新疆农业科学 ›› 2023, Vol. 60 ›› Issue (8): 2046-2054.DOI: 10.6048/j.issn.1001-4330.2023.08.027
收稿日期:
2022-11-03
出版日期:
2023-08-20
发布日期:
2023-08-14
通信作者:
张志东(1977- ),男,新疆乌鲁木齐人,研究员,硕士生导师,研究方向为特殊环境微生物学,(E-mail)zhangzheedong@sohu.com作者简介:
朱静(1981-),女,云南昆明人,研究员,硕士,研究方向为特殊环境微生物学,(E-mail)122543537@qq.com
基金资助:
ZHU Jing(), ZHANG Zhidong(), TANG Qiyong, GU Meiying
Received:
2022-11-03
Online:
2023-08-20
Published:
2023-08-14
Correspondence author:
ZHANG Zhidong (1977-), male, Urumqi of Xinjiang, master degree professor, research field in special environmental microbiology, (E-mail)zhangzheedong@sohu.comSupported by:
摘要:
【目的】分析辐射对短梗霉代谢活性的影响,研究真菌耐辐射机理机制。【方法】分离鉴定一株产黑色素短梗霉,并进行菌株的60Co γ射线照射试验。根据菌株的耐辐射能力,设置60Co γ射线0、2 500、5 000和10 000 Gy 4个辐射剂量,采用Biolog FF 技术检测不同辐射下菌株对FF微平板中碳源的利用程度,分析辐射对产黑色素短梗霉代谢碳源活性的影响。【结果】不同辐射剂量下菌株AWCD值存在明显差异。随着辐射剂量的升高,细胞代谢活性明显下降,羧酸类和氨基酸的利用呈下降趋势,而碳水化合物的利用率呈上升趋势。随着辐射剂量增加,菌株对碳源的利用率发生了变化。在Biolog FF微平板95种碳源中,共有46个碳源被利用,其中7种碳源利用率随辐射剂量上升而增加。其中,D-核糖,蔗糖,D-阿拉伯糖,L-阿拉伯糖的利用率增加最为明显。【结论】辐射能明显降低产黑色素短梗霉的代谢活性,其在高剂量辐射下碳源利用类型会发生明显的改变,可能是该菌辐射应激和适应的一种有效机制。
中图分类号:
朱静, 张志东, 唐琦勇, 顾美英. 基于Biolog FF技术解析辐射对产黑色素短梗霉代谢活性的影响[J]. 新疆农业科学, 2023, 60(8): 2046-2054.
ZHU Jing, ZHANG Zhidong, TANG Qiyong, GU Meiying. Effects of radiation on metabolic activities of Aureobasidium melanogenum based on biolog FF system[J]. Xinjiang Agricultural Sciences, 2023, 60(8): 2046-2054.
