玫瑰黄链霉菌NKZ-259发酵培养基的优化
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摘要
玫瑰黄链霉菌NKZ-259是一株生防菌株,其次级代谢能产生植物生长调节类物质吲哚乙酸(IAA)。为了进一步提高菌株代谢产生IAA的含量,本试验对该菌株的发酵培养基进行了优化。利用单因子试验确定发酵培养基中最适的6种营养成分为葡萄糖、可溶性淀粉、蛋白胨、硝酸钾、磷酸氢二钾和L-色氨酸;通过Plackett-Burman设计筛选出影响菌株发酵产生IAA的主要因素为L-色氨酸、葡萄糖和磷酸氢二钾;采用中心组合试验(CCD)及响应面法分析各因素的交互作用。最终确定菌株代谢产生IAA的最优发酵培养基为:L-色氨酸2.24 g/L,葡萄糖20.7 g/L,磷酸氢二钾0.5 g/L,可溶性淀粉10 g/L,蛋白胨3 g/L,硝酸钾4.5g/L;使用优化后的发酵培养基菌株代谢产生IAA的含量为45.377 4μg/mL,比原始发酵培养基IAA的含量提高了3倍。
Streptomyces roseoflavus NKZ-259 is a biocontrol strain and plant growth regulator indole-3-acetic acid(IAA)is produced in the process of secondary metabolism. The fermentation medium was optimized in this study in order to increase the content of IAA produced by a strain. The 6 optimal nutrients of fermentation medium were determined by single factor experiment,and they were glucose,soluble starch,peptone,potassium nitrate,dipotassium hydrogen phosphate,and L-tryptophan,respectively. Plackett-Burman design method was applied to screen major factors affecting the fermentation production of IAA,which was L-tryptophan,glucose,and dipotassium hydrogen phosphate. Central combination design(CCD)and response surface methodology were used to analyze the interactions of all factors. Finally,the optimized fermentation medium was composed as follow :L-tryptophan 2.24 g/L,glucose 20.7 g/L,dipotassium hydrogen phosphate 0.5 g/L,soluble starch 10 g/L,peptone 3 g/L,potassium nitrate 4.5 g/L. The concentration of IAA reached 45.377 4 μg/mL after the medium optimized,which was increased 3 times compared to the original fermentation medium.
Streptomyces roseoflavus NKZ-259 is a biocontrol strain and plant growth regulator indole-3-acetic acid(IAA)is produced in the process of secondary metabolism. The fermentation medium was optimized in this study in order to increase the content of IAA produced by a strain. The 6 optimal nutrients of fermentation medium were determined by single factor experiment,and they were glucose,soluble starch,peptone,potassium nitrate,dipotassium hydrogen phosphate,and L-tryptophan,respectively. Plackett-Burman design method was applied to screen major factors affecting the fermentation production of IAA,which was L-tryptophan,glucose,and dipotassium hydrogen phosphate. Central combination design(CCD)and response surface methodology were used to analyze the interactions of all factors. Finally,the optimized fermentation medium was composed as follow :L-tryptophan 2.24 g/L,glucose 20.7 g/L,dipotassium hydrogen phosphate 0.5 g/L,soluble starch 10 g/L,peptone 3 g/L,potassium nitrate 4.5 g/L. The concentration of IAA reached 45.377 4 μg/mL after the medium optimized,which was increased 3 times compared to the original fermentation medium.
引文
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[13]王非,王金侠,李强,等. GA_3和IAA处理对4种铁线莲种子萌发的影响[J].草业科学, 2014, 31(4):672-676.
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[15]郭敬华.玫瑰黄链霉菌Men-myco-93-63发酵液防治黄瓜白粉病的效果及作用机理初探[D].保定:河北农业大学, 2007.
[16]葛蓓孛,王家旺,刘彦彦,等.武夷菌素高产基因工程菌发酵培养基的优化[J].中国生物防治学报, 2016, 32(5):642-649.
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[20]王占一,金美花,王玉海,等. Plackett-Burman和Central Composite Design试验设计法优化石榴皮中熊果酸的SFECO_2萃取工艺[J].中药材, 2015, 38(3):610-614.
[21]宋浩,纪兆林,陈夕军,等.地衣芽孢杆菌W10菌株发酵培养基优化[J].扬州大学学报:农业与生命科学版, 2015, 36(1):87-91.
[22]钟小廷.产细菌素蜡样芽孢杆菌的筛选、鉴定及培养基优化[D].成都:西华大学, 2014.
[23]刘志祥,曾超珍.响应面法在发酵培养基优化中的应用[J].北方园艺, 2009(2):127-129.
[24]Wang L, Zhang M, Li Y, et al. Application of response surface methodology to optimize the production of antimicrobial metabolites by Micromonospora Y15[J]. Biotechnology&Biotechnological Equipment, 2017:1-10.
[25]李莉,张赛,何强,等.响应面法在试验设计与优化中的应用[J].实验室研究与探索, 2015, 34(8):41-45.
[26]Li C, Bai J, Cai Z, et al. Optimization of a cultural medium for bacteriocin production by Lactococcus lactis using response surface methodology[J]. Journal of Biotechnology, 2002, 93(1):27-34.
[27]韩兴,刘亚南,甄丹妹,等.玫瑰黄链霉菌Men-myco-93-63产抗生素发酵培养基的优化[J].江苏农业科学, 2017, 45(10):100-103.
[28]Passari AK, Mishra VK, Gupta VK, et al. In vitro and in vivo plant growth promoting activities and DNA fingerprinting of antagonistic endophytic actinomycetes associates with medicinal plants[J].PLoS One, 2015, 10(9):1-18.
