Abstract
Background
Soil salinity is a severe environmental stress, limiting plant growth and sustainable agriculture. Plant growth promoting rhizobacteria (PGPR) and Epichloë gansuensis endophyte can be used to alleviate the adverse effects of soil salinization, promote plant growth and improve resistance to biotic and abiotic stresses.
Aims
The present study was conducted to investigate the diversity of culturable rhizosphere soil bacteria associated with Achnatherum inebrians, as well as the effect of E. gansuensis and PGPR on the germination of A. inebrians seeds under high concentrations of NaCl.
Materials and methods
The PGPR strains were obtained by traditional isolation and culture techniques, and then endophyte-infected and endophyte-free A. inebrians seeds exposed to different concentrations of NaCl were inoculated with PGPR.
Results
A total of 990 bacterial colonies were isolated, and these were assigned to 65 species within 18 genera and 4 phyla. Principal component analysis (PCA) indicated that the rhizosphere soil of A. inebrians plants with Epichloë had differing bacterial diversity from that without Epichloë. When A. inebrians seeds were exposed to NaCl concentrations of 0, 50, 100, 200 mmol/L, seed germination was enhanced at the low salt concentration (50 mmol/L) and inhibited at the highest salt concentration (200 mmol/L). E. gansuensis significantly (P < 0.05) promoted the germination and peroxidase (POD) activity of the seedlings under different NaCl concentrations. Inoculation with three selected isolates of rhizosphere bacteria significantly (P < 0.05) improved seed germination under the four different NaCl concentrations.
Conclusions
This study provides new evidence that inoculation with PGPR of both endophyte-infected and uninfected A. inebrians seeds reduces the impact of NaCl stress on seed germination and provides a basis for further research on the mechanism of PGPR promotion of seed germination of A. inebrians.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data that support the findings of this study are available in tables, figures and supplementary files.
References
Adiputra YT, Hsu TH, Gwo JC (2011) Genetic relationship of milkfish (Chanos chanos) from Indonesia, the Philippines and Taiwan using mitochondrial cytochrome b sequences. J Appl Ichthyol 27(4):1100–1103. https://doi.org/10.1111/j.1439-0426.2010.01629.x
Ansari M, Shekari F, Mohammadi MH, Biró B, Végvári G (2017) Improving germination indices of alfalfa cultivars under saline stress by inoculation with beneficial bacteria. Seed Sci Technol 45(2):475–484. https://doi.org/10.15258/sst.2017.45.2.02
Berg G (2009) Plant-microbe interactions promoting plant growth and health: perspectives for controlled use of microorganisms in agriculture. Appl Microbiol Biot 84(1):11–18. https://doi.org/10.1007/s00253-009-2092-7
Beyer WF, Fridovich I (1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in conditions. Anal Biochem 161(2):559–566. https://doi.org/10.1016/0003-2697(87)90489-1
Bhattacharyya PN, Jha DK (2012) Plant growth-promoting rhizobacteria (PGPR): emergence in agriculture. World J Microbiol Biotechnol 28:1327–1350. https://doi.org/10.1007/s11274-011-0979-9
Brunetti G, Farrag K, Soler-Rovira P, Ferrara M, Nigro F, Senesi N (2012) The effect of compost and Bacillus licheniformis on the phytoextraction of Cr, Cu, Pb and Zn by three brassicaceae species from contaminated soils in the Apulia region, Southern Italy. Geoderma 170:322–330. https://doi.org/10.1016/j.geoderma.2011.11.029