图2 菌株MF1菌丝体显微形态 注:查氏培养基28℃培养7 d,Olympus BX43 显微镜40倍观测
Fig.2 Mycelial characteristics of the strain MF1 Note:cultured on Czapek Dox Agar at 28℃ for 7 days and observed under an Olympus BX43 (40×) microscope
辐射剂量Radiation Dose(Gy) | |||||||
---|---|---|---|---|---|---|---|
0 Gy | 2 500 Gy | 5 000 Gy | 10 000 Gy | ||||
碳源 Carbon source | 吸光值 Absorbance (Ci -R) | 碳源 Carbon source | 吸光值 Absorbance (Ci -R) | 碳源 Carbon source | 吸光值 Absorbance (Ci -R) | 碳源 Carbon source | 吸光值 Absorbance (Ci -R) |
L-苹果酸 L-Malic Acid | 0.716 | 熊果苷 Arbutin | 0.608 | 熊果苷 Arbutin | 0.562 | D-核糖 D-Ribose | 0.188 |
反丁烯二酸 Fumaric Acid | 0.656 | 反丁烯二酸 Fumaric Acid | 0.555 | D-木糖 D-Xylose | 0.391 | 蔗糖 Sucrose | 0.138 |
琥珀酸 Succinic Acid | 0.609 | L-脯氨酸 L-Proline | 0.537 | D-蜜三糖 D-Raffinose | 0.242 | D-阿拉伯糖 D-Arabinose | 0.126 |
L-谷氨酸 L-Glutamic Acidine | 0.602 | L-苹果酸 L-Malic Acid | 0.497 | 6-O-D-吡喃葡萄 糖酰-D-呋喃果糖 Palatinose | 0.239 | D-海藻糖 D-Trehalose | 0.126 |
L-脯氨酸 L-Prol | 0.597 | 琥珀酸 Succinic Acid | 0.444 | D-松三糖 D-Melezitose | 0.232 | 熊果苷 Arbutin | 0.106 |
熊果苷 Arbutin | 0.576 | 水杨苷 Salicin | 0.402 | 糊精 Dextrin | 0.217 | D-木糖 D-Xylose | 0.105 |
6-O-D-吡喃葡萄 糖酰-D-呋喃果糖 Palatinose | 0.531 | D-木糖 D-Xylose | 0.389 | 溴代琥珀酸 Bromosuccinic Acid | 0.215 | L-阿拉伯糖 L-Arabinose | 0.101 |
水杨苷 Salicin | 0.481 | 糊精 Dextrin | 0.37 | D-果糖 D-Fructose | 0.197 | ||
D-葡萄醛酸 D-Glucuronic Acid | 0.476 | L-谷氨酸 L-Glutamic Acid | 0.367 | D-海藻糖 D-Trehalose | 0.195 | ||
D-木糖 D-Xylose | 0.468 | 海藻糖 D-Trehalose | 0.336 | 琥珀酸甲基酯 Succinic Acid Mono-Methyl Ester | 0.192 |
表1 不同辐射剂量下菌株利用最多的10种碳源
Tab.1 Utilization of 10 carbon sources by strain under different radiation doses
辐射剂量Radiation Dose(Gy) | |||||||
---|---|---|---|---|---|---|---|
0 Gy | 2 500 Gy | 5 000 Gy | 10 000 Gy | ||||
碳源 Carbon source | 吸光值 Absorbance (Ci -R) | 碳源 Carbon source | 吸光值 Absorbance (Ci -R) | 碳源 Carbon source | 吸光值 Absorbance (Ci -R) | 碳源 Carbon source | 吸光值 Absorbance (Ci -R) |
L-苹果酸 L-Malic Acid | 0.716 | 熊果苷 Arbutin | 0.608 | 熊果苷 Arbutin | 0.562 | D-核糖 D-Ribose | 0.188 |
反丁烯二酸 Fumaric Acid | 0.656 | 反丁烯二酸 Fumaric Acid | 0.555 | D-木糖 D-Xylose | 0.391 | 蔗糖 Sucrose | 0.138 |
琥珀酸 Succinic Acid | 0.609 | L-脯氨酸 L-Proline | 0.537 | D-蜜三糖 D-Raffinose | 0.242 | D-阿拉伯糖 D-Arabinose | 0.126 |
L-谷氨酸 L-Glutamic Acidine | 0.602 | L-苹果酸 L-Malic Acid | 0.497 | 6-O-D-吡喃葡萄 糖酰-D-呋喃果糖 Palatinose | 0.239 | D-海藻糖 D-Trehalose | 0.126 |
L-脯氨酸 L-Prol | 0.597 | 琥珀酸 Succinic Acid | 0.444 | D-松三糖 D-Melezitose | 0.232 | 熊果苷 Arbutin | 0.106 |
熊果苷 Arbutin | 0.576 | 水杨苷 Salicin | 0.402 | 糊精 Dextrin | 0.217 | D-木糖 D-Xylose | 0.105 |
6-O-D-吡喃葡萄 糖酰-D-呋喃果糖 Palatinose | 0.531 | D-木糖 D-Xylose | 0.389 | 溴代琥珀酸 Bromosuccinic Acid | 0.215 | L-阿拉伯糖 L-Arabinose | 0.101 |
水杨苷 Salicin | 0.481 | 糊精 Dextrin | 0.37 | D-果糖 D-Fructose | 0.197 | ||
D-葡萄醛酸 D-Glucuronic Acid | 0.476 | L-谷氨酸 L-Glutamic Acid | 0.367 | D-海藻糖 D-Trehalose | 0.195 | ||
D-木糖 D-Xylose | 0.468 | 海藻糖 D-Trehalose | 0.336 | 琥珀酸甲基酯 Succinic Acid Mono-Methyl Ester | 0.192 |
[1] |
Cox M M, Battista J R. Deinococcus radiodurans - the consummate survivor[J]. Nature Reviews Microbiology, 2005, 3: 882-892.
