解淀粉芽孢杆菌SQ-2对水稻的促生作用

doi:10.13560/j.cnki.biotech.bull.1985.2023-0770

摘要/Abstract

摘要：

【目的】对实验室分离筛选得到的一株解淀粉芽孢杆菌SQ-2进行促生特性研究，确定该菌株对水稻促生的浓度范围与作用机制，并分析接种菌株前后的土壤菌群结构。【方法】利用钼蓝比色法与固氮酶试剂盒对菌株SQ-2的溶磷能力及固氮酶活性进行检测。将10²、10⁴、10⁶、10⁸和3×10⁸ CFU/mL的菌悬液接种至水培与土培水稻中，分别培养14 d和20 d后测定其水培、土培水稻的根茎干鲜重、茎长与茎粗。采用苯酚-次氯酸钠比色法、茚三酮检测法与3,5-二硝基水杨酸比色法测定土培水稻土壤中脲酶、蛋白酶与蔗糖酶的活性。利用pH计电位法检测土壤pH值，用对应试剂盒检测土壤速效氮磷钾含量及酸性磷酸酶活性，并对接种3×10⁸ CFU/mL组的土培水稻土壤进行根际细菌群落结构分析。【结果】菌株SQ-2对磷酸三钙的溶解量为229.63 mg/L，固氮酶活性为55.07 U/L。与对照相比，接种菌悬液浓度在10⁴CFU/mL时，水培水稻根的干、鲜重增长最为显著；在接种浓度为3×10⁸ CFU/mL时，土培水稻根茎的干、鲜重增长最为显著。随着接种菌悬液浓度的升高，上述土壤酶活性与速效氮磷钾浓度都有着不同程度的增加，而接种菌株3×10⁸ CFU/mL的土壤pH值则由原来的7.83降至7.26。接种菌株SQ-2改变了水稻根际土壤中的菌落构成，显著提高了土壤α多样性的Ace、Chao、Sobs与Shannon指数。【结论】解淀粉芽孢杆菌 SQ-2对土培、水培水稻均有不同程度的促生效果。在土培实验中，能够通过提高土壤酶活性、速效氮磷钾水平及改变土壤菌群结构来起到促生作用，为细菌菌肥的研发提供了新的菌株资源。

关键词: 解淀粉芽孢杆菌, 促生, 溶磷固氮, 土壤酶活性, 根际细菌, 水稻, 细菌群落

Abstract:

【Objective】 To study the growth-promoting characteristics of a strain of Bacillus amyloliquefaciens SQ-2 isolated and screened in the laboratory, and to determine the concentration range, mechanism of action, and changes in soil microbiota induced by this strain to promote rice growth.【Method】 The phosphorus solubility and nitrogenase activity of strain SQ-2 were detected using the molybdenum blue colorimetric method and ELISA nitrogenase kit. Inoculating10², 10⁴, 10⁶, 10⁸, and 3×10⁸ CFU/mL bacterial suspension into hydroponic and soil cultured rice, the dry and fresh weight, length, and thickness of the roots and stems of hydroponic and soil cultured rice were measured, respectively after 14 and 20 d of cultivation. Meanwhile, the activities of urease, protease, and sucrase in soil cultured rice were determined using the phenol sodium hypochlorite colorimetric method, the ninhydrin detection method, and the 3,5-dinitrosalicylic acid colorimetric method. Besides, the soil pH value was determined using the pH meter potential method and the content of nitrogen, phosphorus, and potassium available in soil and the activity of acid phosphatase were detected by utilizing the corresponding reagent kit. Analysis of rhizosphere bacterial community structure in soil cultured rice soil was inoculated with 3×10⁸ CFU/mL components.【Result】 The dissolution of tricalcium phosphate by strain SQ-2 was 229.63 mg/L, and the nitrogenase activity was 55.07 U/L. Compared with the control, the dry and fresh weight of hydroponic rice roots showed the most significant increase at an inoculum suspension concentration of 10⁴ CFU/mL. At a vaccination concentration of 3×10⁸ CFU/mL, the dry and fresh weight of soil cultivated rice rhizomes increased most significantly. As the concentration of the inoculum suspension increased, the activities of various enzymes and the concentration of available nitrogen, phosphorus, and potassium in the soil increased to varying degrees, the soil pH value while inoculating strain 3×10⁸ CFU/mL decreased from 7.83 to 7.26. The inoculant strain SQ-2 changed the colony composition in the rice rhizosphere soil and significantly increased Ace, Chao, Sobs and Shannon indices of soil α-diversity.【Conclusion】 B. amyloliquefaciens SQ-2 had different growth promotion effects on soil culture and hydroculture rice. In soil culture experiments, the growth-promoting effect is notably pronounced through the augmentation of soil enzyme activity, rapid increases in available nitrogen, phosphorus, and potassium levels, and alterations in the soil microbial community structure. Overall, these experimental findings offer valuable insights into new strain resources for the development of bacterial fertilizer.

