固定化生防菌的生物质炭基载体构建与评价

doi:10.16409/j.cnki.2095-039x.2023.02.010

摘要/Abstract

摘要： 解淀粉芽胞杆菌（SZ-60）在人参病害防控及参后地土壤微生物修复方面存在巨大潜力，但直接施用存在活菌数偏少、防效不稳定等问题。拟采用微生物固定化技术，以生物质炭为基础载体，筛选并优化适用于解淀粉芽胞杆菌定殖的炭基载体改性条件，分析载体改性前后性质变化与定殖能力变化的关系，为提升SZ-60防效奠定基础，对该类菌种的开发利用进行研究。结果显示，生物质炭固定化微生物最适改性方法为超声波耦合乙酸改性法。最适改性工艺为：乙酸浓度0.2 mol/L，浸渍时间30 min，料液比1:3，超声时间10 min。改性炭基载体上生防菌定殖数达到原炭的2.88倍。改性前后炭基载体共有16项指标发生了显著变化，以养分元素含量变化最为显著，变幅为33.71%~91.87%。其中铁、锌含量、固定碳占比、挥发分占比是影响定殖效果的关键指标。结果表明，超声波耦合乙酸改性可显著影响生物质炭基载体的养分性质，从而提升解淀粉芽胞杆菌的定殖活菌数。该载体的构建为进一步开展参后地土壤微生物修复实验以及相关生物质炭基微生物菌剂开发奠定了良好的基础。

关键词: 改性, 微生物固定化技术, 生防菌, 人参

Abstract: Bacillus amyloliquefaciens SZ-60 has great potential in the prevention and control of ginseng diseases and the microbial remediation of soil after ginseng planting, but there are problems such as less viable bacteria and unstable control effect when applied directly. Taken biocarbon as the basic carrier, the modification conditions of carbon-based carriers suitable for the colonization of B. amyloliquefaciens was screened and optimized, and the relationship between the property changes of carriers before and after modification and the changes of colonization capacity was analyze, which will lay a foundation for improving the control effect of SZ-60, and development and utilization of this kind of bacteria. The results showed that the most suitable modification method for immobilized microorganisms on biochar was ultrasonic coupling acetic acid modification. The optimum modification process is as follows, acetic acid concentration 0.2 mol/L, immersion time 30 min, material liquid ratio 1:3, and ultrasonic time 10 min. The number of biocontrol bacteria colonization on the modified carbon-based carrier was 2.88 times of that on the original carbon. A total of 16 indicators of carbon-based carrier changed significantly before and after modification, with the most significant change in nutrient element content, ranging from 33.71% to 91.87%. The content of iron and zinc, the proportion of fixed carbon and the proportion of volatile matter are the key indicators affecting the colonization effect. The results showed that ultrasonic coupling acetic acid modification could significantly affect the nutrient properties of biochar carrier, thereby increasing the number of viable bacteria colonized by B. amyloliquefaciens. The construction of the carrier has laid a good foundation for further soil microbial remediation experiments and the development of microbial agents based on relevant biochar.

Key words: modification, microbial immobilization technology, biocontrol bacteria, Panax ginseng

中图分类号:

S476

勾颖, 孙安琪, 张阳, 孙卓, 韩梅, 杨莉. 固定化生防菌的生物质炭基载体构建与评价[J]. 中国生物防治学报, 2023, 39(2): 380-388.

GOU Ying, SUN Anqi, ZHANG Yang, SUN Zhuo, HAN Mei, YANG Li. Improvement and Evaluation of Biochar Carrier for Ginseng Biocontrol Bacteria[J]. Chinese Journal of Biological Control, 2023, 39(2): 380-388.

