岷山红三叶根际优良促生菌对其宿主生长和品质的影响

doi:10.11686/cyxb20140527

摘要/Abstract

摘要：

从岷山红三叶根际分离出高效溶磷菌,并对其分泌生长激素、拮抗病原菌特性进行测定,筛选出优良促生菌6株,根瘤菌1株,按照一定比例混合制成复合菌肥,进行盆栽试验,研究复合菌肥对岷山红三叶产量、品质及异黄酮含量的影响。结果表明:利用筛选的优良促生菌和根瘤菌研制的复合微生物菌肥符合《微生物肥料》标准(NY227-94)。75%化肥+PGPR菌肥处理使岷山红三叶干草总产量较CK增加4.76%,同时,该处理可提高红三叶粗蛋白、粗灰分、钙、磷含量,降低中、酸性洗涤纤维含量;50%化肥+PGPR菌肥处理对岷山红三叶的总干草产量、营养品质以及4种异黄酮的含量均没有显著影响。

Abstract:

Interest in plant growth promoting rhizobacteria (PGPR) has increased due to their potential contribution to sustainable agricultural systems by reducing environmental impact, the consumption of non-renewable resources and input costs. This paper investigates the inoculant effect of PGPR on the hay yield and nutrient quality of Trifolium pratense cv. Minshan. Six PGPR and one rhizobia strains were identified by screening for phosphate solubilization, 3-indoleacetic acid (IAA) secretion and antagonism to plant pathogens. A PGPR inoculant was produced by mixing these strains and pot tests have been conducted to investigate its effects on the growth and quality of T. pratense cv. Minshan when combined with different amounts of chemical fertilizer. The results showed that the nitrogenase activity of the rhizobia strain was 488.2 C₂H₄ nmol/(mL·h). The phosphate solubilization and IAA secreting ability of the five PGPR strains was distributed across the ranges 50.34-229.5 mg/L and 0.86-12.01 μg/mL respectively. The PGPR compound bio-fertilizer demonstrated strong antagonism to the pathogens Fusarium oxysporium f. sp. cucumerinum, Rhizoctonia solani and Fusarium oxysporum f. niveum. The composite microbial inoculant complies with the quality standard for microbiological fertilizer in China (NY227-94). Applying the inoculant in combination with 75% of the control’s chemical fertilizers increased total hay yield by 4.76%. Nutrient quality also improved. The amount of crude protein, crude ash, calcium and phosphorus increased while the amount of neutral detergent fiber and acid detergent fiber declined. Applying the inoculant in combination with 50% of the control’s chemical fertilizers did not increase total hay yield, quality, or the isoflavone content of clover. The experiment has thus shown how the PGPR compound bio-fertilizer can be combined with chemical fertilizers to increase not only the yield and quality of T. pratense cv. Minshan but also the sustainability of agricultural systems.

中图分类号:

S541⁺.2

荣良燕,姚拓,马文彬,李德明,李儒仁,张洁,陆飒. 岷山红三叶根际优良促生菌对其宿主生长和品质的影响[J]. 草业学报, 2014, 23(5): 231-240.

RONG Liang-yan,YAO Tuo,MA Wen-bin,LI De-ming,LI Ru-ren,ZHANG Jie,LU Sa. The inoculant potential of plant growth promoting rhizobacteria strains to improve the yield and quality of Trifolium pratense cv. Minshan[J]. Acta Prataculturae Sinica, 2014, 23(5): 231-240.

参考文献

Reference:

[1] Han W X, Yao T, X L Q, et al. PGPR bio-fertilizers producing and its effect on Arena sativa growth and quality development[J]. Acta Prataculturae Sinica, 2008, 17(2): 75-84.

[2] Meng X J, Yu L P, Cheng W D, et al. Influence of rhizobia inoculation and nitrogen fertilizer application on isoflavone content of minshan red-clover[J]. Prataculture & Animal Husbandry, 2010, 5: 97-100.

[3] Liu J L, Fang F, Shi S H, et al. Isolation and characterization of PGPR from the rhizosphere of the Avena sativa in saline-alkali soil[J]. Acta Prataculturae Sinica, 2013, 22(2): 132-139.[4] Aslanta R, akmak i R, nahin F. Effect of plant growth promoting rhizobacteria on young apple tree growth and fruit yield under orchard conditions[J]. Scientia Horticulturae, 2007, 111(4): 371-377.

[5] Yanni Y G, Rizk R Y, El-Fattah F K A, et al. The beneficial plant growth promoting association of Rhizobium leguminosarum bv. trifolii with rice roots[J]. Functional Plant Biology, 2001, 28(9): 845-870.

