油茶壳堆肥微生物的培养及功能芽孢杆菌的筛选和初步应用

张声峰; 陈硕昌; 梁文凤; 严祥; 冯艺峰; 李桂珍; 杨辉

doi:10.19303/j.issn.1008-0384.2021.07.014

油茶壳堆肥微生物的培养及功能芽孢杆菌的筛选和初步应用

doi: 10.19303/j.issn.1008-0384.2021.07.014

1.
广西大学生命科学与技术学院，广西　南宁　530004
2.
广西壮族自治区林业科学研究院，广西　南宁　530000

基金项目: 广西科技重大专项（桂科 AA20302021-7）；百色市科学研究与技术开发计划（百科20192129）；广西特色经济林培育与利用重点实验室自主课题（JA-20-04-02）

详细信息

作者简介:
张声峰（1990−），男，硕士研究生，研究方向：微生物工程（E-mail：1908391037@st.gxu.edu.cn）

通讯作者:
杨辉（1972−），男，博士，高级工程师，研究方向：食品发酵和酶工程（E-mail：huiy422@gxu.edu.cn）

中图分类号: X 712；X 53
计量
- 文章访问数: 490
- HTML全文浏览量: 135
- PDF下载量: 22
- 被引次数: 0
出版历程
- 收稿日期: 2021-03-02
- 修回日期: 2021-04-25
- 网络出版日期: 2021-07-13
- 刊出日期: 2021-07-28

Functional Bacillus Species in Camellia Seed Shell Compost

1.
College of Life Science and Technology, Guangxi University, Nanning, Guangxi　530004, China
2.
Guangxi Academy of Forestry Sciences, Nanning, Guangxi　530000, China

摘要

摘要: 目的从自然状态下的油茶壳堆肥产品中筛选功能芽孢杆菌，为油茶壳堆肥高效生产、饲料发酵等应用提供菌剂。方法利用高通量测序技术测定广西某油茶壳堆肥中微生物群落分布，筛选适宜堆肥微生物生长的培养基以获得多样性较丰富的堆肥芽孢杆菌菌群，经平板稀释涂布法分离纯化菌株，使用水解圈法和酶活测定方法进行水解酶功能分析，利用菌株形态特征观察、16S rDNA分子鉴定法明确菌株的种属，并运用分子生物学软件 MEGAX构建系统发育树。使用重铬酸钾法测定腐殖酸含量。结果芽孢杆菌科（Bacillaceae）是优势菌科，占比达55.58%；分离到15株芽孢杆菌，同时具有淀粉酶、纤维素酶、蛋白酶3种水解酶活性的菌株有6株；同时具有淀粉酶和纤维素酶活性的有1株；仅具有蛋白酶或纤维素酶一种酶活性的有2株。其中菌株Bacillus sp. YX11的蛋白酶活力达到27.07±3.28 U·mL⁻¹，淀粉酶活力达123.97±3.19 U·mL⁻¹，纤维素酶活力达15.75±0.23 U·mL⁻¹。含有这15株菌的复合菌制剂有利于腐殖酸的生成，提高了堆肥品质。结论鉴定出的Bacillus cereus YX02和 Bacillus flexus FYF01等菌株具有进一步研究开发的价值，可用于开发油茶壳堆肥微生物菌制剂。
- 油茶壳堆肥 /
- 芽孢杆菌 /
- 功能性 /
- 筛选 /
- 应用
Abstract: Objective Bacillus spp that contribute to the fermentation of Camellia oleifera seed shells were isolated for effective composting of the waste material. Method The microbial community in natural camellia seed shell compost found in Guangxi was studied using the high-throughput sequencing technology. Suitable culture medium to foster the growth of richly diverse Bacillus spp from the compost was selected. Flora isolation by dilution with a streaking plate method followed. The hydrolase activities of the isolates were determined by using the hydrolysis circle method and enzyme activity analysis, the species identified by a 16S rDNA analysis, and the phylogeny constructed by MEGAX. The content of humic acid in the compost was measured by a potassium dichromate method. Result Bacillaceae was the dominant family in the compost. It accounted for 55.58% of all isolated flora. Among the 15 Bacillus isolates, 6 exhibited activities of amylase, cellulase, and protease, one of amylase and cellulase, and 2 of protease or protease. Strain YX11 showed a protease activity of 27.07±3.28 U·mL⁻¹, an amylase activity of 123.97±3.19 U·mL⁻¹, and a cellulase activity of 15.75±0.23 U·mL⁻¹. In the presence of numerous Bacillus spp that secreted varieties of hydrolases, formation of humic acid in the compost was enhanced. Conclusion Some of the isolated strains, such as B. cereus YX02 and B. flexus FYF01, might warrant further investigation to develop microbial inoculants for efficient composting camellia seed shells.
- camellia seed shell compost /
- bacillus /
- functional /
- screening /
- application

