Identification of Multi-function Growth-promoting Bacteria in Rice-oilseed Rape Rotation Soil and Its Effect on the Growth and Rhizosphere Bacterial Community of Oilseed Rape (Brassica napus)

doi:10.13560/j.cnki.biotech.bull.1985.2024-0823

Abstract

Abstract:

Objective This study aims to identify efficient phosphate-solubilizing growth-promoting strains to increase the available phosphorus content in soil and promote the growth and healthy development of plants and their root systems. Method A phosphate-solubilizing growth-promoting strain capable of thriving in an alkaline environment was isolated from a long-term rice-oilseed rape rotation in Ningbo, Zhejiang province, China. The strain was identified using a combination of the morphological, physiological and biochemical characteristics, along with 16S rDNA molecular analysis. The incubation and phosphate solubilizing conditions were optimized, and the growth promotion effects on rapes were verified through a pot experiment. Subsequently, high-throughput sequencing techniques were also employed to analyze the structure of the bacterial communities in the rhizosphere soil of oilseed rapes (Brassica napus). Result The isolated strain LM1-2 showed a strong capacity for phosphorus solubilization. The amounts of solubilized phosphorus and indole-3-acetic acid (IAA) synthesized were 493.23 and 9.41 μg/mL, respectively. Strain LM1-2 was identified as Kluyvera intermedia. The shake flask experiments demonstrated that the optimal conditions were an initial pH of 10, a liquid volume 30% of the container's volume, and a culture period of 7-9 d at 25℃, with glucose, tryptone, and tricalcium phosphate serving as carbon, nitrogen, and phosphorus sources, respectively. Strain LM1-2 had the strongest phosphorus-solubilizing ability, reaching 587.03 μg/mL. The pot experiment revealed that, compared to the control group, strain LM1-2 increased available phosphorus by 45.29%. The root length, root diameter, root forks and root crossings of oilseed rape treated with LM1-2 significantly increased by 40.78%, 18.97%, 55.30%, and 87.82%, respectively. Additionally, the fresh and dry weights of the oilseed rape increased by 15.51% and 26.82%, respectively. Inoculation with LM1-2 changed the structure of the rhizosphere bacterial community and significantly increased the Simpson index of alpha diversity, with Acidobacteriota being the dominant phylum and its relative abundance increased by 23.66%. Conclusion Strain LM1-2 demonstrates the excellent performance in phosphorus solubilization and growth promotion of oilseed rape, indicating its potential for diverse applications in agricultural production.

Key words: oilseed rape, phosphate-solubilizing bacteria, Kluyvera intermedia, plant growth promoting rhizobacteria, bacterial community structure

XIA Xin-yuan, XUE Dao-sheng, LI Xin-jing, LONG Jun-jie, LU Kai-xing, DING Wo-na, LI Meng-sha. Identification of Multi-function Growth-promoting Bacteria in Rice-oilseed Rape Rotation Soil and Its Effect on the Growth and Rhizosphere Bacterial Community of Oilseed Rape (Brassica napus)[J]. Biotechnology Bulletin, 2025, 41(4): 289-301.

Figures/Tables 12

Fig. 1 Colony diagram (A) and Gram staining diagram (B) of strain LM1-2

Table 1 Physiological characteristics of strain LM1-2

项目 Item	结果 Result	项目 Item	结果 Result
革兰氏染色	-	V-P试验	-
好氧性试验	好氧	淀粉水解	-
接触酶试验	+	明胶液化	+
甲基红（M.R）试验	+	吲哚试验	+

Fig. 2 Phylogenetic tree of 16S rDNA gene sequence of strain LM1-2

Fig. 3 Effects of different culture conditions on the growth status (OD600) of strain LM1-2Different letters indicate the effects of different conditions on the growth status of LM1-2 (P<0.05)

Fig. 4 Effects of different culture conditions on the phosphorus solubilization ability of strain LM1-2Different letters indicate the effects of different conditions on the phosphate solubilization capacity of LM1-2 (P<0.05)

Fig. 5 Effects of LM1-2 on the soil available phosphorus (A), plants (B) and roots (C) of oilseed rapes in the pot experiment* and ** indicate significant differences between treatments at 0.05 and 0.01 levels, respectively

Fig. 6 Principal component analysis of various indicators among potted oilseed rape seedlings under LM1-2 treatmentSAP: Soil available phosphorus; ABFW: above-ground biomass fresh weight; ABDW: above-ground biomass dry weight; UBFW: underground biomass fresh weight; UBDW: underground biomass dry weight; RL: total root length; RSA: root surface area; RD: root diameter; RV: root volume; RT: number of root tips; RF: number of root forks; RC: number of root crossings. The same below

Fig. 7 Correlation analysis among various indicators of potted oilseed rape seedlings under LM1-2 treatment

Fig. 8 Veen plot (A) and Simpson index (B) of soil bacterial community treated with LM1-2* indicates significant difference between treatments at 0.05 level

Table 2 Alpha diversity indexes of rhizosphere soil bacterial communities under different treatments

处理

Treatment

Ace指数

Ace index

Chao指数

Chao index

Shannon指数

Shannon index

Simpson指数

Simpson index

Coverage指数

Coverage index

PD whole tree指数

PD whole tree index

Fig. 9 Composition of bacterial communities in the rhizosphere soil of oilseed rapes at the phylum (A), class (B), order (C), family (D) and genus (E) levels

Fig. 10 LEfSe analysis of differential species

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