Screening and identification of whole rice surface dominant lactic acid bacteria

doi:10.11686/cyxb2023075

Abstract

Abstract:

In this study， dominant lactic acid bacteria that colonize the leaf and stem surfaces of rice Oryza sativa plants were isolated， screened and identified， in order to develop a collection of potentially bacterial strains potentially useful in silage making as a biological resource. The epiphytic lactic acid bacteria of whole rice plants were isolated by the plate coating method. The growth rate and acid-producing capacity were screened initially， and the antioxidant capacity included H₂O₂ tolerance， 1， 1-diphenyl-2-picrylhydrazyl （DPPH） free radical， hydroxyl free radical and superoxide anion free radical scavenging rate were used as selection criteria for rescreening. The physiological and biochemical characteristics and molecular biological identification of the selected dominant lactic acid bacteria were analyzed. In total 154 strains of lactic acid bacteria were isolated from the rice plants. After 48 h culture， 16 strains were found with optical density at 600 nm （OD_600nm） value greater than 2 and pH value less than 4， among which 9 strains had antioxidant capacity. A strain designated YMA3 had the best overall performance， with scavenging rates of DPPH free radical， hydroxyl free radical and superoxide anion free radical of 72.23%， 72.46% and 86.33%， respectively. YMA3 was identified as Enterococcus faecalis， and showed strong adaptability to high temperature or acidic environments.

Key words: rice, epiphytic lactic acid bacteria, screen, oxidation resistance, physiological-biochemical characteristic, Enterococcus faecalis

Li-juan HUANG, Rong-ji SUN, Wen-jing GAO, Zhi-fei ZHANG, Gui-hua CHEN. Screening and identification of whole rice surface dominant lactic acid bacteria[J]. Acta Prataculturae Sinica, 2024, 33(1): 117-125.

Figures/Tables 8

References 30

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菌株Strain	0 mmol·L^-1	1 mmol·L^-1	2 mmol·L^-1	3 mmol·L^-1	菌株Strain	0 mmol·L^-1	1 mmol·L^-1	2 mmol·L^-1	3 mmol·L^-1
YMA1	1.879±0.01a	1.862±0.01a	1.572±0.01b	0.138±0.01c	BJD3	1.430±0.01a	1.325±0.01b	1.122±0.01c	0.086±0.01d
YMA3	1.889±0.01a	1.871±0.01b	1.830±0.01c	1.575±0.01d	BJD4	1.552±0.01a	1.532±0.01b	1.502±0.01c	1.490±0.01c
YMB4	1.880±0.01a	1.841±0.01b	1.542±0.01c	0.159±0.01d	BJD7	1.665±0.01a	1.642±0.01a	1.561±0.01b	1.322±0.04c
YMC5	1.901±0.01a	1.869±0.01b	1.602±0.01c	0.273±0.01d	BJE1	1.668±0.01a	1.588±0.01b	1.583±0.01c	1.414±0.01d
YMC6	1.879±0.01a	1.752±0.01b	1.643±0.01c	0.318±0.01d	BJE8	1.671±0.01a	1.642±0.01b	1.579±0.02c	1.510±0.01d
YMD10	1.759±0.01a	1.717±0.01a	1.533±0.01b	0.255±0.05c	BJE11	1.659±0.01a	1.641±0.01b	1.588±0.01c	1.016±0.01d
BJA4	1.771±0.01a	1.645±0.01b	1.316±0.01c	1.091±0.01d	CSE9	1.922±0.01a	1.841±0.02b	1.549±0.01c	1.488±0.01d
BJB2	1.956±0.01a	1.932±0.01b	1.633±0.01c	0.287±0.01d	YHE4	1.738±0.03a	1.915±0.01a	1.823±0.01b	1.614±0.03c

菌株Strain	0 mmol·L^-1	1 mmol·L^-1	2 mmol·L^-1	3 mmol·L^-1	菌株Strain	0 mmol·L^-1	1 mmol·L^-1	2 mmol·L^-1	3 mmol·L^-1
YMA1	1.879±0.01a	1.862±0.01a	1.572±0.01b	0.138±0.01c	BJD3	1.430±0.01a	1.325±0.01b	1.122±0.01c	0.086±0.01d
YMA3	1.889±0.01a	1.871±0.01b	1.830±0.01c	1.575±0.01d	BJD4	1.552±0.01a	1.532±0.01b	1.502±0.01c	1.490±0.01c
YMB4	1.880±0.01a	1.841±0.01b	1.542±0.01c	0.159±0.01d	BJD7	1.665±0.01a	1.642±0.01a	1.561±0.01b	1.322±0.04c
YMC5	1.901±0.01a	1.869±0.01b	1.602±0.01c	0.273±0.01d	BJE1	1.668±0.01a	1.588±0.01b	1.583±0.01c	1.414±0.01d
YMC6	1.879±0.01a	1.752±0.01b	1.643±0.01c	0.318±0.01d	BJE8	1.671±0.01a	1.642±0.01b	1.579±0.02c	1.510±0.01d
YMD10	1.759±0.01a	1.717±0.01a	1.533±0.01b	0.255±0.05c	BJE11	1.659±0.01a	1.641±0.01b	1.588±0.01c	1.016±0.01d
BJA4	1.771±0.01a	1.645±0.01b	1.316±0.01c	1.091±0.01d	CSE9	1.922±0.01a	1.841±0.02b	1.549±0.01c	1.488±0.01d
BJB2	1.956±0.01a	1.932±0.01b	1.633±0.01c	0.287±0.01d	YHE4	1.738±0.03a	1.915±0.01a	1.823±0.01b	1.614±0.03c