Bulgarelli D, Schlaeppi K, Spaepen S, van Themaat EVL, Schulze-Lefert P (2013) Structure and functions of the bacterial microbiota of plants. Annu Rev Plant Biol 64(1):807–838. https://doi.org/10.1146/annurev-arplant-050312-120106
Buyer JS, Zuberer DA, Nichols KA, Franzluebbers AJ (2011) Soil microbial community function, structure, and glomalin in response to tall fescue endophyte infection. Plant Soil 339(1–2):401–412. https://doi.org/10.1007/s11104-010-0592-y
Chance B, Maehly AC (1955) Assay of catalases and peroxidases. Method Enzymol 2(55):764–775. https://doi.org/10.1002/9780470110171.ch14
Chen L, Li XZ, Swoboda GA, Young CA, Sugawara K, Leuchtmann A, Schardl CL (2015) Two distinct Epichloë species symbiotic with Achnatherum inebrians, drunken horse grass. Mycologia 107(4):863–873. https://doi.org/10.3852/15-019
Chen N, He RL, Chai Q, Li CJ, Nan ZB (2016) Transcriptomic analyses giving insights into molecular regulation mechanisms involved in cold tolerance by Epichloë endophyte in seed germination of Achnatherum inebrians. Plant Growth Regul 80(3):367–375. https://doi.org/10.1007/s10725-016-0177-8
Christensen MJ, Bennett RJ, Ansari HA, Koga H, Johnson RD, Bryan GT, Simpson WR, Koolaard JP, Nickless EM, Voisey CR (2008) Epichloë endophytes grow by intercalary hyphal extension in elongation grass leaves. Fungal Genet Biol 45(2):84–93. https://doi.org/10.1016/j.fgb.2007.07.013
Chu HY, Fierer N, Lauber CL, Gaporaso JG, Knight R, Grogan P (2010) Soil bacterial diversity in the Arctic is not fundamentally different from that found in other biomes. Environm Microbiol 12(11):2998–3006. https://doi.org/10.1111/j.1462-2920.2010.02277.x
Clairborne A (1985) Catalase activity. In: Greenwald R A, ed. Handbook of methods for oxygen radical research. Boca Raton: CRC Press 283–284. https://doi.org/10.1016/0531-5565(85)90021-X
Cripps MG, Edwards GR, Mckenzie SL (2013) Grass species and their fungal symbionts affect subsequent forage growth. Basic Appl Ecol 14(3):225–234. https://doi.org/10.1016/j.baae.2013.01.008
Devi P, Rodrigues C, Naik CG, Souza LD (2012) Isolation and characterization of antibacterial compound from a mangrove-endophytic fungus, Penicillium chrysogenum MTCC 5108. Indian J Microbiol 52(4):617–623. https://doi.org/10.1007/s12088-012-0277-8
Dimkpa C, Weinand T, Asch F (2009) Plant-rhizobacteria interactions alleviate abiotic stress conditions. Plant Cell Environ 32(12):1682–1694. https://doi.org/10.1111/j.1365-3040.2009.02028.x
Esterbauer HK, Cheeseman H (1990) Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Method Enzymol 186:407–421. https://doi.org/10.1016/0076-6879(90)86134-H
Fan P, Chen D, He Y, Zhou QX, Tian YQ, Gao LH (2016) Alleviating salt stress in tomato seedlings using Arthrobacter and Bacillus megaterium isolated from the rhizosphere of wild plants grown on saline-alkaline lands. Int J Phytoremediat 18(11):1113–1121. https://doi.org/10.1080/15226514.2016.1183583
Flores-Núñez VM, Amora-Lazcano E, Angélica R-D, Cruz-Maya JA, Jan-Roblero J (2018) Comparison of plant growth-promoting rhizobacteria in a pine forest soil and an agricultural soil. Soil Res 56:346–355. https://doi.org/10.1071/SR17227
Francioli D, Schulz E, Buscot F, Reitz T (2018) Dynamics of soil bacterial communities over a vegetation season relate to both soil nutrient status and plant growth phenology. Microb Ecol 75(1):216–227. https://doi.org/10.1007/s00248-017-1012-0
