棉花秸秆纤维素降解菌的筛选鉴定 与降解棉秆效果研究

doi:10.6048/j.issn.1001-4330.2018.01.003

新疆农业科学 ›› 2018, Vol. 55 ›› Issue (1): 16-23.DOI: 10.6048/j.issn.1001-4330.2018.01.003

棉花秸秆纤维素降解菌的筛选鉴定与降解棉秆效果研究

孙美娜¹,张凡凡²,王永强^2,3,魏晓燕^2,4,鲁建江¹

1.石河子大学化学化工学院/新疆兵团化工绿色工程重点实验室,新疆石河子 832000;
2.石河子大学动物科技学院,新疆石河子 832000;
3.河南科技学院动物科技学院,河南新乡 453000;
4.武威市畜牧兽医科学研究院,甘肃武威 733000

出版日期:2018-01-20 发布日期:2018-06-21
通信作者: 鲁建江(1967-),男,副教授,博士,硕士研究生导师,研究方向为生物化工,(E-mail)lujianjiang_xj@ 163.com
作者简介:孙美娜(1989-),女,硕士,研究方向为生物化工,(E-mail)262809861@qq.com
基金资助:
兵团科技援疆项目(2013AB008);新疆维吾尔自治区研究生科研创新项目(XJGR2015037)

Screening and Identification of Cellulolytic Strains from Cotton Straw and Its Effect on Degradation of Cotton Stalk

SUN Mei-na¹, ZHANG Fan-fan², WANG Yong-qiang^{2, 3}, WEI Xiao-yan^2,4, LU Jian-jiang¹

1.School of Chemistry and Chemical Engineering, Shihezi University / Key Laboratory for Green Processing of Chemical Engineering, Xinjiang Production and Construction Corps, Shihezi Xinjiang, 832000 China;
2. College of Animal Science and Technology, Shihezi University, Shihezi Xinjiang 832000, China;
3. College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, He'na Xinxiang 453000, China;
4. Wuwei Academy of Animal Husbandry and Veterinary Sciences, Wuwei Gansu, 733000, China

Online:2018-01-20 Published:2018-06-21
Correspondence author: LU Jian-jiang (1967 -), male, associate professor, PhD, supervisor of postgraduate, engaged in scientific research and teaching work of biological chemical, (E-mail) lujianjiang_xj@ 163.com

摘要/Abstract

摘要： 【目的】 利用微生物学和分子生物学手段筛选和鉴定棉秆表面附着的真菌,获得具有高效纤维素分解能力的真菌。【方法】 通过将棉花秸秆接种至不同选择性培养基中,分离挑选水解圈较大的菌株,进行镜检和分子生物学鉴定;并同时对菌株进行滤纸酶活性(FPA)和羧甲基纤维素钠(CMC-Na)糖化活力的测定,以及连续(25 d)观测棉秆纤维的失重率。【结果】 主要4株菌种水解圈直径大小排序为,SN-1＞SN-4＞SN-3＞SN-4(P=0.013)。对于CMC-Na糖化活力,24~96 h时SN-1菌种显著低于其它3个菌种(P＜0.05),120 h时,各菌种处理间差异不显著(P＞0.05)。对于FPA活性,第24和120 h时,SN-1和SN-4菌种显著高于SN-2和SN-3菌种(P＜0.05);其它时间点,各菌种按活性高低排序为:SN-1、SN-4＞SN-3＞SN-2(P＜0.05)。通过镜检和分子生物学鉴定,得到4株菌种分别为:SN-1为黑曲霉(Aspergillus niger),SN-2为赤霉(Gibberella moniliformis),SN-3为链格孢霉(Alternaria nees),SN-4为青霉菌(Penicillium italicum)。【结论】 黑曲霉降解棉花秸秆的效果最为显著。对于CMC-Na糖化酶活力、滤纸酶活性以及棉花秸秆的实际失重率(降解率),接种黑曲霉或青霉菌时表现均最佳。