参考文献

[1] 孙卓. 人参病害生防细菌的筛选及其防病作用研究[D]. 长春:吉林农业大学, 2014.
[2] 任静, 沈佳敏, 张磊, 等. 生物炭固定化多环芳烃高效降解菌剂的制备及稳定性[J]. 环境科学学报, 2020, 40(12):4517-4523.
[3] 李海玲, 陈丽华, 肖朝虎, 等. 微生物固定化载体材料的研究进展[J]. 现代化工, 2020, 40(8):58-61.
[4] 段淇斌, 姚拓, 韩华雯, 等. 利用几种固体农业废弃物配制生物肥料载体的研究[J]. 干旱区资源与环境, 2016,30(1):147-151.
[5] Ting W, Hongwen S, Xinhao R, et al. Evaluation of biochars from different stock materials as carriers of bacterial strain for remediation of heavy metal-contaminated soil[J]. Scientific Reports, 2017, 7(1):12114.
[6] Jesionowski T, Zdarta J, Krajewska B. Enzyme immobilization by adsorption:a review[J]. Adsorption-Journal of The International Adsorption Society, 2014, 20(5-6):801-821.
[7] 张松, 侯彬, 纪婷婷, 等. 生物质炭固定化融合菌株F14方法的研究及其对芘的去除[J]. 农业环境科学学报, 2018, 37(3):464-470.
[8] Zhou Z, Gao T, Van Zwieten L, et al. Soil microbial community structure shifts induced by biochar and biochar-based fertilizer amendment to karst calcareous soil[J]. Soil Science Society of America Journal, 2019, 83(2):398-408.
[9] Nyambishi T J, Gwenzi W, Chaukura N, et al. Synthesis and nutrient release patterns of a biochar-based N-P-K slow-release fertilizer[J]. International Journal of Environmental Science & Technology, 2017, 15(5):1-10.
[10] Junlin Z, Shujun W, Ruimin W, et al. Ameliorative roles of biochar-based fertilizer on morpho-physiological traits, nutrient uptake and yield in peanut (Arachis hypogaea L.) under water stress[J]. Agricultural Water Management, 2021, 257:107129
[11] Yuan W, Wenqing L I, Binghai D U, et al. Effect of biochar applied with plant growth-promoting rhizobacteria(PGPR)on soil microbial community composition and nitrogen utilization in tomato[J]. Pedosphere, 2021, 31(6):872-881.
[12] 张彦丽, 金志民, 齐虹凌, 等. 一种提高文冠果耐寒能力的炭基肥料及其制备方法:中国, CN201510201395[P]. 2015-04-24
[13] 王孝芳, 梅新兰, 黄大鹏, 等. 生物质炭载体联合有益菌防控番茄土传青枯病的效果研究[J]. 土壤学报, 2022, 59(2):1-10.
[14] 黄茜. 生物质炭固定化微生物对水中壬基酚的去除效果研究[D]. 杭州:浙江大学, 2018.
[15] Nair V, Vinu R. Peroxide-assisted microwave activation of pyrolysis char for adsorption of dyes from wastewater[J]. Bioresource Technology, 2016(5):511-519.
[16] Yu F, Ji J, Xu Z, et al. Effect of ultrasonic power on the structure of activated carbon and the activities of Ru/AC catalyst[J]. Ultrasonics, 2006, 44(8):389-392.
[17] Hu X, Xue Y, Long L, et al. Characteristics and batch experiments of acid- and alkali-modified corncob biomass for nitrate removal from aqueous solution[J]. Environmental Science and Pollution Research, 2018, 25(20):19932-19940
[18] 于俐, 张桐毓, 李琪, 等. 不同病害诱导下土壤性质变化及其对人参皂苷积累的影响[J/Ol]. 吉林农业大学学报, DOI:10.13327/j.jjlau.2021.1262.
[19]Lee S W, Park K H, Lee S H, et al. Effect of green manure crop cultivation on soil chemical properties and root rot disease in continuous cropping field of ginseng[J]. Korean Journal of Medicinal Crop Science, 2017, 25(1):1-9.
[20] 杨莉, 刘宇航, 郝佳, 等. 生物质炭对人参连作土壤微生物组成及功能的影响[J]. 华南农业大学学报, 2022, 43(1):28-36.
[21] 杨莉, 文子伟, 付婧, 等. 生物质炭对连作参地人参种苗与土壤质量的影响[J]. 中药材, 2020, 43(4):791-796.
[22] Zhenyu W, Ying X, Haoyun W, et al. Biodegradation of crude oil in contaminated soils by free and immobilized microorganisms[J]. Pedosphere, 2012, 22(5):717-725.
[23] 孙卓, 杨利民. 人参病原菌拮抗细菌的分离筛选与鉴定[J]. 植物保护学报, 2015, 42(1):79-86.
[24] 中国林业科学研究院林产化学工业研究所. 木炭和木炭试验方法[S]. 国家质量技术监督局.
[25] 鲍士旦主编. 土壤农化分析[M]. 第三版. 北京:中国农业出版社, 2000.
[26] Ok Y S, Chang S X, Gao B, et al. SMART biochar technology-A shifting paradigm towards advanced materials and healthcare research[J]. Environmental Technology & Innovation, 2015, 4:206-209.
[27] 王汉席. 人工湿地玉米秸秆生物炭基质的改性及对城市尾水净化研究[D]. 长春:东北师范大学, 2021.
[28] 龚宇鹏. 芦苇改性生物质炭制备及用于太湖水生态修复的研究[D]. 杭州:浙江大学, 2017.
[29] 周文君. 改性生物质炭及联合苏丹草对石油污染土壤修复效应研究[D]. 乌鲁木齐:新疆农业大学, 2021.
[30] Deng F, Liao C, Yang C, et al. Enhanced biodegradation of pyrene by immobilized bacteria on modified biomass materials[J]. International Biodeterioration & Biodegradation, 2016, 110(2):46-52.
[31] Jing C, Xiaofeng W, Binbin J, et al. Coupling of immobilized photosynthetic bacteria with a graphene oxides/psf composite membrane for textile wastewater treatment:biodegradation performance and membrane anti-fouling behavior[J]. Membranes, 2021,11(3):226-226.
[32] 田秀梅. 固定化石油烃降解菌剂的制备及降解性能研究[D]. 天津:天津理工大学, 2019.
[33] 温嘉伟, 王辉, 张浩, 等. 改性棕榈树纤维生物质炭的制备及其对溶液中Pb~(2+)的吸附性能分析[J]. 农业环境科学学报, 2021, 40(5):1088-1096.
[34] 杨美蓉, 李坤权, 徐恩兵, 等. 基于有效去除铅(Ⅱ)的中孔炭乙二胺改性与影响因素分析[J]. 环境工程学报, 2015, 9(9):4185-4192.
[35] Hale L, Luth M, Crowley D. Biochar characteristics relate to its utility as an alternative soil inoculum carrier to peat and vermiculite[J]. Soil Biology & Biochemistry, 2015, 81(81):228-235.
[36] 严涛, 朱建国, 姜甜, 等. 一株凝结芽胞杆菌的分离筛选及产孢条件优化[J]. 微生物学通报, 2018, 45(2):238-249.
[37] 孙媛媛, 崔树茂, 唐鑫, 等. 发酵乳杆菌的生长限制性因素分析及高密度培养工艺优化[J]. 食品与发酵工业, 2021, 47(6):1-10.
[38] 高欣伟, 崔树茂, 唐鑫, 等. 长双歧杆菌的最适底物解析和高密度发酵工艺优化[J]. 食品与发酵工业, 2021, 47(19):12-20.
[39] 韩华雯. 几种新型植物根际促生菌肥载体筛选及研究[D]. 兰州:甘肃农业大学, 2013.