Gibbs RA, Hayes CR (1988) The use of R2A medium and the spread plate method for the enumeration of heterotrophic bacteria in drinking water. Lett Appl Microbiol 6(2):19–21. https://doi.org/10.1111/j.1472-765X.1988.tb01205.x
Gul B, Ansari R, Flowers TJ, Khan MA (2013) Germination strategies of halophyte seeds under salinity. Environ Exp Bot 92:4–18. https://doi.org/10.1016/j.envexpbot.2012.11.006
Guo J, Mcculley RL, Mcnear DH (2015) Tall fescue cultivar and fungal endophyte combinations influence plant growth and root exudate composition. Front Plant Sci 6:183. https://doi.org/10.3389/fpls.2015.00183
Han QQ, Wu YN, Gao HJ, Xu R, Paré PW, Shi HZ, Zhao Q, Li HR, Khan SA, Wang YQ, Wang SM, Zhang JL (2017) Improved salt tolerance of medicinal plant Codonopsis pilosula by Bacillus amyloliquefaciens GB03. Acta Physiol Plant 39(1):35. https://doi.org/10.1007/s11738-016-2325-1
Johnson LJ, de Bonth AC, Briggs LR, Caradus JR, Finch SC, Fleetwood DJ, Fletcher LR, Hume DE, Johnson RD, Popay AJ, Tapper BA, Simpson WR, Voisey CR, Card SD (2013) The exploitation of epichloae endophytes for agricultural benefit. Fungal Divers 60(1):171–188. https://doi.org/10.1007/s13225-013-0239-4
Jorquera MA, Shaharoona B, Nadeem SM, Mora ML, Crowley DE (2012) Plant growth-promoting rhizobacteria associated with ancient clones of creosote bush (Larrea tridentata). Microb Ecol 64(4):1008–1017. https://doi.org/10.1007/s00248-012-0071-5
Ju YW, Zhong R, Christensen MJ, Zhang XX (2020) Effects of Epichloë gansuensis endophyte on the root and rhizosphere soil bacteria of Achnatherum inebrians under different moisture conditions. Front Microbiol 11:747. https://doi.org/10.3389/fmicb.2020.00747
Khan MA, Weber DJ (2006) [Tasks for Vegetation Science] Ecophysiology of High Salinity Tolerant Plants Volume 40 || Halophyte Seed Germination. 4:55–67. https://doi.org/10.1007/1-4020-4018-0
Kivlin SN, Emery SM, Rudgers JA (2013) Fungal symbionts alter plant responses to global change. Am J Bot 100(7):1445–1457. https://doi.org/10.3732/ajb.1200558
Leuchtmann A, Bacon CW, Schardl CL, White JF, Tadych M (2014) Nomenclatural realignment of Neotyphodium species with genus Epichloë. Mycologia 106(2):202–215. https://doi.org/10.3852/13-251
Li CH, Yan K, Tang LS, Jia ZJ, Li Y (2014) Change in deep soil microbial communities due to long-term fertilization. Soil Biol Biochem 75:264–272. https://doi.org/10.1016/j.soilbio.2014.04.023
Li CJ, Nan ZB, Paul VH, Dapprich PD, Liu Y (2004) A new Neotyphodium species symbiotic with drunken horse grass (Achnatherum inebrians) in China. Mycotaxon 90:141–147
Li CJ, Wang ZF, Chen N, Nan ZB (2009) First report of choke disease caused by Epichloë typhina on orchardgrass (Dactylis glomerata) in China. Plant Dis 93(6):673–673. https://doi.org/10.1094/PDIS-93-6-0673B
Li XZ, Song ML, Yao X, Chai Q, Simpson WR, Li CJ, N ZB, (2017) The effect of seed-borne fungi and Epichloë endophyte on seed germination and biomass of Elymus sibiricus. Front Microbiol 8:2488. https://doi.org/10.3389/fmicb.2017.02488
Liang Y, Wang HC, Li CJ, Nan ZB, Li FD (2017) Effects of feeding drunken horse grass infected with Epichloë gansuensis endophyte on animal performance, clinical symptoms and physiological parameters in sheep. BMC Vet Res 13:223. https://doi.org/10.1186/s12917-017-1120-6
Liu J, Nagabhyru P, Schardl CL (2017) Epichloë festucae endophytic growth in florets, seeds, and seedlings of perennial ryegrass (Lolium perenne). Mycologia 109(5):1–10. https://doi.org/10.1080/00275514.2017.1400305