Key words: Bacillus amyloliquefaciens, promoting growth, dissolving phosphorus and fixing nitrogen, soil enzyme activity, rhizosphere bacteria, rice, bacterial community

李雪, 李容欧, 孔美懿, 黄磊. 解淀粉芽孢杆菌SQ-2对水稻的促生作用[J]. 生物技术通报, 2024, 40(2): 109-119.

LI Xue, LI Rong-ou, KONG Mei-yi, HUANG Lei. The Growth Promoting Effect of Bacillus amyloliquefaciens SQ-2 on Rice[J]. Biotechnology Bulletin, 2024, 40(2): 109-119.

图/表 10

图1 解淀粉芽孢杆菌SQ-2在无机磷培养基中的溶磷作用 A：菌株SQ-2在无机磷固体培养基中的溶磷圈；B：菌株SQ-2在无机磷液体培养基中的溶磷量、pH与菌体数量（1×107）的变化

Fig. 1 Phosphorus dissolution of Bacillus amyloliquefaciens SQ-2 in inorganic phosphorus medium A: Phosphate solubilization zones of B. amyloliquefaciens SQ-2 on inorganic phosphorus solid medium; B: phosphorus dissolution amount, pH and bacterial growth ability（1×107）of B. amyloliquefaciens SQ-2 in inorganic phosphorus liquid medium

图2 接种解淀粉芽孢杆菌SQ-2后钾长石的扫描电子显微照片 A：钾长石对照组（2 μm）；B：接种菌株SQ-2后钾长石的形态（2 μm）；C：钾长石对照组（1 μm）；D：接种菌株SQ-2后钾长石的形态（1 μm）

Fig. 2 Scanning electron micrograph of potassium feldspar after inoculation with B. amyloliquefaciens SQ-2 A: Potassium feldspar control group（2 μm）. B: Morphology of potassium feldspar after inoculation with B. amyloliquefaciens SQ-2（2 μm）. C: Potassium feldspar control group（1 μm）. D: Morphology of potassium feldspar after inoculation with B. amyloliquefaciens SQ-2（1 μm）

表1 不同浓度的解淀粉芽孢杆菌SQ-2对水培水稻根茎干鲜重、茎长与茎粗的影响

Table 1 Effects of B. amyloliquefaciens SQ-2 at different inoculum concentrations on the dry and fresh weight, stem length, and stem diameter of hydroponic rice

处理 Treatment	根干重 Dry weight of root/g	茎干重 Dry weight of stem/g	根鲜重 Fresh weight of root/g	茎鲜重 Fresh weight of stem/g	茎粗 Stem thickness/mm	株高 Height/cm
CK	0.03±0.001c	0.03±0.001c	0.32±0.014b	0.34±0.012ab	0.53±0.020ab	4.92±0.060ab
S1	0.03±0.003c	0.03±0.002bc	0.32±0.002b	0.28±0.010b	0.59±0.020ab	5.28±0.177ab
S2	0.05±0.003a	0.04±0.003a	0.52±0.005a	0.40±0.026a	0.62±0.007a	5.63±0.003a
S3	0.04±0.002ab	0.04±0.001ab	0.42±0.033ab	0.34±0.019ab	0.60±0.007ab	5.71±0.253a
S4	0.04±0.003ab	0.04±0.001abc	0.42±0.007ab	0.36±0.017ab	0.50±0.033b	5.58±0.167a
S5	0.04±0.002bc	0.03±0.001c	0.40±0.024b	0.27±0.004b	0.52±0.010b	4.54±0.180b

