Crops ›› 2019, Vol. 35 ›› Issue (6): 162-167.doi: 10.16035/j.issn.1001-7283.2019.06.026

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Identification of Bacterium HT-6 and Its Antagonistic Stability against Phytophthora infestans

Zhang Congying,Jiang Jizhi,Liang Jiao,Qiao Liu,Huang Jie   

  1. College of Life Sciences, Hebei University, Baoding 071002, Hebei, China
  • Received:2019-04-28 Revised:2019-06-12 Online:2019-12-15 Published:2019-12-11
  • Contact: Jizhi Jiang

Abstract:

In order to clarify the biological characteristics and antagonistic stability of bacterium HT-6 strain on Phytophthora infestans, this experiment was conducted to observe its colony features on different media, detect physiological and biochemical characteristics and analyze 16S rDNA sequence, to determine its classification status. The effects of different temperature, pH, ultraviolet light, and continuous passage on the mycelial growth of Phytophthora infestans were tested. The HT-6 strain was identified as Bacillus subtilis based on results of its colony properties, physiological and biochemical characteristics and 16S rDNA sequence. The inhibition rates of HT-6 strain and its metabolites on P. infestans could keep more than 80% after treatment with 100℃ for 60min, pH 6-8 for 24h, 30W ultraviolet irradiation for 8h, and continuous 10 passages. These results showed that HT-6 strain and its metabolites had thermostability, ultraviolet resistance, and could store for a long time, except toleration acid and alkali. It has great potential value to develop it into a biological control agent for the prevention of potato late blight.

Key words: Potato, Strain identification, 16S rDNA, Bacteriostatic stability, Bacillus subtilis

Fig.1

Phylogenetic tree of HT-6 based on 16S rDNA sequence"

Table 1

Comparison of partial physiological and biochemical characteristics between HT-6 and its similar species"

试验Test HT-6 BS224[16] LXY-6[17] JY-1[18]
淀粉水解试验
Starch hydrolysis test
+ + + +
L-阿拉伯糖利用试验
L-arabionse utilization test
+ + + +
葡萄糖利用试验
Glucose utilization test
+ + + +
乳糖利用试验
Lactose utilization test
+ + ND -
木糖利用试验
Xylose utilization test
+ + + +
V. P试验V. P test + + - +
接触酶试验
Contact enzyme test
+ + + +
H2S试验H2S test + + + ND
溶血试验Hemolysis test + + + +
5% NaCl耐受试验
5% NaCl tolerance test
+ + + +
14% NaCl耐受试验
14% NaCl tolerance test
- - + -
30% NaCl耐受试验
30% NaCl tolerance test
- - + -
蛋白水解试验
Proteolysis test
+ + ND ND
色素试验Pigment test - - ND ND

Fig.2

Test of bacteriostatic stability of HT-6 strain bacterial solution Note: Different lowercase letters are significant difference at 0.05 level"

