Abstract
Purpose
Sugarcane-legume intercrops have gained attention because of their ability to improve crop productivity in an ecofriendly manner. We investigated the roles of rhizospheric and endospheric diazotrophs in soil nutrient transformation.
Materials and methods
We conducted two field experiments with three treatments: sugarcane monoculture, sugarcane + peanut intercrop, and sugarcane + soybean intercrop. Soil and plant samples were collected and the nutrient content, enzymatic activity, and diazotroph population were assessed. The nitrogenase (nifH) gene and 16S rRNA gene were used for molecular characterization. Correlation analysis was performed to discover the relations among the soil variables, plant variables, and microbial communities.
Results and discussion
The multiple variance analysis results indicated that the intercropping systems had a significant (p < 0.05) effect on soil total P, available K, and soil enzyme dehydrogenase. Higher diazotrophic populations in the rhizosphere and plant endosphere were recovered by sugarcane intercropping. The soybean intercrop had the maximum unique operational taxonomic units (OTUs). A total of 64 nifH gene-positive bacteria were identified by 16S rRNA gene sequencing. The BLAST search results classified all diazotrophs into five taxonomic groups: γ-Proteobacteria (28%), Actinobacteria (28%), Firmicutes (25%), β-Proteobacteria (11%), and α-Proteobacteria (8%). All identified bacteria had multiple plant growth-promoting abilities, and only 58% of the bacteria utilized aminocyclopropane-1-carboxylic acid (ACC) as their sole carbon source. Furthermore, the higher yield and γ-Proteobacteria positively correlated with soil organic matter, and urease activity revealed that intercropping improved soil fertility.
Conclusions
Our study concluded that the intercropping system enriched the beneficial diazotrophs in the plant rhizosphere and endosphere and that these microbes could enhance plant growth and serve as effective bioinoculants to sustain sugarcane production. Moreover, higher soil nutrients (NKP) and biological activity showed the significance of our intercropping practice of a short-duration leguminous crop and a long-duration grass crop, and this practice would be a good solution to reduce the overuse of chemical fertilizers.
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Albino U, Saridakis DP, Ferreira MC, Hungria M, Vinuesa P, Andrade G (2006) High diversity of diazotrophic bacteria associated with the carnivorous plant Drosera villosa var. villosa growing in oligotrophic habitats in Brazil. Plant Soil 287:199–207
Ali SZ, Sandhya V, Venkateswar Rao L (2014) Isolation and characterization of drought-tolerant ACC deaminase and exopolysaccharide-producing fluorescent Pseudomonas sp. Ann Microbiol 64:493–502
Ashworth AJ, DeBruyn JM, Allen FL et al (2017) Microbial community structure is affected by cropping sequences and poultry litter under long-term no-tillage. Soil Biol Biochem 114:210–219
Bailly X, Olivieri I, De mita S et al (2006) Recombination and selection shape the molecular diversity pattern of nitrogen-fixing Sinorhizobium sp. associated to Medicago. Mol Ecol 15:2719–2734
Balkcom KS, Wood CW, Adams JF, Meso B (2007) Suitability of peanut residue as a nitrogen source for a rye cover crop. Sci Agric 64:181–186
Bao SD (2002) Soil agricultural chemical analysis, 3rd edition. China Agricultural Press, Beijing
Beneduzi A, Moreira F, Costa PB, Vargas LK, Lisboa BB, Favreto R, Baldani JI, Passaglia LMP (2013) Diversity and plant growth promoting evaluation abilities of bacteria isolated from sugarcane cultivated in the south of Brazil. Appl Soil Ecol 63:94–104