表2 不同浓度的解淀粉芽孢杆菌SQ-2对土培水稻根茎干鲜重、茎长与茎粗的影响

Table 2 Effects of B. amyloliquefaciens SQ-2 at different inoculum concentrations on the dry and fresh weight, stem length, and stem thickness of soil cultured rice

处理 Treatment	根干重 Dry weight of root/g	茎干重 Dry weight of stem/g	根鲜重 Fresh weight of root/g	茎鲜重 Fresh weight of stem/g	茎粗 Stem thickness/mm	株高 Height/cm
CK	0.07±0.001bc	0.03±0.001b	0.81±0.038bc	0.43±0.004c	0.72±0.023bc	10.99±0.243c
S1	0.06±0.001c	0.03±0.001b	0.74±0.031c	0.42±0.006c	0.64±0.003c	11.85±0.180c
S2	0.08±0.002abc	0.03±0.001b	0.94±0.022abc	0.42±0.005c	0.72±0.043bc	11.34±0.013c
S3	0.09±0.002ab	0.03±0.001b	0.99±0.017ab	0.46±0.001bc	0.77±0.037abc	10.67±0.067c
S4	0.10±0.001a	0.04±0.002b	1.11±0.001a	0.51±0.037ab	0.87±0.107ab	12.32±0.377c
S5	0.10±0.005a	0.05±0.001a	1.16±0.009a	0.58±0.003a	0.94±0.001a	12.24±0.493c

图3 解淀粉芽孢杆菌SQ-2在不同浓度下对土培水稻生长的影响 A-F分别添加不同浓度菌液；A：3×108；B：108； C：106；D：104；E：102；F：0（CFU/mL）

Fig. 3 Effects of B. amyloliquefaciens SQ-2 on soil cultured rice under different bacterial inoculum concentrations A-F indicates adding different concentration of bacteria; A: 3×108; B: 108; C: 106; D: 104; E: 102; F: 0（CFU/mL）

图4 解淀粉芽孢杆菌SQ-2在不同浓度下对土壤中速效氮磷钾、pH与酶活性的影响不同小写字母表示显著差异（P<0.05）

Fig. 4 Effects of B. amyloliquefaciens SQ-2 at different concentrations on nitrogen, phosphorus, potassium, pH and enzyme activity in soil The different lowercase letters indicate significant differences (P < 0.05)

图5 接种解淀粉芽孢杆菌SQ-2后土壤营养成分、土壤酶活性与pH的相关性

Fig. 5 Correlation between soil nutrient composition, soil enzyme activity, and pH after inoculation with B. amyloliquefaciens SQ-2（n=18） *P<0.05; ** P<0.01; *** P<0.001

图6 解淀粉芽孢杆菌SQ-2的香农稀释曲线与Venn图 A：Alpha多样性指数香农稀释曲线；B：接种解淀粉芽孢杆菌SQ-2与对照组的根际土菌群Venn图。CK为对照组，S5为3×108 CFU/mL组，下同

Fig. 6 Shannon dilution curve and Venn plot of B. amylo-liquefaciens SQ-2 A: Alpha diversity index Shannon dilution curve. B: Venn plot of rhizosphere soil microbiota of inoculated Bacillus amyloliquefaciens SQ-2 and control group. CK as the control group, S5 at 3×108 CFU/mL concentration. The same below

图7 土壤细菌多样性分析 *** P<0.001; ns表示无显著性差异

Fig. 7 Diversity analysis of soil bacteria *** P<0.001, ns indicate a nonsignificant difference

图8 水稻根际土壤细菌群落组成分析 A：水稻根际土壤细菌门分类水平；B：水稻根际土壤细菌纲分类水平

Fig. 8 Analysis of bacterial community composition in rice rhizosphere soil A: Phylum classification of bacteria in rice rhizosphere soil. B: Class classification of bacteria in rice rhizosphere soil

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