|
|
|
| Screening and Promoting Effects of Cold-adapted PGPB from Melissitus ruthenica and Oxytropis ochrocephala Grown in the Alpine Grassland of Qilian Mountains |
| LI Ming-Yuan1,2, WANG Ji-Lian1,2, YAO Tuo1,*, WANG Zhen-Long1, ZHANG Hui-Rong1, CHAI Jia-Li1, LIU Xiao-Ting1, LI Qing-Pu1 |
1 College of Grassland Science/Key Laboratory of Grassland Ecosystem of Ministry of Education, Gansu Agricultural University, Lanzhou 730070, China;
2 The College of Life and Geographic Sciences / Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region, Kashi University, Kashi 844006, China |
|
|
|
|
Abstract Plant growth-promoting bacteria (PGPB) with low-temperature tolerance are beneficial for developing biofertilizers used in cold environments. To obtain cold-adapted PGPB resources, strains with nitrogen fixation and phosphorus solubilization abilities were screened from 2 leguminous species, Melissitus ruthenica and Oxytropis ochrocephala, which are the dominant leguminous plants grown in the alpine grasslands of Qilian Mountains. They were isolated from the root and rhizosphere of plants using selected medium. Other growth-promoting characteristics, including secretion of indole-3-acetic acid (IAA), production of siderophores and ACC deaminase, were further studied by qualitative and quantitative methods. 16S rRNA PCR-restriction fragment length polymorphism (PCR-RFLP) and 16S rRNA gene sequencing were used to identify the strains. Then the excellent strains were selected to verify their effects on Elymus nutans at low temperatures. Elymus nutans are the main reseeding forage specie of natural grassland in this area. As a result, a total of 87 isolates were screened, of which 45 strains still exert high activity at 4 ℃. The inorganic phosphate solubility was negatively correlated with pH. The lower the pH was, the greater the inorganic phosphate solubility, but there was no linear relationship. The genetic diversity of PGPB in the rhizosphere and root of the two legumes was not rich. They were divided into 5 genera by 16S rRNA gene sequencing and phylogenetic analysis, of which Pseudomonas were the dominant genera, and 63.22% of them had two or more plant growth-promoting characteristics. The inoculation of Elymus nutans. with cold-adapted PGPB possessing different functional characteristics had a significant promoting effect on both the development of root system (root length, root average diameter, root surface area, root dry weight, and the number of root tips) and aboveground parts (aboveground stem height, stem diameter, and stem dry weight), especially the nitrogen-fixing and IAA secreting strains. The cold-adapted PGPB have potential applications for agriculture stock production of cold areas, which is of great significance to the development of microbial fertilizer suitable for the near-natural restoration of degraded alpine grasslands on the Qinghai-Tibet Plateau.
|
|
Received: 19 March 2021
|
|
|
|
Corresponding Authors:
* yaot@gsau.edu.cn
|
|
|
|
[1] 崔伟国, 尹彦舒, 张方博, 等. 2020. 马铃薯细菌多样性解析及促生高产IAA菌株的筛选[J]. 中国土壤与肥料,(01): 223-231.
(Cui W G, Yin Y S, Zhang F B, et al. 2020. Analysis of potato bacterial diversity and screening of high-yielding IAA strains[J]. Soils and Fertilizers Sciences in China, (01): 223-231)
[2] 戴黎聪, 柯浔, 曹莹芳, 等. 2019. 青藏高原矮嵩草草甸地下和地上生物量分配格局及其与气象因子的关系[J]. 生态学报, 39(02): 486-493.
(Dai L C, Ke X, Cao Y F, et al. 2019. Allocation pattern of above-and belowground biomass and its response to meteorological factors on an alpine meadow in Qinghai-Tibet Plateau[J]. Acta Ecologica, 39(02): 486-493.)
[3] 董世魁, 汤琳, 张相锋, 等. 2017. 高寒草地植物物种多样性与功能多样性的关系[J]. 生态学报, 37(05): 1472-1483.
(Dong S K, Tang L, Zhang X F, et al.2017. Relationship between plant species diversity and functional diversity in alpine grasslands[J]. Acta Ecologica, 37(05): 1472-1483.)
[4] 何翔, 张庆, 李楚, 等. 2020. 具铁载体活性病原细菌的筛选及铁摄取干预对其生长影响[J]. 植物保护, 46(03): 85-93.
(He X, Zhang Q, Li C, et al. 2020. Screening of the pathogentic bacteria with siderophore-producing activity and the effect of intervention in iron uptake on its growth[J]. Plant Protection, 46(03): 85-93.)
[5] 贺金生, 卜海燕, 胡小文, 等. 2020a. 退化高寒草地的近自然恢复: 理论基础与技术途径[J]. 科学通报, 65(34): 3898-3908.
(He J S, Bu H Y, Hu X W, et al. 2020a. Close-to nature restoration of degraded alpine grasslands: Theoretical basis and technical approch[J]. Chinese Science Bulletin, 65(34): 3898-3908.)
