水稻根系细菌挥发性有机物对小孢根霉的非接触性抑制作用

doi:10.3864/j.issn.0578-1752.2020.10.006

Abstract

Abstract:

【Objective】The objective of this study is to identify new bacterial volatile organic compounds (VOCs) with potential antimicriobial function by analyzing the composition of VOCs of rice root bacteria and their inhibitory effects on Rhizopus microsporus.【Method】The inhibition rate of different bacteria to R. microsporus was determined by a double Petri dish assay. The VOCs of bacteria were collected by headspace-solid phase microextraction and detected by gas chromatography-mass spectrometry (GC-MS). The detected VOCs were compared and identified in the NIST/EPA/NIH database. According to the inhibitory effect and the peak area of each substance in mass spectrum, the correlation analysis was conducted. The VOCs which have a significant correlation with inhibition rate were purchased and the inhibitory effect of these pure substances on R. microsporum was detected in vitro.【Result】All strains could release VOCs and had different inhibitory effects through non-contact of VOCs. These strains produced a total of 90 VOCs of 14 types, which were esters, olefins, alkanes, ketones, acids, thiophenes, aldehydes, naphthalenes, sulfides, phenols, alcohols, pyrazines, pyrroles and benzenes. All strains could release alcohols. Through correlation analysis, 6 VOCs (2-heptanone, 5-methyl-2-hexanone, 2-nonanone, 5-methyl-3-hexanone, 3-methyl-butanoic acid and methyl isobutyl ketone) were found to have potential antimicriobial activity, there was a significant positive correlation between the release amount and inhibition rate. Among them, 3 of these VOCs (2-heptanone, 2-nonanone and methyl isobutyl ketone) had been reported to have antimicriobial activity in previous studies. The other 3 VOCs (5-methyl-2-hexanone, 5-methyl-3-hexanone and 3-methyl-butanoic acid) had not been reported. In vitro fumigation test was conducted by purchasing two of them (3-methyl-butanoic acid and 5-methyl-2-hexanone). It was found that 3-methyl-butanoic acid seriously inhibited the growth of R. microspores, and 5-methyl-2-hexanone was lethal to the R. microspores. Among them, 5-methyl-2-hexanone was mainly released by the strains of Paenisporosarcina macmurdoensis, Brevibacillus brevis and Bacillus aryabhattai. 5-methyl-3-hexanone was mainly produced by the strains of Pseudomonas, B. aryabhattai and P. macmurdoensis. 3-methyl-butanoic acid was mainly released by strains of Enterobacter kobei, Pseudomonas and B. aryabhattai. 【Conclusion】Rice root bacteria can release a variety of VOCs, some of which have a significant inhibitory effect on rice pathogen R. microsporum. Through correlation analysis and in vitro validation, two new antimicriobial VOCs (5-methyl-2-hexanone and 3-methyl-butanoic acid) and one possible antimicriobial VOC (5-methyl-3-hexanone) were identified. All these strains and VOCs have the potential as biological resources for new drugs and antifungal metabolites.

Key words: rice root, volatile organic compounds (VOCs), inhibitory effect, bacteria, Rhizopus microsporus, headspace-solid phase microextraction-mass combination

WANG EnZhao,FAN FenLiang,LI YanLing,LIU XiongDuo,LU YuQiu,SONG ALin. Noncontact Inhibitory of Volatile Organic Compounds from Rice Root Bacteria on Rhizopus microsporus[J].Scientia Agricultura Sinica, 2020, 53(10): 1986-1996.