Blanco-Canqui H, Lal R (2009) Crop residue removal impacts on soil productivity and environmental quality. CRC Crit Rev Plant Sci 28:139–163
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Bremmer J, Mulvaney C (1982) “Total nitrogen,” In methods of soil analysis—part 2. In: Bluck CA (ed) Chemical and microbiological properties. American Society of Agronomy, Wisconsin, pp 595–624
Carter MR (1993) Soil sampling and methods of analysis. Lewis, Boca Raton
Castro-González R, Martínez-Aguilar L, Ramírez-Trujillo A, Estrada-de los Santos P, Caballero-Mellado J (2011) High diversity of culturable Burkholderia species associated with sugarcane. Plant Soil 345:155–169
Chen P, Du Q, Liu X et al (2017) Effects of reduced nitrogen inputs on crop yield and nitrogen use efficiency in a long-term maize-soybean relay strip intercropping system. PLoS One 12:e0184503. https://doi.org/10.1371/journal.pone.0184503
Dai C-C, Chen Y, Wang X-X, Li P-D (2013) Effects of intercropping of peanut with the medicinal plant Atractylodes lancea on soil microecology and peanut yield in subtropical China. Agrofor Syst 87:417–426
Döbereiner J, Day J (1976) Associative symbiosis in tropical grasses, characterization of microorganisms and dinitrogen fixing sites, In: INTERN. Pullman: Washington State University
Dworkin M, Foster JW (1958) Experiments with some microorganisms which utilize ethane and hydrogen. J Bacteriol 75:592–603
Edwards U, Rogall T, Blöcker H, Emde M, Böttger EC (1989) Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 17:7843–7853
Elkoca E, Kantar F, Sahin F (2007) Influence of nitrogen fixing and phosphorus solubilizing bacteria on the nodulation, plant growth, and yield of chickpea. J Plant Nutr 31:157–171
Fageria NK (2012) Role of soil organic matter in maintaining sustainability of cropping systems. Commun Soil Sci Plant Anal 43:2063–2113
Fischer D, Pfitzner B, Schmid M, Simões-Araújo JL, Reis VM, Pereira W, Ormeño-Orrillo E, Hai B, Hofmann A, Schloter M, Martinez-Romero E, Baldani JI, Hartmann A (2012) Molecular characterisation of the diazotrophic bacterial community in uninoculated and inoculated field-grown sugarcane (Saccharum sp.). Plant Soil 356:83–99
Gordon SA, Paleg LG (1957) Observations on the quantitative determination of indoleacetic acid. Physiol Plant 10:39–47
Guan SY, Zhang D, Zhang Z (1986) Soil enzyme and its research methods. China Agriculture Press, Beijing
Habig J, Swanepoel C (2015) Effects of conservation agriculture and fertilization on soil microbial diversity and activity. Environments 2:358–384
Hammer DAT, Ryan PD, Hammer Ø, Harper DAT (2001) Past: Paleontological Statistics Software Package for Education and Data Analysis. Palaeontol Electron 4:178
Han J, Sun L, Dong X, Cai Z, Sun X, Yang H, Wang Y, Song W (2005) Characterization of a novel plant growth-promoting bacteria strain Delftia tsuruhatensis HR4 both as a diazotroph and a potential biocontrol agent against various plant pathogens. Syst Appl Microbiol 28:66–76
Hardy RW, Holsten RD, Jackson EK, Burns RC (1968) The acetylene-ethylene assay for n(2) fixation: laboratory and field evaluation. Plant Physiol 43:1185–1207
Herridge DF, Peoples MB, Boddey RM (2008) Global inputs of biological nitrogen fixation in agricultural systems. Plant Soil 311:1–18
Jiang Y, Zhou JG, Zou YP (2014) Isolation and primary identification of a new nitrogen-fixation Arthrobacter strain. J Cent China Norm Univ Natural Sci 38:210–214
Kageyama A, Matsumoto A, Omura S, Takahashi Y (2008) Humibacillus xanthopallidus gen. nov., sp. nov. Int J Syst Evol Microbiol 58:1547–1551
Kaur N, Bhullar MS, Gill G (2015) Weed management options for sugarcane-vegetable intercropping systems in north-western India. Crop Prot 74:18–23
