黄褐土玉米秸秆腐解菌株筛选及其促腐能力研究

doi:10.13560/j.cnki.biotech.bull.1985.2022-0100

摘要/Abstract

摘要：

作物秸秆在黏板土壤中腐解慢，严重制约其培肥改土效果，从黏板酸化黄褐土中筛选土著玉米秸秆高效腐解菌株，可弥补外源腐解菌土壤定殖能力弱、腐解效果差及作物生长不良等问题，对秸秆养分高效转化、土壤地力提升具有重要意义。结合平板划线法、刚果红培养基初筛、滤纸条/秸秆崩解复筛等方法筛选黏板酸化黄褐土秸秆土著腐解菌株；通过产酶（滤纸酶、内切葡聚糖酶、外切葡聚糖酶、β-葡萄糖苷酶、木聚糖酶）能力测定等评价菌株的秸秆促腐能力；通过形态学和16S rDNA序列分析进行菌株的种属鉴定；采用土培法对菌株实际促腐效果进行评价。共筛出2株高效秸秆腐解菌株XJ8和XJ3，初步鉴定为蜡样芽孢杆菌（Bacillus cereus）和枯草芽孢杆菌（Bacillus subtilis），将两者接入黄褐土中，培养28 d玉米秸秆腐解率达45.58%、44.33%，高出不接菌对照秸秆腐解率6.33%以上。综上，所筛蜡样芽孢杆菌（B. cereus）、枯草芽孢杆菌（B. subtilis）可用于商品化秸秆腐解菌剂生产，加速黏板土壤的秸秆腐解速率，提高秸秆养分转化效果。

关键词: 黄褐土, 玉米秸秆, 腐解菌, 蜡样芽孢杆菌, 枯草芽孢杆菌

Abstract:

The slow decomposition rate of crop straw largely limits its effect on soil fertility and soil structure improvement. Isolation of efficient native maize straw-decomposing bacteria from clay and acidic yellow-cinnamon soil may make up for the weak colonization ability of exogenous decomposing bacteria，poor decomposing effect and crop growth，which is of great significance about efficient straw transformation and soil fertility improvement. Combined with plate scribing method，initial screening by Congo red medium，and re-screening by filter paper/straw disintegrating，the indigenous straw-decomposing strains of clay acidified yellow-cinnamon soil were isolated. Enzyme activities（filter paper enzyme，endoglucanase，exocellulase，β-glucosidase and xylanase）were used to evaluate the decomposition promoting ability of the isolated strains. The strains were identified by morphology and 16S rDNA sequence analysis. Soil microcosms were established to evaluate the maize straw decomposition promoting effect of the isolated strains. Two strains XJ8 and XJ3 affiliated with Bacillus cereus and Bacillus Subtilis were identified as efficient maize straw decomposers. The decomposition rates of maize straw were 45.58% and 44.33% under two strains inoculated for 28 d in yellow-cinnamon soil，which was over 6.33% improved compared with the control soils with no bacterial strain inoculated. In conclusion，the isolated B. cereus and B. subtilis could be used for the production of commercial straw decomposing agent to accelerate straw decomposition rate and improve the transformation of straw nutrients in cohesive and hardening soil.

Key words: yellow-cinnamon soil, maize straw, microbial decomposers, Bacillus cereus, Bacillus subtilis

张倩, 徐春燕, 张铎, 王亚会, 梁新盈, 李慧. 黄褐土玉米秸秆腐解菌株筛选及其促腐能力研究[J]. 生物技术通报, 2022, 38(12): 233-243.

ZHANG Qian, XU Chun-yan, ZHANG Duo, WANG Ya-hui, LIANG Xin-ying, LI Hui. Isolation of Maize Straw-decomposing Bacteria in Yellow-cinnamon Soil and Its Ability of Promoting Decomposition[J]. Biotechnology Bulletin, 2022, 38(12): 233-243.