关键词: 棉花; 真菌; 酶活性; 降解效果

Abstract: 【Objective】 This study aims to screen and identify fungi which adhere on the surface of the cotton by using microbiology and molecular biology methods in order to obtain fungi with high cellulose decomposition ability.【Method】 The cotton straw was inoculated into different selective media, and the larger hydrolysis circle of strains was separated and selected. At the same time the strains' filter paper enzyme activity (FPA) and sodium carboxymethyl cellulose (CMC-Na) saccharification vitality were determined, and besides, the weight loss rate of cotton stalk fiber was observed continuously for 25 days.【Result】 The diameter of the hydrolysis ring of the 4 main strains was SN-1 > SN-4 > SN-3 > SN-4 (P =0.013). In terms of CMC - Na saccharification vitality, SN - 1 was significantly lower than other three strains (P < 0.05) at 24-96 h, and there was no significant difference between different strains of treatments (P > 0.05) at 120 h. In terms of FPA activity, SN and SN 1-4 strains was significantly higher than SN - 2 and SN - 3 strains (P < 0.05) at 24 and 120 h, at other time points, the strains were sorted according to the activity level: SN-1, SN-4 > SN-3 > SN-2 (P < 0.05). There were 4 bacterial strains obtained by using the method of microscopic and molecular identification, SN - 1 was Aspergillus nige r, SN - 2 was Gibberella moniliformis , SN - 3 was Alternaria nees and SN - 4 was Penicillium italicum . 【Conclusion】 The effect of Aspergillus niger on the degradation of cotton straw was the most remarkable. The activity of CMC-Na saccharifying enzyme, the activity of filter paper enzyme and the actual weight loss rate (degradation rate) of cotton straw were the best when inoculated with Aspergillus niger or Penicillium italicum .

Key words: cotton; fungi; enzymes; degradation effect

中图分类号:

S562

孙美娜,张凡凡,王永强,魏晓燕,鲁建江. 棉花秸秆纤维素降解菌的筛选鉴定与降解棉秆效果研究[J]. 新疆农业科学, 2018, 55(1): 16-23.

SUN Mei-na, ZHANG Fan-fan, WANG Yong-qiang, WEI Xiao-yan, LU Jian-jiang. Screening and Identification of Cellulolytic Strains from Cotton Straw and Its Effect on Degradation of Cotton Stalk[J]. Xinjiang Agricultural Sciences, 2018, 55(1): 16-23.