[6] 贺金生, 刘志鹏, 姚拓, 等. 2020b. 青藏高原退化草地恢复的制约因子及修复技术[J]. 科技导报, 38(17): 66-80.
(He J S, Liu Z P, Yao T, et al. 2020b. Analysis of the main constraints and restoration techniques of degraded grassland on the Tibetan Plateau[J]. Science and Technology Review, 38(17): 66-80.)
[7] 姜勇, 徐柱文, 王汝振, 等. 2019. 长期施肥和增水对半干旱草地土壤性质和植物性状的影响[J]. 应用生态学报, 30(07): 2470-2480.
(Jiang Y, Xu Z W, Wang R Z, et al. 2019. Effect of long-term fertilization and water addition on soil properties and plant community characteristics in a semiarid grassland[J]. Chinese Journal of Applied Ecology, 30(07): 2470-2480.)
[8] 李海云. 2019. 祁连山高寒草地退化过程中“植被-土壤-微生物”互作关系[D]. 博士学位论文, 甘肃农业大学, 导师: 姚拓, pp. 36-43.
(Li H Y. 2019. The interaction of vegetation-soil-microbes in the degradation process of alpine grassland in Qilian Mountains of China [D]. Thesis for Ph.D., Gansu Agricultural University, Suppervisor: Yao T, pp. 36-43.
[9] 李明源, 王继莲, 古丽巴哈尔·萨吾提. 2015. 新疆东帕米尔高原慕士塔格峰洋布拉克冰川雪冰及融水中可培养细菌多样性分析[J]. 冰川冻土, 37(06): 1634-1641.
(Li M Y, Wang J L, Gulibahaer S. 2015. Culturable bacterial diversity in snow, ice and meltwater of the Yangbark Glacier, Muztag[J]. Journal of Glaciology and Geocryology, 37(06): 1634-1641.)
[10] 赵锦梅, 王彦辉, 王紫, 等. 2020. 祁连山东段金强河河谷高寒草地土壤的水文特征[J]. 草业科学, 37(02): 256-265.
(Zhao J M, Wang Y H, Wang Z, et al.2020. Soil hydrological characteristics of alpine grasslands in the Jinqiang River Valley in eastern Qilian Mountains[J]. Pratacultural Science, 37(2): 256-265.)
[11] Adhikari P, Jain R, Sharma A, et al.2021. Plant growth promotion at low temperature by Phosphate-Solubilizing Pseudomonas spp. isolated from high-altitude himalayan soil[J]. Microbial Ecology, 82(3): 677-687.
[12] Agarwal P, Singh P C, Chaudhry V, et al.2019. PGPR-induced OsASR6 improves plant growth and yield by altering root auxin sensitivity and the xylem structure in transgenic Arabidopsis thaliana[J]. Journal of Plant Physiology, 240: 153010.
[13] Basu A, Prasad P, Das S N, et al.2021. Plant growth promoting rhizobacteria (PGPR) as green bioinoculants: Recent developments, constraints, and prospects[J]. Sustainability, 13(3): 1-20.
[14] Berg M, Koskella B.2018. Nutrient- and dose-dependent microbiome-mediated protection against a plant pathogen[J]. Current Biology, 28(15): 2487-2492.
[15] Chen Q Y, Yuan Y L, Hu Y L, et al.2020. Excessive nitrogen addition accelerates N assimilation and P utilization by enhancing organic carbon decomposition in a Tibetan alpine steppe[J]. Science of The Total Environment, 764(5): 142848.
[16] Chrastil J.1976. Colorimetric estimation of indole-3-acetic acid[J]. Analytical Biochemistry, 72(1-2): 134-138.
[17] Collavino M M, Sansberro P A, Mroginski L A, et al.2010. Comparison of in vitro solubilization activity of diverse phosphate-solubilizing bacteria native to acid soil and their ability to promote Phaseolus vulgaris growth[J]. Biology and Fertility of Soils, 46: 727-738.
[18] de Vries F T, Griffiths R I, Knight C G, et al.2020. Harnessing rhizosphere microbiomes for drought-resilient crop production[J]. Science, 368(6488): 270-274.
[19] Doebereiner J.1994. Isolation and identification of aerobic nitrogen fixing bacteria[C]//, Kassem Alef, Paolo Nannipieri (eds). Methods in Applied Soil Microbiology and Biochemistry. Academic Press, Cambridge, pp. 134-141.
[20] Gao J, Luo Y, Wei Y, et al.2019. Screening of plant growth promoting bacteria (PGPB) from rhizosphere and bulk soil of Caragana microphylla in different habitats and their effects on the growth of Arabidopsis seedlings[J]. Taylor & Francis, 33(1): 921-930.
