CN112210509B - 一种郭霍氏细胞杆菌菌株及其用途 - Google Patents
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Abstract
本发明涉及一种郭霍氏细胞杆菌菌株及其应用，所述的菌株郭霍氏细胞杆菌(Cytobacillus kochii)，保藏编号为CGMCC No.20451，菌株的16s rDNA碱基序列如SEQ ID NO：1所示。本发明还涉及将所述菌株用于种植的方法，具体步骤为：(1)发酵培养基配方；(2)发酵方法(包括发酵参数和方法)；(3)发酵液种植应用方法；本发明的有益效果是通过菌株在秸秆中快速繁殖并占据微生态区位优势，抑制致病菌繁殖，避免有害代谢产物积累，加速秸秆降解。
Description
技术领域
本发明涉及一种菌株及其应用，具体涉及一种郭霍氏细胞杆菌菌株及其用途。
背景技术
我国是粮食生产大国，也是秸秆资源大国。秸秆处理是农业生产上的一道难题，以往普遍实行的秸秆直接还田并未能产生预期效应：由于直接还田秸秆分解时间短，腐解不彻底，直接影响下一茬作物的根系生长，造成大量死苗，增加播种量；更重要的是秸秆中大量病原菌和虫卵进入土壤后带来的病虫害问题，不仅加重了农民管理成本，还使农作物普遍减产，降低了生产效益。
秸秆作为农业废弃物，是农业生产的“另一半”，是“放错了地方的资源”，用则利、弃则害。但将秸秆进行回收并资源化利用的手段大多成本高昂，不能产生经济效益，需要依赖政府财政补贴，增加了政府负担。而将秸秆添加腐熟剂就地还田的方式又因为传统腐熟剂中微生物菌种功能的局限性而需要较长的分解时间，因此开发分解能力强，对植物友好的微生物腐熟剂迫在眉睫。
发明内容
本发明的目的之一在于解决上述现有技术的不足，提供一种郭霍氏细胞杆菌。
本发明的目的之二在于解决上述现有技术的不足，提供一种郭霍氏细胞杆菌的用途，将郭霍氏细胞杆菌菌株(CK-006)施至翻耕到地下的秸秆中，所述菌株利用秸秆中养分，快速繁殖并占据微生态区位优势，抑制致病菌繁殖，避免有害代谢产物积累，加速秸秆降解，从而达到绿色种植的目的。
为实现上述目的，本发明采用以下技术方案：
一种郭霍氏细胞杆菌(Cytobacillus kochii)菌株，菌株代码为CK-006，保藏编号为CGMCC No.20451，保藏单位为中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，保藏单位地址为：北京市朝阳区北辰西路1号院3号中国科学院微生物研究所，保藏日期为2020年07月27日。
所述菌株经生理生化及16rDNA序列测定，菌株CK-006为郭霍氏细胞杆菌(Cytobacillus kochii)，菌株的16s rDNA碱基序列如SEQ ID NO：1所示。
本发明还公开了所述郭霍氏细胞杆菌菌株应用于种植业的方法，包括以下内容：
1.配制培养基：蔗糖2％(g/ml)，酵母粉1％(g/ml)，蛋白胨1％(g/ml)，氯化钠0.5％(g/ml)，水补足，灭菌后备用；
2.液体深层发酵培养：将所述菌株单菌落接入上述培养基中，120r/min震荡培养，发酵温度为37℃，发酵时间36h，测得发酵液OD值为7；
3.降解秸秆应用：需要将所述郭霍氏细胞杆菌发酵液与粉碎的秸秆以及速效氮肥一起翻耕，施入土壤，其中郭霍氏细胞杆菌发酵液每亩土地使用量为12.5kg，速效氮肥使用量为秸秆干重的6％。
本发明的优点是：菌株CK-006是环境微生物，适应能力强，安全性好，能够在秸秆中快速繁殖并占据微生态区位优势，广泛抑制致病菌繁殖，避免有害代谢产物积累，快速降解秸秆,从而达到绿色种植养殖的目的，本发明方法简单易行，效果稳定，绿色环保。
附图说明
图1为：郭霍氏细胞杆菌CGMCC No.20451菌株的小麦秸秆降解效果图。
具体实施方式
实施例1：
一种郭霍氏细胞杆菌CGMCC No.20451菌株的应用：
a.配制培养基：蔗糖2％(g/ml)，酵母粉1％(g/ml)，蛋白胨1％(g/ml)，氯化钠0.5％(g/ml)，水补足，高压灭菌后备用；
b.液体深层发酵培养：将所述菌株单菌落接入上述培养基中，120r/min震荡培养，发酵温度为37℃，发酵时间36h，测得发酵液OD值为7；
c.2020年，于浙江省杭州市富阳区中国水稻研究所标准种植地块进行小麦秸秆降解试验，将郭霍氏细胞杆菌发酵液(每亩12.5kg)与粉碎的小麦秸秆(温麦10号，每亩干重300kg,平均长度10cm左右)以及速效氮肥(尿素18kg)一起翻耕，施入土壤。于地下10-15cm处埋一纱网布包，内装小麦茎秆第四节间100根，灌水后一周，于水稻插秧前，收集预先埋好纱网布包中小麦茎秆，清水清洗自然风干(含水率＜55％)后，微控电子万能试验机进行抗拉强度试验，检测秸秆降解情况；
d.实验结果如附图1所示。从图1上可明显看出郭霍氏细胞杆菌CGMCC No.20451菌株处理组小麦秸秆抗拉强度出现显著下降，对照组平均值为18.9MPa，处理组为11.5MPa，抗拉强度下降了39.2％。
以上所述的实施例只是本发明的一种较佳的方案，并非对本发明作任何形式上的限制，在不超出权利要求所记载的技术方案的前提下还有其它的变体及改型。
序列表
<110> 森井生物工程（湖州）有限公司
<120> 一种郭霍氏细胞杆菌及其用途
<130> 2020
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1413
<212> DNA
<213> 郭霍氏细胞杆菌(Cytobacillus kochii)
<400> 1
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agtaacacgt gggcaacctg cctgtaagac tgggataact ccgggaaacc ggggctaata 120
ccggataata tctatttata catataatta gattgaaaga tggttctgct atcacttaca 180
