CN107384840B - 一株抗旱促生复合菌剂及其应用 - Google Patents
一株抗旱促生复合菌剂及其应用 Download PDFInfo
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- CN107384840B CN107384840B CN201710800030.4A CN201710800030A CN107384840B CN 107384840 B CN107384840 B CN 107384840B CN 201710800030 A CN201710800030 A CN 201710800030A CN 107384840 B CN107384840 B CN 107384840B
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Abstract
本发明提供了一种抗旱促生复合菌剂及其应用,该复合菌剂含有保藏编号为CGMCC No.14465的枯草芽孢杆菌和保藏编号为CGMCC No.14466的贪噬菌。本发明的枯草芽孢杆菌有较强的产保水剂聚谷氨酸的能力,聚谷氨酸产量可达6.59g/L;本发明的贪噬菌有较强的生产抗逆促生的ACC酶活性,其酶活可达2.5μM/mg/h。在低中度干旱条件下,使用本发明的复合菌剂能使玉米植株生物量提高50.8%。本发明的复合菌剂适用于干旱退化土壤的修复,可提高土壤的持水性能和水源涵养功能,促进植被抗旱及生长,可为农业节水提供有效措施。
Description
技术领域
本发明属于微生物学领域,具体地说,涉及一种抗旱促生复合菌剂及其应用。
背景技术
我国是水资源严重短缺的国家,是世界上21个贫水和最缺水的国家之一,有资料显示,因干旱损失的粮食占各种自然灾害造成粮食损失的60%以上。微生物土壤修复技术与物理、化学技术相比,具有廉价、高效、不易产生二次污染等特点。因此开发抗旱的微生物菌剂,通过施加菌剂改善土壤的理化性质及植物的抗逆性、促进植物生长,对于大面积干旱退化土壤的修复、提高水源涵养、治理水土流失、发展高效节水农业具有重要意义。
微生物分泌的聚谷氨酸具有良好的保水性,已有研究表明特制的聚谷氨酸的吸水率为化学常用的保水剂聚丙烯酸的2.85倍,其具有较强的保水性,可使水分在土壤中缓慢释放,提高土壤水分的有效性。聚谷氨酸同时具有减渗作用,改变土壤剖面含水率的分布情况,使水分集中在作物根部。因此开发高效的分泌聚谷氨酸的微生物,具有重要的实用价值。
微生物可合成一些活性酶,提高植物的抗逆性。植物在高温、干旱、洪涝、盐胁迫及重金属污染等逆境条件下会加速乙烯的合成,而持续高浓度的乙烯会抑制植物生长。截至目前已经发现有20多种属根际细菌具有植物促生特性,其中很多可产生ACC脱氨酶。ACC脱氨酶是一种有效降低逆境乙烯含量的外源促生物质,可以将植物乙烯前体ACC(1-氨基环丙烷-1-羧酸)水解为α-酮丁酸和氨,从而抑制植物体内乙烯合成,降低其含量进而减少过量乙烯对植物生长发育造成的不利影响。
目前的研究主要是集中在聚谷氨酸产生菌的保水性,产ACC脱氨酶菌的抗盐碱能力、抗重金属及促进小麦、豌豆等生长方面。对于这两类菌能否复合应用,如何应用才能提高其效果,未见报道。因此创制具有抗旱促生功能的复合微生物菌剂,研究菌剂的生态效应和应用技术,是实现土壤的生态修复、节水农业的有效措施。
发明内容
本发明的目的是提供一种抗旱促生复合菌剂及其应用。
本发明提供的一种抗旱促生复合菌剂含有保藏编号为CGMCC No.14465的枯草芽孢杆菌和保藏编号为CGMCC No.14466的贪噬菌。
本发明提供的枯草芽孢杆菌(Bacillus subtilis),它的菌株编号为P1,该菌株已于2017年7月27日保藏在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,中国科学院微生物研究所,邮编100101),分类命名为枯草芽孢杆菌(Bacillus subtilis),保藏编号为CGMCC No.14465。
本发明提供的贪噬菌(Variovorax sp.),它的菌株编号为G1,该菌株已于2017年7月27日保藏在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,中国科学院微生物研究所,邮编100101),分类命名为贪噬菌(Variovorax sp.),保藏编号为CGMCC No.14466。
