花生内生菌的分离及促生长作用初步研究
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摘要
利用表面消毒法从花生中分离出一株具有促生长作用的内生细菌,命名为YX-1,采用形态学特征、生理生化特征结合16SrDNA序列分析对该菌进行鉴定,并采用沙培法研究了花生内生菌对花生幼苗地上部鲜质量、根鲜质量、根长和苗高、根系活力、叶绿素含量及保护酶活力的影响。结果表明,内生细菌YX-1鉴定为苏云金芽孢杆菌(Bacillus thuringiensis)。在10~9cfu/mL菌液浓度下施用1000倍的稀释液,地上部鲜质量、根鲜质量、根长和苗高较对照分别提高44.05%、43.40%、27.76%和25.57%,且均达到显著水平,幼苗叶片中叶绿素含量、根系活力较对照分别提高38.19%、42.08%,叶和根中的SOD、CAT显著高于对照处理,而其中的POD酶活力低于对照处理。结论:花生内生细菌苏云金芽孢杆菌(Bacillus thuringiensis)YX-1能显著促进花生生长,提高花生叶和根中的SOD、CAT酶活力,显示了其在花生生产中良好的应用前景。
YX-1,an endophyte with growth promoting effect was isolated from peanut by using surface sterilization method.The endophyte was identified as Bacillus thuringiensis through morphological characteristics,physiological and biochemical characteristics and 16 SrDNA sequence analysis.The effects of peanut endophyte on fresh weight,root fresh weight,root length and seedling height,root activity,chlorophyll content and protective enzyme activity of peanut seedling were studied by using sand culture method.With the dilution of 1000 times diluted with 109 cfu/mL,the over-ground fresh mass,fresh root mass,root length and seedling height of the plant were increased by 44.05%,43.40%,27.76%and 25.57%than the control treatment respectively,and reached significant level.The chlorophyll content and root activity in the leaves of the seedlings were increased by 38.19%,42.08%,respectively.The SOD and CAT in roots and leaves were significantly higher than the control treatment,while the activities of POD in leaves and roots were lower than the control treatment.The results showed that the endophyte Bacillus thuringiensis YX-1 can promote the growth of peanut plant and increase the enzyme activity of SOD and CAT,and also showed its potential application prospect in peanut production.
YX-1,an endophyte with growth promoting effect was isolated from peanut by using surface sterilization method.The endophyte was identified as Bacillus thuringiensis through morphological characteristics,physiological and biochemical characteristics and 16 SrDNA sequence analysis.The effects of peanut endophyte on fresh weight,root fresh weight,root length and seedling height,root activity,chlorophyll content and protective enzyme activity of peanut seedling were studied by using sand culture method.With the dilution of 1000 times diluted with 109 cfu/mL,the over-ground fresh mass,fresh root mass,root length and seedling height of the plant were increased by 44.05%,43.40%,27.76%and 25.57%than the control treatment respectively,and reached significant level.The chlorophyll content and root activity in the leaves of the seedlings were increased by 38.19%,42.08%,respectively.The SOD and CAT in roots and leaves were significantly higher than the control treatment,while the activities of POD in leaves and roots were lower than the control treatment.The results showed that the endophyte Bacillus thuringiensis YX-1 can promote the growth of peanut plant and increase the enzyme activity of SOD and CAT,and also showed its potential application prospect in peanut production.
引文
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[25]赵颖,于飞,郭明敏,等.碱蓬内生真菌JP3的分离、鉴定及促生作用研究[J].沈阳师范大学学报(自然科学版),2015,33(1):116-120.
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[29]张欣琪.废弃烟叶有机肥对Cd胁迫下甘蓝叶片POD,CAT,SOD的活性的影响[J].中国资源综合利用,2018,136(4):32-34.
[30]韩涛,赵宏伟,贾淡,等.孕穗期低温胁迫对寒地粳稻功能叶片SOD,POD和CAT活性的影响[J].吉林农业科学,2015,40(6):21-25.
[31]Donghee L,Chinbum L.Chilling stress-induced changes of antioxidant enzymes in the leaves of cucumber:in gel enzyme activity assays[J].Plant Science,2000,159(1):75-85.
[2]刘娟,汤丰收,张俊,等.国内花生生产技术现状及发展趋势研究[J].中国农学通报,2017,33(22):13-18.
[3]黄玉茜,韩立思,韩晓日,等.辽宁风沙土区连作年限对花生光合特性和产量的影响[J].沈阳农业大学学报,2011(4):438-442.
