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Study on Biological Characteristics and Optimization of Culture Conditions of Two Efficient Phosphate-Solubilizing Bacteria
HUMingzhu, ZHANGJunjie, SHIXuerui, WANGWei, DAIJinxia
Chin Agric Sci Bull ›› 2026, Vol. 42 ›› Issue (5) : 42-49.
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Abbreviation (ISO4): Chin Agric Sci Bull
Editor in chief: Yulong YIN
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Study on Biological Characteristics and Optimization of Culture Conditions of Two Efficient Phosphate-Solubilizing Bacteria
This study aims to investigate the biological characteristics of phosphate-solubilizing strains B23 and S10, optimize their cultivation conditions, and provide basic data for exploring the phosphate-solubilizing mechanism and preparing phosphate-solubilizing bacterial fertilizers. The Salkowski colorimetric method and the phenol-sulfuric acid method were used to quantitatively determine the strain's IAA production and exopolysaccharide production; single-factor level experiments were adopted to optimize phosphate-solubilizing culture conditions for strains; the colorimetric method was used to determine phosphatase and glucose dehydrogenase in strains; and the effects of the strains on the germination of Avena sativa, Triticum aestivum, and Medicago sativa seeds were investigated through pot experiments. The results indicate that both S10 and B23 have the abilities to produce IAA, dissolve organophosphorus, produce siderophores, produce HCN, and produce exopolysaccharide, with IAA yieds of 18.875 mg/L and 7.603 mg/L respectively; exopolysaccharide yields of 16.290 μg/mL and 16.941 μg/mL respectively,S10 also has the ability to produce NH3. 16S rRNA gene sequence analysis indicates that they belong to the Serratia and Acinetobacter, respectively. The strain exhibits the strongest phosphate-solubilizing effect when the carbon and nitrogen sources are sucrose and urea, with the optimal initial pH values being pH 6.0 and pH 7.0, respectively. Compared with before optimization, the activities of alkaline phosphatase, acid phosphatase, and glucose dehydrogenase in S10 were increased by 33.97%, 47.20%, and 20.94%, respectively; in B23, they were increased by 12.47%, 99.19%, and 54.63%, respectively. Both B23 and S10 can significantly improve the germination vigor, germination rate, and sprout length of Avena sativa, Triticum aestivum, and Medicago sativa. They also have a significant promoting effect on the root length of Avena sativa and Triticum aestivum. The above results indicate that strains S10 and B23 have strong phosphate-solubilizing abilities and multiple plant growth-promoting potentials. They can promote seed germination and may serve as candidate strains for preparing phosphate-solubilizing bio-fertilizers.
phosphate soluble microorganisms / exopolysaccharides / phosphatase / glucose dehydrogenase / phosphate-solubilizing ability / growth-promoting characteristics / seed germination
| [1] |
杨剑锋, 王乐, 张园园, 等. 呼和浩特市苜蓿根腐病病原菌鉴定及致病性测定[J]. 植物保护学报, 2022, 49(4):1093-1101.
|
| [2] |
梁锦锋, 于红卫, 叶国平, 等. 有机磷化物对磷细菌生长和解磷的影响[J]. 江西农业学报, 2007, 19(8):89-90,93.
|
| [3] |
杨顺, 杨婷, 林斌, 等. 两株溶磷真菌的筛选、鉴定及溶磷效果的评价[J]. 微生物学报, 2018, 58(2):264-273.
|
| [4] |
李琦. 产植酸酶促生菌特性、解磷机理及植酸酶基因功能鉴定[D]. 兰州: 甘肃农业大学, 2024.
|
| [5] |
|
| [6] |
林燕青, 吴承祯, 洪伟, 等. 解磷菌的研究进展[J]. 武夷科学, 2015(31):161-169.
|
| [7] |
林英, 司春灿, 韩文华, 等. 解磷微生物研究进展[J]. 江西农业学报, 2017, 29(2):99-103.
|
| [8] |
钟传青, 黄为一. 不同种类解磷微生物的溶磷效果及其磷酸酶活性的变化[J]. 土壤学报, 2005, 42(12):286-290.
|
| [9] |
曾汇文, 李倩如, 王雅士, 等. 土壤解磷细菌解磷机制及其促生作用综述[J]. 湘南学院学报, 2022, 43(2):12-20.