枯草芽孢杆菌CGMCC No.14465的16s rRNA序列如SEQ ID NO.3所示,革兰氏染色为阳性;贪噬菌CGMCC No.14466的16s rRNA序列如SEQ ID NO.4,革兰氏染色为阳性。
进一步地,本发明的抗旱促生复合菌剂中CGMCC No.14465的枯草芽孢杆菌和保藏编号为CGMCC No.14466的贪噬菌的有效活菌数均为108~109cfu/mL。且P1用量是G1用量的2倍时,抗旱促生效果最佳。
所述复合菌剂还可包括载体。所述载体可为固体载体或液体载体。所述固体载体为矿物材料、植物材料或高分子化合物;所述矿物材料为粘土、滑石、高岭土、蒙脱石、白碳、沸石、硅石和硅藻土中的至少一种;所述植物材料为玉米粉、豆粉和淀粉中的至少一种;所述高分子化合物为聚乙烯醇和/或聚二醇。所述液体载体可为植物油、矿物油或水;所述有机溶剂为癸烷和/或十二烷。所述菌剂中,所述活性成分可以以被培养的活细胞、活细胞的发酵液、细胞培养物的滤液或细胞与滤液的混合物的形式存在。所述菌剂的剂型可为多种剂型,如液剂、悬浮剂、粉剂、颗粒剂、可湿性粉剂或水分散粒剂。根据需要,所述复合菌剂中还可添加表面活性剂(如吐温20、吐温80等)、粘合剂、稳定剂(如抗氧化剂)、pH调节剂等。
本发明提供了上述抗旱促生复合菌剂在促进植物生长中的应用。
本发明提供了上述抗旱促生复合菌剂在提高土壤保水性和/或修复植被中的应用。
本发明提供了上述抗旱促生复合菌剂在提高植物抗旱能力中的应用。
因此,所述复合菌剂可为下述A1)、A2)或A3):
A1)抗旱和促生的菌剂;
A2)抗旱的菌剂;
A3)促生的菌剂。
所述枯草芽孢杆菌(Bacillus subtilis)P1 CGMCC No.14465与贪噬菌(Variovorax sp.)G1 CGMCC No.14465在制备B1)、B2)或B3)菌剂中的应用也属于本发明的保护范围:
B1)抗旱和促生的菌剂;
B2)抗旱的菌剂;
B3)促生的菌剂。
上述菌剂在干旱及轻中度干旱土壤中的应用,或在植被修复以及植被促生方面的应用也属于本发明的保护范围。
上述应用中,菌剂的保水及促生性能可用于干旱土壤及植被的修复,促进植物抗旱及生长。所述贪噬菌(Variovorax sp.)G1 CGMCC No.14466,可使玉米发芽时间缩短24h(本发明G1发酵液对玉米浸种后,玉米发芽时间是48h,而未施加G1发酵液的玉米发芽时间72h),G1可使玉米种子发芽率达到97%,提高5%。所述枯草芽孢杆菌(Bacillus subtilis)P1 CGMCC No.14465与贪噬菌(Variovorax sp.)G1CGMCC No.14466耦合应用,与未施加菌剂相比可使玉米植株的生物量较未使用复合菌剂的对照组提高50.8%。所述枯草芽孢杆菌(Bacillus subtilis)P1 CGMCC No.14465与贪噬菌(Variovorax sp.)G1CGMCC No.14466可在用于培养枯草芽孢杆菌和贪噬菌的培养基中培养。其适宜培养条件为:pH 7.0,温度20-40℃。
本发明枯草芽孢杆菌(Bacillus subtilis)P1 CGMCC No.14465产聚谷氨酸的发酵培养基为:葡萄糖20g·L-1,酵母粉1g·L-1,谷氨酸钠18g·L-1,(NH4)2SO4 1g·L-1,CaCl20.173g·L-1,K2HPO4 0.5g·L-1,MgSO4 0.1g·L-1。
本发明贪噬菌(Variovorax sp.)G1 CGMCC No.14466产ACC脱氨酶的富集培养基为TSB培养基:胰蛋白胨15g·L-1,大豆蛋白胨5g·L-1,NaCl 5g·L-1,H2O,pH值7.2。
本发明贪噬菌(Variovorax sp.)G1 CGMCC No.14466产ACC脱氨酶的分离培养基为DF培养基:KH2PO4 4g·L-1,Na2HPO4 6g·L-1,MgSO4·7H2O 0.2g·L-1,葡萄糖2g·L-1,葡萄糖酸钠2g·L-1,柠檬酸2g·L-1,组分1、组分2溶液各0.1mL(组分1:H3BO3 10mg,MnSO4·H2O 11.19mg,ZnSO4·7H2O 124.6mg,CuSO4·5H2O 78.22mg,MoO3 10mg溶于100mL灭菌蒸馏水中,4℃保存。组分2:FeSO4·7H2O 100mg溶于10mL灭菌蒸馏水中,4℃保存),筛选产ACC脱氨酶的ADF培养基:超纯水溶解ACC,细菌过滤器(d=0.22μm)抽滤灭菌,加到不含(NH4)2SO4且预先灭菌的DF培养基中,ACC最终浓度为3.0mM·L-1。