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Figures/Tables 6

Fig. 1

Table 1

Composition of mVOCs"

挥发性有机物名称VOCs name	2-60	34	35	62	146	2-18	26	50	3
(E)-1-甲基-2-(丙-1-烯-1-基)二硫 (E)-1-Methyl-2-(prop-1-en-1-yl) disulfane	-	-	-	-	-	-	+	-	-
β-苯基乙基丁酸酯beta-Phenylethyl butyrate	-	-	-	-	-	-	+	-	-
11-十二烯-2-酮11-Dodecen-2-one	-	-	-	-	-	+	-	-	-
2-甲基-1-丁醇2-Methyl-1-butanol	-	+	-	-	-	-	-	-	-
3-甲基-1-丁醇3-Methyl-1-butanol	+	+	+	+	+	-	+	-	-
1-丁醇,3-甲基-乙酸1-Butanol, 3-methyl-acetate	-	+	+	-	-	-	-	-	-
2-乙基-1-己醇2-Ethyl-1-hexanol	+	-	+	+	+	+	-	-	+
1-十八碳烯1-Octadecene	+	-	-	-	-	-	-	-	-
1-辛醇1-Octanol	+	-	-	-	-	-	-	-	-
7-甲基-1-辛烯7-Methyl-1-octene	+	-	-	-	-	-	-	-	-
1-十五碳烯1-Pentadecene	-	-	-	-	-	-	+	-	-
2-氯丙酸,十六烷基酯2- Chloropropionic acid, hexadecyl ester	-	-	-	-	-	-	+	-	-
5-庚基二氢-2(3H)-呋喃酮5-Heptyldihydro-2(3H)-furanone	-	-	+	-	+	-	-	-	-
2,4,6-环庚三烯-1-酮2,4,6-Cycloheptatrien-1-one	+	-	-	-	-	-	-	-	-
2,4-二硫戊烷2,4-Dithiapentane	-	-	-	-	-	-	-	-	+
6-甲基-2,4-庚二酮6-Methyl-2,4-heptanedione	-	-	-	-	-	+	-	-	-
3,7,11-三甲基-2,6,10-十二碳三烯-1-醇 3,7,11-Trimethyl-2,6,10-dodecatrien-1-ol	-	-	-	-	-	-	-	-	+
2-癸酮2-Decanone	+	+	+	+	-	+	+	-	-
2-十二烷酮2-Dodecanone	-	-	+	-	-	+	-	-	-
2-乙基-1-己醇2-Ethyl-1-hexanol	-	-	-	-	-	-	-	+	-
2-庚酮2-Heptanone	+	+	+	+	-	+	+	-	-
3-甲基-2-庚酮3-Methyl-2-heptanone	-	-	-	+	-	-	-	-	-
5-甲基-2-庚酮5-Methyl-2-heptanone	+	-	-	+	-	+	-	-	-
6-甲基-2-庚酮6-Methyl-2-heptanone	+	-	+	+	-	+	-	-	-
2-十六烷酮2-Hexadecanone	-	-	-	-	-	+	-	-	-
2-己酮2-Hexanone	+	-	+	+	-	-	-	-	-
3,4-二甲基-2-己酮3,4-Dimethyl-2-hexanone	+	-	-	-	-	-	-	-	-
5-甲基-2-己酮5-Methyl-2-hexanone	+	-	-	+	-	+	-	-	-
2-壬基酮2-Nonanone	-	+	+	+	-	+	+	-	-
2-辛酮2-Octanone	-	-	+	+	-	-	-	-	-
2-十五烷酮2-Pentadecanone	-	+	-	-	-	-	-	-	-
2-戊酮2-Pentanone	-	+	+	-	-	-	-	-	-
3-甲基-2-戊酮3-Methyl-2-pentanone	-	-	-	-	-	+	-	-	-
2-十四烷酮2-Tetradecanone	-	-	+	+	-	+	-	-	-
2-十三烷酮2-Tridecanone	-	-	-	+	-	-	-	-	-
2-十一烷酮2-Undecanone	-	+	+	-	-	+	+	-	-
3-十二烷酮3-Dodecanone	-	-	-	+	-	-	-	-	-
7-苯基-3-庚烯7-Phenyl-3-heptene	-	+	-	-	-	-	-	-	-
5-甲基-3-己酮5-Methyl-3-hexanone	+	-	+	+	-	-	-	-	-
3-十五烷酮3-Pentadecanone	-	-	-	+	-	-	-	-	-
3-戊酮3-Pentanone	-	-	-	+	-	-	-	-	-
3-十三烷酮3-Tridecanone	+	-	-	-	-	+	-	-	-
5-甲基-4-己烯-3-酮5-Methyl-4-hexen-3-one	+	-	-	-	-	-	-	-	-
6,10,14-三甲基-5,9,13-戊三烯-2-酮6,10,14-Trimethyl-5,9,13-pentadecatrien-2-one	-	-	-	-	-	-	-	+	-
6-甲基-5-庚烯-2-酮6-Methyl-5-hepten-2-one	+	-	-	-	-	-	-	+	-
6-叔丁基-2,4-二甲基苯酚6-Tert-butyl-2,4-dimethylphenol	-	+	+	-	-	-	-	-	-
