CN115369042A - 一株枝顶孢菌320及其应用 - Google Patents
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Abstract
本发明属于微生物促进植物生长领域，本申请提供了一株枝顶孢菌(Acremonium strictum)320，其保藏编号为CGMCC No.21935，该菌株已于2021年3月25日保藏于中国微生物菌种保藏管理委员会普通微生物中心，其简称CGMCC，其地址为北京市朝阳区北辰西路1号院3号，其保藏号为CGMCC No.21935。本申请的菌株能促进植物生长，植株鲜重、根重、株高、茎粗都显著增加，叶片的叶绿素含量也明显增加。
Description
技术领域
本发明属于微生物促进植物生长领域。
背景技术
目前各类化肥农药的施加依然是农作物增产保收的主要手段，但其产的不良后果是破坏了生态环境系统，严重威胁着人类健康。降低化肥农药施用已刻不容缓，这也是当今绿色农业发展必经之路。利用有益微生物促进作物生长是化肥农药减施增效的有效途径之一。已有研究表明促生微生物能够通过改善土壤结构、分解有机质和矿物质、固氮、解磷、解钾、降解重金属等多种方式促进植物生长发育和抵御环境胁迫。
发明内容
为了解决本申请中技术问题，本申请提供了一株枝顶孢菌(Acremoniumstrictum)320，其保藏编号为CGMCC No.21935，该菌株已于2021年3月25日保藏于中国微生物菌种保藏管理委员会普通微生物中心，其简称CGMCC，其地址为北京市朝阳区北辰西路1号院3号，其保藏号为CGMCC No.21935。
本申请的菌株能促进植物生长，植株鲜重、根重、株高、茎粗都显著增加，叶片的叶绿素含量也明显增加。
附图说明
图1是枝顶孢菌320的菌落形态图。
图2是枝顶孢菌320促进拟南芥侧根生长图。
图3是枝顶孢菌320代谢产物促进小白菜生长图。
图4是枝顶孢菌320代谢产物促进普通烟草生长图。
图5直立枝顶孢霉代谢产物促进本氏烟生长图。
具体实施方式
实施例1
菌种获得
1)该菌株是我们进行植物组织培养时从培养基上采集，并在马铃薯琼脂培养基PDA(马铃薯200g，葡萄糖20g，琼脂20g)上分离。
2)菌株鉴定
首先提取了真菌的DNA，通过真菌通用引物ITS1(TCCGTAGGTGAACCTGCGG)和ITS4(TCCTCCGCTTATTGATATGC)进行PCR扩增，对PCR产物进行核苷酸序列测定，如序列表中SEQID NO.1所示,所获得的序列在NCBI(https://blast.ncbi.nlm.nih.gov/Blast.cgi)上进行比对，发现它与枝顶孢属(Acremonium sp.)G4同源性高达99.66％。
单菌落培养后观察，该菌株菌落颜色呈淡橘黄色，表面微微湿润，菌丝呈分枝状，菌丝中间有隔，其孢子呈椭圆形直径约为1.5μm，长约4μm(图1)。通过18S rRNA序列形态学分析，鉴定其分类命名Acremonium strictum，中文对应名称为枝顶孢菌，命名为320，在本专利中简称为菌株320。枝顶孢菌(Acremonium strictum)320已于2021年3月25日保藏于中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC，地址：北京市朝阳区北辰西路1号院3号)，保藏号为CGMCC No.21935。
实施例2
菌株功能鉴定：通过和植物共培养以及利用其代谢产物浇灌证明该菌具有显著促进植物生长的特性。
将萌发后的拟南芥幼苗移到1/2MS固体培养基(MS盐4.3g/L，蔗糖10g/L，琼脂粉7.5g/L，pH 5.8)上，在培养皿下部接种枝顶孢菌(Acremonium strictum)320，放置到23℃培养箱中培养。共培养15天后发现，枝顶孢菌处理后的拟南芥侧根和叶片生长旺盛，侧根长度增加，约为对照的2倍；叶片明显变大、变绿(图2)。
实施例3
将在固体培养基中培养好的枝顶孢菌(Acremonium strictum)320菌落挑取到MS液体培养基(MS盐4.3g/L，蔗糖10g/L，pH 5.8)中，放置28℃恒温摇床中，200rpm振荡培养。培养7天的液体培养物经8层纱布过滤去除菌丝体后，6000rpm离心5min收集培养液，通过0.45μm微孔滤膜过滤，收集代谢物。将真菌的代谢物直接浇灌于培养在土壤中的小白菜(Brassica campestris L.ssp.chinensis Makino)中，每隔一周浇灌一次。自第一次浇灌开始，每隔一段时间拍照记录它们的生长变化。以MS液体培养基为对照。
结果表明，与对照相比，浇灌代谢产物的小白菜植株生长更为旺盛。代谢物处理10天后小白菜的叶片明显增大。代谢物处理25天后的小白菜植株生长显著增加，植株重量和叶片面积约为对照的2倍。(图3)。
实施例4
将在固体培养基中培养好的枝顶孢菌(Acremonium strictum)320菌落挑取到MS液体培养基中，放置28℃恒温摇床中，200rpm振荡培养。培养7天的液体培养物经8层纱布过滤去除菌丝体后，6000rpm离心5min收集培养液，通过0.45μm微孔滤膜过滤，收集代谢物。将真菌的代谢物直接浇灌于培养在土壤中的本氏烟草(Nicotiana benthamiana)中，每隔一周浇灌一次。自第一次浇灌开始，每隔一段时间拍照记录它们的生长变化。以MS液体培养基为对照。
结果表明，与对照相比，浇灌代谢产物的本氏烟草植株生长更为旺盛，植株鲜重、根重、株高、茎粗都显著增加，处理25天其中植株重量和根重量大约是对照的2.5倍，植株高度和茎粗度大约是对照的1.5倍。叶片的叶绿素含量也明显增加(图4)。
