Screening, Identification of Plant Growth Promoting Rhizobacteria and Its Effect on Reducing Fertilization While Increasing Efficiency in Wheat (Triticum aestivum)
LI Yong-Bin, LI Yun-Long, GUAN Guo-Hua*, CHEN San-Feng*
State Key Laboratory of Agricultural Biotechnology/College of Biology, China Agricultural University, Beijing 100193, China
Abstract Plant growth-promoting rhizobacteria (PGPR) are microorganisms that live in the rhizosphere and root surface of plants, and can directly or indirectly promote plant growth and development. The application of microbial fertilizers containing plant growth-promoting rhizobacteria can significantly improve the nutritional status and growth of plants. The demand of modern ecological agriculture for non-chemical fertilizers and the improvement of people's awareness of environmental protection make it great potential for the development and research of microbial fertilizers. The collection and preservation of bacterial resources is fundamental to the development of microbial fertilizers, and thus it is extremely important to screen for excellent strains. Therefore, this study collected rhizosphere soil of wheat (Triticum aestivum), maize (Zea mays), tomato (Solanum lycopersicum) and cucumber (Cucumis sativus) grown in Beijing. A total of 500 bacterial strains were isolated and screened with spread plate method, and their plant growth-promoting properties were tested. Through pot experiment in the greenhouse, 15 bacterial strains with excellent growth-promoting effects on tomato seedlings were identified. The 16S rRNA gene sequence analysis revealed that 11 strains 4-L, 14-L, z4-3, z7-2, z13-5, S6, 1-18, Z20-2, Z23-3, Z28-4, Z30-2 were Paenibacillus sp., 2 strains z13-2 and z13-3 were Pseudomonas azotoformans, strain z14-1 was Alcaligenes faecalis, and strain 56 was Bacillus subtilis. On this basis, Paenibacillus sp. S6, 1-18 and B. subtilis 56 with good plant growth-promoting ability were inoculated onto wheat for field plot experiment. In this experiment, 5 kinds of treatment were set, including control 1 (without strain inoculation and the amount of urea was reduced by 10%); control 2 (without strain inoculation and the amount of urea was not reduced), treatment 3 (Paenibacillus sp. S6 was inoculated and the amount of urea was reduced by 10%), treatment 4 (B. subtilis 56 was inoculated and the amount of urea was reduced by 10%), and treatment 5 (Paenibacillus sp. 1-18 was inoculated and the amount of urea was reduced by 10%). At the returning green stage of wheat, the effects of different treatments on the growth of wheat were evaluated. The results showed that the plant height of wheat in treatments 3, 4, 5 was significantly higher than that in control 1 (P<0.05), but had no significant difference from control 2. Similarly, the fresh weight of wheat in treatments 3, 4, 5 was significantly higher than that in control 1 (P<0.05), but had no significant difference from control 2. In addition, wheat yield was counted at maturity. The results showed that under the condition of reducing 10% urea, the yield of wheat inoculated with Paenibacillus sp. S6, Paenibacillus sp. 1-18 and B. subtilis 56 increased by 6.9%, 8.8%, and 10.4% (P<0.05), respectively, compared with the control 1; while there was no significant difference compared with the control 2. The above results indicated that it was feasible to replace 10% of chemical fertilizers with microbial fertilizers containing above 3 strains. Among them, B. subtilis 56 performed best in reducing fertilizer use and increasing yield, and thus it could be demonstrated and promoted on a large scale. This study could provide excellent strain resources for the development of microbial fertilizers, and provide reference for spread and application of the technique using microbial fertilizers replacing some chemical fertilizers.
LI Yong-Bin,LI Yun-Long,GUAN Guo-Hua, et al. Screening, Identification of Plant Growth Promoting Rhizobacteria and Its Effect on Reducing Fertilization While Increasing Efficiency in Wheat (Triticum aestivum)[J]. 农业生物技术学报, 2020, 28(8): 1471-1476.
[1] 金京京,齐永志,甄文超.2016.枯草芽胞杆菌B1514可湿性粉剂对小麦纹枯病的防效及对土壤微生物区系和小麦产量的影响[J].农药学学报,18(5):596-604.
