烟草根际高效解磷菌的筛选鉴定及促生作用研究

doi:10.13560/j.cnki.biotech.bull.1985.2021-1511

摘要/Abstract

摘要：

为获得优质的解磷促生菌种资源,助力农业生产减肥增效,对山东省高密市方市乡烟草根际土壤解磷菌进行筛选,将筛选得到的高效解磷菌3P29进行分子生物学鉴定,并研究其解磷性能及促生能力。菌株3P29鉴定为皮特不动杆菌,其对卵磷脂的转化量为13.38 mg/L,磷酸钙的转化量为19.83 mg/L,具有高效分解有机磷及无机磷的功能。烟草盆栽试验表明,烟草主根变长且有更多侧根产生,从而增强根对营养元素的吸收,烟草叶片全氮磷钾含量在无磷营养液条件下提高了71%、49%和134%,可为解磷菌肥的研发与利用提供优良菌株资源。

关键词: 解磷作用, 根系发育, 烟草, 皮特不动杆菌

Abstract:

In order to obtain high-quality phosphate-solubilizing bacterial resources and enhance agricultural production efficiency,we screened phosphorus-solubilizing bacteria from the tobacco rhizosphere soil of Fangshi township,Gaomi city,Shandong province,among which the highly efficient phosphate-solubilizing bacterium 3P29 was identified by molecular biology,then the ability of phosphate-solubilizing and growth-promoting effect were further studied. The strain 3P29 was identified as Acinetobacter pittobacter and the transformation amount of lecithin and calcium phosphate were 13.38 mg/L and 19.83 mg/L,respectively,indicating that the strain 3P29 efficiently decomposed organic and inorganic phosphorus. Tobacco pot experiment showed that the primary root of tobacco became longer and more lateral roots were produced,thereby enhancing the absorption of nutrient elements by the roots. The total nitrogen,phosphorus and potassium content of tobacco leaves increased by 71%,49% and 134% under the conditions of phosphorus-free nutrient solution,which may provide superior strain resources for the development and utilization of phosphate solubilizing bacterial fertilizers.

Key words: phosphate solubilizing, root development, tobacco, Acinetobacter pittii

刘广超, 叶青, 车永梅, 李雅华, 安东, 刘新. 烟草根际高效解磷菌的筛选鉴定及促生作用研究[J]. 生物技术通报, 2022, 38(8): 179-187.

LIU Guang-chao, YE Qing, CHE yong-mei, LI Ya-hua, AN Dong, LIU Xin. Screening and Identification of High-efficiency Phosphate Solubilizing Bacteria in Tobacco Rhizosphere and Its Growth-promoting Effects[J]. Biotechnology Bulletin, 2022, 38(8): 179-187.

图/表 10

表1 解磷菌3P29的解磷能力

Table 1 Phosphorus solubilizing ability of strain 3P29

菌株编号 Strain No.	有机磷含量Organic phosphorus content /（mg·L^-1）	无机磷含量Inorganic phosphorus content /（mg·L^-1）
2P1	11.210	9.260
2P10	9.260	14.490
2P14	2.620	23.250
2P23	10.210	10.690
2P24	7.750	23.920
3P25	3.040	25.330
3P28	2.890	26.190
3P29	13.380	19.830
3P33	0.000	24.870
3P37	0.000	23.520

图1 解磷菌3P29的菌落形态、革兰氏染色及分泌生长素的能力

Fig.1 Colony morphology,Gram staining and auxin secretion ability of strain 3P29

图2 基于解磷菌3P29和相关菌株的16S rDNA 序列采用邻接法建立的系统发育树

Fig. 2 Phylogenetic tree established using the neighbor-joining method,based on 16S rDNA sequences of strain 3P29 and related strains

图3 基于解磷菌3P29和相关菌株的gyrB序列采用邻接法建立的系统发育树

Fig. 3 Phylogenetic tree established using the neighbor-joining method,based on gyrB sequences of strain 3P29 and related strains

图4 解磷菌3P29对土壤矿质元素含量的影响

Fig. 4 Effects of strain 3P29 on mineral element contents in soil

图5 解磷菌3P29对土壤酶活的影响

Fig.5 Effects of strain 3P29 on soil enzyme activity

图6 解磷菌3P29对烟草生长的影响 CKt：无菌处理,CTt：接种3P29,1代表无磷霍格兰营养液培养,2代表花无缺全营养液培养,bar=10 cm

Fig. 6 Effects of strain 3P29 on tobacco growth CKt：Aseptic processing. CTt：Inoculation with 3P29. 1 represents phosphorus-free Hoagland nutrient solution culture,and 2 represents flower-free whole nutrient solution culture,bar=10 cm

表2 解磷菌3P29对烟草生长指标的影响

Table 2 Effects of strain 3P29 on the growth index of tobacco

处理Treatment	株高Height/cm	叶面积Leaf area/cm²	叶片数Number of blades	茎粗Stem thick/cm
CKt1	10.70±0.40	304±0.42	6.67±0.58	0.83±0.03
CTt1	13.40±0.46**	370.67±0.55**	8.67±0.57**	1.13±0.06**
CKt2	12.23±0.25	342.67±0.15	8.00±0.35	1.03±0.15
CTt2	17.13±0.32**	489±0.49**	11.33±0.49**	1.33±0.03**

表3 解磷菌3P29对烟草地上部分和地下部分生物量的影响

Table 3 Effects of strain 3P29 on the biomass of aerial and underground parts of tobacco

处理 Treatment	地上鲜重 Fresh weight on the ground/g	地上干重 Dry weight on the ground/g	地下鲜重 Underground fresh weight/g	地下干重 Underground dry weight/g
CKt1	34.37±0.42	3.12±0.03	1.30±0.10	0.12±0.01
CTt1	59.63±0.55**	6.05±0.05**	2.60±0.10**	0.27±0.01**
CKt2	53.83±0.15	5.45±0.04	1.80±0.10	0.18±0.01
CTt2	92.93±0.49**	9.46±0.02**	2.70±0.17**	0.29±0.01**

图7 解磷菌3P29对烟草叶片氮、磷和钾元素含量的影响图中**表示差异达到显著水平（P<0.01）

Fig. 7 Effects of strain 3P29 on contents of nitrogen,phosphorus and potassium in tobacco leaves **in the figure indicates significant differences at P<0.01 level

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