[6] Turan M, Gulluce M, Karadayi M, et al. Role of soil enzymes produced by PGPR strains in wheat growth and nutrient uptake parameters in the filed conditions[J]. Current Opinion in Biotechnology, 2011, 22(supplement 1): 133.

[7] Rong L Y, Yao T, Huang G B, et al. Screening of plant growth promoting rhizobacteria strains and effects of inoculant on growth of maize by replacing part of chemical fertilizers[J]. Agricultural Research In The Arid Areas, 2013, 2: 59-65.

[8] Myresiotis C K, Vryzas Z, Papadopoulou-Mourkidou E. Enhanced root uptake of acibenzolar Smethyl (ASM) by tomato plants inoculated with selected Bacillus plant growth promoting rhizobacteria (PGPR)[J]. Applied Soil Ecology, 2014, 77: 26-33.

[9] Udayashankar A, Chandra Nayaka S, Reddy M, et al. Plant growth promoting rhizobacteria mediate induced systemic resistance in rice against bacterial leaf blight caused by Xanthomonas oryzae pv. Oryzae[J]. Biological Control, 2011, 59(2): 114-122.

[10] Zhang Y. Screening plant growth promoting rhizobacteria resources and thrie promotion mechanisms from rhizosphere of four forages in Ali Alpine grassland of Tibet[D].Lanzhou: Gansu Agricultural University, 2013.

[11] Zhang Y, Zhu Y, Yao T, et al. Interactions of four PGPRs isolated from pasture rhizosphere[J]. Acta Prataculturae Sinica, 2013, 22(1): 29-37.

[12] Rong L Y, Yao T, Zhao G Q, et al. Screening of siderophore-producing PGPR bacteria and their antagonism against the pathogens[J]. Plant Protection, 2011, (1): 59-64.

[13] Han H W, Sun L N, Yao T, et al. Effects of effective microbial inoculants on alfalfa growth character究[J]. cAta Agrectir Sinica, 2013, (2): 353-359.

[14] The Ministry of Agriculture microbial fertilizer Quality Supervision and Testing Center. Agricultural microbial agents GB 20287-2006[S]. Beijing: China Standard Press, 2006.

[15] Gansu Agricultural University. Grassland ecological chemistry experiment guide[M]. Beijing: Agricultural Press, 1987.

[16] Chen H Q, Jin Z Y, Tao G J, et al. Determination of isoflavones in different parts of trifolium pratense by high performance liquid chromatography[J]. Food Science, 2004, (6): 150-153.

[17] The Ministry of Agriculture microbial fertilizer quality wupervision and testing center．Microbial fertilizer N / Y 227-1994[S]. Beijing: China Standard Press, 1994.

[18] Ma X, Wang L L, Li W J, et al. Effects of different nitrogen levels on nitrogen fixation and seed production of alfalfa inoculated with rhizobia[J]. Acta Prataculturae Sinica, 2013, 22(1): 95-102.

[19] Li Z D, Chen S R, Yang C D, et al. Screening, identification and solubilizing properties of the endophytic phosphate-solubilizing bacteria isolated from Ana phalis lactea[J]. Acta Prataculturae Sinica, 2013, 22(6): 150-158.

[20] Karlidag H, Esitken A, Turan M, et al. Effects of root inoculation of plant growth promoting rhizobacteria (PGPR) on yield, growth and nutrient element contents of leaves of apple[J]. Scientia Horticulturae, 2007, 114(1): 16-20.

[21] Pe akovi M, Karaklaji Staji, Milenkovi S, et al. Biofertilizer affecting yield related characteristics of strawberry (Fragaria×ananassa Duch.) and soil micro-organisms[J]. Scientia Horticulturae, 2013, 150: 238-243.

[22] Zhang K, Yao T, Zhang D G, et al. Effect of associative nitrogen-fixing biofertilizer produced with different garriers on growth and nitrogen fixing ability of Hordeum vulgare var.nudum Hook.f.[J]. Acta Agredtir Sinica, 2010, (3): 426-430.

[23] Govindarajan M, Balandreau J, Muthukumarasamy R, et al. Improved yield of micropropagated sugarcane following inoculation by endophytic Burkholderia vietnamiensis[J]. Plant and Soil, 2006, 280(2): 239-252.

[24] Yao T, Pu X P, Zhang D G, et al. Associative nitrogen-fixing bacteria in the rhizosphere of Avena sativa in an alpine region Ⅲ Effect on Avena sativa growth and quantification of nitrogen fixed[J]. Acta Prataculturae Sinica, 2004, 13(5): 101-105.

