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1.
从土壤中分离筛选出7株解有机磷的微生物。对这7株解磷细菌进行了形态、生理生化性状测定及16S rDNA序列分析(GenBank accession No: S2, AY651922; S3, AY661923; X1, AY651925; Y1, AY651924; H1, AY663435; H2,AY663436 and He, AY663436)。其中S2、S3、X1和He属于假单胞菌属(Pseudomonas),Y1属于芽孢杆菌属(Bacillus),H1属于不动杆菌属(Acinetobacter),H2属于寡养单胞菌属(Stenotrophomonas)。进一步通过G+C含量和DNA-DNA杂交研究, 结果表明,S2、S3和X1为产碱假单胞菌(Pseudomonas alcaligenes),Y1为蜡状芽孢杆菌(Bacillus cereus)。  相似文献   

2.
采用稀释涂布平板、分离培养和16S r DNA序列分析法对我国甘肃白银地区半干旱荒漠草原土壤可培养细菌、放线菌、真菌数量及群落分布特征进行了分析,比较了荒漠草原和耕地土壤微生物多样性。发现荒漠草原土壤可培养细菌、放线菌、真菌数量分别为1.23×106、0.19×106、0.18×106cfu·g-1,耕地三类微生物数量分别是3.03×106、0.53×106、0.05×106cfu·g-1。荒漠草原可培养细菌、放线菌数量明显低于耕地,而真菌数量高于耕地。从荒漠草原分离出14株细菌,分别属于γ-变形菌纲(γ-Proteobacteria)噬冷杆菌属(Psychrobacter),放线菌门(Actinobacteria)皮球菌属(Kytococcus),厚壁菌门(Firmicutes)芽孢杆菌属(Bacillus)、亮氨酸芽孢杆菌属(Lysinibacillus)、土壤芽孢杆菌属(Solibacillus)、气球菌属(Aerococcus),优势菌为芽孢杆菌属和噬冷杆菌属。耕地分离出可培养细菌19株,分别属于ɑ-变形菌纲(ɑ-Proteobacteri)根瘤菌属(Rhizobium)、中华根瘤菌属(Sinorhizobium),γ-变形菌纲(γ-Proteobacteria)假单胞菌属(Pseudomonas),厚壁菌门(Firmicutes)芽孢杆菌属(Bacillus),放线菌门(Actinobacteria)微杆菌属(Microbacterium)、节杆菌属(Arthrobacter)、微球菌属(Micrococcus)、考克氏菌属(Kocuria),以放线菌门细菌为主(占57.9%)。从荒漠草原分离放线菌共8株,分别属于链霉菌属(Atreptomyces)、小单孢菌属(Micromonspora)、间孢囊菌属(Intrasporangium),而耕地主要为链霉菌属(Atreptomyces)、小单孢菌属(Micromonspora)。荒漠草原真菌主要是交链孢霉属(Alternaria)、芽枝霉属(Cladosporium),耕地土壤真菌包括青霉属(Penicillium)、交链孢霉属(Alternaria)、曲霉属(Aspergillus)、毛霉属(Mucor)、链孢霉属(Coniothecium)。试验结果表明,荒漠草原与耕地土壤微生物都具有较丰富的多样性,但微生物群落结构存在一定差异,同一区域不同深度土壤中微生物数量和种类也存在差异,耕地土壤微生物多样性明显高于荒漠草原。  相似文献   

3.
为获得对植物真菌病害有稳定作用效果的生防细菌,采用平板对峙培养法从大豆和玉米根部内生细菌中筛选获得12株菌株,这些细菌对大豆立枯丝核菌(Rhizoctonia solani)、大豆尖孢镰刀菌(Fusarium oxysporum f.sp.glycine)和番茄枯萎病菌(Fusarium oxgsporum f.sp.lycopersici)均具有明显拮抗作用。16S rRNA基因鉴定表明12株内生拮抗细菌分别属于芽孢杆菌属(Bacillus)、类芽孢杆菌属(Paenibacillus)和假单胞菌属(Pseudomonas)。选取5株细菌进行盆栽接种试验,结果表明,供试细菌均对大豆根腐病具有一定的防治效果,其中,菌株J1和J2防效分别达到20.4%和32.1%。  相似文献   