HTML全文

图 1 油茶壳堆肥原核微生物群落组成（科类）

Figure 1. Composition of prokaryotic microbial community at family level in camellia seed shell compost

下载: 全尺寸图片幻灯片

图 2 六组培养基中的细菌数量和OD₆₀₀值的变化趋势

注：Y：厌氧培养，HJ：好氧静置培养，HY：好氧摇床培养

Figure 2. Bacteria CFU by plate count and OD₆₀₀ measurement on 6 culture media

Notes: Y: anaerobic culture; HJ: aerobic static culture; HY: aerobic culture on shaking table.

下载: 全尺寸图片幻灯片

图 3 光学显微镜下部分菌株革兰氏染色图（×4 000倍）

注：a：菌株YX07；b：菌株FYF01；c：菌株YX12；d：菌株FYF07

Figure 3. Gram staining bacteria morphology under microscope (×4 000)

Notes: a: Strain YX07; b: Strain FYF01; c: Strain YX12; d: Strain FYF07.

下载: 全尺寸图片幻灯片

图 4 16S rDNA序列PCR产物琼脂凝胶电泳结果

注：M：DL2000 （DNA Marker）；1~8：部分菌株基因组 PCR产物

Figure 4. Agarose gel electrophoresis of PCR products from 16S rDNA sequence

Notes: M: DL2000 (DNA marker); 1-8: genomic PCR products of strains.

下载: 全尺寸图片幻灯片

图 5 基于16S rDNA序列的系统发育树

Figure 5. Phylogenetic tree of strains based on 16S rDNA sequence

下载: 全尺寸图片幻灯片

图 6 菌株的蛋白酶、淀粉酶、纤维素酶活力

Figure 6. Activities of protease, amylase, and cellulase of strains

下载: 全尺寸图片幻灯片

图 7 应用试验腐殖酸含量

Figure 7. Humic acid content in compost at application trial

下载: 全尺寸图片幻灯片

表 1 菌制剂应用试验配方

Table 1. Proposed formulations of microbial inoculants

组别 Group	油茶壳 Camellia shell/g	菌液 microbial inoculants/mL	糖蜜 Molasses/mL	花生枯 Peanut cake/g	尿素 Urea/g	水 H₂O/mL
菌制剂 experiment group	15	15	6	1.2	1.2	100
对照组 control group	15	0	6	1.2	1.2	115

下载: 导出CSV

表 2 菌株形态特征

Table 2. Morphological characteristics of isolated Bacillus spp

菌株编号 Strains	菌落质地 Colony texture	菌落颜色 Colony color	菌落形态 Colony morphology
YX01	干燥、皱褶 dry and wrinkled	半透明色 translucent color	不规则圆形，有毛边 irregular round with rough edges
YX02	干燥、皱褶 dry and wrinkled	乳白色 milky white	较规则圆形，中间隆起 regular round, middle uplift
YX03	光滑、湿润 smooth and moist	半透明色 translucent color	较规则圆形 regular round
YX07	光滑、湿润 smooth and moist	乳白色 milky white	圆形 round
YX08	光滑、湿润 smooth and moist	半透明色 translucent color	圆形 round
YX11	湿润、黏稠 wet and sticky	乳白色 milky white	圆形 round
YX12	光滑、湿润 smooth and moist	乳白色 milky white	圆形 round
YX13	干燥、皱褶 dry and wrinkled	枯黄色 withered yellow	不规则圆形，有毛边 irregular round with rough edges
YX15	光滑、湿润 smooth and moist	乳白色 milky white	圆形 round
YX16	光滑、湿润 smooth and moist	乳白色 milky white	圆形 round
FYF01	干燥、皱褶 dry and wrinkled	乳白色 milky white	不规则圆形，有毛边 irregular round with rough edges
FYF04	光滑、湿润 smooth and moist	乳白色 milky white	较规则圆形 regular round
FYF07	光滑、湿润 smooth and moist	乳白色 milky white	较规则圆形，中间隆起 regular round, middle uplift
FYF08	干燥、皱褶 dry and wrinkled	半透明色 translucent color	不规则圆形，有毛边 irregular round with rough edges
FYF10	光滑、湿润 smooth and moist	乳白色 milky white	较规则圆形 regular round