Liu JY, Wang XR, Li MN, Du Q, Li QY, Ma PD (2015) Jilinibacillus soli gen. nov., sp. nov., a novel member of the family Bacillaceae. Arch Microbiol 197(1):11–16. https://doi.org/10.1007/s00203-014-1032-9
Ma M, Christensen M, Nan Z (2015) Effects of the endophyte epichloe festucae var. lolii of perennial ryegrass (lolium perenne) on indicators of oxidative stress from pathogenic fungi during seed germination and seedling growth. Eur J Plant Pathol 141(3):571–583. https://doi.org/10.1007/s10658-014-0563-x
Marsh DR, Galliard T (1986) Measurement of polyphenol oxidase activity in wheat-milling fractions. J Cereal Sci 4(3):241–248. https://doi.org/10.1016/S0733-5210(86)80026-1
Martínez-Viveros O, Jorquera MA, Crowley DE, Gajardo G, Mora ML (2010) Mechanisms and practical considerations involved in plant growth promotion by rhizobacteria. J Soil Sci Plant Nutr 10(3):293–319. https://doi.org/10.4067/S0718-95162010000100006
Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7(9):405–410. https://doi.org/10.1016/S1360-1385(02)02312-9
Munns R, Gilliham M (2015) Salinity tolerance of crops-what is the cost? New Phytol 208(3):668–673. https://doi.org/10.1111/nph.13519
Nan ZB, Li CJ (2000) Neotyphodium in native grasses in China and observations on endophyte/host interactions. Proceedings of the 4th international Neotyphodium/grass interactions symposium, Soest, Germany.
Novas MV, Gentile A, Cabral D (2003) Comparative study of growth parameters on diaspores and seedlings between populations of Bromus setifolius from Patagonia, differing in Neotyphodium endophyte infection. Flora 198(6):421–426. https://doi.org/10.1078/0367-2530-00115
Parvaiz M (2014) Response of maize to salt stress a critical review. Int J Healthc Sci (IJHS) 1(1):13–25
Paul D, Lade H (2014) Plant-growth-promoting rhizobacteria to improve crop growth in saline soils: a review. Agron Sustain Dev 34(4):737–752. https://doi.org/10.1007/s13593-014-0233-6
Radhakrishnan R, Shim KB, Lee BW, Hwang CD, Pae SB, Park CH, Kim SU, Lee CK, Baek IU (2013) IAA-producing Penicillium sp. NICS01 triggers plant growth and suppresses Fusarium sp.-induced oxidative stress in sesame (Sesamum indicum L.). J Microbiol Biotechn 23(6):856–863. https://doi.org/10.4014/jmb.1209.09045
Ramadoss D, Lakkineni VK, Bose P, Ali S, Annapurna K (2013) Mitigation of salt stress in wheat seedlings by halotolerant bacteria isolated from saline habitats. Springerplus 2:6. https://doi.org/10.1186/2193-1801-2-6
Rančić A, Soković M, Karioti A, Vukojević J, Skaltsa H (2006) Isolation and structural elucidation of two secondary metabolites from the filamentous fungus Penicillium ochrochloron with antimicrobial activity. Environ Toxicol Phar 22(1):80–84. https://doi.org/10.1016/j.etap.2005.12.003
Rajendran G, Sing F, Desai AJ, Archana G (2008) Enhanced growth and nodulation of pigeon pea by co-inoculation of Bacillus strains with Rhizobium spp. Bioresource Technol 99(11):4544–4550. https://doi.org/10.1016/j.biortech.2007.06.057
Rojas X, Guo JQ, Leff JW, McNear DH Jr, Fierer N, McCulley RL (2016) Infection with a shoot-specific fungal endophyte (Epichloë) alters tall fescue soil microbial communities. Microb Ecol 72(1):197–206. https://doi.org/10.1007/s00248-016-0750-8
Sanchez-Azofeifa A, Oki Y, Fernandes GW, Ball RA, Gamon J (2012) Relationships between endophyte diversity and leaf optical properties. Trees (berlin) 26(2):291–299. https://doi.org/10.1007/s00468-011-0591-5