本发明的枯草芽孢杆菌(Bacillus subtilis)P1 CGMCC No.14465有较强的产保水剂(聚谷氨酸)的能力,本发明贪噬菌(Variovorax sp.)G1 CGMCC No.14466,具有较强的分泌抗逆活性酶(ACC脱氨酶)的能力。本发明的上述两种菌制成的复合菌剂适用于干旱和轻中度干旱地区土壤的保水、植被的修复及促生,可以有效提高生态涵养区的生态保护和水源涵养功能,促进高效节水农业的发展,具有重要意义。本发明解决了施用化学保水剂费用高且容易造成二次污染的问题,降低了生产和使用成本,对于保护生态环境及人类健康具有重要的意义。
附图说明
图1为枯草芽孢杆菌P1与贪噬菌G1的16S rRNA的系统发育树。
图2为聚谷氨酸水解液的紫外扫描峰图。
图3为贪噬菌G1接种在ADF平板上,指示该菌株可利用ACC为唯一氮源。
图4为在40℃恒温蒸发条件下枯草芽孢杆菌(Bacillus subtilis)P1产聚谷氨酸发酵液对土壤失水率的影响。“水”表示空白对照;“培养基”表示未接种枯草芽孢杆菌P1的培养基溶液,“P1”表示接种枯草芽孢杆菌P1产聚谷氨酸的发酵液。
图5为枯草芽孢杆菌P1与贪噬菌G1制成的复合菌剂对玉米幼苗抗旱的作用。
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。在不背离本发明精神和实质的情况下,对本发明方法、步骤或条件所作的修改或替换,均属于本发明的范围。
若未特别指明,实施例中所用的化学试剂均为常规市售试剂,实施例中所用的技术手段为本领域技术人员所熟知的常规手段。
实施例1、枯草芽孢杆菌P1和贪噬菌G1的分离和鉴定
一、枯草芽孢杆菌的分离
1、初筛取2g豆豉,放置于10ml无菌水中,振荡2min,再静置30min,然后放在水浴锅中,100℃维持5min。冷却后吸取上清液1mL,采用平板培养法进行系列稀释涂布于LB培养基上,37℃培养24h,挑取生长迅速并且能够拉丝的单菌落,分离纯化后,用LB试管斜面培养基,4℃保存。
2、复筛将初筛得到的菌株接种到种子LB培养基中,37℃,150r·min-1摇瓶20h,然后按2%的接种量接种到摇瓶发酵培养基中,37℃,200r·min-1培养48h,发酵液用黏度计测量表观黏度,选择黏度较大的菌株作为复筛菌株,将分离纯化所得的一株具有明显拉丝的菌株命名为菌株P1。
二、贪噬菌G1的分离
称取玉米根际土壤样品10g,悬浮于90mL TSB培养液中,28℃,200r·min-1振荡培养24h后,吸取1mL悬浮液至50mL TSB培养液中,同条件培养24h后,吸取1mL悬浮液至50mLDF培养基中,相同条件下培养24h后,吸取1mL悬浮液至50mL ADF培养基中,相同条件下培养48h。将ADF培养液制成10-1-10-6稀释液,选取合适稀释度在ADF固体平板培养基上涂布,28℃培养48h。选取不同形态的单菌落于ADF平板上反复划线纯化,获得产ACC脱氨酶菌株G1。
三、菌株生理生化指标分析
对枯草芽孢杆菌(Bacillus subtilis)P1与贪噬菌(Variovorax sp.)G1进行革兰氏染色和碳氮源利用情况分析。结果表明菌株P1为革兰氏染色阳性,利用葡萄糖,蔗糖,山梨醇,天门冬氨酸,L-苹果酸,甲酸;菌株G1为革兰氏染色阳性,利用葡萄糖,果糖,半乳糖,山梨醇,葡萄糖醛,葡萄糖酸,天门冬氨酸,L-苹果酸,α-酮戊二酸(表1)。
表1菌株P1与菌株G1的生理生化试验结果
注:“+”表示反应为阳性;“-”表示反应为阴性。
四、菌株的鉴定