下载: 导出CSV

表 3 菌株16S rDNA鉴定结果

Table 3. Identifications by 16S rDNA

序号 Serial number	编号 Number	菌种分类 Species classification
1	YX01	Bacillus sp.
2	YX02	Bacillus cereus
3	YX03	Bacillus sp.
4	YX07	Bacillus sp.
5	YX08	Bacillus sp.
6	YX11	Bacillus sp.
7	YX12	Bacillus sp.
8	YX13	Bacillus sp.
9	YX15	Brevibacterium oceani
10	YX16	Bacillus sp.
11	FYF01	Bacillus flexus
12	FYF04	Bacillus sp.
13	FYF07	Bacillus sp.
14	FYF08	Bacillus sp.
15	FYF10	Bacillus sp.

下载: 导出CSV

表 4 不同培养基上菌落直径与透明圈直径比

Table 4. Ratio of transparent ring diameter to colony diameter on different mediums

菌株编号 Strains	脱脂乳糖培养基 Skim milk medium			淀粉培养基 Starch medium			刚果红培养基 Congo red medium
菌株编号 Strains	透明圈直径D Transparent ring diameter/mm	菌落直径d Colony diameter/mm	直径比D/d Diameter ratio	透明圈直径D Transparent ring diameter/mm	菌落直径d Colony diameter/mm	直径比D/d Diameter ratio	透明圈直径D Transparent ring diameter/mm	菌落直径d Colony diameter/mm	直径比D/d Diameter ratio
YX01	0.0	7.5	0.00	13.0	7.5	1.73	18.0	5.0	3.60
YX02	28.0	26.5	1.06	0.0	16.0	0.00	0.0	11.0	0.00
YX03	0.0	0.0	0.00	0.0	4.0	0.00	20.0	8.0	2.50
YX07	0.0	0.0	0.00	0.0	6.5	0.00	0.0	8.5	0.00
YX08	0.0	0.0	0.00	0.0	11.0	0.00	0.0	2.0	0.00
YX11	25.0	20.0	1.25	19.5	6.0	3.25	30.0	11.0	2.73
YX12	0.0	0.0	0.00	0.0	5.0	0.00	0.0	8.0	0.00
YX13	29.0	23.0	1.26	20.0	8.0	2.50	18.0	8.5	2.12
YX15	0.0	0.0	0.00	0.0	6.5	0.00	0.0	10.0	0.00
YX16	0.0	0.0	0.00	0.0	4.0	0.00	0.0	7.0	0.00
FYF01	14.0	12.0	1.17	16.0	6.0	2.67	32.0	6.5	4.92
FYF04	30.0	21.5	1.39	14.0	7.0	2.00	24.5	6.0	4.08
FYF07	26.0	19.0	1.37	20.0	11.5	1.74	24.0	9.0	2.67
FYF08	0.0	0.0	0.00	0.0	7.0	0.00	0.0	2.0	0.00
FYF10	25.5	19.5	1.31	18.0	12.5	1.44	25.0	9.0	2.78

下载: 导出CSV

参考文献(52)