参考文献 23

[1]	张鹏忠, 王新江, 托乎提. 新疆棉花产业发展现状、存在问题及对策建议[J]. 新疆农业科学, 2008, 45(S2): 174-176.ZHANG Peng-zhong, WANG Xin-jiang, TUO Hu-ti. (2008). The Development Present, Problems, Countermeasures and Suggestions in Xinjiang Cotton Industry [J]. Xinjiang Agricultural Sciences , 45(S2): 174-176. (in Chinese)
[2]	孔庆平, 孔杰, 徐海江, 等. 新疆棉花集约高效生产技术研发策略[J]. 新疆农业科学, 2015, 52(7): 1 352-1 358.KONG Qing-ping, KONG Jie, XU Hai-jiang, et al. (2015). Research and Development Strategy of Intensive and Efficient Production Technology in Xinjiang Cotton [J]. Xinjiang Agricultural Sciences , 52(7): 1,352-1,358. (in Chinese)
[3]	黄乐平. 新疆棉花生物技术研究现状与发展前景探讨[J]. 新疆农业科学, 2007, 44(S3): 1-5.HUANG Le-ping. (2007).The Research of Cotton Biotechnology on Present Situation and Development Prospects in Xinjiang [J]. Xinjiang Agricultural Sciences , 44(S3): 1-5. (in Chinese)
[4]	杨丽娟. 农作物秸秆还田的方式及技术要求[J]. 现代农业, 2011, (5): 165.YANG Li-juan. (2011). The Crop Straw Counters-Field Way and Technical Requirements [J]. Modern Agriculture , 165(5): 165. (in Chinese)
[5]	白宝伟, 张琴, 滕立平, 等. 一株棉秆纤维素分解真菌的分离筛选及酶学性质研究[J]. 新疆农业科学, 2012, 49(4): 701-707.BAI Bao-wei, ZHANG Qin, TENG Li-ping, et al. (2012).Isolation of a Cotton Stalk Cellulose Decomposing Fungus and Its cellulase Properties [J]. Xinjiang Agricultural Sciences , 49(4): 701-707. (in Chinese)
[6]	穆小民, 吴显荣. 纤维素酶分子生物学研究进展及趋向[J]. 草食家畜, 1994, 14(3):25-27.MU Xiao-min, WU Xian-rong. (1994).The Research Progress and Trends of Cellulose Enzyme Molecular Biology [J]. Grass- Feeding Livestock , 14(3):25-27. (in Chinese)
[7]	刘娣. 秸秆纤维素高效降解真菌的筛选、鉴定及其纤维素酶基因克隆[D]. 北京: 中国农业科学院, 2008.LIU Di. (2008). Screening, Identification and Cellulase Gene Cloning of High Effective Straw Cellulose- Degrading Fungus Strains [D]. Master Thesis. Chinese Academy of Agricultural Sciences, Beijing. (in Chinese)
[8]	齐西珍, 赵军旗, 田朝光, 等. 真菌来源中碱性纤维素酶研究[C]//. 中国酶工程学术研讨会, 2013.QI Xi-zhen, ZHAO Jun-qi, TIAN Chao-guang, et al. (2013). The Research of Source of Alkaline Cellulase in Fungi [C]//. Proceedings of Chinese Enzyme Engineering Academic Seminar, Naning, . (in Chinese)
[9]	高静, 席琳乔, 马春晖. 土壤中棉秆纤维素降解菌的筛选[J]. 塔里木大学学报, 2010, 22(4): 6-10.GAO Jing, XI Lin-qiao, MA Chun-hui. (2010). Solation of Cotton Straw Cellulose-degrading Fungi from Soil [J]. Journal of Tarim University , 22(4): 6-10. (in Chinese)
[10]	王璐, 张海红, 杨柳, 等. 3株纤维素分解真菌的分子鉴定[J]. 江苏农业科学, 2013, 41(8): 51-53.WANG Lu, ZHANG Hai-hong, Yang Liu, et al. (2013). Molecular Identification of 3 strains of Cellulolytic Fungi [J]. Jiangsu Agricultural Sciences , 41(8): 51-53. (in Chinese)
[11]	向殿军, 满丽莉, 张春凤,等. 玉米秸秆纤维素降解菌的分离及发酵条件优化[J]. 安徽农业科学, 2014, 42(4):1 159-1 161.XIANG Dian-jun, MAN Li-li, ZHANG Chun-feng, et al. (2014). Optimization of the Separation and Fermentation Conditions of Corn Straw Cellulose-Degrading Fungus [J]. Journal of Anhui Agricultural Sciences , 42(4): 1,159-1,161. (in Chinese)