DOI |
[2] |
McGee E J, Synnott H J, Johanson K J, et al. Chernobyl fallout in a Swedish spruce forest ecosystem[J]. Journal of Environmental Radioactivity, 2000, 48:59-78.
DOI URL |
[3] |
Baeza A, Guillén J. Influence of the soil bioavailability of radionuclides on the transfer of uranium and thorium to mushrooms[J]. Applied Radiation and Isotopes, 2006, 64:1020-1026.
PMID |
[4] |
Dighton J, Tugay T, Zhdanova N. Fungi and ionizing radiation from radionuclides[J]. FEMS microbiology letters, 2008, 281: 109-120.
DOI PMID |
[5] |
Mironenko N V, Alekhina I A, Zhdanova N N, et al. Intraspecific variation in gamma-radiation resistance and genomic structure in the filamentous fungus Alternaria alternata: A case study of strains inhabiting Chernobyl reactor No.4[J]. Ecotoxicology and Environmental Safety, 2000, 45:177-187.
PMID |
[6] | 王戈林, 宁华, 沈萍, 等. 酪氨酸酶基因工程菌产黑色素的发酵条件研究[J]. 中国医药工业杂志, 1999, 30(4):150-154. |
WANG Gelin, NING Hua, SHEN Ping, et al. Study on production of melanin by tyrosinase gene engineering bacteria[J]. Chinese Journal of Pharmaceuticals, 1999, 30(4):150-154. | |
[7] |
Zalar P, Gostinĉar C, de Hoog Gauth, et al. Redefinition of Aureobasidium pullulans and its varieties[J]. Studies in Mycology, 2008, 61(1): 21-38.
DOI URL |
[8] | 张志东, 谢玉清, 王玮, 等. 耐辐射黑色酵母状真菌的筛选和特性研究[J]. 微生物学通报, 2012, 39(5): 724-731. |
ZHANG Zhidong, XIE Yuqing, WANG Wei, et al. Isolation and character of radio-resistant blackyeast-like fungus[J]. Microbiology China, 2012, 39(5): 724-731. | |
[9] | 周宁一. 耐辐射的产黑色素酵母状真菌[J]. 微生物学通报, 2012, 39(5):722-723. |
ZHOU Ningyi. Radio resistant melanin-producing yeast-like fungi[J]. Microbiology China, 2012, 39(5):722-723. | |
[10] | 王丽敏, 李军, 胡小松. 苹果原料中酵母菌的分离鉴定[J]. 中国农业大学学报, 2004, 9(4): 14-17. |
WANG Limin, LI Jun, HU Xiaosong. Isolation and identification of yeastsfrom apple[J]. Journal of China Agricultural University, 2004, 9(4): 14-17. | |
[11] | 李运, 盛慧, 赵荣华. Biolog微生物鉴定系统在菌种鉴定中的应用[J]. 酿酒科技, 2005, 26(7): 84-85. |
LI Yun, SHENG Hui, ZHAO Ronghua. Utilization of biolog microbesidentification system in the identification of microbial species[J]. Liquor-making Science & Technology, 2005, 26(7): 84-85. | |
[12] | Buyer J S, Roberts D P, Millner P, et al. Analysis of fungal communities by sole carbon source utilization profiles[J]. Journal of Microbiological Methods, 2001,(45): 53-60. |
[13] | 朱静, 顾美英, 王玮, 等. 一株耐辐射真菌的鉴定及黑色素分离提取[J]. 新疆农业科学, 2013, 50(10):1858-1864. |