[1]	HU J B, SHI Y, LIU Y, et al. Anatomical structure of Camellia oleifera shell [J]. Protoplasma, 2018, 255(6): 1777−1784. doi: 10.1007/s00709-018-1271-8
[2]	覃佐东, 谢吉勇, 黄生辉, 等. 油茶壳综合利用研究进展 [J]. 生物加工过程, 2016, 14(5):74−78. doi: 10.3969/j.issn.1672-3678.2016.05.014 QIN Z D, XIE J Y, HUANG S H, et al. Progress in utilization of camellia shells [J]. Chinese Journal of Bioprocess Engineering, 2016, 14(5): 74−78.(in Chinese) doi: 10.3969/j.issn.1672-3678.2016.05.014
[3]	SHRESTHA K, SHRESTHA P, WALSH K B, et al. Microbial enhancement of compost extracts based on cattle rumen content compost - characterisation of a system [J]. Bioresource Technology, 2011, 102(17): 8027−8034. doi: 10.1016/j.biortech.2011.06.076
[4]	SONG C, ZHANG Y, XIA X, et al. Effect of inoculation with a microbial consortium that degrades organic acids on the composting efficiency of food waste [J]. Microbial Biotechnology, 2018, 11(6): 1124−1136. doi: 10.1111/1751-7915.13294
[5]	FANG Y, JIA X B, CHEN L J, et al. Effect of thermotolerant bacterial inoculation on the microbial community during sludge composting [J]. Canadian Journal of Microbiology, 2019, 65(10): 750−761. doi: 10.1139/cjm-2019-0107
[6]	THATOI H, BEHERA B C, MISHRA R R, et al. Biodiversity and biotechnological potential of microorganisms from mangrove ecosystems: A review [J]. Annals of Microbiology, 2013, 63(1): 1−19. doi: 10.1007/s13213-012-0442-7
[7]	胡亚杰, 韦建玉, 卢健, 等. 枯草芽孢杆菌在农作物生产上的应用研究进展 [J]. 作物研究, 2019, 33(2):167−172. HU Y J, WEI J Y, LU J, et al. Research prog ress of Bacillus subtilis application in crops production [J]. Crop Research, 2019, 33(2): 167−172.(in Chinese)
[8]	MAYENDE L, WILHELMI B, PLETSCHKE B. Cellulases (CMCases) and polyphenol oxidases from thermophilic Bacillus spp. isolated from compost [J]. Soil Biology and Biochemistry, 2006, 38(9): 2963−2966. doi: 10.1016/j.soilbio.2006.03.019
[9]	JOSÉ V C. Inoculating composted pine bark with beneficial organisms to make a disease suppressive compost for container production in mexican forest nurseries [J]. Plants Journal, 2004, 2(5): 181−185.
[10]	KRAUSE M S, DE CEUSTER T J J, TIQUIA S M, et al. Isolation and characterization of rhizobacteria from composts that suppress the severity of bacterial leaf spot of radish [J]. Phytopathology, 2003, 93(10): 1292−1300. doi: 10.1094/PHYTO.2003.93.10.1292
[11]	付冰妍, 孙向阳, 余克非, 等. 芽孢杆菌B01固态发酵及其对园林废弃物堆肥的影响 [J]. 环境科学研究, 2020(2):450−457. FU B Y, SUN X Y, YU K F, et al. Solid state fermentation of Bacillus B01 and its effect on green waste composting [J]. Research of Environmental Sciences, 2020(2): 450−457.(in Chinese)
[12]	余培斌, 杜晶, 陈建新. 高温好氧堆肥过程中芽孢杆菌的筛选、鉴定及应用 [J]. 食品与发酵工业, 2020, 46(12):199−205, 212. YU P B, DU J, CHEN J X. Study on screening and identification of Bacillus in the process of high-temperature aerobic composting and its application [J]. Food and Fermentation Industries, 2020, 46(12): 199−205, 212.(in Chinese)
[13]	ZHANG J P, YING Y, LI X B, et al. Physical and chemical properties of Camellia oleifera shell composts with different additives and its maturity evaluation system [J]. Environmental Science and Pollution Research, 2020, 27(28): 35294−35302. doi: 10.1007/s11356-020-09861-3
[14]	詹孝慈, 罗在柒, 武忠亮, 等. 不同氮源及微生物菌剂能提高油茶壳堆肥效果 [J]. 分子植物育种, 2019, 17(12):4153−4160. ZHAN X C, LUO Z Q, WU Z L, et al. Different nitrogen sources and microbial inoculants could improve the composting of Camellia oleifera shell [J]. Fenzi Zhiwu Yuzhong (Molecular Plant Breeding), 2019, 17(12): 4153−4160.(in Chinese)
