芽孢杆菌生物特性及其机理研究进展.中国农学通报,29(11):158-163.
Wang Ying-ying,Zhang Zi-xin,Xie Xue-wen,Guo Xian-kun,Jin Zhi-wen,Han Zhi-qi,Li Bao-ju.2014.Diagnosis and control of pepper anthracnose.Chinea Vegetables,(11):74-76.(in Chinese)王莹莹,张自心,谢学文,郭现坤,晋知文,韩之琪,李宝聚.2014.辣椒炭疽病的诊断与防治.中国蔬菜,(11):74-76.
Xia Hua,Zhu Hong-jian,Zhou Qian,Gao Bi-da.2012.Pathogen identification of a new anthracnose of pepper in Zhijiang,Hunan.Acta Phytopathologica Sinica,42(2):120-125.(in Chinese)夏花,朱宏建,周倩,高必达.2012.湖南芷江辣椒上一种新炭疽病的病原鉴定.植物病理学报,42(2):120-125.
Yang Hong-feng,Yu Xiang-yang,Xue Ya-rong,Liu Chang-hong.2014.Colonization of endophytic Bacillus amyloliquefaciens CC09 in wheat roots observed under transmission electron microscopy and its biocontrol efficiency against diseases.Chinese Journal of Biological Control,30(6):839-844.(in Chinese)杨洪凤,余向阳,薛雅蓉,刘常宏.2014.内生解淀粉芽孢杆菌CC09在小麦根部定殖的电镜观察及防病效果.中国生物防治学报,30(6):839-844.
Yang Qing,Yi Tu-yong.2009.Research progressin chilli anthracnose and its control.Acta Agriculturae Jiangxi,21(7):107-109.(in Chinese)杨青,易图永.2009.辣椒炭疽病及其防治研究进展.江西农业学报,21(7):107-109.
Yang Shao-bo,Liu Xun-li.2008.Research advances in Paenibacillus polymyxa and their bioactive substances.Microbiology,35(10):1621-1625.(in Chinese)杨少波,刘训理.2008.多粘类芽孢杆菌及其产生的生物活性物质研究进展.微生物学通报,35(10):1621-1625.
Zeng Da-xing,Zhang Xiao-yang,Jia Shu-juan,La Wen-jun,Tu Guo-quan.2015.Bacillus subtilis C-D6 as a potential biocontrol agent against appressorium formation of Colletotrichum capsici.Microbiology China,42(12):2377-2385.(in Chinese)曾大兴,张晓阳,贾书娟,喇文军,涂国全.2015.枯草芽孢杆菌C-D6对辣椒炭疽菌附着胞形成的抑制作用研究.微生物学通报,42(12):2377-2385.
Zhao Shuang,Liu Wei-cheng,Qiu Ji-yan,Wang Jing-guo,Zhou Li-gang.2008.Research progress of antimicrobial compounds and disease resistant mechanism of Paenibacillus polymyxa.Chinese Agricultural Science Bulletin,24(7):347-350.(in Chinese)赵爽,刘伟成,裘季燕,王敬国,周立刚.2008.多粘类芽孢杆菌抗菌物质和防病机制之研究进展.中国农学通报,24(7):347-350.
Zhu Hong-jian,Ouyang Xiao-yan,Zhou Qian,Gao Bi-da.2012.Isolation,identification and optimizing fermentation conditions of an antagonistic strain against Colletotrichum acutata.Acta Phytopathologica Sinica,42(4):418-424.(in Chinese)朱宏建,欧阳小燕,周倩,高必达.2012.一株辣椒尖孢炭疽病菌拮抗菌株的分离鉴定与发酵条件优化.植物病理学报,42(4):418-424.
Buchanan R E,Gibbons N E.1994.Bergey’s manual of systematic bacteriology 9thed.Baltimore:Williams&Wilkins Company.
Hao T,Chen S.2017.Colonization of wheat,maize and cucumber by Paenibacillus polymyxa WLY78.PLoS ONE,12(1):e0169980.
He Hong,Shen Zhao-chang,Qiu Si-xin,Cai Xue-qing,Guan Xiong,Hu Fang-ping.2004.Fermentation conditions of endophytic antagonistic strain BS-2 of Bacillus subtilis.Chinese Journal of Biological Control,20(1):38-41.(in Chinese)何红,沈兆昌,邱思鑫,蔡学清,关雄,胡方平.2004.内生拮抗枯草芽孢杆菌BS-2菌株的发酵条件.中国生物防治,20(1):38-41.
Jiang Gui-fang,Song Li.2014.Advances on biologically controling pepper anthracnose.Hubei Agricultural Sciences,53(11):2481-2485.(in Chinese)蒋桂芳,宋力.2014.辣椒炭疽病生物防治技术的研究与展望.湖北农业科学,53(11):2481-2485.
Kloepper J W.1992.A review of issues related to messunring colonization of plant roots by bacteria.Canadian Journal of Microbiology,38(6):667-672.
Mao Ai-jun,Hu Qia,Geng San-sheng.2004.Studies on the inoculation technique for resistant varieties of pepper to anthracnose and the application.Acta Agriculturae Boreali-Sinica,(2):87-91.(in Chinese)毛爱军,胡洽,耿三省.2004.辣椒炭疽病抗病性鉴定技术及利用.华北农学报,(2):87-91.
Qin Gang,Wu Qing-li.2012.The intial screen of biological control and study of antimicrobial activity of capsicum anthracnose.Northern Horticulture,(6):124-128.(in Chinese)秦刚,吴庆丽.2012.辣椒炭疽病生防菌筛选及抗菌活性初步研究.北方园艺,(6):124-128.