ZHU Jing, GU Meiying, WANG Wei, et al. Study on the tolerance and adsorption of heavy metal ions by bacteria isolated from radiation-polluted soil[J]. Xinjiang Agricultural Sciences, 2013, 50(6): 1101-1107. | |
[14] | 朱静, 房世杰, 王玮, 等. 耐辐射短梗霉黑色素的发酵条件优化及稳定性研究[J]. 新疆农业科学, 2016, 53(9): 1692-1699. |
ZHU Jing, Fang Shijie, WANG Wei, et al. Study on the optimization of fermentation conditions and stability of melanin from a radiation-resistant Aureobasidium pullulans[J]. Xinjiang Agricultural Sciences, 2016, 53(9):1692-1699. | |
[15] | Gerrits van der Ende AHG, de Hoog G S. Variability and molecular diagnostics of the neurotropic species Cladophialophora bantiana[J]. Studies in Mycology, 1999, (43): 151-162. |
[16] |
Zalar P, Gostinĉar C, de Hoog G S, et al. Redefinitionof Aureobasidium pullulans and its varieties[J]. Studies in Mycology, 2008, 61(1): 21-38.
DOI URL |
[17] |
Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution, 1987,(4): 406-425.
DOI PMID |
[18] | Tamura K, Peterson D, Peterson N, et al. MEGA5: Molecular evolutionary genetics analysis usingmaximum likelihood, evolutionary distance, andmaximum parsimony methods[J]. MolecularBiology and Evolution, 2011, 28(10): 2731-2739. |
[19] | 邢华铭, 杜海涛, 张黎黎, 等. PCR-DGGE 与Biolog技术在土壤微生物多样性研究的比较[J]. 农业开发与装备, 2013, (10):48-49. |
XING Huaming, DU Haitao, ZHANG Lili, et al. Comparison of PCR-DGGEand Biolog technology research in soil microbial diversity[J]. Agricultural Development and Equipments, 2013, (10): 48-49. | |
[20] | Jiang S Y, Wang WX, Xue X X. Diversity in Microbial Carbon Metabolism of the Oil Shale at the Western Open Group in Fushun Basin[J]. Advanced Materials Research, 2014, 864-867:140-144. |
[21] | 杨永华, 姚健, 华晓梅. 农药污染对土壤微生物群落功能多样性的影响[J]. 微生物学杂志, 2000, 20(2): 23-25. |
YANG Yonghua, YAO Jian, HUA Xiaomei. Effect of pesticide pollution against functional microbial diversity in soil[J]. Journal of Microbiology, 2000, 20(2): 23-25. | |
[22] | Tugai T, Zhdanova N N, Zheltonozhskii V A, et al. Development of radioadaptive properties for microscopic fungi, long time located on terrains with a heightened background radiation after emergency on Chernobyl NPP[J]. Radiats Biol Radioecol, 2007, 47(5):543-549. |
[23] |
Gu MY, Zhang ZD, Wang W, et al. The Effects of Radiation Pollution on the PopulationDiversities and Metabolic Characteristics of Soil Microorganisms[J]. Water Air Soil Pollut., 2014, 225: 2133.