[21] Gómez-Godínez L J, Fernandez-Valverde S L, Romero J C M, et al.2019. Metatranscriptomics and nitrogen fixation from the rhizoplane of maize plantlets inoculated with a group of PGPRs[J]. Systematic and Applied Microbiology. 42(4): 517-525.
[22] Jain R, Pandey A.2016. A phenazine-1-carboxylic acid producing polyextremophilic Pseudomonas chlororaphis (MCC2693) strain, isolated from mountain ecosystem, possesses biocontrol and plant growth promotion abilities[J]. Microbiol Research. 190: 63-71.
[23] Li H, Qiu Y, Yao T, et al.2020. Effects of PGPR microbial inoculants on the growth and soil properties of Avena sativa, Medicago sativa, and Cucumis sativus seedlings[J]. Soil & Tillage Research. 199: 104577.
[24] Li Z, Zeng Z, Song Z, et al.2021. Vital roles of soil microbes in driving terrestrial nitrogen immobilization[J]. Global Change Biology. 27(9): 1848-1858.
[25] Majeed A, Abbasi M K, Hameed S, et al.2015. Isolation and characterization of plant growth-promoting rhizobacteria from wheat rhizosphere and their effect on plant growth promotion[J]. Frontiers in microbiology, 6(17): 198.
[26] Malhotra H, Vandana, Sharma S, et al.2018. Phosphorus nutrition: Plant growth in response to deficiency and excess[C]//. M. Hasanuzzaman, M. Fujita, H. Oku, et al.,(eds). Plant Nutrients and Abiotic Stress Tolerance. Springer Singapore, pp. 171-190.
[27] Molina-Romero D, Baez A, Quintero-Hernández V, et al.2017. Compatible bacterial mixture, tolerant to desiccation, improves maize plant growth[J]. PLOS ONE, 12(11): e0187913.
[28] Pandey A, Yarzábal L A.2019. Bioprospecting cold-adapted plant growth promoting microorganisms from mountain environments[J]. Applied Microbiology and Biotechnology, 103(2): 643-657.
[29] Penrose D M, Glick B R.2010. Methods for isolating and characterizing ACC deaminase-containing plant growth-promoting rhizobacteria[J]. Physiologia Plantarum, 118(1): 10-15.
[30] Sarkar J, Chakraborty B, Chakraborty U.2018. Plant growth promoting rhizobacteria protect wheat plants against temperature stress through antioxidant signalling and reducing chloroplast and membrane injury[J]. Journal of Plant Growth Regulation, 37(4): 1396-1412.
[31] Shekhar N C.1999. An efficient microbiological growth medium for screening phosphate solubilizing microorganisms[J]. Fems Microbiology Letters, 170(1): 265-270.
[32] Tao C, Li R, Xiong W, et al.2020. Bio-organic fertilizers stimulate indigenous soil Pseudomonas populations to enhance plant disease suppression[J]. Microbiome, DOI:10.21203/rs.3.rs-18216/v2
[33] Tiepo A N, Hertel M F, Rocha S S, et al.2018. Enhanced drought tolerance in seedlings of Neotropical tree species inoculated with plant growth-promoting bacteria[J]. Plant Physiology and Biochemistry, 130: 277-288.
[34] Vyas P, Gulati A.2009. Organic acid production in vitro and plant growth promotion in maize under controlled environment by phosphate-solubilizing fluorescent Pseudomonas[J]. BMC Microbiology, 9: 174-189.
[35] Wagg C, Schlaeppi K, Banerjee S, et al.2019. Fungal-bacterial diversity and microbiome complexity predict ecosystem functioning[J]. Nature Communications, 10: 4841.
[36] Wu H J, Gu Q, Xie Y L, et al.2019. Cold-adapted Bacilli isolated from the Qinghai-Tibetan Plateau are able to promote plant growth in extreme environments[J]. Environmental Microbiology, 21(9): 3505-3526.
[37] Xie J, Yan Z, Wang G, et al.2021. A bacterium isolated from soil in a karst rocky desertification region has efficient phosphate-solubilizing and plant growth-promoting ability[J]. Frontiers in Microbiology, 11: 625450.
[38] Yarzábal L A.2020. Perspectives for using glacial and periglacial microorganisms for plant growth promotion at low temperatures[J]. Applied microbiology and biotechnology, 104(8): 3267-3278.
[39] Zhang R F, MVivanco J, Shen Q.2017. The unseen rhizosphere root-soil-microbe interactions for crop production[J]. Current Opinion in Microbiology, 37: 8-14.
[40] Zhang W J, Xue X, Peng F, et al.2019. Meta-analysis of the effects of grassland degradation on plant and soil properties in the alpine meadows of the Qinghai-Tibetan Plateau[J]. Global Ecology and Conservation, 20: e00774.
[41] Zhou D, Huang X F, Chaparro J M, et al.2016. Root and bacterial secretions regulate the interaction between plants and PGPR leading to distinct plant growth promotion effects[J]. Plant and Soil, 401(1-2): 259-272. |
|
|
|
