Screening of Cellulose-degrading Bacteria in Pleurotus ostreatus Cultivation Substrate and Evaluation of Degradation Effect of Microbial Consortium

doi:10.13560/j.cnki.biotech.bull.1985.2024-1167

Abstract

Abstract:

Objective To screen cellulose-degrading bacteria from the cultivation substrate of Pleurotus ostreatus and construct an efficient microbial consortium for the degradation of agricultural waste. Method Bacterial strains were isolated and purified using sodium carboxymethyl cellulose (CMC-Na) as the sole carbon source from the cultivation substrate of P. ostreatus, and those with cellulose-degrading capabilities were preliminarily identified through Congo red staining. Molecular biology techniques were employed for their identification and to construct a phylogenetic tree. Eleven microbial consortiums were established from strains without mutual antagonism and the cellulase activities of these communities were measured at different fermentation times. The optimal cellulose-degrading microbial consortium was then used in degradation experiments with rice straw and cottonseed hulls to determine its cellulose-degrading capability. Result Four symbiotic bacteria with efficient cellulose-degrading capabilities were identified: Streptomyces sp., Paenibacillus lautus, Microbacterium arborescens and Microbacterium aoyamense. The results indicated that composite microbial system H, composed of Streptomyces sp. Q5 and P. lautus Q6, showed as the most effective cellulose-degrading strain. On the fourth day of culture, its filter paper cellulase (FPase) enzyme activity increased by 229.97% and 134.29%, respectively, compared to the single strains. Additionally, the carboxymethyl cellulase (CMCase) enzyme activity increased by 92.81% and 21.94%, respectively, when compared to the individual strains. After 15 d of treatment with composite microbial system H, the degradation rate was 38.72% for rice straw and 35.76% for cottonseed hulls. Specifically, the degradation rates of lignin, cellulose, and hemicellulose in the rice straw were 33.94%, 31.17%, and 22.43%, respectively, while the corresponding rates for cottonseed hulls were 27.95%, 25.56%, and 53.86%. Further analysis of the surface morphological changes of both rice straw and cottonseed hulls before and after degradation confirmed that composite microbial system H possessed a superior cellulose-degrading capability. Conclusion The microbial consortium constructed in this study can efficiently degrade straw and cottonseed hulls, providing a theoretical basis for improving the effective utilization of agricultural waste resources.

Key words: microbial consortium, Pleurotus ostreatus cultivation substrate, cellulose degradation, degradation of agricultural waste

LIU Jiao-jiao, MU De-mei, XIA Li-ming, FANG Yong, LIU Zuo-jun. Screening of Cellulose-degrading Bacteria in Pleurotus ostreatus Cultivation Substrate and Evaluation of Degradation Effect of Microbial Consortium[J]. Biotechnology Bulletin, 2025, 41(7): 326-335.

Figures/Tables 9

Table 1 Experimental group design

复合菌系 Microbial consortium	菌株名称 Name of strain
A	Q3
B	Q5
C	Q6
D	Q7
E	Q3、Q5
F	Q3、Q6
G	Q3、Q7
H	Q5、Q6
I	Q5、Q7
J	Q6、Q7
K	Q3、Q5、Q6
L	Q3、Q5、Q7
M	Q3、Q6、Q7
N	Q5、Q6、Q7、
O	Q3、Q5、Q6、Q7

Table 2 Ratio of hyaline circle diameter to colony diameter of cellulose-degrading strains

菌株编号 Strain No.	透明圈直径 Transparent circle diameter （D）（cm）	菌落直径 Colony diameter （d）（cm）	D/d
Q1	0.44±0.06 d	0.19±0.01 f	2.40±0.36 e
Q2	0.72±0.21 d	0.26±0.01 d	2.83±0.93 de
Q3	1.47±0.13 c	0.21±0.01 ef	7.18±0.61 b
Q4	/	0.07±0.01 g	/
Q5	2.72±0.21 b	0.65±0.15 a	4.19±0.35 c
Q6	2.97±0.37 b	0.38±0.01 c	7.81±1.19 b
Q7	0.75±0.09 d	0.22±0.01 e	3.42±0.50 cd
Q8	/	0.06±0.01 g	/
Q9	0.57±0.19 d	0.39±0.08 c	1.47±0.55 f
糙皮侧耳	4.39±0.11 a	0.44±0.03 b	10.08±0.31 a

Fig. 1 Congo red clearance zones of cellulose-degrading bacterial strainsCK: Control group; Q1-Q9: cellulose-degrading bacteria; P: Pleurotus ostreatus

Fig. 2 Phylogenetic tree based on the 16S rRNA sequences of the bacteriaThe strain marked with T is the model strain

Table 3 Stain relations between the antagonism

菌株编号 Strain No.	Q3	Q5	Q6	Q7
Q3	/	-	-	-
Q5	-	/	-	-
Q6	-	-	/	-
Q7	-	-	-	/

Fig. 3 Enzyme activities of single strains and microbial consortiumA: Filter paper enzyme activity of single and compound bacterial systems; B: CMCase activity of single and compound bacterial systems

Fig. 4 Growth curve of bacterial strain

Fig. 5 Degradation rates of rice straw and cottonseed hulls by microbial consortium

Fig. 6 Graph of rice straw (A) and cottonseed hulls (B) before and after degradation by microbial consortiumsRS-1: Rice straw before degradation; RS-2: rice straw after degradation; CH-1: cottonseed hulls before degradation; CH-1: cottonseed hulls after degradation

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