DOI URL |
[24] | 张志东, 张丽娟, 朱静, 等. 核辐射污染区真菌的分布及多样性研究[J]. 微生物学杂志, 2018, 38(1):50-57. |
ZHANG Zhidong, ZHANG Lijuan, ZHU Jing, et al. Preliminary study on fungi distribution and diversity in nuclearradiation polluted area[J]. Journal of Microbiology, 2018, 38(1): 50-57. | |
[25] |
Taylor TN, Hass H, Kerp H, et al. Perithecial ascomycetes from the 400 million year old Rhyniechert: an example of ancestral polymorphism[J]. Mycologia, 2005, 97: 269-285.
PMID |
[26] | Jansonius J, Kalgutkar RM. Redescription of some fossil fungal spores[J]. Palynology, 2000, (24): 37-47. |
[27] |
Robinson CH. Cold adaptation in Arctic and Antarctic fungi[J]. New Phytologist, 2001, 151: 341-353.
DOI URL |
[28] |
Durrell L, Shields LM. Fungi Isolated in Culture from Soils of the Nevada Test Site[J]. Mycologia, 1960, 52: 636-641.
DOI URL |
[29] |
Gochenaur S, Woodwell G M. The Soil Microfungi of a Chronically Irradiated Oak-Pine Forest[J]. Ecology, 1974, 55: 1004-1016.
DOI URL |
[30] |
Andrea S, Luz M M, Ramón de Anda, et al. A novel plasmid vector designed for chromosomal gene integration and expression: Use for developing a genetically stable Escherichia coli melanin production strain[J]. Plasmid, 2013, 69: 16-23.
DOI PMID |
[31] |
Charles E T, Amy A E, Charles E M, et al. Gamma radiation interacts with melanin to alter its oxidation-reduction potential and results in electric current production[J]. Bioelectrochemistry, 2011, 82: 69-73.
DOI PMID |
[32] |
Ravella, S R Quinones, T S. Ret1er, et al. Extracellular polysacchatide (EPS) production by a novel strain of yeast-like fungus Aureobasidium pullulans[J]. Carbohydrate Polymers, 2010, 82(3):728-732.
DOI URL |
[33] |
Zheng Weifa, Bradley S M, Barbara M S, et al. Effects of melanin on the accumulation of exopolysaccharides byAureobasidium pullulans grown on Nitrate[J]. Bioresource Technology, 2008, 99(16):7480-7486.
DOI PMID |
[34] |
Zhang D P, Spadaro D V, Garibaldi S, et al. Cloning, characterization, expression and antifungal activity of an alkaline serine protease of Aureobasidium pullulans PL5 involved in the biological control of postharvest pathogens[J]. International Journal of Food Microbiology, 2012, 153(3):453-464.
DOI URL |
[35] | 陈波, 蒲刚军. 出芽短梗霉的发酵性能研究[J]. 食品科技, 2002, (11):15-17. |
CHEN Bo, PU Gangjun. Study on Fermentation Properties of Aureobasidium pullulans[J]. Food Science and Technology, 2002, (11):15-17. | |
[36] |
Schu T M, Dick R. Shifts in substrate utilization potential and structure of soil microbial communities in response to carbon substrates[J]. Soil Biology and Biochemistry, 2001, 33(11):1481-1491.
DOI URL |
[37] |
Sangyong L, Jong-Hyun J, Laurence B, et al. Conservation and diversity of radiation and oxidativestress resistance mechanisms in Deinococcus species[J]. FEMS Microbiology Reviews, 2019, 43:19-52.
DOI PMID |
[38] |
易星, 莫远亮, 姜冬梅, 等. 多胺的生物学功能及其调控机制[J]. 动物营养学报, 2014, 26(2): 348-352.
DOI |
YI Xing, MO Yuanliang, JIANG Dongmei, et al. Biological Functions of polyamine and its regulatory mechanisms[J]. Chinese Journal of Animal Nutrition, 2014, 26(2): 348-352. | |
[39] |
Wang H, Wang J, Li L, et al. Metabolic activities of five botryticides against Botrytis cinerea examined using the Biolog FF MicroPlate[J]. Scientific Reports, 2016, 6:31025.
DOI PMID |
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