|
| [10] |
The presence of toxic metals in natural environments presents a potential health hazard for humans. Metal contaminants in these environments are usually tightly bound to colloidal particles and organic matter. This represents a major constraint to their removal using currently available in situ remediation technologies. One technique that has shown potential for facilitated metal removal from soil is treatment with an anionic microbial surfactant, rhamnolipid. Successful application of rhamnolipid in metal removal requires knowledge of the rhamnolipid–metal complexation reaction. Therefore, our objective was to evaluate the biosurfactant complexation affinity for the most common natural soil and water cations and for various metal contaminants. The conditional stability constant (log K) for each of these metals was determined using an ion‐exchange resin technique. Results show the measured stability constants follow the order (from strongest to weakest): Al3+ > Cu2+ > Pb2+ > Cd2+ > Zn2+ > Fe3+ > Hg2+ > Ca2+ > Co2+ > Ni2+ > Mn2+ >Mg2+ > K+ These data indicate that rhamnolipid will preferentially complex metal contaminants such as lead, cadmium, and mercury in the presence of common soil or water cations. The measured rhamnolipid–metal stability constants were found in most cases to be similar or higher than conditional stability constants reported in the literature for metal complexation with acetic acid, oxalic acid, citric acid, and fulvic acids. These results help delineate the conditions under which rhamnolipid may be successfully applied as a remediation agent in the removal of metal contaminants from soil, as well as surface waters, ground water, and wastestreams.
|
| [11] |
刘合凤. 栓皮栎根际高效解磷菌的筛选鉴定及促生特性和效应[D]. 曲阜: 曲阜师范大学, 2024.
|
| [12] |
马连杰, 张慧, 尹学伟, 等. 高效解磷菌RL-7的鉴定、解磷能力及促生效应[J]. 江苏农业科学, 2024, 52(14):280-284.
|
| [13] |
刘泽平, 王志刚, 徐伟慧, 等. 水稻根际促生菌的筛选鉴定及促生能力分析[J]. 农业资源与环境学报, 2018, 35(2):119-125.
|
| [14] |
于鑫焱. 高产铁载体假单胞菌HYS铁载体合成调控机理研究[D]. 武汉: 武汉大学, 2014.
|
| [15] |
We examined the sensitivity and the specificity of three versions of the Salkowski colorimetric technique. Two of these allowed the detection of indoleacetic acid (IAA) over a low range of concentrations (0.5 to 20 (mu)g/ml), while the third permitted the detection of IAA over a range of higher concentrations (5 to 200 (mu)g/ml). Overall, the three formulations reacted not only with auxin (IAA) but also with indolepyruvic acid and indoleacetamide. Therefore, these techniques appear to be specific for IAA, indolepyruvic acid, and indoleacetamide rather than for IAA alone.
|
| [16] |
黄承敏, 肖茜, 王蓉蓉, 等. 一株高产胞外多糖乳酸菌的分离鉴定及其产胞外多糖的研究[J]. 中国酿造, 2019, 38(1):80-83.
该研究从自然发酵剁辣椒中分离筛选一株高产胞外多糖的乳酸菌，经形态观察、生理生化试验及分子生物学对其进行鉴定，并探索菌株产胞外多糖的最佳碳源和最佳培养时间。结果表明，筛选并鉴定得到一株高产胞外多糖的植物乳杆菌（Lactobacillus plantarum）Y-20。菌株Y-20产胞外多糖的最佳碳源为麦芽糖，且在MRS培养基中培养16 h时，OD600 nm值达到最大（3.62），胞外多糖产量达到最高，为242.95 mg/L。
|
| [17] |
刘尚, 张红, 张乾申, 等. 烟区土壤高效氨化芽孢杆菌菌株的筛选鉴定及特性研究[J]. 中国烟草科学, 2020, 41(2):15-20.
|
| [18] |
陈庆河, 翁启勇, 何玉仙, 等. 大白菜健、病株根际细菌荧光反应及产HCN能力比较研究[J]. 福建农业学报, 1998(2):31-33.
|
| [19] |
中华人民共和国农业部. 植株全磷含量测定钼锑抗比色法:NY/T2421—2013[S]. 北京: 中国农业出版社, 2014.
|
| [20] |
闵九康, 郝心仁, 严慧峻, 等(译). 土壤分析法[M]. 北京: 中国农业科技出版社,1991:592-599.
|
| [21] |
|
| [22] |
李慧萍. 梭梭和云杉根际溶磷细菌的特性研究及溶磷机制解析[D]. 兰州: 兰州大学, 2023.
|
| [23] |
赵为容. 小麦亲和性根际解磷菌解磷机理及促生效果研究[D]. 合肥: 安徽农业大学, 2018.
|
| [24] |
侯莹莹, 胡小梅. 3株玉米根际解磷菌筛选鉴定及促生作用研究[J]. 东北农业大学学报, 2024, 55(5):46-55.
|
| [25] |
吕俊, 潘洪祥, 于存. 马尾松根际溶磷细菌Paraburkholderia sp.的筛选、鉴定及溶磷特性研究[J]. 生物技术通报, 2020, 36(9):147-156.