[15]	ZHANG J P, YING Y, YAO X H. Effects of turning frequency on the nutrients of Camellia oleifera shell co-compost with goat dung and evaluation of co-compost maturity [J]. PLoS One, 2019, 14(9): e0222841. doi: 10.1371/journal.pone.0222841
[16]	秦楠, 栗东芳, 杨瑞馥. 高通量测序技术及其在微生物学研究中的应用 [J]. 微生物学报, 2011, 51(4):445−457. QIN N, LI D F, YANG R F. Next-generation sequencing technologies and the application in microbiology-A review [J]. Acta Microbiologica Sinica, 2011, 51(4): 445−457.(in Chinese)
[17]	葛慈斌, 蓝江林, 刘波, 等. 解淀粉芽胞杆菌FJAT-8754产纤维素酶和淀粉酶发酵动力学模型的构建 [J]. 福建农业学报, 2019, 34(6):697−704. GE C B, LAN J L, LIU B, et al. Kinetics of cellulase and amylase-producing fermentation of Bacillus amylolique faciens FJAT-8754 [J]. Fujian Journal of Agricultural Sciences, 2019, 34(6): 697−704.(in Chinese)
[18]	赵国群, 牛梦天, 卢士康, 等. 梨渣固态发酵培养多粘类芽孢杆菌的工艺 [J]. 农业工程学报, 2016, 32(7):303−308. ZHAO G Q, NIU M T, LU S K, et al. Cultivation of Paenibacillus polymyxa by solid-state fermentation of pear residues [J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2016, 32(7): 303−308.(in Chinese)
[19]	胡瑞萍, 丁贤, 李来好, 等. 响应面法优化枯草芽孢杆菌NHS1产芽孢发酵培养 [J]. 生态学杂志, 2018, 37(2):605−612. HU R P, DING X, LI L H, et al. Optimization of fermentation medium composition by response surface methodology for the spore production of Bacillus subtilis [J]. Chinese Journal of Ecology, 2018, 37(2): 605−612.(in Chinese)
[20]	董佩佩, 汪祥燕, 刘元香, 等. 一株凝结芽孢杆菌(Bacillus coagulans)发酵培养基的优化 [J]. 中国酿造, 2018, 37(4):28−32. doi: 10.11882/j.issn.0254-5071.2018.04.006 DONG P P, WANG X Y, LIU Y X, et al. Optimization of fermentation medium of Bacillus coagulans [J]. China Brewing, 2018, 37(4): 28−32.(in Chinese) doi: 10.11882/j.issn.0254-5071.2018.04.006
[21]	MATHAKIYA R A, ROY A, NANDASANA K N, et al. Evaluation of a rapid molecular method for detection of Listeria monocytogenes directly from broth culture[J]. Veterinary World, 2009, 2(5): 177-178.
[22]	何深宏, 程方俊, 罗干, 等. 解淀粉芽孢杆菌高产纤维素酶菌株的筛选与鉴定 [J]. 福建农业学报, 2020, 35(7):781−787. HE S H, CHENG F J, LUO G, et al. Screening and identifying cellulase-producing Bacillus amyloliquefaciens [J]. Fujian Journal of Agricultural Sciences, 2020, 35(7): 781−787.(in Chinese)
[23]	YU D, TONG W, CHEN Y, et al. Improvement of bacterial genomic DNA extraction in clinical specimens [J]. Chinese Journal of Microecology, 2007, 6(19): 519−520.
[24]	王佳楠, 石妍云, 郑力燕, 等. 石油降解菌的分离鉴定及4株芽胞杆菌种间效应 [J]. 环境科学, 2015, 36(6):2245−2251. WANG J N, SHI Y Y, ZHENG L Y, et al. Isolation and identification of petroleum degradation bacteria and interspecific interactions among four Bacillus strains [J]. Environmental Science, 2015, 36(6): 2245−2251.(in Chinese)
[25]	DUNLAP C A, SCHISLER D A, PERRY E B, et al. Bacillus swezeyi sp. nov. and Bacillus haynesii sp. nov., isolated from desert soil [J]. International Journal of Systematic and Evolutionary Microbiology, 2017, 67(8): 2720−2725. doi: 10.1099/ijsem.0.002007
[26]	ADAWAREN E O, DU PLESSIS M, SULEMAN E, et al. The complete mitochondrial genome of Gyps coprotheres (Aves, Accipitridae, Accipitriformes): Phylogenetic analysis of mitogenome among raptors [J]. PeerJ, 2020, 8: e10034. doi: 10.7717/peerj.10034
