一株溶磷菌的分离鉴定及溶磷促生作用
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摘要
为了提高土壤磷素利用率,为微生物磷肥开发提供高效菌种资源,采用平板分离法从荒漠绿洲过渡带白刺根际土壤分离筛选出一株溶磷能力强的真菌P-1407,对其溶磷特性、产酸特性和耐盐碱特性进行研究。结果表明,液体条件下培养10 d,菌株P-1407对Ca_3(PO_4)_2表现出高效的溶解能力,最大溶磷量为557μg·mL~(-1);其溶磷量和pH值变化呈显著负相关,溶磷过程中分泌的有机酸种类主要为葡萄糖酸、草酸及甲酸;在高浓度NaCl和较高的pH值条件下仍保持较高的溶磷能力;结合培养性状、显微形态特征和ITS rDNA序列分析,鉴定菌株P-1407为草酸青霉(Penicillium oxalicum);盆栽试验表明,P-1407菌株对番茄的促生作用较对照显著提高,番茄种子发芽率提高了2.53%,番茄植株的株高、根长、地上部鲜重、地上部干重、地下部鲜重和地下部干重分别增加了109.27%、78.41%、82.95%、121.69%、94.74%和64.71%。因此,P-1407是一株展现良好应用前景的高效溶磷菌种,有望为土壤的改良和生物菌肥的开发提供种质资源。
In order to improve phosphorus of soil and provide P-solubilizing microbial resource for bio-fertilizer production,a fungi P-1407 with strong phosphate-solubilizing ability was isolated from rhizosphere soil of Nitraria tangutorum in Minqin desert-oasis ecotone by using flat plate separation method. The dissolving characteristics, acid production, and resistance to salt-alkaline of strain P-1407 were also studied.The results showed that the strain P-1407 had high-efficiency of phosphate-solubilizing capability for Ca_3(PO_4)_2 during a 10 d shaking incubation. The maximum activity of phosphate-dissolving was reached at 557 μg·mL~(-1). Also, Its P solubilizing capacity was negatively related to soil pH.It was revealed that the strain P-1407 mainly secreted gluconic acid, oxalic acid, and formic acid using HPLC analysis.The strain P-1407 was stable under alkaline and high salt conditions. According to its cultural characteristics, micromorphological characteristics, and ITS rDNA sequence, the strain P-1407 was identified as Penicillium oxalicum.The pot culture experiment results showed that the germination rate of tomato seed treated with P-1407 increased by 2.53% compared with that of the control, and the plant height, root length, fresh weight of aboveground parts, dry weight of aboveground parts, fresh weight of underground parts and dry weight of underground parts were increased by 109.27%, 109.27%, 82.95%, 121.69%, 82.95%, and 121.69%, respectively.Therefore,the strain P-1407 is a promising microbial resource for development of high efficiency biological phosphorus fertilizer of soil in the future.
In order to improve phosphorus of soil and provide P-solubilizing microbial resource for bio-fertilizer production,a fungi P-1407 with strong phosphate-solubilizing ability was isolated from rhizosphere soil of Nitraria tangutorum in Minqin desert-oasis ecotone by using flat plate separation method. The dissolving characteristics, acid production, and resistance to salt-alkaline of strain P-1407 were also studied.The results showed that the strain P-1407 had high-efficiency of phosphate-solubilizing capability for Ca_3(PO_4)_2 during a 10 d shaking incubation. The maximum activity of phosphate-dissolving was reached at 557 μg·mL~(-1). Also, Its P solubilizing capacity was negatively related to soil pH.It was revealed that the strain P-1407 mainly secreted gluconic acid, oxalic acid, and formic acid using HPLC analysis.The strain P-1407 was stable under alkaline and high salt conditions. According to its cultural characteristics, micromorphological characteristics, and ITS rDNA sequence, the strain P-1407 was identified as Penicillium oxalicum.The pot culture experiment results showed that the germination rate of tomato seed treated with P-1407 increased by 2.53% compared with that of the control, and the plant height, root length, fresh weight of aboveground parts, dry weight of aboveground parts, fresh weight of underground parts and dry weight of underground parts were increased by 109.27%, 109.27%, 82.95%, 121.69%, 82.95%, and 121.69%, respectively.Therefore,the strain P-1407 is a promising microbial resource for development of high efficiency biological phosphorus fertilizer of soil in the future.