[4]Chen M N,Li X,Yang Q L,et al.Soil eukaryotic microorgansm succession as affected by continuous cropping of peanut-athogenic and beneficial fungi were selected[J].Plos One,2012,7(7):e40659.
[5]黄玉茜,韩晓日,杨劲峰,等.花生连作土壤微生物区系变化研究[J].土壤通报,2011(3):552-556.
[6]Huang Y Q,Han X R,Yang J F,et al.Autotoxioity of peanut and identification of phytotoxic substances in rhizosphere soil[J].Allelopathy Journa1,2013,31(2):297-308.
[7]滕应,任文杰,李振高,等.花生连作障碍发生机理研究进展[J].土壤学报,2015(2):259-265.
[8]VStone J K,Bacon C W,White J F Jr.An overview of endophytic microbes:Endophytism defined[M].Microbial Endophytes.New York:Marcel Dekker,Inc,2000:3-29.
[9]Abdul L K,Muhammad W,et al.Fungal endophyte Penicillium janthinellum LKS improves growth of ABA-deficient tomato under salinity[J].World J Microbiol Biotechnol,2013,29:2133-2144.
[10]郜晨,黄淑芬,胡莉,等.尼瓦拉野生稻内生菌多样性和促生作用[J].应用与环境生物学报,2018(1):33-38.
[11]赵龙飞,徐亚军,曹冬建,等.溶磷性大豆根瘤内生菌的筛选、抗性及系统发育和促生[J].生态学报,2015,35(13):4425-4435.
[12]宋子红,丁立孝,马伯军,等.花生内生菌的种群及动态分析[J].植物保护学报,1999,26(4):309-313.
[13]丁立孝,倪新江,王东昌,等.花生内生抗虫工程菌的应用研究[J].吉林农业大学学报,2001,23(3):41-45,57.
[14]杨文博.微生物学实验[M].北京:化学工业出版社,2004.
[15]杜连祥.工业微生物学实验技术[M].天津:天津科学技术出版社,1992.
[16]陈泽斌,代方平,寸林江,等.烟草内生细菌分离方法的优化研究[J].中国烟草学报,2014(1):90-95.
[17]韩旭,杨衍,孙继华,等.砂培黄灯笼辣椒根系分泌物化学成分鉴定[J].热带农业科学,2015,35(12):73-79.
[18]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:62-64.
[19]布坎南R E,吉本斯N E.伯杰细菌鉴定手册[M].北京:科学出版社,1984:732-735.
[20]舒展,张晓素,陈娟,等.叶绿素含量测定的简化[J].植物生理学通讯,2010,46(4):399-402.
[21]李仲芳.植物生理学实验指导[M].西安:西安交通大学出版社,2012.
[22]史树德.植物生理学实验指导[M].北京:中国林业出版社,2011.
[23]Bak K,Gaj R.Effectt of differentiated phosphorus and potassium fertilization on maize grain yield and plant nutritional status at a critical growth stage[J].J Elementtol,2016,21(2):337-348.
[24]武利勤,顾海科,王青,等.石斛内生甲基营养芽胞杆菌的拮抗和促生作用研究[J].生物技术通报,2016,32(8):200-206.
[25]赵颖,于飞,郭明敏,等.碱蓬内生真菌JP3的分离、鉴定及促生作用研究[J].沈阳师范大学学报(自然科学版),2015,33(1):116-120.
[26]叶小梅,常志州,季国军,等.番茄内生菌102菌株的促生作用研究[J].土壤肥料,2005(5):41-42,45.
[27]Yang H,Puri A,Padda K P,et al.Effects of Paenibacillus polymyxainoculation and different soil nitrogen treatments on lodgepole pine seedling growth[J].Can J Forest Res,2016,46(6):816-821.
[28]Bowler C,And M V M,Inze D.Superoxide dismutase and stress tolerance[J].Annu rev plant Physiol plant Mol biol,1992,43(1):83-116.
[29]张欣琪.废弃烟叶有机肥对Cd胁迫下甘蓝叶片POD,CAT,SOD的活性的影响[J].中国资源综合利用,2018,136(4):32-34.
[30]韩涛,赵宏伟,贾淡,等.孕穗期低温胁迫对寒地粳稻功能叶片SOD,POD和CAT活性的影响[J].吉林农业科学,2015,40(6):21-25.
[31]Donghee L,Chinbum L.Chilling stress-induced changes of antioxidant enzymes in the leaves of cucumber:in gel enzyme activity assays[J].Plant Science,2000,159(1):75-85.