图/表 7

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菌株编号 Strain No.	水解圈直径（D） Hydrolytic circle diameter/mm	菌落直径（d） Colony diameter/mm	水解圈直径/菌落直径D/d	滤纸条崩解程度 Degradation degree of filter strips	秸秆崩解程度 Degradation degree of straw	分离方案 Isolation scheme
XJ1	6.16±0.34	6.14±0.34	1.00±0.00	+	+++	方案1
XJ2	9.75±4.56	4.18±0.09	2.29±1.02	++	+++	方案1
XJ3	37.71±0.67	7.78±0.73	4.93±0.42	++	+	方案1
XJ4	38.58±0.65	8.40±0.91	4.66±0.43	+	+	方案1
XJ5	5.83±0.03	4.41±0.30	1.34±0.10	++	+++	方案1
XJ6	17.63±0.42	5.06±0.13	3.49±0.16	+	+	方案1
XJ7	17.83±0.44	4.41±0.27	4.09±0.34	+	+	方案1
XJ8	35.05±0.09	7.74±0.56	4.57±0.30	+	+++	方案1
XJ9	34.37±1.01	3.68±0.41	9.53±0.85	+++	++	方案1
XJ10	5.25±0.21	4.62±0.07	1.14±0.03	+++	+	方案1
XJ11	5.68±0.33	3.77±0.12	1.51±0.08	++	+	方案1
XJ12	32.45±0.68	3.84±0.18	8.50±0.57	+++	++	方案1
XJ13	35.28±0.57	3.69±0.38	9.80±1.13	++	+++	方案1
XJ14	18.36±0.76	4.88±0.41	3.81±0.29	+++	+++	方案1
XJ15	18.20±0.61	4.78±0.13	3.81±0.15	++	+++	方案1
XJ16	18.07±0.37	4.38±0.34	4.17±0.26	+++	+++	方案1
XJ17	30.87±0.59	8.48±0.52	3.66±0.21	+	++	方案1
XJ18	33.75±0.45	7.14±0.29	4.75±0.22	++	+	方案1
XJ19	14.30±7.36	5.52±0.30	2.75±1.56	+	++	方案1
XJ20	17.45±0.26	6.33±0.06	2.76±0.07	+	++	方案1
XJ21	18.24±0.27	4.16±0.26	4.42±0.28	+	+++	方案1
XJ22	7.79±0.20	5.35±0.02	1.46±0.03	+	+++	方案3
XJ23	17.27±1.00	5.79±0.22	2.98±0.07	++	++	方案3
XJ24	15.57±0.33	4.55±0.24	3.45±0.25	+	+	方案3
XJ25	15.24±0.57	5.51±0.12	2.76±0.09	+	++	方案3
XJ26	5.39±0.04	4.69±0.03	1.15±0.01	+	++	方案3
XJ27	36.81±0.61	7.42±0.19	4.97±0.21	++	+++	方案3
XJ28	4.32±0.24	2.77±0.08	1.55±0.04	+	+++	方案3
XJ29	4.41±0.19	3.32±0.13	1.33±0.01	++	++	方案3
XJ30	4.02±0.15	2.79±0.22	1.46±0.14	++	++	方案3
XJ31	34.00±0.09	5.15±0.66	6.81±0.79	+++	+	方案3
XJ32	19.69±0.43	5.55±0.08	3.55±0.12	++	+	方案3
XJ33	19.70±0.48	5.58±0.06	3.53±0.06	+	+	方案3
XJ34	35.88±1.14	6.76±1.51	5.97±1.48	+++	++	方案3
XJ35	4.61±0.22	2.16±0.09	2.14±0.14	+++	++	方案3
XJ36	19.62±0.23	5.47±0.05	3.59±0.07	+++	++	方案1
XJ37	33.96±0.32	3.91±0.29	8.78±0.65	+++	++	方案3
XJ38	7.98±0.62	6.13±0.32	1.30±0.06	+++	++	方案3
XJ39	13.37±0.58	3.87±0.16	3.46±0.07	+++	++	方案3
XJ40	37.27±0.20	10.11±0.23	3.69±0.07	++	++	方案3
XJ41	5.53±0.28	3.86±0.20	1.43±0.03	+	++	方案3
XJ42	37.67±0.58	7.16±0.71	5.37±0.54	+	+	方案3
XJ43	19.66±0.20	5.30±0.24	3.73±0.19	+	+++	方案3
XJ44	5.28±0.21	7.04±0.69	0.73±0.09	+	++	方案2
XJ45	4.07±0.29	3.85±0.21	1.06±0.02	++	++	方案2
XJ46	7.97±0.66	5.74±0.32	1.38±0.04	+	++	方案2
XJ47	6.73±0.58	5.10±0.33	1.33±0.12	+++	++	方案2
XJ48	5.85±0.14	5.49±0.13	1.07±0.01	+	+	方案2
XJ49	20.25±0.95	5.23±0.22	3.87±0.04	+++	+++	方案2
XJ50	6.07±0.27	6.04±0.27	1.01±0.00	+	++	方案2
XJ51	6.95±0.30	5.38±0.16	1.29±0.02	++	+	方案2
XJ52	18.24±0.70	6.09±0.14	2.99±0.05	+	++	方案2
XJ53	33.64±0.20	9.22±0.31	3.66±0.14	+	+	方案2
XJ54	21.58±0.47	5.84±0.15	3.70±0.18	++	+++	方案2
XJ55	4.28±0.27	3.32±0.29	1.30±0.04	++	+++	方案2
XJ56	5.28±0.97	3.58±0.16	1.51±0.35	+	++	方案2
XJ57	7.05±0.21	5.33±0.43	1.34±0.09	+	+++	方案2
XJ58	4.56±0.07	4.19±0.09	1.09±0.02	+	+	方案2