引文
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[38] 王光华,周克琴,金剑,等.黑土区高效溶磷真菌筛选及其溶解磷矿粉效果的研究[J].中国生态农业学报,2004,12(3):143-145.
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[40] Behbahani M.Investigation of biological behavior and colonization ability of Iranian indigenous phosphate solubilizing bacteria [J].Sci.Hort.2010,12(4):393-399.
[41] 赵小蓉,林启美.微生物解磷的研究进展[J].土壤肥料,2001,5(3):7-11.
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[43] Chen G C,He Z L,Wang Y J.Impact of pH on microbial biomass carbon and microbial biomass phosphorus in red soils [J].Pedosphere,2004,14(1):9-15.
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[2] Hongyang Sun,Yanhong Wu,Jun Zhou,et al.Incubation experiment demonstrates effects of carbon and nitrogen on microbial phosphate-solubilizing function[J].Science China,2017,60(4):436-438.
[3] Fernondez L A,Zalba P,Gomez M A.Phosphate solubilization activity of bacterial strains in soil and their effect on soybean growth under greenhouse conditions [J].Biology and Fertility of Soils,2007,43(6):805-809.
[4] 余旋,朱天辉,刘旭.不同解磷菌剂对美国山核桃根际微生物和酶活性的影响[J].林业科学,2012,48(2):118-123.
[5] Stalstrom VA.Beitrag Zur Kenntrusder einwinsking sterilizer and in garung befindlieher striffe any dil loslieshkeit der phosphorus are destrical cum phosphours [J].Zbt Bakt Abt II,1903,11:724-732.
[6] Rivas R,Peix A,Mateos P F,et al.Biodiversity of populations of phosphate solubilizing rhizobia that nodulates chickpea in different Spanish soils [J].Plant and Soil,2006,287(1-2):23-33.
[7] 胡山,牛世全,龙洋,等.河西走廊盐碱土壤中一株高效溶磷菌的鉴定及条件优化[J].微生物学通报,2017,44(2):358-365.
[8] 江红梅,殷中伟,史发超,等.一株耐盐日本曲霉的筛选及其溶磷促生作用[J].微生物学报,2018,58(5):862-881.
[9] 易艳梅,黄为一.不同生态区土壤溶磷微生物的分布特征及影响因子[J].生态与农村环境学报,2010,26(5):448-453.
[10] 尹瑞龄.我国旱地土壤的溶磷微生物[J].土壤,1988,20( 5) :243-246.
[11] 黄伟,黄欠如,胡锋,等.红壤溶磷菌的筛选及溶磷能力的比较[J].生态与农村环境学报,2006,22(3):37-40.
[12] 林启美,孙焱鑫.四种不同生态系统的土壤溶磷细菌数量及种群分布[J].生态环境学报,2000,9(1):34-37.
[13] Guzel R.Kudoyarova,Lidiya B.Vysotskaya,Tatiana N.Arkhipova,et al.Effect of auxin producing and phosphate solubilizing bacteria on mobility of soil phosphorus,growth rate,and P acquisition by wheat plants [J].Acta Physiologiae Plantarum,2017,39(11):253-260.
[14] 陶涛,叶明,刘冬,等.无机解磷细菌的筛选、鉴定及其溶磷能力研究[J].合肥工业大学学报(自然科学版),2011,34( 2) :304-308.
[15] Jesús A.Ibarra-Galeana,Claudia Castro-Martínez,Rosario A.Fierro-Coronado,et al.Characterization of phosphate-solubilizing bacteria exhibiting the potential for growth promotion and phosphorus nutrition improvement in maize(Zea mays L.)in calcareous soils of Sinaloa,Mexico [J].Annals of Microbiology,2017,67:801-811.
[16] 马文文,姚拓,蒲小鹏,等.东祁连山7 种禾草根际溶磷菌筛选及其溶磷特性[J].草业科学,2015,32(4) :515-523.
[17] 江红梅,殷中伟,史发超,等.一株耐盐溶磷真菌的筛选、鉴定及其生物肥料的应用效果[J].植物营养与肥料学报 2018,24(3):728-742.
[18] Chatli A,Beri V,Sidhu B.Isolation and characterisation of phosphate solubilising microorganisms from the cold desert habitat of Salix alba Linn in trans Himalayan region of Himachal Pradesh [J].Indian Journal of Microbiology,2008,48(2):267-273.