利用PCR对菌株P1与菌株G1的16s rRNA进行扩增,上游引物为27F(序列如SEQ IDNO:1所示),下游引物为1492R(序列如SEQ ID NO:2所示),扩增条件为95℃,5min;95℃,1min,50℃,1min,72℃,2min,25个循环;72℃,10min。菌株P1的16s rRNA的DNA序列如SEQID NO.3 7所示。菌株G1的16s rRNA的DNA序列如SEQ ID NO.4所示。将该序列在Genbank数据库进行BLAST(网址:http://blast.ncbi.nlm.nih.gov/Blast.cgi)比对,结果菌株P1的序列与Bacillus subtilis subsp.subtilis str.168(NC000964)的相似性为99%。根据菌株P1的16s rRNA序列同源性分析结果,将菌株P1鉴定为枯草芽孢杆菌(Bacillussubtilis)。菌株G1的序列与贪噬菌(Variovorax paradoxus)菌株的相似性为99%。根据菌株G1的16s rRNA序列同源性分析结果,将菌株G1鉴定为贪噬菌属(Variovorax sp.)。菌株P1与菌株G1的系统发育树如图1所示。
枯草芽孢杆菌(Bacillus subtilis)P1于2017年7月27日保藏在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,中国科学院微生物研究所,邮编100101),分类命名为枯草芽孢杆菌(Bacillus subtilis),保藏编号为CGMCC No.14465。
贪噬菌(Variovorax sp.)G1于2017年7月27日保藏在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,中国科学院微生物研究所,邮编100101),分类命名为贪噬菌(Variovorax sp.),保藏编号为CGMCCNo.14466。
实施例2枯草芽孢杆菌P1产聚谷氨酸(γ-PGA)的试验
一、菌株P1产聚谷氨酸的提取纯化
1.初步纯化聚谷氨酸(γ-PGA)
将菌株P1的发酵液8000r·min-1离心15min,去除菌体,取上清液用HCl调pH至3-4,加3-4倍的无水乙醇,低温静置过夜。去上清液得粗品,透析脱盐过夜。将透析后的样品进行冷冻干燥,得到纯化粗品。
二、聚谷氨酸产物的分析鉴定
1.聚谷氨酸的水解
取0.05g纯化样品,放入水解管中,加入10mL 6mol·L-1的HCl,通入氮气,维持5min后封口,120℃水解12h,冷却后过滤至小烧杯中备用。
2.紫外扫描光谱分析
将水解后的溶液稀释适当倍数,在190-600nm的波长下进行紫外扫描,与聚谷氨酸标准品进行对比(图2)。样品水解液与聚谷氨酸标准品在264nm处均出现吸收峰,因此可以确定为发酵产物由谷氨酸聚合而成。此外在218nm处出现吸收峰,据资料显示聚谷氨酸的吸收峰为216nm,由紫外扫描的光谱图鉴定菌株P1的产物为聚谷氨酸。
三、菌株P1聚谷氨酸产量的测定
对菌株P1发酵液用黏度计测量表观黏度同时对其产量进行测定,结果表明菌株P1的聚谷氨酸产量为6.59g·L-1。
表2菌株P1产聚谷氨酸发酵液粘度及产量
实施例3贪噬菌G1产ACC脱氨酶的试验
一、菌株G1产ACC脱氨酶的鉴定及产量测试
1.菌株G1产ACC脱氨酶的鉴定
菌株G1可以在ADF平板上生长,表明菌株G1可利用ACC为唯一氮源(图3)。
2.菌株G1产ACC脱氨酶的活性测定
采用α-酮丁酸测酶活法对菌株G1菌株产ACC脱氨酶的活性进行测定。将粗酶提取物(0.2mL)和ACC(0.5M)混合,置于30℃水浴15min。培养结束后,加入1mL 0.56M盐酸(HCl)终止反应,向其中加入0.3mL 0.2%2,4-二硝基苯肼溶液,在30℃下放置30分钟。然后,加入2mL 2M氢氧化钠,在540nm下测吸光度,根据α酮丁酸标准曲线计算ACCD活性。ACC脱氨酶活力以反应体系中每毫克酶蛋白每小时生成的α-酮丁酸μM量表示,酶活力单位为α-酮丁酸μM/mg·h。经计算得出菌株G1的ACC脱氨酶活性为2.5μM/mg·h。
实施例4含枯草芽孢杆菌P1与贪噬菌G1的复合菌剂抗旱保与植被修复效果试验
一、枯草芽孢杆菌P1的发酵液对土壤保水性能的研究
用P1菌的聚谷氨酸发酵液进行实验(产聚谷氨酸的发酵培养基为葡萄糖20g·L-1,酵母粉1g·L-1,谷氨酸钠18g·L-1,(NH4)2SO41g·L-1,CaCl2 0.173g·L-1,K2HPO40.5g·L-1,MgSO4 0.1g·L-1。),40℃恒温蒸发30h,加聚谷氨酸发酵液的样品土壤(16g土壤加6mLP1菌的发酵液,有效活菌数大概为108-109cfu/mL)失水率始终低于加蒸馏水和培养基(未接种P1的发酵培养基)的样品,蒸发3小时,P1发酵液、培养基和水处理组失水率,失水率基本一致,分别为8.06%、7.57%和7.5%,可能是由于泥土表面的水分相对比较均匀的蒸发;到第6h发酵液处理组的失水率明显低于对照组,添加P1发酵液的土壤失水率为16.93%,明显低于加蒸馏水对照土壤的失水率(20.61%),见图4。说明聚谷氨酸发酵液可以减缓土壤中水分的蒸发,对土壤有一定保水效果。