Kaushal M, Kaushal R (2015) Acetylene reductase activity and molecular characterization of plant growth promoting rhizobacteria to know efficacy in integrated nutrient management system. Indian J Biotechnol 14:221–227
Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: molecular evolutionary geaetics analysis across computing platforms. Mol Biol Evol 35:1547–1549
Kushwah SS, Reddy DD, Somasundaram J, Srivastava S, Khamparia RS (2016) Crop residue retention and nutrient management practices on stratification of phosphorus and soil organic carbon in the soybean–wheat system in vertisols of central India. Commun Soil Sci Plant Anal 47:2387–2395
Lacava PT, Araujo WL, Marcon J, Maccheroni W, Azevedo JL (2004) Interaction between endophytic bacteria from citrus plants and the phytopathogenic bacteria Xylella fastidiosa, causal agent of citrus-variegated chlorosis. Lett Appl Microbiol 39:55–59
Li Y-R, Yang L-T (2015) Sugarcane agriculture and sugar industry in China. Sugar Tech 17:1–8
Li Z, Luo Y, Teng Y (2008) Research methods for soil and environmental microbiology. Science Press, Beijing
Li X, Mu Y, Cheng Y, Liu X, Nian H (2013) Effects of intercropping sugarcane and soybean on growth, rhizosphere soil microbes, nitrogen and phosphorus availability. Acta Physiol Plant 35:1113–1119
Li Y-R, Zhou X-Z, Yang L-T (2015) Biological nitrogen fixation in sugarcane and nitrogen transfer from sugarcane to cassava in an intercropping system. 6:214–218
Li Q, Chen J, Wu L, Luo X, Li N, Arafat Y, Lin S, Lin W (2018) Belowground interactions impact the soil bacterial community, soil fertility, and crop yield in maize/peanut intercropping systems. Int J Mol Sci 19. https://doi.org/10.3390/ijms19020622
Lin L, Guo W, Xing Y, Zhang X, Li Z, Hu C, Li S, Li Y, An Q (2012a) The actinobacterium Microbacterium sp. 16SH accepts pBBR1-based pPROBE vectors, forms biofilms, invades roots, and fixes N2 associated with micropropagated sugarcane plants. Appl Microbiol Biotechnol 93:1185–1195
Lin L, Li Z, Hu C, Zhang X, Chang S, Yang L, Li Y, An Q (2012b) Plant growth-promoting nitrogen-fixing enterobacteria are in association with sugarcane plants growing in Guangxi, China. Microbes Environ 27:391–398
Loiret FG, Ortega E, Kleiner D, Ortega-Rodes P, Rodes R, Dong Z (2004) A putative new endophytic nitrogen-fixing bacterium Pantoea sp. from sugarcane. J Appl Microbiol 97:504–511
Makinde EA, Oluwatoyinbo FI, Ayoola OT (2006) Intercropping and crop residue incorporation: effects on soil nutrient status. J Plant Nutr 29:235–244
Malik KA, Jung C, Claus D, Schlegel HG (1981) Nitrogen fixation by the hydrogen-oxidizing bacterium Alcaligenes latus. Arch Microbiol 129:254–256
Mehnaz S, Deeba B, Lazarovits G (2010) Genetic and phenotypic diversity of plant growth promoting rhizobacteria isolated from sugarcane plants growing in Pakistan. J Microbiol Biotechnol 20:1614–1623
Mendes R, Pizzirani-Kleiner AA, Araujo WL, Raaijmakers JM (2007) Diversity of cultivated endophytic bacteria from sugarcane: genetic and biochemical characterization of Burkholderia cepacia complex isolates. Appl Environ Microbiol 73:7259–7567
Neugschwandtner RW, Kaul H-P (2015) Nitrogen uptake, use and utilization efficiency by oat–pea intercrops. Field Crop Res 179:113–119
Pan I, Dam B, Sen SK (2012) Composting of common organic wastes using microbial inoculants. 3 Biotech 2:127–134
Patil HJ, Solanki MK (2016) Microbial inoculant: modern era of fertilizers and pesticides. In: Singh D, Singh H, Prabha R (eds) Microbial inoculants in sustainable agricultural productivity: vol. 1: research perspectives. Springer, New Delhi