由马尾松根际筛选高效溶磷细菌,为马尾松幼苗的人工培育及微生物菌肥提供菌种资源。利用稀释涂布平板法和钼锑抗比色法从马尾松根际土壤中分离、筛选溶磷细菌,根据形态特征、生理生化及16S rDNA序列分析对菌株进行鉴定;通过检测菌株对4种不同难溶性磷酸盐的溶解量,筛选出较易被溶解的磷酸盐后,探究其在不同条件下对该磷酸盐的溶解特性;通过测定接种菌株的马尾松幼苗生物量、磷素含量以及根际土壤有效磷含量等指标分析其对马尾松幼苗生长的影响。从马尾松根际分离得到一株具有较好溶磷能力的菌株WJ2,该菌株在无机磷液体培养基内培养5 d后,其溶磷量达244.13 mg/L,经综合鉴定该菌株属于伯克霍尔德菌属（Paraburkholderia sp.）。菌株WJ2对4种不同无机磷酸盐的溶解能力依次为Ca<sub>3</sub>（PO<sub>4</sub>）<sub>2</sub>（204.42 mg/L）>AlPO<sub>4</sub>（177.70 mg/L）>FePO<sub>4</sub>·2H<sub>2</sub>O（151.82 mg/L）>CaHPO<sub>4</sub>·2H<sub>2</sub>O（108.19 mg/L）。Pearson相关分析表明,菌株WJ2对这4种磷酸盐的溶解量与培养液pH值均呈显著负相关关系。该菌株在以乳糖、葡萄糖作为碳源,以尿素、硫酸铵、硝酸铵等作为氮源时对Ca<sub>3</sub>（PO<sub>4</sub>）<sub>2</sub>表现出较强的溶解能力,在C/N为10∶1、NaCl含量为0 g/L、初始pH为8.0的条件下溶磷能力较高。在盆栽试验中,与对照相比,接种菌株WJ2能显著提高马尾松幼苗的苗高、主根长、鲜重,增加量分别为11.74%、45.73%、46.00%。此外,接种菌株WJ2的土壤有效磷含量和植株磷素含量与对照相比也显著增加,增加量分别为18.32%和13.82%。菌株WJ2能够溶解多种难溶性无机磷酸盐,对提高土壤有效磷含量和促进马尾松幼苗生长的效果显著,有望为开发高效微生物磷肥提供种质资源。
|
| [26] |
|
| [27] |
李翯, 王鹏宇, 陈子轩, 等. 黏质沙雷氏菌H04的有机磷降解活性及促生能力[J]. 江苏农业科学, 2023, 51(6):194-199.
|
| [28] |
孙合美. 水稻根际溶磷菌的溶磷效应及对植物的促生作用[D]. 长春: 吉林农业大学, 2016.
|
| [29] |
钟传青, 黄为一. 不同种类解磷微生物的溶磷效果及其磷酸酶活性的变化[J]. 土壤学报, 2005, 42(2):286-294.
|
| [30] |
潘林, 弥春霞, 徐青山, 等. 解磷细菌解磷机制研究进展及其在水稻上的应用[J]. 中国稻米, 2024, 30(3):1-9.
磷是植物生长不可缺少的元素。土壤中总磷含量丰富，但可被植物直接吸收利用的有效磷含量却很低。传统上主要通过施用磷肥提高缺磷地区的作物产量。然而，施入土壤中的磷不仅易被固定，还会转化成难以被植物直接吸收利用的有机磷，从而降低植物对磷的吸收利用率，造成环境污染。解磷细菌具有将土壤中难溶态磷转化为可被植物直接吸收利用的有效磷的功能，提高植物对磷的吸收利用率。解磷细菌种类繁多，作用机制复杂且影响因素众多。本文综述了解磷细菌解磷机制的研究进展，总结了解磷细菌在水稻生产上的应用情况，以为提高稻田土壤难溶态磷的利用效率提供理论依据。
|
| [31] |
李青青, 张芮, 高彦婷, 等. 耐盐碱解磷菌的溶磷效果及其对黄豆萌发的影响[J]. 微生物学通报, 2024, 51(11):4574-4589.
|
| [32] |
何朋杰, 罗李琦, 程晓睿, 等. 生姜根际解磷芽孢杆菌的筛选及其促生效应评价[J]. 南方农业学报, 2024, 55(6):1608-1618.
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