[12]	陈天寿. 微生物培养基的制造与应用[M].北京: 中国农业出版社, 1995.CHEN Tian-shou. (1995). Manufacture and Application of Microbial Culture Medium [M]. Beijing: China Agriculture Press, 1995. (in Chinese)
[13]	房兴堂, 陈宏, 赵雪锋, 等. 秸秆纤维素分解菌的酶活力测定[J]. 生物技术通讯, 2007, 18(4): 39-44.FANG Xing-tang, CHEN Hong, ZHAO Xue-feng, et al. (2007). Determination of Enzyme Activity of Straw Cellulose- Decomposing Microorganisms [J]. Letters in Biotechnology , 18(4): 39-44. (in Chinese)
[14]	傅力, 丁友昉, 张篪. 纤维素酶测定方法的研究[J]. 新疆农业大学学报, 2000, 23(2): 45-48.FU Li, DING You-fang, ZHANG Chi. (2000). The Study of Cellulase Method [J]. Journal of Xinjiang Agricultural University , 23(2): 45-48. (in Chinese)
[15]	张凡凡, 于磊, 张前兵,等. 沙尔套山天然割草场主要豆科牧草营养价值综合评价研究[J]. 新疆农业科学, 2014, 51(10): 1 907-1 915.ZHANG Fan-fan, YU Lei, ZHANG Qian-bing, et al. (2014). Comprehensive Assessment of the Main Legume Forages'Nutritional Value of Natural Mowing Steppe in Shaertao Mountain, Zhaosu, Xinjiang [J]. Xinjiang Agricultural Sciences, 51(10): 1,907-1,915. (in Chinese)
[16]	Benson, D. A., Boguski, M. S., Lipman, D. J., Ostell, J., & Ouellette, B. F. (1998). Genbank. Nucleic Acids Research , 26(1): 1.
[17]	张莉莉, 张苓花. 利用氯化苄提取真菌基因组DNA及其分子生物学分析[J]. 大连工业大学学报, 2000, 19(1):36-39.ZHANG Li-li, ZHANG Ling-hua. (2000). Isolation and molecular biological analyses of genomic DNAs from fungi using benzyl chloride [J]. Journal of Dalian Institute of Light Industry , 19(1): 36-39. (in Chinese)
[18]	吴明隆. SPSS统计应用实务[M]. 北京: 科学出版社, 2003.WU Min-long. (2003). The SPSS Statistical Practice [M]. Beijing: Science Press. (in Chinese)
[19]	Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., & Higgins, D. G. (1997). The clustal_x windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research , 25(25): 4,876-4,882.
[20]	Kumar, S., Tamura, K., & Nei, M. (2004). Mega3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Briefings in Bioinformatics , 5(2): 150-163.
[21]	东秀珠, 蔡妙英. 常见细菌系统鉴定手册[K]. 北京: 科学出版社, 2001: 353-370.DONG Xiu-zhu, CAI Miao-ying. (2001). Manual of the Identification of Common Bacterial System [K]. Beijing: Science Press: 353-370. (in Chinese)
[22]	张恒芳, 李林, 史喜林, 等. 玉米秸秆低温纤维素分解菌的筛选及分解效果测定[J]. 玉米科学, 2013,(5): 147-150.ZHANG Heng-fang, LI Lin, SHI Xi-lin, et al. (2013). Screening of Cellulose Degradation Bacteria Decomposing Maize Straw under Low Temperature and the Function Determining [J]. Journal of Maize Sciences , (5): 147-150. (in Chinese)
[23]	郭艳, 张进良, 王会平, 等. 玉米秸秆还田土壤中纤维素分解菌的分离筛选和鉴定[J]. 农业科学与技术, 2010, 11(4): 134-137.GUO Yan, ZHANG Jin-liang, WANG Hui-ping, et al. (2010). Isolation, Screening and Identification of Cellulose Decomposing Strains in Straw amended Soil [J]. Agricultural Science and Technology , 11(4): 134-137. (in Chinese)

