Growth Promoting of Pinus massoniana Seedlings Regulated by Rhizosphere Phosphate-solubilizing Paraburkholderia spp.

doi:10.13560/j.cnki.biotech.bull.1985.2022-1226

Abstract

Abstract:

It is aimed to screen phosphate-solubilizing rhizobacteria（PSB）and to clarify its growth-promoting effect on Pinus massoniana seedlings and the mechanism. The soil-dilution plate method was used to isolate and purify the PSB strains. The PSB strains were identified by morphology and 16s rDNA sequencing methods. Finally, the masson pine seedlings were inoculated with PSB strains for 60 d, then the seedling growth and physiological indexes, soil physical and chemical properties, and rhizosphere bacterial community were determined. The results showed that three PSB strains（WJ10, WJ25 and WJ41）with strong phosphate-solubilizing abilities were isolated from the P. massoniana rhizosphere soil and belonged to Paraburkholderia spp. The three PSB strains had the strongest phosphate-solubilizing abilities to aluminum phosphate, followed by tricalcium phosphate, calcium monophosphate and ferric phosphate. Based on the pot experiment, the three PSB strains effectively promoted seedlings growth, with the best effects on the seedling height and root length promotion for WJ25, followed by WJ41 and WJ10. The main mechanisms of seedling promotion of the three PSB strains include: 1）The PSB strains increased the root activity, chlorophyll b, soluble protein, nitrogen, phosphorus and potassium contents in masson pine seedlings; 2）meanwhile, PSBs enhanced the contents of available phosphorus, available potassium, active nitrogen, soil urease and phosphatase in rhizosphere soil; 3）in addition, the addition of three PSB strains also affected the composition and diversity of the rhizosphere bacterial community, and promoted the significant enrichment of beneficial bacteria such as Bacillus, Nitrosospira, Gemmata and Cytophaga in the rhizosphere soil. In conclusion, the three Paraburkholderia spp. strains could promote masson pine seedlings growth by regulating plant physiology and changing rhizosphere micro-environment. This study provides a theoretical basis for the development and application of rhizosphere PSB fertilizer of P. massoniana.

Key words: phosphate-solubilizing bacteria, growth promotion, Paraburkholderia spp., Pinus massoniana

XU Hong-Yun, LV Jun, YU Cun. Growth Promoting of Pinus massoniana Seedlings Regulated by Rhizosphere Phosphate-solubilizing Paraburkholderia spp.[J]. Biotechnology Bulletin, 2023, 39(6): 274-285.

Figures/Tables 10

Fig. 1 Phosphate solubilization zones on PVK agar plates A: WJ10; B: WJ25; C: WJ41

Fig. 2 Morphological structure of PSB strain under electron microscope A: WJ10; B: WJ25; C: WJ41

Fig. 3 Phylogenetic tree based on 16S rDNA sequence in PSB strains using Neighbor Joining method

Fig. 4 P solubilizing capacity of three PSB in four different phosphate medium and acidification of medium during the phosphate solubilization A and B: Strain WJ10; C and D: strain WJ25; E and F: strain WJ41. The data in the figure are means, and the error line is the SE. The same blow

Fig. 5 Effects of three PSB strain inoculations on the growths of P. massoniana seedlings for 60 d

Fig. 6 Effects of PSB strain inoculations on the growth, nutrient uptake and physiological indexes of P. massoniana seedlings A: Seedling height; B: root length; C: lateral root number; D: fresh weight; E: dry weight; F: soluble protein content; G: root activity; H: chlorophyll content; I: plant nutrient. Different letters indicate siginificant difference（P<0.05）. The same below

Fig. 7 Effects of PSB strains on soil nutrient A: Available nutrient content of soil. B: Total nutrient content of soil. C: Soil urease. D: Soil catalase. E: Soil phosphatase

Table 1 Alpha diversity index of bacteria in rhizosphere soil of P. massoniana with different treatments

Treatment	OTUs	Shannon	ACE	Chao1	Simpson
CK	2181±100.00ab	5.67±0.07b	3126.18±107.24a	2968.00±175.58a	0.0134±0.002a
WJ10	1967±75.48bc	5.82±0.14ab	2713.49±60.82b	2636.03±78.64b	0.0090±0.001a
WJ25	2244±179.85a	5.86±0.18a	3109.78±98.45a	2960.25±57.96a	0.0116±0.002a
WJ41	1877±73.75c	5.66±0.17b	2604.44±45.00b	2474.54±47.17b	0.0121±0.002a

Fig. 8 Relative abundance of bacteria at phyla levels and genus levels in the rhizosphere soils under different treatments A: Phyla levels; B: genus levels

Table 2 Correlation between the rhizosphere soil available nutrients, enzyme activity and bacterial alpha diversity index of P. massoniana

α多样性指数 Alpha diversity index	水解N Available N	有效P Available P	速效K Available K	尿素酶 Urease	过氧化氢酶 Catalase	磷酸酶 Phosphatase
Shannon	0.883	0.250	0.426	0.548	0.903	0.426
ACE	-0.139	-0.884	-0.441	-0.519	-0.157	-0.512
Chao1	-0.091	-0.850	-0.452	-0.504	-0.191	-0.516
Simpson	-0.689	-0.773	-0.378	-0.605	-0.464	-0.449

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