菌株 Strain	DPPH自由基清除率 DPPH free radical scavenging rate			羟自由基清除率 Hydroxyl free radical scavenging rate			超氧阴离子自由基清除率 Superoxide anion free radical scavenging rate
菌株 Strain	完整菌体 IC	无菌体提取物 CFE	发酵上清液 CFS	完整菌体 IC	无菌体提取物 CFE	发酵上清液 CFS	完整菌体 IC	无菌体提取物 CFE	发酵上清液 CFS
YMA3	7.89±0.86a	18.07±2.23d	72.23±0.50c	62.12±0.92a	21.31±2.92a	72.46±9.45a	86.33±0.19a	2.65±0.98c	ND
BJA4	6.91±1.50ab	19.49±1.59cd	72.82±0.37bc	19.37±7.10d	14.49±4.02ab	36.31±6.85bc	75.13±7.96a	6.40±1.93a	ND
BJD4	8.78±2.05a	20.37±2.22bcd	73.51±0.82b	34.08±9.28c	12.61±4.25b	35.41±4.84c	84.77±0.38a	5.38±1.52ab	ND
BJD7	6.87±1.84abc	17.53±2.60d	72.64±0.94bc	47.75±5.46b	ND	25.21±3.21d	29.44±16.74b	5.16±1.38ab	ND
BJE1	4.52±1.04bcd	18.76±7.88d	73.67±1.63b	34.64±7.87c	10.23±0.40b	19.26±5.60de	81.47±6.32a	5.97±1.33ab	ND
BJE8	8.47±1.59a	34.67±3.70a	72.21±0.75c	46.41±4.63b	11.02±4.02b	43.63±2.40bc	5.57±1.65c	6.18±1.95ab	ND
BJE11	9.23±3.79a	24.96±3.85b	75.56±0.63a	9.24±6.03e	ND	27.29±8.33cd	76.32±10.60a	4.39±1.57bc	ND
CSE9	7.09±1.58a	24.41±5.65b	73.56±0.95b	24.41±4.55cd	8.69±3.93b	36.92±3.53bc	ND	4.78±1.07ab	3.33±0.93
YHE4	8.17±1.82a	24.06±4.09bc	72.90±1.22bc	28.95±9.32c	11.22±5.47b	13.17±3.00e	79.04±6.28a	2.71±1.49c	ND

菌株 Strain	DPPH自由基清除率 DPPH free radical scavenging rate			羟自由基清除率 Hydroxyl free radical scavenging rate			超氧阴离子自由基清除率 Superoxide anion free radical scavenging rate
菌株 Strain	完整菌体 IC	无菌体提取物 CFE	发酵上清液 CFS	完整菌体 IC	无菌体提取物 CFE	发酵上清液 CFS	完整菌体 IC	无菌体提取物 CFE	发酵上清液 CFS
YMA3	7.89±0.86a	18.07±2.23d	72.23±0.50c	62.12±0.92a	21.31±2.92a	72.46±9.45a	86.33±0.19a	2.65±0.98c	ND
BJA4	6.91±1.50ab	19.49±1.59cd	72.82±0.37bc	19.37±7.10d	14.49±4.02ab	36.31±6.85bc	75.13±7.96a	6.40±1.93a	ND
BJD4	8.78±2.05a	20.37±2.22bcd	73.51±0.82b	34.08±9.28c	12.61±4.25b	35.41±4.84c	84.77±0.38a	5.38±1.52ab	ND
BJD7	6.87±1.84abc	17.53±2.60d	72.64±0.94bc	47.75±5.46b	ND	25.21±3.21d	29.44±16.74b	5.16±1.38ab	ND
BJE1	4.52±1.04bcd	18.76±7.88d	73.67±1.63b	34.64±7.87c	10.23±0.40b	19.26±5.60de	81.47±6.32a	5.97±1.33ab	ND
BJE8	8.47±1.59a	34.67±3.70a	72.21±0.75c	46.41±4.63b	11.02±4.02b	43.63±2.40bc	5.57±1.65c	6.18±1.95ab	ND
BJE11	9.23±3.79a	24.96±3.85b	75.56±0.63a	9.24±6.03e	ND	27.29±8.33cd	76.32±10.60a	4.39±1.57bc	ND
CSE9	7.09±1.58a	24.41±5.65b	73.56±0.95b	24.41±4.55cd	8.69±3.93b	36.92±3.53bc	ND	4.78±1.07ab	3.33±0.93
YHE4	8.17±1.82a	24.06±4.09bc	72.90±1.22bc	28.95±9.32c	11.22±5.47b	13.17±3.00e	79.04±6.28a	2.71±1.49c	ND

项目Item	结果Result
革兰氏染色Gram stain	阳性Positive
过氧化氢酶Catalase	阴性Negative
葡萄糖产气Glucose gas production	阴性Negative
发酵类型Fermentation type	同型Homo
耐温Temperature resistance （℃）
5	+
10	++
37	++
45	++
50	++
耐酸Acid resistance （pH）
3.0	+
3.5	++
4.0	++
5.0	++
6.0	++
7.0	++
耐盐Salt tolerance （NaCl， %）
3.0	++
6.8	++
10.0	+