5株纤维菌属菌株的分离、鉴定及产酶活性研究

doi:10.11924/j.issn.1000-6850.casb2021-0013

中国农学通报 ›› 2021, Vol. 37 ›› Issue (15): 112-119.doi: 10.11924/j.issn.1000-6850.casb2021-0013

所属专题：生物技术

5株纤维菌属菌株的分离、鉴定及产酶活性研究

王光琴¹^,²(), 黄莺¹^,²(), 温明霞³, 芶剑渝³, 代飞⁴, 丁梦娇¹^,²

¹贵州大学烟草学院,贵阳 550025
²贵州省烟草品质研究重点实验室,贵阳 550025
³贵州省烟草公司遵义市公司,贵州遵义 563000
⁴贵州省烟草公司安顺市公司,贵州安顺 561000

收稿日期:2021-01-07 修回日期:2021-02-08 出版日期:2021-05-25 发布日期:2021-05-18
通讯作者: 黄莺
作者简介:王光琴,女,1994年出生,贵州福泉人,在读硕士,研究方向：土壤营养与施肥。通信地址：550025 贵州贵阳花溪贵州大学西校区,Tel：18798297604,E-mail： 18798297604@163.com。
基金资助:
贵州省烟草公司科技项目“原位氮代谢微生物菌剂的研发与应用”(201707);贵州省烟草公司科技项目“基于有机质空间变量的烤烟施肥模型研究”(201909)

Five Strains of Cellulosimicrobium: Isolation, Identification and Enzymatic Activity

Wang Guangqin¹^,²(), Huang Ying¹^,²(), Wen Mingxia³, Gou Jianyu³, Dai Fei⁴, Ding Mengjiao¹^,²

¹Guizhou University, Guiyang 550025
²Key Laboratory of Tobacco Quality Research, Guiyang 550025
³Guizhou Tobacco Company, Zunyi Company, Guizhou Zunyi 563000
⁴Guizhou Tobacco Company, Anshun Company, Guizhou Anshun 561000

Received:2021-01-07 Revised:2021-02-08 Online:2021-05-25 Published:2021-05-18
Contact: Huang Ying

摘要/Abstract

摘要：

旨在将贵州的玉米秸秆生物质资源通过微生物降解还田,应用到烤烟轮作生产上。以植烟土壤为材料,采用刚果红染色法以及CMCase活性测定进行菌株的筛选,通过失重法测定秸秆降解率。实验共获得44株分解纤维素菌株;初筛获得20株纤维素分解菌。选择D/d较大的5株菌株,通过拮抗实验共获得复合菌剂10株。5株高效纤维素分解菌初步鉴定为Cellulosimicrobium cellulans与Cellulosimicrobium funkei（同源性都达100%）;通过CMCase活性测定,复合菌剂酶活高于单一菌剂,选择酶活高(314.39 U/mL)且作用时间短（5天）的菌剂进行秸秆降解实验,25天降解率达53.35%,是对照的1.87倍。因此,5株菌株可作为贵州烟区玉米秸秆还田的潜在开发菌种,这对于资源有效利用、环境友好和植烟土壤连作障碍改良具有重要意义。

关键词: 纤维素分解菌, 贵州烟区, 玉米秸秆, 植烟土壤, 纤维菌属菌株

Abstract:

The aim is to apply the corn stalk biomass resources of Guizhou in tobacco crop rotation by using the microbial degradation method. The tobacco planting soil was used as material, the Congo red staining method and CMCase enzyme activity determination were used to screen the bacteria strains, and the straw degradation rate of corn stalk was determined by weight loss method. The results showed that 44 cellulose decomposing bacteria strains were obtained in the experiment, 20 cellulose decomposing bacteria strains were obtained in the preliminary screening. Five strains with larger D/d value were selected and 10 compound bacteria strains were obtained through antagonistic experiment. In addition, the five high efficient cellulose decomposing bacterial strains were preliminarily identified as Cellulosimicrobium cellulans or Cellulosimicrobium funkei (100% homology). The CMCase enzyme activity of the compound bacteria strains was higher than that of the single bacteria strain, and the bacteria with high enzyme activity value (i.e., 314.39 U/mL) and short time of achieving the enzyme activity (i.e., 5 days) were selected to do corn stalk degradation experiments. The degradation rate of corn stalk was about 53.35% after 25 days, which was 1.87 times of the control group. Therefore, the five strains can be used as potential developing strains for corn stalk returning to field in tobacco growing areas of Guizhou Province, and could play a significant role in the effective utilization of resources, environmental protection and improvement of continuous cropping obstacles in tobacco-growing soil.

Key words: cellulolytic bacteria, Guizhou tobacco area, corn stalk, tobacco-growing soil, Cellulosimicrobium

中图分类号:

S572

王光琴, 黄莺, 温明霞, 芶剑渝, 代飞, 丁梦娇. 5株纤维菌属菌株的分离、鉴定及产酶活性研究[J]. 中国农学通报, 2021, 37(15): 112-119.