Pikovskaya R (1948) Mobilization of phosphorus in soil in connection with the vital activity of some microbial species. Mikrobiologiya 17:362–370
Ponnuswamy K, Santhi P, Shanmugasundaram V (1994) Economics of sugarcane based cropping system under various moisture regimes. Bharatiya Sugar 20:57–58
Prakash O, Green SJ, Jasrotia P, Overholt WA, Canion A, Watson DB, Brooks SC, Kostka JE (2012) Rhodanobacter denitrificans sp. nov., isolated from nitrate-rich zones of a contaminated aquifer. Int J Syst Evol Microbiol 62:2457–2462
Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glöckner FO (2012) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41:D590–D596
Robertson GP, Vitousek PM (2009) Nitrogen in agriculture: balancing the cost of an essential resource. Annu Rev Environ Resour 34:97–125
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Sambrook J, Russell DW. (2001) Molecular Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press, New York
Sarr PS, Yamakawa T, Asatsuma S et al (2010) Investigation of endophytic and symbiotic features of Ralstonia sp. TSC1 isolated from cowpea nodules Papa. African J Microbiol Res 4:1959–1963
Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56
Shukla S, Pandey M (1999) Feasibility of inclusion of mustard (Brassica juncea) genotypes in intercropping and sequential cropping systems with sugarcane (Saccharum officinarum). Indian J Agric Sci 69:247–250
Siddikee MA, Chauhan PS, Anandham R et al (2010) Isolation, characterization, and use for plant growth promotion under salt stress, of ACC deaminase-producing halotolerant bacteria derived from coastal soil. J Microbiol Biotechnol 20:1577–1584
Singh J, Singh DK (2005) Dehydrogenase and phosphomonoesterase activities in groundnut (Arachis hypogaea L.) field after diazinon, imidacloprid and lindane treatments. Chemosphere 60:32–42
Singh SN, Yadav RL, Yadav DV et al (2010) Introducing autumn sugarcane as a relay intercrop in skipped row planted rice–potato cropping system for enhanced productivity and profitability in the Indian sub-tropics. Exp Agric 46:519–530
Solanki MK, Kumar S, Pandey AK, Srivastava S, Singh RK, Kashyap PL, Srivastava AK, Arora DK (2012) Diversity and antagonistic potential of Bacillus spp. associated to the rhizosphere of tomato for the management of Rhizoctonia solani. Biocontrol Sci Tech 22:203–217
Solanki MK, Singh RK, Srivastava S, Kumar S, Kashyap PL, Srivastava AK, Arora DK (2014) Isolation and characterization of siderophore producing antagonistic rhizobacteria against Rhizoctonia solani. J Basic Microbiol 54:585–597
Solanki MK, Wang Z, Wang F-Y, Li CN, Lan TJ, Singh RK, Singh P, Yang LT, Li YR (2017) Intercropping in sugarcane cultivation influenced the soil properties and enhanced the diversity of vital diazotrophic bacteria. Sugar Tech 19:136–147
Tamura K, Nei M, Kumar S (2004) Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci 101:11030–11035
Topp E, Mulbry WM, Zhu H, Nour SM, Cuppels D (2000) Characterization of S-triazine herbicide metabolism by a Nocardioides sp. isolated from agricultural soils. Appl Environ Microbiol 66:3134–3141
Waclawovsky AJ, Sato PM, Lembke CG, Moore PH, Souza GM (2010) Sugarcane for bioenergy production: an assessment of yield and regulation of sucrose content. Plant Biotechnol J 8:263–276
Walkley A, Black IA (1934) An examination of the Degtjareff method for determining soil organic matter, and a proposed modification of the chromic acid titration method. Soil Sci 37:29–38
Wang Z-G, Jin X, Bao X-G, Li XF, Zhao JH, Sun JH, Christie P, Li L (2014) Intercropping enhances productivity and maintains the most soil fertility properties relative to sole cropping. PLoS One 9:e113984. https://doi.org/10.1371/journal.pone.0113984