[29]刘伟.利用地衣芽孢杆菌发酵生产细胞分裂素[D].武汉:华中农业大学, 2005.
[2]徐长宝,任晓亮,朱桂玲.打破柿树种子休眠和促进发芽的方法[J].林业科技开发, 2009, 23(2):109-112.
[3]宋文,肖景义,赵莹,等.植物激素萘乙酸在果树生产中的应用[J].特种经济动植物, 2012, 15(12):48-49.
[4]Mao X, Tang L, Tan T, et al. Determination of plant growth regulators in pears by microwave-assisted extraction and liquid chromatography with electrospray ionization mass spectrometry.[J]. Journal of Separation Science, 2014, 37(11):1352-8.
[5]史晓梅,金芬,黄玉婷,等.水果中常用植物生长调节剂的研究进展[J].食品工业科技, 2012, 33(4):417-422+426.
[6]张兴,马志卿,冯俊涛,等.植物源农药研究进展[J].中国生物防治学报, 2015, 31(5):685-698.
[7]Goudjal Y, Zamoum M, Sabaou N, et al. Potential of endophytic Streptomyces spp. for biocontrol of Fusarium root rot disease and growth promotion of tomato seedlings[J]. Biocontrol Science&Technology, 2016, 26(12):1691-1705.
[8]Narayana K J, Peddikotla P, Krishna P S J, et al. Indole-3-acetic acid production by Streptomyces albidoflavus[J]. Journal of Biological Research-Thessaloniki, 2009, 11:49-55.
[9]Abd-Alla MH, El-Sayed ESA, Rasmey AHM. Indole-3-acetic acid(IAA)production by Streptomyces atrovirens isolated from rhizospheric soil in Egypt[J]. Journal of Biology&Earth Sciences, 2013(3):182-193.
[10]Shi LM, Thin TN, Ge BB, et al. Antifungal and plant growthpromoting activities of Streptomyces roseoflavus strain NKZ-259[J]. Biological Control, 2018, 125:57-64.
[11]Lin L, Xu X. Indole-3-acetic acid production by endophytic Streptomyces sp. En-1 isolated from medicinal plants[J]. Current Microbiology, 2013, 67(2):209-217.
[12]尤桂春,武竞超,林文忠. IAA不同浓度处理对柠檬枝条扦插生根的影响[J].东南园艺, 2016, 4(1):8-9.
[13]王非,王金侠,李强,等. GA_3和IAA处理对4种铁线莲种子萌发的影响[J].草业科学, 2014, 31(4):672-676.
[14]Aminullah, Zada K, Jamal M, et al. Effect of indole acetic acid(IAA)on yield and yield contributing parameters of soybean[J]. Pakistan Journal of Biological Sciences, 2000, 3(5):856-857.
[15]郭敬华.玫瑰黄链霉菌Men-myco-93-63发酵液防治黄瓜白粉病的效果及作用机理初探[D].保定:河北农业大学, 2007.
[16]葛蓓孛,王家旺,刘彦彦,等.武夷菌素高产基因工程菌发酵培养基的优化[J].中国生物防治学报, 2016, 32(5):642-649.
[17]张东艳,刘晔,吴越,等.花生根际产IAA菌的筛选鉴定及其效应研究[J].中国油料作物学报, 2016, 38(1):104-110.
[18]Glickmann E, Dessaux Y. A critical examination of the specificity of the salkowski reagent for indolic compounds produced by phytopathogenic bacteria[J].Applied&Environmental Microbiology, 1995, 61(2):793-6.
[19]吴翔,甘炳成,黄忠乾,等.一株产IAA菌株的筛选、鉴定及培养条件优化[J].四川农业大学学报, 2014, 32(4):432-435+461.
[20]王占一,金美花,王玉海,等. Plackett-Burman和Central Composite Design试验设计法优化石榴皮中熊果酸的SFECO_2萃取工艺[J].中药材, 2015, 38(3):610-614.
[21]宋浩,纪兆林,陈夕军,等.地衣芽孢杆菌W10菌株发酵培养基优化[J].扬州大学学报:农业与生命科学版, 2015, 36(1):87-91.
[22]钟小廷.产细菌素蜡样芽孢杆菌的筛选、鉴定及培养基优化[D].成都:西华大学, 2014.
[23]刘志祥,曾超珍.响应面法在发酵培养基优化中的应用[J].北方园艺, 2009(2):127-129.
[24]Wang L, Zhang M, Li Y, et al. Application of response surface methodology to optimize the production of antimicrobial metabolites by Micromonospora Y15[J]. Biotechnology&Biotechnological Equipment, 2017:1-10.
[25]李莉,张赛,何强,等.响应面法在试验设计与优化中的应用[J].实验室研究与探索, 2015, 34(8):41-45.
[26]Li C, Bai J, Cai Z, et al. Optimization of a cultural medium for bacteriocin production by Lactococcus lactis using response surface methodology[J]. Journal of Biotechnology, 2002, 93(1):27-34.
[27]韩兴,刘亚南,甄丹妹,等.玫瑰黄链霉菌Men-myco-93-63产抗生素发酵培养基的优化[J].江苏农业科学, 2017, 45(10):100-103.
[28]Passari AK, Mishra VK, Gupta VK, et al. In vitro and in vivo plant growth promoting activities and DNA fingerprinting of antagonistic endophytic actinomycetes associates with medicinal plants[J].PLoS One, 2015, 10(9):1-18.
[29]刘伟.利用地衣芽孢杆菌发酵生产细胞分裂素[D].武汉:华中农业大学, 2005.