[27]	QIAN Y, SUN Y, ZHONG L, et al. The GATA-Type Transcriptional Factor Are1 Modulates the Expression of Extracellular Proteases and Cellulases in Trichoderma reesei [J]. International Journal of Molecular Sciences, 2019, 20(17): 4100. doi: 10.3390/ijms20174100
[28]	ELAMARY R, SALEM W M. Optimizing and purifying extracellular amylase from soil bacteria to inhibit clinical biofilm-forming bacteria [J]. PeerJ, 2020, 8: e10288. doi: 10.7717/peerj.10288
[29]	HAJIABADI S, MASHREGHI M, REZA BAHRAMI A, et al. Isolation and molecular identification of cellulolytic bacteria from Dig Rostam hot spring and study of their cellulase activity [J]. BIOCELL, 2020, 44(1): 63−71. doi: 10.32604/biocell.2020.08171
[30]	WU C L, LIU D, YANG X H, et al. Improving production of protease from Pseudoalteromonas sp. CSN423 by random mutagenesis [J]. Marine Biotechnology (New York, N Y), 2016, 18(5): 610−618. doi: 10.1007/s10126-016-9721-9
[31]	曹丹, 彭浩, 兰阿峰, 等. 一株α-淀粉酶产生菌的分离、鉴定及产酶条件研究 [J]. 食品研究与开发, 2020, 41(6):169−174. CAO D, PENG H, LAN A F, et al. Isolation, identification and enzyme production conditions of an α-amylase producing strain [J]. Food Research And Developmen, 2020, 41(6): 169−174.(in Chinese)
[32]	冯红梅, 秦永胜, 李筱帆, 等. 高温纤维素降解菌群筛选及产酶特性 [J]. 环境科学, 2016, 37(4):1546−1552. FENG H M, QIN Y S, LI X F, et al. Screening and enzyme production characteristics of thermophilic cellulase-producing strains [J]. Environmental Science, 2016, 37(4): 1546−1552.(in Chinese)
[33]	附录B（规范性附录）总腐植酸含量的测定方法[J]. 腐植酸, 2016(2): 47−48. Appendix B (normative appendix) determination method of total humic acid content[J]. Humic acid, 2016 (2): 47 − 48. (in Chinese).
[34]	JAIN S, SALUJA B, GUPTA A, et al. Validation of arsenic resistance in Bacillus cereus strain AG27 by comparative protein modeling of arsC gene product [J]. The Protein Journal, 2011, 30(2): 91−101. doi: 10.1007/s10930-011-9305-5
[35]	BHADRA B, RAGHUKUMAR C, PINDI P K, et al. Brevibacterium oceani sp. nov., isolated from deep-sea sediment of the Chagos Trench, Indian Ocean [J]. International Journal of Systematic and Evolutionary Microbiology, 2008, 58(1): 57−60. doi: 10.1099/ijs.0.64869-0
[36]	JEBELI M A, MALEKI A, AMOOZEGAR M A, et al. Bacillus flexus strain As-12, a new arsenic transformer bacterium isolated from contaminated water resources [J]. Chemosphere, 2017, 169: 636−641. doi: 10.1016/j.chemosphere.2016.11.129
[37]	LIU D, LI M, XI B, et al. Metaproteomics reveals major microbial players and their biodegradation functions in a large‐scale aerobic composting plant [J]. Microbial Biotechnology, 2015, 8(6): 950−960. doi: 10.1111/1751-7915.12290
[38]	聂文翰, 戚志萍, 冯海玮, 等. 复合菌剂秸秆堆肥对土壤碳氮含量和酶活性的影响 [J]. 环境科学, 2017, 38(2):783−791. NIE W H, QI Z P, FENG H W, et al. Steaw composts with composite inoculants and theie effects on soil caebon and niteogen contents and enzyme activity [J]. Environmental Science, 2017, 38(2): 783−791.(in Chinese)
[39]	WANG M, MIAO J, WANG X, et al. Genomic and transcriptome analyses of a thermophilic bacterium Geobacillus stearothermophilus B5 isolated from compost reveal its enzymatic basis for lignocellulose degradation [J]. Microorganisms, 2020, 8(9): 1−18.
[40]	王腾浩, 潘岳龙, 沈炜, 等. 蜡样芽孢杆菌与粪肠球菌协同发酵豆粕工艺条件优化 [J]. 饲料研究, 2020, 43(3):74−77. WANG T H, PAN Y L, SHEN W, et al. The optimization of fermentation conditions of soybean meal by using Bacillus Cereus and Enterococcus Faecalis [J]. Feed Research, 2020, 43(3): 74−77.(in Chinese)