菌株编号 Strain No.	水解圈直径（D） Hydrolytic circle diameter/mm	菌落直径（d） Colony diameter/mm	水解圈直径/菌落直径D/d	滤纸条崩解程度 Degradation degree of filter strips	秸秆崩解程度 Degradation degree of straw	分离方案 Isolation scheme
XJ1	6.16±0.34	6.14±0.34	1.00±0.00	+	+++	方案1
XJ2	9.75±4.56	4.18±0.09	2.29±1.02	++	+++	方案1
XJ3	37.71±0.67	7.78±0.73	4.93±0.42	++	+	方案1
XJ4	38.58±0.65	8.40±0.91	4.66±0.43	+	+	方案1
XJ5	5.83±0.03	4.41±0.30	1.34±0.10	++	+++	方案1
XJ6	17.63±0.42	5.06±0.13	3.49±0.16	+	+	方案1
XJ7	17.83±0.44	4.41±0.27	4.09±0.34	+	+	方案1
XJ8	35.05±0.09	7.74±0.56	4.57±0.30	+	+++	方案1
XJ9	34.37±1.01	3.68±0.41	9.53±0.85	+++	++	方案1
XJ10	5.25±0.21	4.62±0.07	1.14±0.03	+++	+	方案1
XJ11	5.68±0.33	3.77±0.12	1.51±0.08	++	+	方案1
XJ12	32.45±0.68	3.84±0.18	8.50±0.57	+++	++	方案1
XJ13	35.28±0.57	3.69±0.38	9.80±1.13	++	+++	方案1
XJ14	18.36±0.76	4.88±0.41	3.81±0.29	+++	+++	方案1
XJ15	18.20±0.61	4.78±0.13	3.81±0.15	++	+++	方案1
XJ16	18.07±0.37	4.38±0.34	4.17±0.26	+++	+++	方案1
XJ17	30.87±0.59	8.48±0.52	3.66±0.21	+	++	方案1
XJ18	33.75±0.45	7.14±0.29	4.75±0.22	++	+	方案1
XJ19	14.30±7.36	5.52±0.30	2.75±1.56	+	++	方案1
XJ20	17.45±0.26	6.33±0.06	2.76±0.07	+	++	方案1
XJ21	18.24±0.27	4.16±0.26	4.42±0.28	+	+++	方案1
XJ22	7.79±0.20	5.35±0.02	1.46±0.03	+	+++	方案3
XJ23	17.27±1.00	5.79±0.22	2.98±0.07	++	++	方案3
XJ24	15.57±0.33	4.55±0.24	3.45±0.25	+	+	方案3
XJ25	15.24±0.57	5.51±0.12	2.76±0.09	+	++	方案3
XJ26	5.39±0.04	4.69±0.03	1.15±0.01	+	++	方案3
XJ27	36.81±0.61	7.42±0.19	4.97±0.21	++	+++	方案3
XJ28	4.32±0.24	2.77±0.08	1.55±0.04	+	+++	方案3
XJ29	4.41±0.19	3.32±0.13	1.33±0.01	++	++	方案3
XJ30	4.02±0.15	2.79±0.22	1.46±0.14	++	++	方案3
XJ31	34.00±0.09	5.15±0.66	6.81±0.79	+++	+	方案3
XJ32	19.69±0.43	5.55±0.08	3.55±0.12	++	+	方案3
XJ33	19.70±0.48	5.58±0.06	3.53±0.06	+	+	方案3
XJ34	35.88±1.14	6.76±1.51	5.97±1.48	+++	++	方案3
XJ35	4.61±0.22	2.16±0.09	2.14±0.14	+++	++	方案3
XJ36	19.62±0.23	5.47±0.05	3.59±0.07	+++	++	方案1
XJ37	33.96±0.32	3.91±0.29	8.78±0.65	+++	++	方案3
XJ38	7.98±0.62	6.13±0.32	1.30±0.06	+++	++	方案3
XJ39	13.37±0.58	3.87±0.16	3.46±0.07	+++	++	方案3
XJ40	37.27±0.20	10.11±0.23	3.69±0.07	++	++	方案3
XJ41	5.53±0.28	3.86±0.20	1.43±0.03	+	++	方案3
XJ42	37.67±0.58	7.16±0.71	5.37±0.54	+	+	方案3
XJ43	19.66±0.20	5.30±0.24	3.73±0.19	+	+++	方案3
XJ44	5.28±0.21	7.04±0.69	0.73±0.09	+	++	方案2
XJ45	4.07±0.29	3.85±0.21	1.06±0.02	++	++	方案2
XJ46	7.97±0.66	5.74±0.32	1.38±0.04	+	++	方案2
XJ47	6.73±0.58	5.10±0.33	1.33±0.12	+++	++	方案2
XJ48	5.85±0.14	5.49±0.13	1.07±0.01	+	+	方案2
XJ49	20.25±0.95	5.23±0.22	3.87±0.04	+++	+++	方案2
XJ50	6.07±0.27	6.04±0.27	1.01±0.00	+	++	方案2
XJ51	6.95±0.30	5.38±0.16	1.29±0.02	++	+	方案2
XJ52	18.24±0.70	6.09±0.14	2.99±0.05	+	++	方案2
XJ53	33.64±0.20	9.22±0.31	3.66±0.14	+	+	方案2
XJ54	21.58±0.47	5.84±0.15	3.70±0.18	++	+++	方案2
XJ55	4.28±0.27	3.32±0.29	1.30±0.04	++	+++	方案2
XJ56	5.28±0.97	3.58±0.16	1.51±0.35	+	++	方案2
XJ57	7.05±0.21	5.33±0.43	1.34±0.09	+	+++	方案2
XJ58	4.56±0.07	4.19±0.09	1.09±0.02	+	+	方案2