[25] Gao X X, Man M Y, Chen X R, et al. Identification and determination of biological characteristics of Kobresia capillifolia endophytic bacteria X4 in the East Qilian Mountain Alpine grasslands[J]. Acta Prataculturae Sinica, 2013, 22(4): 137-146.

[26] Shahab S, Ahmed N, Khan N S. Indole acetic acid production and enhanced plant growth promotion by indigenous PSBs[J]. African Journal of Agricultural Research, 2009, 4(11): 1312-1316.

[27] Wahyudi A T, Astuti R P, Widyawati A, et al. Characterization of Bacillus sp. strains isolated from rhizosphere of soybean plants for their use as potential plant growth for promoting rhizobacteria[J]. Journal of Microbiology and Antimicrobials, 2011, 3(2): 34-40.

[28] Recep K, Fikrettin S, Erkol D, et al. Biological control of the potato dry rot caused by Fusarium species using PGPR strains[J]. Biological Control, 2009, 50(2): 194-198.

[29] Abbasi M, Sharif S, Kazmi M, et al. Isolation of plant growth promoting rhizobacteria from wheat rhizosphere and their effect on improving growth, yield and nutrient uptake of plants[J]. Plant Biosystems, 2011, 145(1): 159-168.

[30] Domenech J, Reddy M, Kloepper J, et al. Combined application of the biological product LS213 with Bacillus, Pseudomonas or Chryseobacterium for growth promotion and biological control of soil-borne diseases in pepper and tomato[J]. Biocontrol, 2006, 51(2): 245-258.

[31] Sandhya V, Grover M, Reddy G, et al. Alleviation of drought stress effects in sunflower seedlings by the exopolysaccharides producing Pseudomonas putida strain GAP-P45[J]. Biology and Fertility of Soils, 2009, 46(1): 17-26.

[32] Vijayan R, Palaniappan P, Tongmin S A, et al. Rhizobitoxine enhances nodulation by inhibiting ethylene synthesis of Bradyrhizobium elkanii from Lespedeza species: validation by homology modeling and molecular docking study[J]. World Journal of Pharmaceutical Sciences, 2013, 2: 4079-4094.

[33] Beneduzi A, Moreira F, Costa P B, et al. Diversity and plant growth promoting evaluation abilities of bacteria isolated from sugarcane cultivated in the south of Brazil[J]. Applied Soil Ecology, 2013, 63: 94-104.

[34] Dawwam G, Elbeltagy A, Emara H, et al. Beneficial effect of plant growth promoting bacteria isolated from the roots of potato plant[J]. Annals of Agricultural Sciences, 2013, 58(2): 195-201.

[35] Shen J, Li R, Zhang F, et al. Crop yields, soil fertility and phosphorus fractions in response to long-term fertilization under the rice monoculture system on a calcareous soil[J]. Field Crops Research, 2004, 86(2): 225-238.

[36] Nadeem S M, Ahmad M, Zahir Z A, et al. The role of mycorrhizae and plant growth promoting rhizobacteria (PGPR) in improving crop productivity under stressful environments[J]. Biotechnology Advances, 2014, 32(2): 429-448.

[37] Yue H, Mo W, Li C, et al. The salt stress relief and growth promotion effect of RS 5 on cotton[J]. Plant and Soil, 2007, 297(12): 139-145.

[38] Zhang Y, Yao T, Zhu Y. Effects of compound inoculum on growth characteristics and quality of Trifolium[J]. Plant Nutrition and Fertilizer Science, 2012, 5: 1277-1285.

[39] Orhan E, Esitken A, Ercisli S, et al. Effects of plant growth promoting rhizobacteria (PGPR) on yield, growth and nutrient contents in organically growing raspberry[J]. Scientia Horticulturae, 2006, 111(1): 38-43.

[40] De-Bashan L E, Hernandez J P, Bashan Y. The potential contribution of plant growth-promoting bacteria to reduce environmental degradation-A comprehensive evaluation[J]. Applied Soil Ecology, 2012, 61: 171-189.

[41] Karag z K, Ate F, Karag z H, et al. Characterization of plant growth promoting traits of bacteria isolated from the rhizosphere of grapevine grown in alkaline and acidic soils[J]. European Journal of Soil Biology, 2012, 50: 144-150.

参考文献：

[1] 韩文星, 姚拓, 席琳乔, 等. PGPR菌肥制作及其对燕麦生长和品质影响的研究[J]. 草业学报, 2008, 17(2): 75-84.