Ren X L,Zhang N,Cao M H,Wu K,Shen Q L,Huang Q W.2012.Biological control of tobacco black shank and colonization of tobacco roots by Paenibacillus polymyxa strain C5.Biology and Fertility of Soil,48:618-620.
Ryu C M,Kim J W,Choi O H,Kim S H,Park C S.2006.Improvement of biological control capacity of Paenibacillus polymyxa E681 by seed pelleting on sesame.Biological Control,39(3):282-289.
Song Yong-yan,Li Ping,Zheng Ai-ping,Li Shu-jin,Tan Fu-rong.2006.Taxonomy of biocontrolling bacterial strain LM-3 and study on its antifungal protein.Journal of Sichuan University,43(5):1110-1115.(in Chinese)宋永燕,李平,郑爱萍,李姝晋,谭芙蓉.2006.生防细菌LM-3的鉴定及其抗菌蛋白的研究.四川大学学报,43(5):1110-1115.
Than P P,Haryudian P,Sitthisack P,Paul W J,Kevin D H.2008.Chili anthracnose disease caused by Colletotrichum species.Journal of Zhejiang University Science B,9(10):764-778.
Tian Jian,Zhou Hong-li,Yi You-jin,Zhou Jin-wei,Bo Lian-yang,Hu Xie-xin,Luo Cheng-yin,Xia Bo.2016.Screening of an antagonistic endophytic bacterium against Colletotrichum capsici and its fresh-keeping effects on postharvest Capsicum.Modern Food Science and Technology,32(5):151-160.(in Chinese)田建,周红丽,易有金,周金伟,柏连阳,胡谢馨,罗程印,夏菠.2016.辣椒炭疽病菌拮抗内生菌筛选及其对辣椒采后保鲜效果的研究.现代食品科技,32(5):151-160.
Wang Liu-qing,Wang Qiu-ying,Liao Mei-de.2013.The progress of biological properties and mechanisms of Paenibacillus polymyxa.Chinese Agricultural Science Bulletin,29(11):158-163.(in Chinese)王刘庆,王秋影,廖美德.2013.多粘类芽孢杆菌生物特性及其机理研究进展.中国农学通报,29(11):158-163.
Wang Ying-ying,Zhang Zi-xin,Xie Xue-wen,Guo Xian-kun,Jin Zhi-wen,Han Zhi-qi,Li Bao-ju.2014.Diagnosis and control of pepper anthracnose.Chinea Vegetables,(11):74-76.(in Chinese)王莹莹,张自心,谢学文,郭现坤,晋知文,韩之琪,李宝聚.2014.辣椒炭疽病的诊断与防治.中国蔬菜,(11):74-76.
Xia Hua,Zhu Hong-jian,Zhou Qian,Gao Bi-da.2012.Pathogen identification of a new anthracnose of pepper in Zhijiang,Hunan.Acta Phytopathologica Sinica,42(2):120-125.(in Chinese)夏花,朱宏建,周倩,高必达.2012.湖南芷江辣椒上一种新炭疽病的病原鉴定.植物病理学报,42(2):120-125.
Yang Hong-feng,Yu Xiang-yang,Xue Ya-rong,Liu Chang-hong.2014.Colonization of endophytic Bacillus amyloliquefaciens CC09 in wheat roots observed under transmission electron microscopy and its biocontrol efficiency against diseases.Chinese Journal of Biological Control,30(6):839-844.(in Chinese)杨洪凤,余向阳,薛雅蓉,刘常宏.2014.内生解淀粉芽孢杆菌CC09在小麦根部定殖的电镜观察及防病效果.中国生物防治学报,30(6):839-844.
Yang Qing,Yi Tu-yong.2009.Research progressin chilli anthracnose and its control.Acta Agriculturae Jiangxi,21(7):107-109.(in Chinese)杨青,易图永.2009.辣椒炭疽病及其防治研究进展.江西农业学报,21(7):107-109.
Yang Shao-bo,Liu Xun-li.2008.Research advances in Paenibacillus polymyxa and their bioactive substances.Microbiology,35(10):1621-1625.(in Chinese)杨少波,刘训理.2008.多粘类芽孢杆菌及其产生的生物活性物质研究进展.微生物学通报,35(10):1621-1625.
Zeng Da-xing,Zhang Xiao-yang,Jia Shu-juan,La Wen-jun,Tu Guo-quan.2015.Bacillus subtilis C-D6 as a potential biocontrol agent against appressorium formation of Colletotrichum capsici.Microbiology China,42(12):2377-2385.(in Chinese)曾大兴,张晓阳,贾书娟,喇文军,涂国全.2015.枯草芽孢杆菌C-D6对辣椒炭疽菌附着胞形成的抑制作用研究.微生物学通报,42(12):2377-2385.
Zhao Shuang,Liu Wei-cheng,Qiu Ji-yan,Wang Jing-guo,Zhou Li-gang.2008.Research progress of antimicrobial compounds and disease resistant mechanism of Paenibacillus polymyxa.Chinese Agricultural Science Bulletin,24(7):347-350.(in Chinese)赵爽,刘伟成,裘季燕,王敬国,周立刚.2008.多粘类芽孢杆菌抗菌物质和防病机制之研究进展.中国农学通报,24(7):347-350.
Zhu Hong-jian,Ouyang Xiao-yan,Zhou Qian,Gao Bi-da.2012.Isolation,identification and optimizing fermentation conditions of an antagonistic strain against Colletotrichum acutata.Acta Phytopathologica Sinica,42(4):418-424.(in Chinese)朱宏建,欧阳小燕,周倩,高必达.2012.一株辣椒尖孢炭疽病菌拮抗菌株的分离鉴定与发酵条件优化.植物病理学报,42(4):418-424.