(Jin J J,Qi Y Z,Zhen W C.2016.Effects of Bacillus subtilis B1514 wettable powder on the control effects of wheat sharp eyespot,the soil microflora communities and the yield of wheat[J].Chinese Journal of Pesticide Science,18(5):596-604.)
[2] 罗菲,汪涯,曾庆桂,等.2011.东乡野生稻根际可培养细菌多样性及其植物促生活性分析[J].生物多样性,19(4):476-484.
(Luo F,Wang Y,Zeng Q G,et al.2011.Diversity and plant growth promoting activities of the cultivable rhizobacteria of Dongxiang wild rice (Oryza rufipogon)[J].Biodiversity Science,19(4):476-484.)
[3] 邢芳芳,高明夫,禚优优,等.2016.大麦根际高效溶磷菌的筛选、鉴定及促生效果研究[J].华北农学报,31(S1):252-257.
(Xing F F,Gao M F,Zhuo Y Y,et al.2016.Screening and identification of phosphate solubilizing bacteria in Hordeum vulgare rhizosphere and its growth promoting effect[J].Acta Agriculture Boreali-Simica,31(S1):252-257.)
[4] 张霞,唐文华,张立群.2007.枯草芽孢杆菌B931防治植物病害和促进植物生长的作用[J].作物学报,33(2):236-241.
(Zhang X,Tang W H,Zhang L Q.2007.Biological control of plant diseases and plant growth promotion by Bacillus subtilis B931[J].Acta Agronomia Sinica,33(2):236-241.)
[5] 朱金英,张书良,郭建军,等.2013.枯草芽孢杆菌菌株AMCC100001对棉花的促生效应[J].中国棉花,40(9):16-18.
(Zhu J Y,Zhang S L,Guo J J.et al.2013.Effects of Bacillus subtilis AMCC100001 on the promotion of cotton growth[J].China Cotton,40(9):16-18.)
[6] Anand R,Grayston S,Chanway C.2013.N2 fixation and seedling growth promotion of lodgepole pine by endophytic Paenibacillus polymyxa[J].Microbial Ecology,66(2):369-374.
[7] Backer R,Rokem J S,Ilangumaran,G,et al.2018.Plant growth-promoting rhizobacteria:Context,mechanisms of action,and roadmap to commercialization of biostimulants for sustainable agriculture[J].Frontiers in Plant Science,9:1-17.
[8] Han H,Sheng X,Hu J,He L,Wang Q.2018.Metal-immobilizing Serratia liquefaciens CL-1 and Bacillus thuringiensis X30 increase biomass and reduce heavy metal accumulation of radish under field conditions[J].Ecotoxicology and Environmental Safety,161:526-533.
[9] Kloepper J W,Leong J,Teintze M,et al.1980.Enhanced plant growth by siderophores produced by plant growth-promoting rhizobacteria[J].Nature,286(5776):885-886.
[10] Majeed A,Muhammad Z,Ahmad,H.2018.Plant growth promoting bacteria:Role in soil improvement,abiotic and biotic stress management of crops[J].Plant cell Reports,37(12):1599-1609.
[11] Marmur J.1961.A procedure for the isolation of DNA from microorganisms[J].Journal of Molecular Biology,3(2):208-218.
[12] Stein T,Borchert S,Conrad B,et a1.2002.Two different antibiotic-like peptides originate from the ericin gene cluster of Bacillus subtilis A1/3[J].Journal of Bacteriology,184(6):1703-1711.
[13] Tamehiro N,Okamoto Y,Okamoto S,et a1.2002.Bacilysocin,a novel phospholipids antibiotic produced by Bacillus subtilis 168[J].Antimicrobial Agents & Chemotherapy,46(2):315-320.
[14] Weselowski B,Nathoo N,Eastman A W,et al.2016.Isolation,identification and characterization of Paenibacillus polymyxa CR1 with potentials for biopesticide,biofertilization,biomass degradation and biofuel production[J].BMC Microbiology,16:244.