[2] 孟祥君, 俞联平, 程文定, 等. 接种根瘤菌与施肥对岷山红三叶异黄酮含量的影响[J]. 草业科学, 2010, 5: 97-100.

[3] 刘佳莉, 方芳, 史煦涵, 等. 2株盐碱地燕麦根际促生菌的筛选及其促生作用研究[J]. 草业学报, 2013, 22(2): 132-139.

[4] Aslanta塂 R, akmaki R, 塁ahin F. Effect of plant growth promoting rhizobacteria on young apple tree growth and fruit yield under orchard conditions[J]. Scientia Horticulturae, 2007, 111(4): 371-377.

[5] Yanni Y G, Rizk R Y, El-Fattah F K A, et al. The beneficial plant growth-promoting association of Rhizobium leguminosarum bv. trifolii with rice roots[J]. Functional Plant Biology, 2001, 28(9): 845-870.

[6] Turan M, Gulluce M, Karadayi M, et al. Role of soil enzymes produced by PGPR strains in wheat growth and nutrient uptake parameters in the filed conditions[J]. Current Opinion in Biotechnology, 2011, 22(supplement 1): 133.

[7] 荣良燕, 姚拓, 黄高宝, 等. 植物根际优良促生菌(PGPR)筛选及其接种剂部分替代化肥对玉米生长影响研究[J]. 干旱地区农业研究, 2013, 2: 59-65.

[8] Myresiotis C K, Vryzas Z, Papadopoulou-Mourkidou E. Enhanced root uptake of acibenzolar-S-methyl (ASM) by tomato plants inoculated with selected Bacillus plant growth promoting rhizobacteria (PGPR)[J]. Applied Soil Ecology, 2014, 77: 26-33.

[9] Udayashankar A, Chandra Nayaka S, Reddy M, et al. Plant growth-promoting rhizobacteria mediate induced systemic resistance in rice against bacterial leaf blight caused by Xanthomonas oryzae pv. Oryzae[J]. Biological Control, 2011, 59(2): 114-122.

[10] 张英. 西藏阿里高寒草原四种牧草根际促生菌资源筛选及促生机理研究[D].兰州：甘肃农业大学, 2013.

[11] 张英, 朱颖, 姚拓, 等. 分离自牧草根际四株促生菌株(PGPR)互作效应研究[J]. 草业学报, 2013, 22(1): 29-37.

[12] 荣良燕, 姚拓, 赵桂琴, 等. 产铁载体PGPR菌筛选及其对病原菌的拮抗作用[J]. 植物保护, 2011, （1）: 59-64.

[13] 韩华雯, 孙丽娜, 姚拓, 等. 苜蓿根际有益菌接种剂对苜蓿生长特性影响的研究[J]. 草地学报, 2013, (2): 353-359.

[14] 农业部微生物肥料质量监督检验测试中心. 农用微生物菌剂GB 20287-2006[S]. 北京：中国标准出版社， 2006.

[15] 甘肃农业大学. 草原生态化学实验指导[M]. 北京: 农业出版社, 1987.

[16] 陈寒青, 金征宇, 陶冠军, 等. 红车轴草不同部位中异黄酮含量的测定[J]. 食品科学, 2004, （6）: 150-153.

[17] 农业部微生物肥料质量监督检验测试中心．微生物肥料N/Y 227-1994[S]. 北京：中国标准出版社， 1994.

[18] 马霞, 王丽丽, 李卫军, 等. 不同施氮水平下接种根瘤菌对苜蓿固氮效能及种子生产的影响[J]. 草业学报, 2013, 22(1): 95-102.

[19] 李振东, 陈秀蓉, 杨成德, 等. 乳白香青内生解磷菌的筛选鉴定及解磷特性研究[J]. 草业学报, 2013, 22(6): 150-158.

[20] Karlidag H, Esitken A, Turan M, et al. Effects of root inoculation of plant growth promoting rhizobacteria (PGPR) on yield, growth and nutrient element contents of leaves of apple[J]. Scientia Horticulturae, 2007, 114(1): 16-20.

[21] Peakovi M, Karaklaji-Staji , Milenkovi S, et al. Biofertilizer affecting yield related characteristics of strawberry (Fragaria×ananassa Duch.) and soil micro-organisms[J]. Scientia Horticulturae, 2013, 150: 238-243.

[22] 张堃, 姚拓, 张德罡, 等. 不同剂型联合固氮菌肥对青稞促生效应和固氮能力研究[J]. 草地学报, 2010, （3）: 426-430.

[23] Govindarajan M, Balandreau J, Muthukumarasamy R, et al. Improved yield of micropropagated sugarcane following inoculation by endophytic Burkholderia vietnamiensis[J]. Plant and Soil, 2006, 280(2): 239-252.