Wang Z, Solanki MK, Pang F, Singh RK, Yang LT, Li YR, Li HB, Zhu K, Xing YX (2017) Identification and efficiency of a nitrogen-fixing endophytic actinobacterial strain from sugarcane. Sugar Tech 19:492–500
Wei C-Y, Lin L, Luo L-J, Xing YX, Hu CJ, Yang LT, Li YR, An Q (2014) Endophytic nitrogen-fixing Klebsiella variicola strain DX120E promotes sugarcane growth. Biol Fertil Soils 50:657–666
Wilson P, Knight SG (1952) Experiments in bacterial physiology. Burgess, Minneapolis
Witte C-P (2011) Urea metabolism in plants. Plant Sci 180:431–438
Xia H-Y, Wang Z-G, Zhao J-H, Sun JH, Bao XG, Christie P, Zhang FS, Li L (2013) Contribution of interspecific interactions and phosphorus application to sustainable and productive intercropping systems. Field Crop Res 154:53–64
Xing Y-X, Wei C-Y, Mo Y, Yang LT, Huang SL, Li YR (2016) Nitrogen-fixing and plant growth-promoting ability of two endophytic bacterial strains isolated from sugarcane stalks. Sugar Tech 18:373–379
Yang W, Li Z, Wang J, Wu P, Zhang Y (2013) Crop yield, nitrogen acquisition and sugarcane quality as affected by interspecific competition and nitrogen application. Field Crop Res 146:44–50
Yang F, Liao D, Fan Y, Gao R, Wu X, Rahman T, Yong T, Liu W, Liu J, du J, Shu K, Wang X, Yang W (2017) Effect of narrow-row planting patterns on crop competitive and economic advantage in maize–soybean relay strip intercropping system. Plant Prod Sci 20:1–11
Zahir ZA, Zafar-ul-Hye M, Sajjad S, Naveed M (2011) Comparative effectiveness of Pseudomonas and Serratia sp. containing ACC-deaminase for coinoculation with Rhizobium leguminosarum to improve growth, nodulation, and yield of lentil. Biol Fertil Soils 47:457–465
Zehr JP, Mellon MT, Zani S (1998) New nitrogen-fixing microorganisms detected in oligotrophic oceans by amplification of nitrogenase (nifH) genes. Appl Environ Microbiol 64:3444–3450
Zhang F, Li L (2003) Using competitive and facilitative interactions in intercropping systems enhances crop productivity and nutrient-use efficiency. Plant Soil 248:305–312
Zhang J, Hu F, Li H, Gao Q, Song X, Ke X, Wang L (2011) Effects of earthworm activity on humus composition and humic acid characteristics of soil in a maize residue amended rice–wheat rotation agroecosystem. Appl Soil Ecol 51:1–8
Zhang W-F, Dou Z-X, He P, Ju XT, Powlson D, Chadwick D, Norse D, Lu YL, Zhang Y, Wu L, Chen XP, Cassman KG, Zhang FS (2013) New technologies reduce greenhouse gas emissions from nitrogenous fertilizer in China. Proc Natl Acad Sci 110:8375–8380
Funding
The present research work was supported by grants from GXAAS (No. GNKB2014021), National High Technology Research and Development Program (“863” Program) of China (2013AA102604), National Natural Science Foundation of China (31171504, 31101122, 31471449), Guangxi Special Funds for Bagui Scholars’ and Distinguished Experts, Guangxi Natural Science Foundation, and Guangxi Academy of Agriculture Sciences Fund (2011GXNSFF018002, 2012GXNSFDA053011, 2013NXNSFAA019073, and GuiNongKe2014YD01).
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Y.R.L., L.T.Y. designed the study. M.K.S., F.Y.W., and C.N.L. conducted the experiments. M.K.S. and Z.W. did the isolation and characterization of microbes. T.J.L. supported data collection and analysis. M.K.S., C.N.L., R.K.S., P.S., L.T.Y, and Y.R.L. wrote the manuscript.
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Solanki, M.K., Wang, FY., Wang, Z. et al. Rhizospheric and endospheric diazotrophs mediated soil fertility intensification in sugarcane-legume intercropping systems. J Soils Sediments 19, 1911–1927 (2019). https://doi.org/10.1007/s11368-018-2156-3
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DOI: https://doi.org/10.1007/s11368-018-2156-3