Wang Guangqin, Huang Ying, Wen Mingxia, Gou Jianyu, Dai Fei, Ding Mengjiao. Five Strains of Cellulosimicrobium: Isolation, Identification and Enzymatic Activity[J]. Chinese Agricultural Science Bulletin, 2021, 37(15): 112-119.

图/表 8

参考文献 27

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菌株	菌源	透明圈直径D/cm	菌株直径d/cm	D/d
FCM-1	遵义湄潭土壤	0.75±0.05ab	0.22±0.06a	3.4
FCM-10	遵义湄潭土壤	0.83±0.05a	0.20±0.05a	4.2
FCM-11	遵义湄潭土壤	0.76±0.01ab	0.20±0.03a	3.8
FCY-2	安顺杨武土壤	0.69±0.06b	0.25±0.05a	2.8
FCW-2	毕节威宁土壤	0.83±0.05a	0.21±0.01a	4.0

菌株	菌源	透明圈直径D/cm	菌株直径d/cm	D/d
FCM-1	遵义湄潭土壤	0.75±0.05ab	0.22±0.06a	3.4
FCM-10	遵义湄潭土壤	0.83±0.05a	0.20±0.05a	4.2
FCM-11	遵义湄潭土壤	0.76±0.01ab	0.20±0.03a	3.8
FCY-2	安顺杨武土壤	0.69±0.06b	0.25±0.05a	2.8
FCW-2	毕节威宁土壤	0.83±0.05a	0.21±0.01a	4.0

菌株/复合菌剂编号	菌株/复合菌剂命名	CMCase活性/(U/mL)
菌株/复合菌剂编号	菌株/复合菌剂命名	2 d	5 d	7 d
1	FCM-1	152.20±3.11hi	146.00±11.89i	142.15±3.73l
2	FCM-10	179.72±4.78g	241.48±0.00de	260.45±4.20i
3	FCM-11	166.38±0.52h	243.85±10.07de	204.03±3.93j
4	FCY-2	158.18±2.58h	153.65±3.36hi	172.10±1.05k
5	FCW-2	146.56±1.56i	189.26±2.24fg	174.33±4.20k
6	FCM-1+FCM-10	258.08±0.00b	251.67±6.13d	358.11±2.74a
7	FCM-1+FCM-11	235.86±7.73d	230.18±3.22e	291.05±2.74df
8	FCM-1+FCY-2	209.49±7.73f	245.35±9.66de	283.27±4.75efg
9	FCM-1+FCW-2	230.77±5.26de	285.56±4.29c	298.23±8.10d
10	FCM-10+FCM-11	242.34±8.02c	281.51±14.10c	297.04±2.74d
11	FCM-10+FCY-2	224.76±10.01e	314.39±14.94a	346.13±5.08bc
12	FCM-10+FCW-2	255.76±2.12b	310.85±12.90ab	343.74±5.77c
13	FCM-11+FCY-2	203.47±4.46f	204.64±10.10f	273.39±8.90h
14	FCM-11+FCW-2	228.92±2.78de	173.58±1.70gh	292.25±2.54de
15	FCY-2+FCW-2	290.38±11.90a	292.82±20.44bc	290.45±10.16dg

菌株/复合菌剂编号	菌株/复合菌剂命名	CMCase活性/(U/mL)
菌株/复合菌剂编号	菌株/复合菌剂命名	2 d	5 d	7 d
1	FCM-1	152.20±3.11hi	146.00±11.89i	142.15±3.73l
2	FCM-10	179.72±4.78g	241.48±0.00de	260.45±4.20i
3	FCM-11	166.38±0.52h	243.85±10.07de	204.03±3.93j
4	FCY-2	158.18±2.58h	153.65±3.36hi	172.10±1.05k
5	FCW-2	146.56±1.56i	189.26±2.24fg	174.33±4.20k
6	FCM-1+FCM-10	258.08±0.00b	251.67±6.13d	358.11±2.74a
7	FCM-1+FCM-11	235.86±7.73d	230.18±3.22e	291.05±2.74df
8	FCM-1+FCY-2	209.49±7.73f	245.35±9.66de	283.27±4.75efg
9	FCM-1+FCW-2	230.77±5.26de	285.56±4.29c	298.23±8.10d
10	FCM-10+FCM-11	242.34±8.02c	281.51±14.10c	297.04±2.74d
11	FCM-10+FCY-2	224.76±10.01e	314.39±14.94a	346.13±5.08bc
12	FCM-10+FCW-2	255.76±2.12b	310.85±12.90ab	343.74±5.77c
13	FCM-11+FCY-2	203.47±4.46f	204.64±10.10f	273.39±8.90h
14	FCM-11+FCW-2	228.92±2.78de	173.58±1.70gh	292.25±2.54de
15	FCY-2+FCW-2	290.38±11.90a	292.82±20.44bc	290.45±10.16dg

5株纤维菌属菌株的分离、鉴定及产酶活性研究

Five Strains of Cellulosimicrobium: Isolation, Identification and Enzymatic Activity

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