测试项目 Test item	菌株编号 Strain No.
测试项目 Test item	XJ3	XJ8
形态学 Morphology	杆状，菌落白色，表面光滑	杆状，菌落白色，表面粗糙
过氧化氢酶反应 Catalase test	阳性	阳性
M.R. 反应 Methyl red reaction	阳性	阳性
乙酰甲基甲醇反应 Acetyl methyl carbinol	阴性	阴性
淀粉水解 Starch hydrolysis	阳性	阳性
明胶水解 Gelatin hydrolyzation	阳性	阳性
硝酸盐还原 Nitrate reduction	阳性	阳性
柠檬酸盐利用 Utilization of citrate	阳性	阳性
好氧性试验 Aerobic test	好氧	兼性厌氧
革兰氏染色 Gram stain	阴性	阴性

测试项目 Test item	菌株编号 Strain No.
测试项目 Test item	XJ3	XJ8
形态学 Morphology	杆状，菌落白色，表面光滑	杆状，菌落白色，表面粗糙
过氧化氢酶反应 Catalase test	阳性	阳性
M.R. 反应 Methyl red reaction	阳性	阳性
乙酰甲基甲醇反应 Acetyl methyl carbinol	阴性	阴性
淀粉水解 Starch hydrolysis	阳性	阳性
明胶水解 Gelatin hydrolyzation	阳性	阳性
硝酸盐还原 Nitrate reduction	阳性	阳性
柠檬酸盐利用 Utilization of citrate	阳性	阳性
好氧性试验 Aerobic test	好氧	兼性厌氧
革兰氏染色 Gram stain	阴性	阴性