Phytophthora infestans could produce some signal molecules which stimulated or induced Streptomyces microflavus Sy11 strain to produce anti-oomycetes substances when it was cultured against the S. microflavus Sy11 strain. To clarify in which culture mode P. infestans could more likely produce these signal molecules and what kinds of substance they are, the growth rate method was adopted to compare the induction activities of the signal molecules produced by the pathogen between individual culture(solid culture and liquid culture)and dual culture(solid culture and liquid culture)with S. microflavus Sy11 strain, and different organic solvents were used to analyze the substance type of the signal moleculars. The results showed- DOI:10.3969/j.issn.1000-1565.2020.01.010对放线菌Sy11有诱导作用的致病疫霉信号分子研究郝志伟,蒋继志,吴路芳,张红霞,张荷花(河北大学 生命科学学院,河北 保定 071002)摘 要:致病疫霉(Phytophthora infestans)能产生一种信号分子,诱导放线菌Sy11菌株产生抑菌物质从而抑制致病疫霉.为明确在哪种培养方式下致病疫霉更易产生这种信号分子以及信号分子的物质类别,本文采用生长速率法对致病疫霉单独培养(单独固体培养和单独液体培养)以及与Sy11共培养(对峙培养和混合培养)产生的信号分子诱导Sy11抑菌作用的活性进行了比较,并以不同有机溶剂对信号分子的物质类别进行了初步分析.结果显示,致病疫霉无论单独培养还是与Sy11共培养,均可产生诱导Sy11抑菌作用的信号分子,其中以在固体黑麦培养基上与Sy11对峙培养产生的信号分子的诱导活性最强,菌落直径为24.79 mm,但与其他3种培养方式所产生的信号分子的诱导活性并无显著性差异;对致病疫霉单独液体培养后无菌体发酵液的分析表明,其中脂类粗提物的诱导活性(菌落直径32.8 mm)远高于糖类(菌落直径61.5 mm)和蛋白质类(菌落直径60 mm)粗提物,进一步对其中脂类物质的萃取显示,乙酸乙酯的萃取效果明显优于乙醇和石油醚.乙酸乙酯萃取物诱导Sy11后对致病疫霉菌丝生长有显著的抑制作用(菌落直径为35 mm)与乙醇萃取液的处理达到显著性差异.以上结果表明,致病疫霉生长过程中无论单独培养还是与Sy11共培养均可产生能诱导Sy11抑菌作用的信号分子,该信号分子以脂类物质为主,可用乙酸乙酯萃取得到.关键词:致病疫霉;Sy11菌株;诱导感应现象;信号分子中图分类号:S435.32 文献标志码:A 文章编号:1000-1565(2020)01-0063-08Signal molecule produced by Phytophthora infestans induced actinomyces Sy11HAO Zhiwei, JIANG Jizhi, WU Lufang, ZHANG Hongxia, ZHANG Hehua(College of Life Sciences, Hebei University, Baoding 071002,China)Abstract: Phytophthora infestans could produce some signal molecules which stimulated or induced Streptomyces microflavus Sy11 strain to produce anti-oomycetes substances when it was cultured against the S. microflavus Sy11 strain. To clarify in which culture mode P. infestans could more likely produce these signal molecules and what kinds of substance they are, the growth rate method was adopted to compare the induction activities of the signal molecules produced by the pathogen between individual culture(solid culture and liquid culture)and dual culture(solid culture and liquid culture)with S. microflavus Sy11 strain, and different organic solvents were used to analyze the substance type of the signal moleculars. The results showed- 收稿日期:2019-05-31 基金项目:河北省自然科学基金资助项目(C2015201231);河北省研究生创新项目(CXZZSS2018002) 第一作者:郝志伟(1993—),男,河北大学在读硕士研究生,主要从事植物与微生物相互作用的分子机理研究.E-mail:2942236040@qq.com 通信作者:蒋继志(1960—),男,河北大学教授,博士生导师,主要从事植物病害生物防治及其分子机理研究.E-mail:jizhijiang909@163.com第1期郝志伟等:对放线菌Sy11有诱导作用的致病疫霉信号分子研究that P.infestans could produce signal molecules inducing Sy11 either alone or in co-culture with Sy11. In which, the most induction activity was found in co-cultured of P.infestans with Sy11 on solid rye medium, the colony diameter was 24.79 mm, but there were no significant differences comparing with other three culture modes. Analysis of free cell fermentation broth of P.infestans culture alone by different organic solvents showed that induction activity of crude lipid extracts or lipid-containing substances(colony diameter 32.8 mm)was much higher than that of polysaccharide crude extracts(colony diameter 61.5 mm)and protein crude extracts(colony diameter 60 mm). Futher experimental results indicated that the extraction effect of the ethyl acetate was better than that of ethanol and petroleum ether. The colony diameter of P.infestans was 35 mm after induction by the ethyl acetate extracts and the significant difference was found between ethyl acetate and ethanol extracts treatments. These findings suggested that P.infestans could produce some signal molecules which induce the anti-oomycetes of Sy11 strain in both individual culture and co-culture mode with Sy11, these signal molecules are lipid substances and easily extracted with ethyl acetate.