实施例5
将在固体培养基中培养好的枝顶孢菌(Acremonium strictum)320菌落挑取到MS液体培养基中，放置28℃恒温摇床中，200rpm振荡培养。培养7天的液体培养物经8层纱布过滤去除菌丝体后，6000rpm离心5min收集培养液，通过0.45μm微孔滤膜过滤，收集代谢物。将真菌的代谢物直接浇灌于培养在土壤中的普通烟草(N.tabacum L.)中，每隔一周浇灌一次。自第一次浇灌开始，每隔一段时间拍照记录它们的生长变化。以MS液体培养基为对照。
结果表明，外施直立枝顶孢菌代谢物的烟草植株重量和根重量大约是对照的2.5倍，植株高度约为对照的2倍，茎粗度约为对照的1.5倍。叶片的叶绿素含量也显著增加(图5)。
序列表
<110> 中国科学院微生物研究所
<120> 一株枝顶孢菌320及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 580
<212> DNA
<213> 枝顶孢菌(Acremonium strictum)
<400> 1
ctccgtaggt gaacctgcgg agggatcatt acaaaacgag tgcttcggct ctccaaccca 60
ttgtgaacat acctatcgtt ccctcggcgg gctcagcgcg cggccggcct ccgggccacc 120
gggcgtccgc cggggacaac caaacccgaa tttttcgtgt atatctgagg ggcgaaagcc 180
cgaaaacaaa atgaatcaaa actttcaaca acggatctct tggctctggc atcgatgaag 240
aacgcagcga aatgcgataa gtaatgtgaa ttgcagaatt cagtgaatca tcgaatcttt 300
gaacgcacat tgcgcccgcc ggcactccgg cgggcatgcc tgtccgagcg tcatttcaac 360
cctcagggcc ccctttcggg ggcggcacct ggttctgggg atcagcggcc cttcggggcc 420
cctgtccccc aaattgagtg gcggtcgcgc cgcagcctcc cctgcgtagt agcacacctc 480
gcaccggaga gcggctcggc cacgccgtaa aacccccaat ttttacaggt tgacctcgga 540
tcaggtagga atacccgctg aacttaagca tatcaataag 598
Claims (5)
1.一株枝顶孢菌(Acremonium strictum)320，其保藏编号为CGMCC No.21935。
2.根据权利要求1所述枝顶孢菌320及其代谢产物在制作促进植物生长助剂中的应用。
3.根据权利要求2所述的应用，其特征在于，所述的植物包括拟南芥、小白菜和烟草植物。
4.根据权利要求3所述的应用，其特征在于，所述的烟草植物包括本氏烟草植物和普通烟草植物。
5.根据权利要求2所述的应用，其特征在于，所述的助剂包括肥料。
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19990026602A (ko) * | 1997-09-25 | 1999-04-15 | 서주원 | 신규의 Acremonium sp. MJN1 균주와 그추출물의 무공해 식물성장 촉진제로서의 용도 |
| CN102363748A (zh) * | 2011-09-30 | 2012-02-29 | 中国科学院南海海洋研究所 | 一种高效纤维素降解真菌新种Acremonium sp.DPZ-SYz-2-3及其应用 |
| US20160262400A1 (en) * | 2013-11-08 | 2016-09-15 | Symborg S.L. | A bacillus subtilis strain deposited under deposit number cect 8258 and method for protecting or treating plants |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19990026602A (ko) * | 1997-09-25 | 1999-04-15 | 서주원 | 신규의 Acremonium sp. MJN1 균주와 그추출물의 무공해 식물성장 촉진제로서의 용도 |
| CN102363748A (zh) * | 2011-09-30 | 2012-02-29 | 中国科学院南海海洋研究所 | 一种高效纤维素降解真菌新种Acremonium sp.DPZ-SYz-2-3及其应用 |
| US20160262400A1 (en) * | 2013-11-08 | 2016-09-15 | Symborg S.L. | A bacillus subtilis strain deposited under deposit number cect 8258 and method for protecting or treating plants |
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