4.
光合细菌对鱼病原细菌生长的影响   总被引:2,自引:0,他引:2  
从发病的棒花鱼(Abbottina rivularis)和鲫鱼(Carassius auratus)分离得到7株菌,经人工感染证实其中的B3、J1和J2等3株菌均为病原细菌。B3和J1鉴定为嗜水气单胞菌(Aeromonas hydrophila),J2鉴定为豚鼠气单胞菌(Aeromonas caviae)。采用平板扩散法测定了光合细菌嗜酸红假单胞菌Y7(Rhodopseudomonas acidophila Y7)、球形红假单胞菌X3(R.spheroids X3)和桃红荚硫菌S(Tniocapsa roseopersicina S)对病原细菌B3和J2生长的影响。结果表明:在营养琼脂平板上3种光合细菌对两种病原细菌(B3和J2)无明显拮抗作用,但光合细菌能覆盖在病原细菌上生长;3种光合细菌对病原细菌生长有明显的抑制作用,其中球形红假单胞菌X3抑制作用最大,但X3代谢产物对病原细菌生长无明显抑制作用,说明X3对病原细菌的抑制作用来自菌体细胞。X3还能明显抑制水体中的病原菌B3和J2的生长。  相似文献   

5.
刘冬晖  李凤巧  靳志丽  李孝刚 《土壤》2022,54(4):750-755
植物微生物组是维护植物生长发育、提升抗逆防病的重要调控因素。为发挥植物微生物促进烟草生长、改善烟草根区微生态功能作用,本研究从烟草根表分离筛选可培养细菌组,并对不同菌株的促生能力进行测定。结果表明:(1)从烟草根表分离并鉴定出可培养菌株310株,隶属于31个属,其中主要为芽孢杆菌属(Bacillus)、假单胞菌属(Pseudomonas);(2)对比分析发现假单胞菌属、芽孢杆菌属、寡养单胞菌属(Stenotrophomonas)和成对杆菌属(Dyadobacter)为4种供试土壤烟草根表共有的细菌类群;(3)对进一步筛选得到的16株菌株进行促生能力的测定,发现6株菌具有固氮能力,5株菌产铁载体,4株菌可溶解无机磷,4株菌产IAA;(4)盆栽试验验证16株菌株的促生效果,其中37.5%的菌株对烟草生长具有显著促进作用,烟草株高、总鲜物质量和地下部干物质量分别比对照提高35.1%、27.9%和30.7%。总之,从烟草根表分离获得多种具有促生能力的菌株,为未来构建促进烟草健康生长的复合菌剂提供了理论基础。  相似文献   