[1] Kamoun S, Furzer O, Jones J D , et al. The top 10 oomycete pathogens in molecular plant pathology. Molecular Plant Pathology, 2015,16(4):413-434.
doi: 10.1111/mpp.12190 pmid: 25178392
[2] 高康, 何蒲明 . 马铃薯主粮化战略研究. 合作经济与科技, 2018,7(14):31-33.
[3] Mosquera T, Alvarez M F, Jiménez-Gómez J M ,et al. Targeted and untargeted approaches unravel novel candidate genes and diagnostic SNPs for quantitative resistance of the potato (Solanum tuberosum L.) to Phytophthora infestans causing the late blight disease. PLoS ONE, 2016,11(6):e0156254.
doi: 10.1371/journal.pone.0156254 pmid: 27281327
[4] Miao J, Dong X, Lin D , et al. Activity of the novel fungicide oxathiapiprolin against plant-pathogenic oomycetes. Pest Management Science, 2016,72(8):1572-1577.
doi: 10.1002/ps.4189 pmid: 26577849
[5] 李成斌, 张红霞, 李岩 , 等. 2015-2017年北方5省(区)致病疫霉抗药性监测及与嘧菌酯交互抗性. 河北农业大学学报, 2018,41(6):75-79,103.
[6] Lazazzara V, Perazzolli M, Pertot I , et al. Growth media affect the volatilome and antimicrobial activity against Phytophthora infestans in four Lysobacter type strains. Microbiological Research, 2017,201(4):52-62.
doi: 10.1016/j.micres.2017.04.015 pmid: 28602402
[7] 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. 河北农业大学学报, 2015,38(2):76-81.
[8] 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. 河北农业大学学报, 2014,37(4):80-85.
[9] Cai G H, Myers K, Fry W E , et al. Phytophthora infestans RNA virus 2,a novel RNA virus from Phytophthora infestans,does not belong to any known virus group. Archives of Virology, 2019,164(2):567-572.
doi: 10.1007/s00705-018-4050-0 pmid: 30343382
[10] 武志华, 郭维维, 董晔 , 等. 抗马铃薯晚疫病菌的粘细菌菌株X6-Ⅱ-1的分离鉴定、拮抗活性及发酵条件的优化. 农业生物技术学报, 2018,26(9):1467-1479.
[11] Christopher K M, Tanya A, Amy N , et al. Phenazine-1-carboxylic acid production by Pseudomonas fluorescens LBUM636 alters Phytophthora infestans growth and late blight development. Phytopathology, 2017,107(5):273-279.
doi: 10.1094/PHYTO-06-16-0247-R pmid: 27827009
[12] 李丽艳, 蒋继志, 郭文 . 致病疫霉拮抗细菌的筛选及抑菌作用研究. 安徽农业科学, 2011,39(24):14671-14672,14675.
[13] 侯宁, 张荷花, 申芬 , 等. 核桃内生细菌HT-6对致病疫霉的持续抑制作用及其抑菌特性研究. 中国植保导刊, 2018,38(8):5-12,54.
[14] 布坎南, 吉本斯 . 伯杰氏细菌鉴定手册. 北京: 科学出版社, 1984.
[15] 东秀珠, 蔡妙英 . 常见细菌系统鉴定手册. 北京: 科学出版社, 2001.
[16] 喻江 . 枯草芽孢杆菌224 yplQ、ytjAa基因缺失菌株的构建及对其溶血性影响. 哈尔滨:东北农业大学, 2008.
[17] 李晓宇 . 解淀粉芽孢杆菌LXY-6-2筛选、鉴定、诱变与发酵条件优化及其对辣椒根腐病的生物防治研究. 杨凌:西北农林科技大学, 2008.
[18] 欧阳慧 . 抗猕猴桃果腐病西姆芽孢杆菌的分离鉴定及抑菌活性的初步研究. 南昌:江西农业大学, 2017.
[19] 梁允刚, 孟晶, 谭兰玉 , 等. 马铃薯晚疫病生防菌的分离鉴定与防治效果. 北方农业学报, 2017,45(4):54-57.
[20] Simon C, Annika G, Gil C , et al. Versatile antagonistic activities of soil-borne Bacillus spp. and Pseudomonas spp. against Phytophthora infestans and other potato pathogens. Frontiers in Microbiology, 2018,9:143.
doi: 10.3389/fmicb.2018.00143 pmid: 29487574
[21] 吴真真, 蒋继志, 汪洁 , 等. 致病疫霉颉颃细菌SR13-2菌液稳定性研究. 中国植保导刊, 2013,33(8):9-12,8.
[22] 王游游, 蒋继志, 王雪宁 , 等. 致病疫霉拮抗细菌WL2鉴定及其发酵液稳定性分析. 河北农业大学学报, 2017,40(4):90-95.
[23] 吴艳清, 王游游, 王畅 , 等. 枯草芽孢杆菌WL2脂肽粗提物对致病疫霉的抑制作用及其分离鉴定. 河北大学学报(自然科学版), 2018,38(6):632-639.
[24] 马佳, 李颖, 胡栋 , 等. 芽胞杆菌生物防治作用机理与应用研究进展. 中国生物防治学报, 2018,34(4):639-648.
[25] Zhao H, Xu X, Lei S , et al. Iturin A-like lipopeptides from Bacillus subtilis trigger apoptosis,paraptosis,and autophagy in Caco-2 cells. Journal of Cellular Physiology, 2019,234(5):6414-6427.
doi: 10.1002/jcp.27377 pmid: 30238995
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