7-甲基辛烷-2,4-二酮,烯醇式7-Methyloctane-2,4-dione, enol form	-	-	-	-	-	+	-	-	-
乙酸,2-苯乙基酯Acetic acid, 2-phenylethyl ester	-	+	-	-	-	-	-	-	-
乙酸,氯-十六烷基酯Acetic acid, chloro-hexadecyl ester	-	+	-	-	-	-	-	-	-
顺式乙酸,非3-烯基酯cis-Acetic acid, non-3-enyl ester	-	-	+	-	-	-	-	-	-
(2-甲氧基乙基)-苯 (2-Methoxyethyl)-benzene	-	-	-	-	-	-	+	-	-
(甲氧基甲基)-苯 (Methoxymethyl)-benzene	-	-	-	-	-	-	+	-	-
苯乙酸,乙酯Benzeneacetic acid, ethyl ester	+	-	-	-	-	-	-	-	-
苯甲酸,乙酯Benzoic acid, ethyl ester	-	-	-	-	-	-	-	+	-
苯甲醇Benzyl alcohol	-	-	-	+	+	-	-	-	-
苄基甲基酮Benzyl methyl ketone	+	-	-	+	+	+	+	-	-
1-甲氧基-3-甲基丁烷1-Methoxy-3-methyl-butane	-	-	-	-	-	-	+	-	-
2-甲基-2-(甲硫基)-丁烷2-Methyl-2-(methylthio)-butane	-	-	-	-	-	-	-	+	-
丁烷磺酸,硫-甲酯Butanethioic acid, S-methyl ester	+	-	-	-	-	-	-	-	-
丁酸,1-乙烯基己酯Butanoic acid, 1-ethenylhexyl ester	-	-	+	-	-	-	-	-	-
3-甲基丁酸3-Methyl-butanoic acid	-	+	+	+	-	-	-	-	-
丁酸,3-甲基-乙酯Butanoic acid, 3-methyl-ethyl ester	+	-	-	+	-	-	-	-	-
顺式双环[3.3.0]辛-2-烯cis-Bicyclo[3.3.0]oct-2-ene	+	-	-	-	-	-	-	-	-
环庚烯Cycloheptene	-	-	-	-	-	-	+	-	-
1-甲基环己烯1-Methyl-cyclohexene	-	-	-	+	-	-	-	-	-
3-乙烯基-环戊烯3-Ethenyl-cyclopentene	+	-	-	-	-	-	-	-	-
双环戊二烯Dicyclopentadiene	+	+	+	+	-	+	-	+	-
三硫化二甲基Dimethyl trisulfide	+	+	+	+	+	-	-	+	+
二硫化二甲基Dimethyl disulfide	+	+	+	+	+	-	+	+	+
十二酸,乙酯Dodecanoic acid, ethyl ester	+	-	-	-	-	-	-	-	-
1-(2-氨基苯基)-乙酮1-(2-Aminophenyl)-ethanone	-	+	-	-	-	-	+	-	-
13-甲基十四烷酸乙酯Ethyl 13-methyl-tetradecanoate	+	-	-	-	-	-	-	-	-
3-(甲硫基)-(E)-2-丙酸乙酯Ethyl 3-(methylthio)-(E)-2-propenoate	-	-	-	-	-	-	-	+	-
3-(甲硫基)-(Z)-2-丙酸乙酯Ethyl 3-(methylthio)-(Z)-2-propenoate	-	-	-	-	-	-	-	+	-
十三烷酸乙酯Ethyl tridecanoate	+	-	-	-	-	-	-	-	-
甲基异丁基酮Methyl isobutyl ketone	+	-	+	+	-	-	-	-	-
异戊酸甲酯Methyl isovalerate	-	-	+	-	-	-	-	-	-
硫羟乙酸甲酯Methyl thiolacetate	-	-	-	-	-	-	-	-	+
十氢萘Decahydro-naphthalene	-	+	+	-	-	-	-	-	-
十八烷Octadecanal	-	-	+	-	-	-	-	-	-
苯乙醇Phenylethyl alcohol	-	+	+	+	-	-	+	-	-
丙酸,2-苯乙基酯Propanoic acid, 2-phenylethyl ester	-	+	-	-	-	-	-	-	-
2-乙基-5-甲基吡嗪2-Ethyl-5-methyl-pyrazine	-	-	+	-	-	-	-	-	-
三甲基吡嗪Trimethyl-pyrazine	-	-	+	-	-	-	-	-	-
吡咯Pyrrole	-	-	-	+	-	-	-	-	-
3-甲基丁硫醇甲酯S-Methyl 3-methylbutanethioate	+	-	+	+	+	-	-	-	+
丙酮三氧化物TATP	-	-	-	-	+	-	-	-	-
十四烷酸,乙酯Tetradecanoic acid, ethyl ester	+	-	-	-	-	-	-	-	-
2-甲氧基-5-甲基-噻吩2-Methoxy-5-methyl-thiophene	-	-	-	-	+	-	-	-	-
十一酸,乙酯Undecanoic acid, ethyl ester	+	-	-	-	-	-	-	-	-