二、菌株P1产聚谷氨酸发酵液对玉米苗期抗旱性作用
为试验P1菌发酵的聚谷氨酸发酵液在玉米幼苗生长过程的作用,两周施用一次P1聚谷氨酸发酵液,聚谷氨酸浓度为6.6g·L-1(有效活菌数为108-109cfu/mL),共施加两次。控制土壤含水量为30%-40%,相当于中度干旱胁迫。移栽30天后,测定植株生长状况,利用R语言统计软件进行统计分析、并计算其各项指标的增长情况,结果见表3。聚谷氨酸发酵液处理组在叶面积和生物量等方面与对照组有明显差异,并且显著提高土壤含水率。由此可见,在干旱胁迫下,聚谷氨酸发酵液可提高土壤含水率,对玉米幼苗期生长具有良好的促进作用。
表3聚谷氨酸发酵液对玉米幼苗的影响
注:CK表示水;CM表示培养基
三、菌株G1产ACC脱氨酶对玉米发芽及抗旱的作用
为分析产ACC脱氨酶菌株G1对玉米发芽及抗旱的作用,以菌株G1发酵液(G1菌液OD=1.0,有效活菌数为108-109cfu/mL)。对玉米进行浸种,分别考察了菌株G1发酵液、去除菌体上清液以及培养基浸种与不浸种处理对玉米发芽及后期幼苗抗旱的作用。在玉米幼苗生长的过程中,两周施用一次菌株G1发酵液(OD=1.0),共施加两次。控制土壤含水量为30%-40%,相当中度干旱胁迫,移栽30天后,测定植株生长状况,利用R语言进行统计分析、并计算其各项指标的增长率。结果表明,与对照水浸种相比,应用菌株G1发酵液浸种可使发芽时间缩短24h,发芽率提高5%。菌株G1浸种联合后续施加G1发酵液处理与对照水处理相比可以使玉米植株的生物量(地上+地下总和)可提高13.2%,表明产ACC脱氨酶菌株可增强玉米幼苗的抗旱能力。
表4菌株G1产ACC脱氨酶对玉米幼苗发芽率及抗旱生长的影响
注:CK为水;CM为培养基
四、菌株P1与菌株G1联合应用对玉米发芽及抗旱的作用
为进一步提高菌剂的保水及抗旱效果,在上述实验基础上,将菌株G1与P1联合应用。以菌株G1的菌液进行常规浸种,以缩短发芽时间同时提高发芽率;在后续玉米幼苗施加抗旱菌剂时,将菌液P1与G1(两种菌液中有效活菌数均在108-109cfu·mL-1),进行不同的体积配比,分别设置CK,P1;G1;P1:G1=1:1;P1:G1=2:1;P1:G1=3:1共六个不同菌剂施加处理组。P1:G1=2:1组即为每400g土壤中施加约50mL聚谷氨酸发酵液(6.6g·L-1)和25mL G1(109cfu·mL-1)菌液。其他处理组除配比不同外,处理方式同此组。
结果表明,P1:G1=2:1时,复合菌剂的保水抗旱效果较好,不用复合菌剂时,玉米幼苗生长一个月地上与地下生物量总和为452mg,应用P1:G1=2:1体积配比的菌剂后,玉米幼苗的生物量为682mg,可以使玉米植株的生物量提高50.8%(图5)。由此得出,在应用G1和P1的复合菌剂时,首先利用菌株G1的发酵液进行浸种处理,在田间施用时,将两种菌株的发酵液按照P1:G1=2:1进行配比,制成复合菌剂施用,以提高植株的抗旱效果。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
序列表
<110> 北京农业生物技术研究中心
<120> 一株抗旱促生复合菌剂及其应用
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
agagtttgat cctggctcag 20
<210> 2
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
tacggctacc ttgttacgac tt 22
<210> 3
<211> 1449
<212> DNA
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 3
agtttcggct gctaataatg caagtcgagc ggacagatgg gagcttgctc cctgatgtta 60
gcggcggacg ggtgagtaac acgtgggtaa cctgcctgta agactgggat aactccggga 120