[41]	LI Y, CHI M, GE X, et al. Identification of a novel hydrolase encoded by hy-1 from Bacillus amyloliquefaciens for bioremediation of carbendazim contaminated soil and food [J]. International journal of agricultural and biological engineering, 2019, 12(2): 218−224. doi: 10.25165/j.ijabe.20191202.4190
[42]	阚洪媛, 杨世鑫, 孙梁伦, 等. 一株耐铅、锌、铬菌株的分离鉴定及其吸附能力 [J]. 微生物学通报, 2020(12):3974−3986. KAN H Y, YANG S X, SUN L L, et al. Isolation, identification and adsorption capacity of a strain resistant to lead, zinc and chromium [J]. Microbiology China, 2020(12): 3974−3986.(in Chinese)
[43]	潘建华, 刘瑞霞. 蜡状芽孢杆菌Bacillus cereus吸附铅的研究 [J]. 环境科学, 2004, 25(2):166−169. doi: 10.3321/j.issn:0250-3301.2004.02.034 PAN J H, LIU R X. Biosorption of Pb ²⁺ by Bacillus cereus Biomass [J]. Environmental Science, 2004, 25(2): 166−169.(in Chinese) doi: 10.3321/j.issn:0250-3301.2004.02.034
[44]	呼庆, 齐鸿雁, 窦敏娜, 等. 强抗镉蜡状芽孢杆菌的分离鉴定及其抗性机理 [J]. 环境科学, 2007, 28(2):427−430. doi: 10.3321/j.issn:0250-3301.2007.02.038 HU Q, QI H Y, DOU M N, et al. Isolation, molecular characterization and resistance mechanism study on a cadmium hyperresistant Bacillus cereus [J]. Environmental Science, 2007, 28(2): 427−430.(in Chinese) doi: 10.3321/j.issn:0250-3301.2007.02.038
[45]	段海明. 两株蜡状芽孢杆菌对毒死蜱的降解动力学研究 [J]. 中国生态农业学报, 2013, 21(2):207−211. DUAN H M. Kinetics of chlorpyrifos degradation by Bacillus cereus strains [J]. Chinese Journal of Eco-Agriculture, 2013, 21(2): 207−211.(in Chinese)
[46]	LU M, ZHANG Z Z. Phytoremediation of soil co-contaminated with heavy metals and deca-BDE by co-planting of Sedum alfredii with tall fescue associated with Bacillus cereus JP12 [J]. Plant and Soil, 2014, 382(1/2): 89−102.
[47]	NAYAK A K, PANDA S S, BASU A, et al. Enhancement of toxic Cr (VI), Fe, and other heavy metals phytoremediation by the synergistic combination of native Bacillus cereus strain and Vetiveria zizanioides L [J]. International Journal of Phytoremediation, 2018, 20(7): 682−691. doi: 10.1080/15226514.2017.1413332
[48]	XIONG Y W, LI X W, WANG T T, et al. Root exudates-driven rhizosphere recruitment of the plant growth-promoting rhizobacterium Bacillus flexus KLBMP 4941 and its growth-promoting effect on the coastal halophyte Limonium sinense under salt stress [J]. Ecotoxicology and Environmental Safety, 2020, 194: 110374. doi: 10.1016/j.ecoenv.2020.110374
[49]	KUMAR A, PRIYADARSHINEE R, SINGHA S, et al. Biodegradation of alkali lignin by Bacillus flexus RMWW II: Analyzing performance for abatement of rice mill wastewater [J]. Water Science and Technology, 2019, 80(9): 1623−1632. doi: 10.2166/wst.2020.005
[50]	MOHANTY S S, KUMAR A. Response surface methodology mediated optimization of Indanthrene Blue RS by a novel isolated bacterial strain Bacillus flexus TS8 [J]. Water Environment Research, 2020, 92(4): 569−578. doi: 10.1002/wer.1246
[51]	REDA F M, HASSANEIN W A, MOABED S, et al. Potential exploitation of Bacillus flexus biofilm against the cowpea weevil, Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) [J]. Egyptian Journal of Biological Pest Control, 2020, 30(1): 1−7. doi: 10.1186/s41938-020-0205-x
[52]	NAM J H, BAE W, LEE D H. Oceanobacillus caeni sp. nov., isolated from a Bacillus-dominated wastewater treatment system in Korea [J]. International Journal of Systematic and Evolutionary Microbiology, 2008, 58: 1109−1113. doi: 10.1099/ijs.0.65335-0