Table 1

Fig. 2

Fig. 3

Table 2

Fig. 4

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挥发性有机物名称 VOCs name	相关性系数 Correlation coefficient	挥发性有机物名称 VOCs name	相关性系数 Correlation coefficient
3-甲基-1-丁醇3-Methyl-1-butanol (+)	0.093	2-十一烷酮2-Undecanone (+)	0.485
2-乙基-1-己醇2-Ethyl-1-hexanol	-0.396	5-甲基-3-己酮5-Methyl-3-hexanone (++)	0.863**
2-癸酮2-Decanone	0.074	苄基甲基酮Benzyl methyl ketone	-0.169
2-庚酮2-Heptanone (+)	0.544**	3-甲基丁酸3-Methyl-butanoic acid (++)	0.691*
5-甲基-2-庚酮5-Methyl-2-heptanone	-0.705*	双环戊二烯Dicyclopentadiene	0.025
6-甲基-2-庚酮6-Methyl-2-heptanone	0.146	三硫化二甲基Dimethyl trisulfide (+)	-0.494*
2-己酮2-Hexanone	-0.251	二硫化二甲基Dimethyl disulfide (+)	-0.074
5-甲基-2-己酮5-Methyl-2-hexanone (++)	0.710**	甲基异丁基酮Methyl isobutyl ketone (+)	0.781**
2-壬酮2-Nonanone (+)	0.514*	苯乙醇Phenylethyl alcohol	-0.576*
2-十四烷酮2-Tetradecanone	-0.882**	3-甲基丁硫醇甲酯S-Methyl 3-methylbutanethioate	0.346

Noncontact Inhibitory of Volatile Organic Compounds from Rice Root Bacteria on Rhizopus microsporus

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