Schardl CL, Leuchtmann A, Spiering MJ (2004) Symbioses of grasses with seedborne fungal endophytes. Annu Rev Plant Biol 55(1):315–340. https://doi.org/10.1146/annurev.arplant.55.031903.141735
Shahbaz M, Ashraf M (2013) Improving salinity tolerance in cereals. Criti Rev Plant Sci 32(4):237–249. https://doi.org/10.1080/07352689.2013.758544
Sharma V, Salwan R, Shanmugam V (2018) Unraveling the multilevel aspects of least explored plant beneficial Trichoderma saturnisporum isolate GITX-Panog (C). Eur J Plant Pathol 152:169–183. https://doi.org/10.1007/s10658-018-1461-4
Shen CC, Xiong JB, Zhang HY, Feng YZ, Lin XG, Li XY, Liang WJ, Chu HY (2013) Soil pH drives the spatial distribution of bacterial communities along elevation on Changbai Mountain. Soil Biol Biochem 57:204–211. https://doi.org/10.1016/j.soilbio.2012.07.013
Soto-Barajas MC, Zabalgogeazcoa I, Gómez-Fuertes J, Gómez-Fuertes J, González-Blanco V, Vázquez-de-Aldana BR (2015) Epichloë endophytes affect the nutrient and fiber content of lolium perenne, regardless of plant genotype. Plant Soil 405:265–277. https://doi.org/10.1007/s11104-015-2617-z
Tan L, Chen S, Wang T, Dai S (2013) Proteomic insights into seed germination in response to environmental factors. Proteomics 13:1850–1870. https://doi.org/10.1002/pmic.201200394
Tan TS, Syed HS, Yap WB (2017) Expression of surface-bound non-structural 1 (NS1) protein of influenza virus A H5N1 on Lactobacillus casei strain C1. Lett Appl Microbiol 64:446–451. https://doi.org/10.1111/lam.12738
Theocharis A, Bordiec S, Fernandez O, Paquis S, Dhondt-Cordelier S, Baillieul F, Clement C, Barka EA (2012) Burkholderia phytofirmans PsJN primes Vitis vinifera L. and confers a better tolerance to low non-freezing temperatures. Mol Plant Microbe in 25:241–249. https://doi.org/10.1094/MPMI-05-11-0124
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The clustal_x windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882. https://doi.org/10.1093/nar/25.24.4876
Vandegrift R, Roy BA, Pfeifer-Meister L, Johnson BR, Bridgham SD (2015) The herbaceous landlord: integrating the effects of symbiont consortia within a single host. Peer J 3:e1379. https://doi.org/10.7717/peerj.1379
Wang JF, Tian P, Christensen M, Zhang XX, Li CJ, Nan ZB (2018) Effect of Epichloë gansuensis endophyte on the activity of enzymes of nitrogen metabolism, nitrogen use efficiency and photosynthetic ability of Achnatherum inebrians under various NaCl concentrations. Plant Soil 422:267–281. https://doi.org/10.1007/s11104-018-3868-2
Waqas M, Khan AL, Kamran M, Hamayun M, Kang SM, Kim YH, Lee IJ (2012) Endophytic fungi produce gibberellins and indoleacetic acid and promotes host-plant growth during stress. Molecules 17:10754–10773. https://doi.org/10.3390/molecules170910754
Xia C, Christensen MJ, Zhang XX, Nan ZB (2018) Effect of Epichloë gansuensis endophyte and transgenerational effects on the water use efficiency, nutrient and biomass accumulation of Achnatherum inebrians under soil water deficit. Plant Soil 424:555–571. https://doi.org/10.1007/s11104-018-3561-5
Xia C, Li NN, Zhang XX, Feng Y, Christensen MJ, Nan ZB (2016) An Epichloë endophyte improves photosynthetic ability and dry matter production of its host Achnatherum inebrians infected by Blumeria graminis under various soil water conditions. Fungal Ecol 16:26–34. https://doi.org/10.1016/j.funeco.2016.04.002
Xia C, Zhang XX, Christensen MJ, Nan ZZ, Li CJ (2015) Epichloë endophyte affects the ability of powdery mildew (Blumeria graminis) to colonise drunken horse grass (Achnatherum inebrians). Fungal Ecol 16:26–33. https://doi.org/10.1016/j.funeco.2015.02.003