棉花秸秆纤维素降解菌的筛选鉴定与降解棉秆效果研究

Screening and Identification of Cellulolytic Strains from Cotton Straw and Its Effect on Degradation of Cotton Stalk

PDF下载

可视化

摘要/Abstract

引用本文

使用本文

参考文献 23

相关文章 15

编辑推荐

Metrics

[1]	陈茂光, 林涛, 张昊, 刘海军, 王一帆, 汤秋香. 地膜类型对棉花生长的影响及自身降解和回收特性分析[J]. 新疆农业科学, 2023, 60(9): 2101-2108.
[2]	杨川, 张凯, 陈冰, 张慧, 柳萍, 常松, 盛建东. 棉花植株形态特征对不同水分状况的响应[J]. 新疆农业科学, 2023, 60(9): 2120-2127.
[3]	杨国江, 陈云, 林祥群, 何江勇, 刘盛林, 曲永清. 氮肥减施下有机肥替代对滴灌棉花产量、氮素吸收利用及土壤硝态氮的影响[J]. 新疆农业科学, 2023, 60(9): 2138-2145.
[4]	李雪玲, 郭俊先, 陈莉, 宋鹤岭, 张众. 不同覆膜宽度对棉花农田环境的影响[J]. 新疆农业科学, 2023, 60(8): 1840-1847.
[5]	阳妮, 玛依拉·玉素音, 杨延龙, 李春平, 张大伟, 徐海江, 赖成霞. 黄萎病枯斑型与黄化型病症棉花叶片的植物挥发物对比[J]. 新疆农业科学, 2023, 60(8): 1975-1986.
[6]	米尔扎提·木塔力甫, 石秀楠, 柏军兵, 祖拜代·阿布都克日木, 吾勒加勒哈斯·阿扎提, 石书兵. 不同脱绒方式及PEG胁迫下对棉花种子活力及幼苗性状的影响[J]. 新疆农业科学, 2023, 60(7): 1561-1568.
[7]	江柱, 张江辉, 白云岗, 杨鹏年, 刘洪波, 肖军, 刘旭辉. 膜下咸水滴灌水肥盐调控对棉花生长及产量的影响[J]. 新疆农业科学, 2023, 60(6): 1389-1397.
[8]	王文涛, 吴博, 邰红忠, 练文明, 戴翠荣, 李双江, 蒲艳梅. 新疆阿拉尔垦区不同播期对棉花生长的影响[J]. 新疆农业科学, 2023, 60(6): 1413-1422.
[9]	桑志伟, 梁亚军, 龚照龙, 郑巨云, 王俊铎, 李雪源, 陈全家. 不同陆地棉种质资源机采性状分析[J]. 新疆农业科学, 2023, 60(5): 1088-1098.
[10]	逯涛, 曾庆涛, 张文, 王文博, 王政洋, 杨芮, 孙玉岩. 主成分分析及灰色关联度分析综合评价棉花产量与品质[J]. 新疆农业科学, 2023, 60(5): 1099-1109.
[11]	王宁, 史应武, 牛新湘, 杨红梅, 楚敏, 詹发强, 包慧芳, 杨蓉, 龙宣杞, 丁荣荣. 棉花根际溶磷菌WJP-7发酵培养基优化及防病增产效果[J]. 新疆农业科学, 2023, 60(5): 1263-1270.
[12]	张泓, 吐尔逊江·买买提, 张少民, 张军高, 周小云. 近30 a全国棉花生产区域时空变化及区域优势分析[J]. 新疆农业科学, 2023, 60(4): 1028-1040.
[13]	代健敏, 张巨松, 徐新龙, 李始鑫, 翟梦华, 孙明辉. 氮肥运筹对雹后重播受旱棉花生长特性及产量影响[J]. 新疆农业科学, 2023, 60(4): 798-809.
[14]	丁宇, 张江辉, 白云岗, 刘洪波, 郑明, 赵经华, 肖军, 韩政宇. 双膜条件下不同干播湿出水分处理对棉花生理、生长特性的影响[J]. 新疆农业科学, 2023, 60(4): 810-822.
[15]	张宸, 梁悦, 殷昊, 张应榕, 陈波浪. 氮肥形态和品种对棉花根系形态与氮素积累的影响[J]. 新疆农业科学, 2023, 60(4): 823-831.

棉花秸秆纤维素降解菌的筛选鉴定 与降解棉秆效果研究

Screening and Identification of Cellulolytic Strains from Cotton Straw and Its Effect on Degradation of Cotton Stalk

PDF下载

可视化

摘要/Abstract

引用本文

使用本文

参考文献 23

相关文章 15

编辑推荐

Metrics

棉花秸秆纤维素降解菌的筛选鉴定与降解棉秆效果研究