Screening of rhizosphere growth promoting bacteria and their application in tailings improvement

doi:10.11686/cyxb2021233

Abstract

Abstract:

This research was aimed at developing economic and effective methods which have strong applicability and adoption prospects for tailings improvement and vegetation restoration using plant growth-promoting rhizobacteria （PGPR）. PGPR from tailings primary environments were screened and a single factor pot experiment with a control and eight treatments was conducted to test their effects on seed germination and growth of Achillea millefolium. The treatments were： an assembly of mixed PGPR （B）， biochar （C）， manure （F） and their combinations ［BC， BF， CF， a compound treatment of B， C and F （BⅠCF）， and BⅡCF （the same as BⅠCF， but with doubled mixed PGPR content）］. Measurements included rhizosphere microbial community composition， tailings nutrients， and for plant tissues， enzyme activities and the total amount and distribution of tailings nutrients including total nitrogen and available phosphorus and potassium. The screened PGPR comprised KSB1 （Pseudomonas sp.）， KSB2 （Pseudomonas sp.）， KSB7 （Pseudomonas sp.） and KSB21 （Rhizobium sp.） selected for their ability to fix nitrogen， dissolve phosphorus， hydrolyze potassium， synthesize indoleacetic acid or secrete 1-aminocyclopropane carboxylic acid deaminase. The pot experiment showed that： 1） The abundance of Pseudomonas and Rhizobium in the rhizosphere environment and the phosphatase activity in tailings can be increased by inoculating mixed PGPR. 2） The effect on tailings nutrients and the growth of A. millefolium of adding biochar alone， was not obvious. 3） The application of manure promoted the growth of A. millefolium， enhanced the activities of tailings enzymes， and significantly increased the abundance of Cyanobacteria （P<0.01）， the total amount of nitrogen in the potted system， and the total amount of available phosphorus and potassium. 4） Compared with F alone， both the BF and CF treatments further improved various measures of A. millefolium growth and tailings enzyme activity. In addition， the total nitrogen in the potted system in the BF treatment increased by 11.18%， and the total available phosphorus and potassium increased by 22.54% and 22.00% respectively， while the total available potassium only increased by 8.32% in CF. 5） The various measures for BICF and BIICF treatments were generally higher than other treatments. Compared with BICF， BIICF had significantly improved indicators including alkaline hydrolysable nitrogen， phosphatase activity and A. millefolium biomass and the total nitrogen in the pot system increased by 9.54%， the total potassium content of the A. millefolium plant tissues increased by 19.12%， and the relative abundances of Cyanobacteria and Pseudomonas were increased by 51.96% and 13.62%， respectively. 6） Under these experimental conditions， neither inoculating with mixed PGPR nor adding biochar alone resulted in any significant effect on tailings improvement or vegetation restoration； manure played a key role in improving the nutrient supply of tailings and promoting the growth of A. millefolium.This work indicates the benefits of mixed PGPR and biochar are directly related to the application of manure. The BIICF treatment is effective in tailings improvement and the mixed PGPR at least play the role in enhancing nitrogen fixation， phosphorus solubilization and potassium solubilization， and strengthening the potassium absorption and utilization ability of A. millefolium.

Key words: tailings improvement, plant growth promoting rhizobacteria, biochar, Achillea millefolium

Yi-chao CHEN, Xiao-ying SUN, Zhi-jie XIE, Pan ZHOU, Lu ZHANG, Xue-li GAO, Dong LI, Xiao-feng LIU. Screening of rhizosphere growth promoting bacteria and their application in tailings improvement[J]. Acta Prataculturae Sinica, 2022, 31(7): 50-63.

Figures/Tables 12

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处理 Treatments	铁尾矿砂 Iron tailings （kg）	LB培养基 Luria-bertani medium （mL）	混合促生菌液 Plant growth promoting rhizobacteria （mL）	生物炭 Biochar （g）	粪肥 Manure （g）
空白对照CK	5	2.4	/	/	/
接种促生菌液B	5	1.2	1.2	/	/
添加生物炭C	5	2.4	/	5	/
施用粪肥F	5	2.4	/	/	30
接种促生菌液并添加生物炭BC	5	1.2	1.2	5	/
接种促生菌液并施用粪肥BF	5	1.2	1.2	/	30
添加生物炭并施用粪肥CF	5	2.4	/	5	30
接种促生菌液、添加生物炭并施用粪肥BⅠCF	5	1.2	1.2	5	30
接种2倍剂量的促生菌液、添加生物炭并施用粪肥BⅡCF	5	/	2.4	5	30