Yang B, Wang XM, Ma HY, Jia Y, Li X, Dai CC (2014) Effects of the fungal endophyte Phomopsis Liquidambari on nitrogen uptake and metabolism in rice. Plant Growth Regul 73(2):165–179. https://doi.org/10.1007/s10725-013-9878-4
Yang HJ, Sung Y (2011) Biocontrol of mildew with Bacillus subtilis in bitter gourd (momordica charantia L.) seeds during germination. Sci Horticulturae (amsterdam) 130:38–42. https://doi.org/10.1016/j.scienta.2011.05.017
Yao LH, Wang DJ, Kang L, Wang DK, Zhang Y, Hou XY, Guo YJ (2018) Effects of fertilizations on soil bacteria and fungi communities in a degraded arid steppe revealed by high through-put sequencing. PeerJ 6:e4623. https://doi.org/10.7717/peerj.4623
Zhang W, Card SD, Mace WJ, Christensen MJ, McGill CR, Matthew C (2017) Defining the pathways of symbiotic Epichloë colonization in grass embryos with confocal microscopy. Mycologia 109:153–161. https://doi.org/10.1080/00275514.2016.1277469
Zhang XM, Wei HW, Chen QS, Han XG (2014) The counteractive effects of nitrogen addition and watering on soil bacterial communities in a steppe ecosystem. Soil Biol Biochem 72:26–34. https://doi.org/10.1016/j.soilbio.2014.01.034
Zhang XX, Fan XM, Li CJ, Nan ZB (2010a) Effects of cadmium stress on seed germination, seedling growth and antioxidative enzymes in Achnatherum inebrians plants infected with a Neotyphodium endophyte. Plant Growth Regul 60:91–97. https://doi.org/10.1007/s10725-009-9422-8
Zhang XX, Li CJ, Nan ZB (2010b) Effects of cadmium stress on growth and anti-oxidative systems in Achnatherum inebrians symbiotic with Neotyphodium gansuense. J Hazard Mater 175:703–709. https://doi.org/10.1016/j.jhazmat.2009.10.066
Zhang XX, Li CJ, Nan ZB, Matthew C (2012) Neotyphodium endophyte increases drunken horse grass (Achnatherum inebrians) resistance to herbivores and seed predators. Weed Res 52:70–78. https://doi.org/10.1111/j.1365-3180.2011.00887.x
Zhong R, Xia C, Ju YW, Li NN, Zhang XX, Nan ZB, Christensen MJ (2018) Effects of Epichloë gansuensis on root-associated fungal communities of Achnatherum inebrians under different growth conditions. Fungal Ecol 31:29–36. https://doi.org/10.1016/j.funeco.2017.10.005
Zhong R, Xia C, Ju YW, Zhang XX, Duan TY, Nan ZB (2019) A foliar Epichloë endophyte and soil moisture modified belowground arbuscular mycorrhizal fungal biodiversity associated with Achnatherum inebrians. Plant Soil. https://doi.org/10.1007/s11104-019-04365-7
Acknowledgements
We wish to thank the editor and anonymous reviewers for their valuable comments. This work was financially supported by the National Nature Science Foundation of China (31772665 and 32061123004), National Basic Research Program of China (2014CB138702), and the Fundamental Research Funds for the Central Universities (lzujbky-2020-cd04), Lanzhou University.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing financial interest.
Additional information
Responsible Editor: Stijn Spaepen
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Michael J. Christensen is retired from AgResearch
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Ju, Y., Kou, M., Zhong, R. et al. Alleviating salt stress on seedings using plant growth promoting rhizobacteria isolated from the rhizosphere soil of Achnatherum inebrians infected with Epichloë gansuensis endophyte. Plant Soil 465, 349–366 (2021). https://doi.org/10.1007/s11104-021-05002-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11104-021-05002-y