HAO Zhiwei
,
JIANG Jizhi
,
WU Lufang
,
ZHANG Hongxia
,
ZHANG Hehua
. Signal molecule produced by Phytophthora infestans induced actinomyces Sy11[J]. Journal of Hebei University(Natural Science Edition), 2020
, 40(1)
: 63
-70
.
DOI: 10.3969/j.issn.1000-1565.2020.01.010
[1] 李明春,魏东盛,邢来君.关于卵菌纲分类地位演变的教学体会[J].菌物研究,2006,4(3):70-74.DOI:10.13341/j.jfr.2006.03.016.
[2] NOWICKI M, FOOLAD M R, NOWAKOWSKA M, et al. Potato and tomato late blight caused by Phytophthora infestans: an overview of pathology and resistance breeding[J]. Plant Disease, 2012, 96(1): 4-17. DOI:10.1094/PDIS-05-11-0458.
[3] 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[J].农业生物技术学报,2018,26(9):1467-1479.DOI:10.3969/j.issn.1674-7968.2018.09.002.
[4] 任兴波,武志华,崔海辰,等. 致病疫霉拮抗菌株YR-7的分离鉴定及其活性物质[J]. 微生物学通报, 2016, 43(7): 1513-1523. DOI:10.13344/j.microbiol.china.150617.
[5] NAGAOKA T,NAKATA K,KOUNO K, et al. Antifungal activity of oosporein from an antagonistic fungus against Phytophthora infestans[J]. Z Naturforsch C, 2004, 59(3-4): 302-304. DOI:10.1515/znc-2004-3-432.
[6] 赵璞钰. 阿拉善地区粘细菌的分离鉴定及其抗致病疫霉活性的初步研究[D]. 呼和浩特: 内蒙古农业大学, 2018.
[7] 何烈干,汤洁,熊正葵,等.马铃薯种质资源对晚疫病的室内及田间抗性鉴定[J].南方农业学报,2018,49(5):898-904.DOI:10.3969/j.issn.2095-1191.2018.05.11.
[8] 刘月,蒋继志,郭俊亭,等.致病疫霉拮抗放线菌的筛选及抑菌作用初步研究[J].河北农业大学学报,2012,35(3):82-85.DOI:10.3969/j.issn.1000-1573.2012.03.015.
[9] 姚期凤,漆树华.微生物群体效应信号分子研究进展[J].微生物学杂志,2015,35(4):63-71.DOI:10.3969/j.issn.1005-7021.2015.04.012.
[10] 王洪洋,秦丽娟,唐唯,等.致病疫霉RXLR效应蛋白相关研究进展[J]. 生物技术通报, 2018,34(2): 102-111. DOI:10.13560/j.cnki.biotech.bull.1985.2017-0981.
[11] 王游游,蒋继志,田荟遥,等.拮抗放线菌Sy11菌株感应的致病疫霉信号分子初步研究[J]. 作物杂志, 2017(4):150-154. DOI:10.16035/j.issn.1001-7283.2017.04.025.
[12] 王雪宁. 致病疫霉对放线菌Sy11抑菌作用的诱导[D]. 保定: 河北大学, 2018.
[13] 陶申傲. 榆耳液体发酵工艺及其抑菌活性的研究[D]. 北京: 北京化工大学, 2006.
[14] KRZYCZKOWSKA J, KOZLOWSKA M. Effect of oils extracted from plant seeds on the growth and lipolytic activity of yarrowia lipolytica yeast[J]. Journal of the American Oil Chemists' Society,2017,94(5):661-671.DOI:10.1007/s11746-017-2975-1.
[15] 李鹏. 杨树菇产抗菌活性物质的研究[D]. 济南: 山东大学, 2005.
[16] WANG S M,BOEVINK P C,WELSH L,et al. Delivery of cytoplasmic and apoplastic effectors from Phytophthora infestans haustoria by distinct secretion pathways[J]. New Phytologist, 2017, 216(1):205-215. DOI:10.1111/nph.14696.
[17] ANDERSON R G,DEB D,FEDKENHEUER K, et al. Recent progress in RXLR effector research[J]. Molecular Plant-Microbe Interactions, 2015, 28(10): 1063-1072. DOI:10.1094/MPMI-01-15-0022-CR.
[18] MARVIN W, STEPHEN P D, GREENBERG E P. Bacterial quorum sensing: the progress and promise of an emerging research area[J]. Nature, 2017,551(7680): 313-320.DOI:10.1038/nature24624.