[19] Bhattacharya S,Barman S,Ghosh R.Phosphate solubilizing activity of Emericella nidulans V1 isolated from vermicompost [J].Indian Journal of Experimental Biology,2013,51(10):840-848.
[20] Naveenkumar K,Thippeswamy B,Banakar S.Lignolytic and phosphate solubilizing efficiency of fungal species isolated from Arecanut husk waste [J].Journal of Research Biology,2012,2(2):143-151.
[21] Rinu K,Malviya M,Sati P,et al.Response of cold-tolerant Aspergillus spp.to solubilization of Fe and Al phosphate in presence of different nutritional sources [J].Isrn Soil Science,2013:135-143.
[22] 李海云,朱世全,孔维宝,等.猪粪堆肥中一株溶磷菌的筛选鉴定及溶磷能力初步测定[J].环境科学学报,2015,35(05):1464-1470.
[23] 任嘉红,刘辉,吴晓蕙,等.南方红豆杉根际溶无机磷细菌的筛选、鉴定及其促生效果[J].微生物学报,2012,52(3):295-303.
[24] 乔志伟,洪坚平,谢英荷,等.一株石灰性土壤强溶磷真菌的分离鉴定及溶磷特性[J].应用与环境生物学报,2013,19(5):873-877.
[25] 乔欢,吴小芹,王早.一株嗜松青霉JP-NJ4的解磷特性[J].微生物学通报,2014,41(9):1741-1748.
[26] 吴楠,王飞,田治国,等.NaCl胁迫对变异紫叶三叶草种子萌发及保护酶活性的影响[J].西北农林科技大学学报(自然科学版),2013,41(2):117-122.
[27] 刘建新,王瑞娟,王鑫,等.Na(NO3)3 对盐胁迫下黑麦草幼苗生长及抗逆生理特性的影响[J].中国生态农业学报,2011,19(2):353-357.
[28] 陈毓荃.生物化学实验方法和技术[M].北京:科学出版社,2002.
[29] 李合生,孙群,赵世杰.植物生理生化试验原理与技术[M].北京:高等教育出版社,2000.
[30] 贺梦醒 ,高毅 ,胡正雪,等.解磷菌株B25的筛选、鉴定及其解磷能力[J].应用生态学报,2012,23(1):235-239.
[31] 赵小蓉,林启美,李保国.溶磷菌对4种难溶性磷酸盐溶解能力的初步研究[J].微生物学报,2002,42(2):236-241.
[32] 刘文干,何园球,张坤,等.一株红壤溶磷菌的分离、鉴定及溶磷特性[J].微生物学报,2012,52(3):326-333.
[33] 赵小蓉.微生物的解磷机理及其影响因素 [D].北京:中国农业大学,2001.
[34] 范丙全,金继运,葛诚.溶磷草酸青霉筛选及其溶磷效果的初步研究[J].中国农业科学,2002,35(5):525-530.
[35] 王莉晶,高晓荣,吕军,等.解磷真菌 C2的分离鉴定及其在土壤中实际解磷效果的研究[J].土壤通报,2009,40(4):771-775.
[36] 万璐,康丽华,廖宝文,等.红树林根际解磷菌分离、培养及解磷能力的研究[J].林业科学研究 2004,17(1):89-94.
[37] 张丽珍,樊晶晶,牛伟,等.盐碱地柠条根围土中黑曲霉的分离鉴定及解磷能力测定[J].生态学报,2011,31(24):7571-7578.
[38] 王光华,周克琴,金剑,等.黑土区高效溶磷真菌筛选及其溶解磷矿粉效果的研究[J].中国生态农业学报,2004,12(3):143-145.
[39] Villegas J,Fortin JA.Phosphorus solubilization and pH changes as a result of the interactions between soil bacteria and arbuscular mycorrhizal fungi on a medium containing NO-3 as nitrogen source [J].Botany,2002,80(5):571-576.
[40] Behbahani M.Investigation of biological behavior and colonization ability of Iranian indigenous phosphate solubilizing bacteria [J].Sci.Hort.2010,12(4):393-399.
[41] 赵小蓉,林启美.微生物解磷的研究进展[J].土壤肥料,2001,5(3):7-11.
[42] Patel DK,Archana G,Kumar GN.Variation in the nature of organic acid secretion and mineral phosphate solubilization by Citrobacter sp.DHRSS in the presence of different sugars [J].Current Microbiology,2008,56(2):168-174.
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