aaccggggct aataccggat ggttgtttga accgcatggt tcaaacataa aaggtggctt 180
cggctaccac ttacagatgg acccgcggcg cattagctag ttggtgaggt aacggctcac 240
caaggcgacg atgcgtagcc gacctgagag ggtgatcggc cacactggga ctgagacacg 300
gcccagactc ctacgggagg cagcagtagg gaatcttccg caatggacga aagtctgacg 360
gagcaacgcc gcgtgagtga tgaaggtttt cggatcgtaa agctctgttg ttagggaaga 420
acaagtaccg ttcgaatagg gcggtacctt gacggtacct aaccagaaag ccacggctaa 480
ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggaa ttattgggcg 540
taaagggctc gcaggcggtt tcttaagtct gatgtgaaag cccccggctc aaccggggag 600
ggtcattgga aactggggaa cttgagtgca gaagaggaga gtggaattcc acgtgtagcg 660
gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag gcgactctct ggtctgtaac 720
tgacgctgag gagcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780
cgtaaacgat gagtgctaag tgttaggggg tttccgcccc ttagtgctgc agctaacgca 840
ttaagcactc cgcctgggga gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg 900
cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt 960
cttgacatcc tctgacaatc ctagagatag gacgtcccct tcgggggcag agtgacaggt 1020
ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc 1080
aacccttgat cttagttgcc agcattcagt tgggcactct aaggtgactg ccggtgacaa 1140
accggaggaa ggtggggatg acgtcaaatc atcatgcccc ttatgacctg ggctacacac 1200
gtgctacaat ggacagaaca aagggcagcg aaaccgcgag gttaagccaa tcccacaaat 1260
ctgttctcag ttcggatcgc agtctgcaac tcgactgcgt gaagctggaa tcgctagtaa 1320
tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca 1380
ccacgagagt ttgtaacacc cgaagtcggt gaggtaacct ttaggagcca gccgccgaag 1440
gtgaaccca 1449
<210> 4
<211> 1393
<212> DNA
<213> 贪噬菌(Variovorax sp)
<400> 4
gtatcgccct ccttgcggtt aagctaacta cttctggcag aacccgctcc catggtgtga 60
cgggcggtgt gtacaagacc cgggaacgta ttcaccgtga cattctgatc cacgattact 120
agcgattccg acttcacgca gtcgagttgc agactgcgat ccggactacg actggtttta 180