gatgggcccg cggcgcatta gctagttggt gaggtaacgg ctcaccaagg cgacgatgcg 240
tagccgacct gagagggtga tcggccacac tgggactgag acacggccca gactcctacg 300
ggaggcagca gtagggaatc ttccgcaatg gacgaaagtc tgacggagca acgccgcgtg 360
agtgatgaag gttttcggat cgtaaaactc tgttgttagg gaagaacaag tatcggagta 420
actgccggta ccttgacggt acctaaccag aaagccacgg ctaactacgt gccagcagcc 480
gcggtaatac gtaggtggca agcgttgtcc ggaattattg ggcgtaaagc gcgcgcaggc 540
ggttccttaa gtctgatgtg aaagcccacg gctcaaccgt ggagggtcat tggaaactgg 600
ggaacttgag tgcagaagag gaaagtggaa ttccaagtgt agcggtgaaa tgcgtagaga 660
tttggaggaa caccagtggc gaaggcgact ttctggtctg taactgacgc tgaggcgcga 720
aagcgtgggg agcaaacagg attagatacc ctggtagtcc acgccgtaaa cgatgagtgc 780
taagtgttag agggtttccg ccctttagtg ctgcagcaaa cgcattaagc actccgcctg 840
gggagtacga ccgcaaggtt gaaactcaaa ggaattgacg ggggcccgca caagcggtgg 900
agcatgtggt ttaattcgaa gcaacgcgaa gaaccttacc aggtcttgac atcctctgac 960
aatcctagag ataggacttt ccccttcggg ggacagagtg acaggtggtg catggttgtc 1020
gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgatctta 1080
gttgccagca tttagttggg cactctaagg tgactgccgg tgacaaaccg gaggaaggtg 1140
gggatgacgt caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggat 1200
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gccgcggtga atacgttccc gggccttgta cacaccgccc gtcacaccac gagagtttgt 1380
aacacccgaa gtcggtgggg taaccttttg gac 1413
Claims (4)
1.一种郭霍氏细胞杆菌(Cytobacilluskochii)菌株，菌株代码为CK-006，其特征在于，保藏编号为CGMCC No.20451，保藏单位为中国微生物菌种保藏管理委员会普通微生物中心，保藏单位地址为：北京市朝阳区北辰西路1号院3号中国科学院微生物研究所，保藏日期为2020年07月27日。
2.如权利要求1所述的郭霍氏细胞杆菌菌株在降解秸秆中的应用。
3.如权利要求2所述的应用，其特征在于，将郭霍氏细胞杆菌的发酵液与粉碎的秸秆以及速效氮肥一起翻耕，施入土壤，其中发酵液每亩土地使用量为12.5kg，速效氮肥使用量为秸秆干重的6％。
4.如权利要求3所述的应用，其特征在于，所述郭霍氏细胞杆菌的发酵液的培养方法为：
(1)培养基配方：蔗糖质量浓度2％，酵母粉质量浓度1％，蛋白胨质量浓度1％，氯化钠质量浓度0.5％，水补足，灭菌备用；
(2)液体深层发酵培养：将所述菌株单菌落接入上述培养基中，120r/min振荡培养，发酵温度为37℃，发酵时间36h，测得发酵液OD值为7。
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| CN110167350A (zh) * | 2016-08-30 | 2019-08-23 | 阿坤纳斯公司 | 确定的微生物组合物 |
| CN110951650A (zh) * | 2019-12-27 | 2020-04-03 | 原起科技(杭州)有限公司 | 柯赫氏芽孢杆菌及其用途 |
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| A phylogenomic and comparative genomic framework for resolving the polyphyly of the genus Bacillus: Proposal for six new genera of Bacillus species, Peribacillus gen. nov., ......, Metabacillus gen. nov. and Alkalihalobacillus gen. nov.;Sudip Patel 等;《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY》;20200203;第70卷(第1期);参见全文 * |
| Cytobacillus kochii strain CK-006 16S ribosomal RNA gene, partial sequence，Genbank登录号：MT798822.1;Kancheng, L.;《Genbank》;20200731;参见序列表 * |
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