[24] 姚拓, 蒲小鹏, 张德罡, 等. 高寒地区燕麦根际联合固氮菌研究Ⅲ-固氮菌对燕麦生长的影响及其固氮量测定[J]. 草业学报, 2004, 13(5): 101-105.

[25] 高晓星, 满百膺, 陈秀蓉, 等. 东祁连山线叶嵩草内生细菌X4的产吲哚乙酸、解磷、抗菌和耐盐特性研究及分子鉴定[J]. 草业学报, 2013, 22(4): 137-146.

[26] Shahab S, Ahmed N, Khan N S. Indole acetic acid production and enhanced plant growth promotion by indigenous PSBs[J]. African Journal of Agricultural Research, 2009, 4(11): 1312-1316.

[27] Wahyudi A T, Astuti R P, Widyawati A, et al. Characterization of Bacillus sp. strains isolated from rhizosphere of soybean plants for their use as potential plant growth for promoting rhizobacteria[J]. Journal of Microbiology and Antimicrobials, 2011, 3(2): 34-40.

[28] Recep K, Fikrettin S, Erkol D, et al. Biological control of the potato dry rot caused by Fusarium species using PGPR strains[J]. Biological Control, 2009, 50(2): 194-198.

[29] Abbasi M, Sharif S, Kazmi M, et al. Isolation of plant growth promoting rhizobacteria from wheat rhizosphere and their effect on improving growth, yield and nutrient uptake of plants[J]. Plant Biosystems, 2011, 145(1): 159-168.

[30] Domenech J, Reddy M, Kloepper J, et al. Combined application of the biological product LS213 with Bacillus, Pseudomonas or Chryseobacterium for growth promotion and biological control of soil-borne diseases in pepper and tomato[J]. Biocontrol, 2006, 51(2): 245-258.

[31] Sandhya V, Grover M, Reddy G, et al. Alleviation of drought stress effects in sunflower seedlings by the exopolysaccharides producing Pseudomonas putida strain GAP-P45[J]. Biology and Fertility of Soils, 2009, 46(1): 17-26.

[32] Vijayan R, Palaniappan P, Tongmin S A, et al. Rhizobitoxine enhances nodulation by inhibiting ethylene synthesis of Bradyrhizobium elkanii from Lespedeza species: validation by homology modeling and molecular docking study[J]. World Journal of Pharmaceutical Sciences, 2013, 2: 4079-4094.

[33] Beneduzi A, Moreira F, Costa P B, et al. Diversity and plant growth promoting evaluation abilities of bacteria isolated from sugarcane cultivated in the south of Brazil[J]. Applied Soil Ecology, 2013, 63: 94-104.

[34] Dawwam G, Elbeltagy A, Emara H, et al. Beneficial effect of plant growth promoting bacteria isolated from the roots of potato plant[J]. Annals of Agricultural Sciences, 2013, 58(2): 195-201.

[35] Shen J, Li R, Zhang F, et al. Crop yields, soil fertility and phosphorus fractions in response to long-term fertilization under the rice monoculture system on a calcareous soil[J]. Field Crops Research, 2004, 86(2): 225-238.

[36] Nadeem S M, Ahmad M, Zahir Z A, et al. The role of mycorrhizae and plant growth promoting rhizobacteria (PGPR) in improving crop productivity under stressful environments[J]. Biotechnology Advances, 2014, 32(2): 429-448.

[37] Yue H, Mo W, Li C, et al. The salt stress relief and growth promotion effect of RS-5 on cotton[J]. Plant and Soil, 2007, 297(1-2): 139-145.

[38] 张英, 姚拓, 朱颖. 复合接种剂对三叶草生长特性和品质的影响[J]. 植物营养与肥料学报, 2012, 5: 1277-1285.

[39] Orhan E, Esitken A, Ercisli S, et al. Effects of plant growth promoting rhizobacteria (PGPR) on yield, growth and nutrient contents in organically growing raspberry[J]. Scientia Horticulturae, 2006, 111(1): 38-43.

[40] De-Bashan L E, Hernandez J P, Bashan Y. The potential contribution of plant growth-promoting bacteria to reduce environmental degradation-A comprehensive evaluation[J]. Applied Soil Ecology, 2012, 61: 171-189.

[41] Karagz K, Ate塂 F, Karagz H, et al. Characterization of plant growth-promoting traits of bacteria isolated from the rhizosphere of grapevine grown in alkaline and acidic soils[J]. European Journal of Soil Biology, 2012, 50: 144-150.