tgggattagc tccccctcgc gggttggcaa ccctttgtac cagccattgt atgacgtgtg 240
tagccccacc tataagggcc atgaggactt gacgtcatcc ccaccttcct ccggtttgtc 300
accggcagtc tcattagagt gcccaactga atgtagcaac taatgacaag ggttgcgctc 360
gttgcgggac ttaacccaac atctcacgac acgagctgac gacagccatg cagcacctgt 420
gttacggctc tctttcgagc actaagccat ctctggcaaa ttccgtacat gtcaaaggtg 480
ggtaaggttt ttcgcgttgc atcgaattaa accacatcat ccaccgcttg tgcgggtccc 540
cgtcaattcc tttgagtttc aaccttgcgg ccgtactccc caggcggtca acttcacgcg 600
ttagcttcgt tactgagtca gtgaagaccc aacaaccagt tgacatcgtt tagggcgtgg 660
actaccaggg tatctaatcc tgtttgctcc ccacgctttc gtgcatgagc gtcagtacag 720
gcccagggga ttgccttcgc catcggtgtt cctccgcata tctacgcatt tcactgctac 780
acgcggaatt ccatccccct ctgccgtact ccagcaatgc agtcacagat gcagttccca 840
ggttgagccc ggggatttca caactgtctt acatcaccgc ctgcgcacgc tttacgccca 900
gtaattccga ttaacgcttg caccctacgt attaccgcgg ctgctggcac gtagttagcc 960
ggtgcttatt cttacggtac cgtcattagc cctctttatt agaaagagcc gtttcgttcc 1020
gtacaaaagc agtttacaac ccgaaggcct tcatcctgca cgcggcatgg ctggatcagg 1080
cttgcgccca ttgtccaaaa ttccccactg ctgcctcccg taggagtctg ggccgtgtct 1140
cagtcccagt gtggctggtc gtcctctcag accagctaca gatcgaaggc ttggtgagcc 1200
tttacctcac caactaccta atctgccatc ggccgctcca ttcgcgcaag gtcttgcgat 1260
cccctgcttt catccgtaga tcgtatgcgg tattagcaca gctttcgctg cgttatcccc 1320
cacgattggg cacgttccga tgtattactc acccgttcgc cactcgccgc caggattgct 1380
cccgcgctgc cgt 1393
Claims (9)
1.一种抗旱促生复合菌剂,其特征在于,该菌剂含有活性菌,所述活性菌由有效活菌数均为108~109cfu/mL的保藏编号为CGMCC No.14465的枯草芽孢杆菌和保藏编号为CGMCCNo.14466的贪噬菌以体积比2:1配比制成。
2.权利要求1所述的抗旱促生复合菌剂在促进植物生长中的应用。
3.权利要求1所述的抗旱促生复合菌剂在提高土壤保水性和/或修复植被中的应用。
4.权利要求1所述的抗旱促生复合菌剂在提高植物抗旱能力中的应用。
5.权利要求1所述的抗旱促生复合菌剂,其中还包括载体。
6.权利要求1所述的抗旱促生复合菌剂在提高植物抗旱能力的同时促进植物生长中的应用。
7.权利要求1所述的抗旱促生复合菌剂在修复植被的同时提高土壤保水性中的应用。
8.权利要求2-7任意一项所述的用途,其特征在于,用于干旱土壤中。
9.权利要求2-7任意一项所述的用途,其特征在于,用于轻中度干旱土壤中。
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