处理 Treatments	铁尾矿砂 Iron tailings （kg）	LB培养基 Luria-bertani medium （mL）	混合促生菌液 Plant growth promoting rhizobacteria （mL）	生物炭 Biochar （g）	粪肥 Manure （g）
空白对照CK	5	2.4	/	/	/
接种促生菌液B	5	1.2	1.2	/	/
添加生物炭C	5	2.4	/	5	/
施用粪肥F	5	2.4	/	/	30
接种促生菌液并添加生物炭BC	5	1.2	1.2	5	/
接种促生菌液并施用粪肥BF	5	1.2	1.2	/	30
添加生物炭并施用粪肥CF	5	2.4	/	5	30
接种促生菌液、添加生物炭并施用粪肥BⅠCF	5	1.2	1.2	5	30
接种2倍剂量的促生菌液、添加生物炭并施用粪肥BⅡCF	5	/	2.4	5	30

菌株 Strain	固氮能力 Nitrogen fixation ability （nmol C₂H₄·mg^-1 Pr·h^-1）	溶磷能力 Phosphate dissolving capacity （mg·L^-1·7 d^-1）		解钾能力 Potassium dissolving ability （mg·L^-1·7 d^-1）		3-IAA产率 3-IAA yield （mg·L^-1·3 d^-1）		ACC脱氨酶活性 ACC deaminase activity （μmol α-KA·mg^-1 Pr·h^-1）
菌株 Strain	固氮能力 Nitrogen fixation ability （nmol C₂H₄·mg^-1 Pr·h^-1）	Ca₃（PO₄）₂	矿砂 Tailings	钾矿石 Potassium ore	矿砂 Tailings	加入L‐色氨酸 Add L-tryptophan	不加L‐色氨酸 Without L-tryptophan	ACC脱氨酶活性 ACC deaminase activity （μmol α-KA·mg^-1 Pr·h^-1）
KSB1	ND	25.85±4.15	6.98 ± 1.91	ND	ND	57.58±5.49	7.64±0.42	2.39±0.38
KSB2	ND	ND	ND	39.18±4.57	21.11±1.53	6.36±1.05	ND	3.01±0.36
KSB7	ND	ND	ND	ND	ND	41.67±4.94	0.91±0.32	3.79±0.50
KSB21	1016.89±3 9.21	1.64±0.34	ND	16.57±0.54	3.31±0.26	9.85±1.05	0.65±0.11	ND

菌株 Strain	固氮能力 Nitrogen fixation ability （nmol C₂H₄·mg^-1 Pr·h^-1）	溶磷能力 Phosphate dissolving capacity （mg·L^-1·7 d^-1）		解钾能力 Potassium dissolving ability （mg·L^-1·7 d^-1）		3-IAA产率 3-IAA yield （mg·L^-1·3 d^-1）		ACC脱氨酶活性 ACC deaminase activity （μmol α-KA·mg^-1 Pr·h^-1）
菌株 Strain	固氮能力 Nitrogen fixation ability （nmol C₂H₄·mg^-1 Pr·h^-1）	Ca₃（PO₄）₂	矿砂 Tailings	钾矿石 Potassium ore	矿砂 Tailings	加入L‐色氨酸 Add L-tryptophan	不加L‐色氨酸 Without L-tryptophan	ACC脱氨酶活性 ACC deaminase activity （μmol α-KA·mg^-1 Pr·h^-1）
KSB1	ND	25.85±4.15	6.98 ± 1.91	ND	ND	57.58±5.49	7.64±0.42	2.39±0.38
KSB2	ND	ND	ND	39.18±4.57	21.11±1.53	6.36±1.05	ND	3.01±0.36
KSB7	ND	ND	ND	ND	ND	41.67±4.94	0.91±0.32	3.79±0.50
KSB21	1016.89±3 9.21	1.64±0.34	ND	16.57±0.54	3.31±0.26	9.85±1.05	0.65±0.11	ND

处理 Treatments	碱解氮 Alkali hydrolyzable nitrogen （mg·kg^-1）	有机质 Organic matter （g·kg^-1）	脲酶 Urease （NH₄⁺-N mg·g^-1·24 h^-1）	磷酸酶 Phosphatase （PhOH mg·g^-1·24 h^-1）	蔗糖转化酶 Saccharase （glu， mg·g^-1·24 h^-1）
CK	1.62±0.08c	3.14±0.12de	5.01±0.16d	1.24±0.08e	0.14±0.01de
B	1.56±0.12c	2.95±0.18e	4.94±0.13d	1.48±0.05d	0.13±0.01de
C	1.51±0.09c	3.39±0.19cd	4.83±0.20d	1.21±0.06e	0.12±0.01e
F	2.32±0.06b	3.65±0.13c	5.35±0.15c	1.66±0.09c	0.23±0.02c
BC	1.65±0.11c	3.40±0.16cd	5.06±0.14d	1.51±0.03d	0.16±0.01d
BF	2.41±0.08b	3.90±0.10b	5.61±0.10bc	1.92±0.12b	0.27±0.02b
CF	2.36±0.05b	4.01±0.15b	5.72±0.17ab	1.71±0.04c	0.24±0.02c
BⅠCF	2.43±0.07b	4.16±0.11ab	5.95±0.19a	2.03±0.12b	0.29±0.03ab
BⅡCF	2.56±0.06a	4.30±0.20a	6.03±0.38a	2.17±0.15a	0.31±0.03a