6.
山西矿区复垦土壤中解磷细菌的筛选及鉴定   总被引:5,自引:1,他引:4  
【目的】矿区复垦土壤贫瘠、 有效磷含量低。解磷细菌能够将有机磷和难溶性无机磷转化为可溶性磷,促进植物对磷素的利用。因此筛选和鉴定具有解磷能力的菌株,可为解决矿区生态恢复使用的微生物肥料提供菌种资源。【方法】采用平板分离法初筛菌株,得到D/d1.5的菌株,然后以磷酸钙为磷源,通过液体发酵试验复筛菌株,挑选出解磷率高于巨大芽孢杆菌(Bacillus megaterium)As1.223的菌株。以磷矿粉和卵磷脂为磷源,液体发酵试验测定菌株的解磷能力及磷酸酶活性。进行菌株的生长试验以测定菌株温度适宜性、 耐盐性及耐酸碱性。通过形态学、 基因序列分析及脂肪酸组成分析综合进行菌株鉴定。 菌落形态观察用营养琼脂平板培养基培养;菌体形态即细胞形态及其大小采用扫描电镜观察;基因序列分析采用16S rDNA序列测定,基因在线比对采用EzTaxon数据库;使用美国MIDI公司的Sherolock全自动细菌鉴定系统对菌株进行脂肪酸组成分析。【结果】利用无机磷和有机磷平板培养基,从山西省矿区复垦区土壤样品中筛选出19株解磷微生物,其中D/d1.5的有7株。在以磷酸钙为磷源的液体培养试验中,4株菌的解磷率高于巨大芽孢杆菌As1.223,解磷率为7.89%~12.61%,最高的为菌株Y14。4株菌对磷矿粉的解磷率为0.81%~1.21%,最高的为菌株Y14。在以卵磷脂为磷源的液体培养试验中,4株菌的解磷率与酸性磷酸酶活性分别为1.79%~3.07%和24.3~28.4U/L,均高于巨大芽孢杆菌As1.223; 碱性磷酸酶活性为11.9~50.2U/L;菌株Y14的解磷率与磷酸酶活性均最高。4株菌均有较强的环境适应能力,以Y14的适应性最强。H22、 Y11和Y34与假单胞菌属(Pseudomonas sp.)同源性在99%以上,Y14与泛菌属(Pantoea sp.)有99.79%的同源性; H22、 Y11和Y34的细胞脂肪酸组成特征峰与假单胞菌属(Pseudomonas sp.)相一致,Y14与泛菌属(Pantoea sp.)相一致;H22、 Y11和Y34被鉴定为假单胞菌(Pseudomonas sp.),Y14为泛菌属(Pantoea sp.)。【结论】分离、 筛选到4株高效解磷菌,对于磷酸钙和卵磷脂的解磷率均高于巨大芽孢杆菌As1.223。4株菌分别隶属于假单胞菌属(Pseudomonas sp.)和泛菌属(Pantoea sp.)。菌株Y14无机磷与有机磷平板的D/d值分别为3.28与1.59,降解磷酸钙、 磷矿粉、 卵磷脂的解磷率分别为12.61%、 1.21%、 3.07%,酸性与碱性磷酸酶活性分别为28.4 U/L和50.2 U/L,均为4株菌里最高的,且环境适应能力最强,生长温度为20~60℃,能耐受pH 4~11的酸碱梯度和2%~7%的盐分梯度,Y14被鉴定为泛菌属(Pantoea sp.)。4株菌均具有良好的解磷能力及较强的环境适应能力,可望进一步研发成为微生物肥料生产菌种。综合D/d值、 解磷率、 磷酸酶活性和生长试验,本试验最终确定适合山西矿区复垦农田推广的高效解磷菌菌株为Y14。  相似文献   

7.
几株固氮芽孢杆菌的分离与鉴定   总被引:6,自引:0,他引:6  
摘要: 从小麦(Triticum aestivum)、玉米(Zea mays)、黑麦草(Lolium sp.)和柳树(Salix sp.)的根际土壤中分离得到能在无氮培养基上生长的29株芽孢杆菌(Bacilli),通过固氮酶活的测定以及固氮酶结构基因 nifH 的PCR扩增得到7株固氮芽孢杆菌。对这7株固氮菌株进行了生理生化性状测定、16S rDNA序列分析(GenBank accession No. AY373358, AY373360,~AY373364和AY376876)、G+C mol%含量的测定及DNA-DNA杂交实验,结果表明,其中5株菌属于芽孢杆菌,另外2株菌属于类芽孢杆菌。在这7株被鉴定的菌株中,菌株T1被鉴定为Bacillus cereus;菌株G1、C4和C5被鉴定为B. megaterium;菌株W5的生理生化性状、16S rDNA和G+C mol%与Bacillus marisflavi 相近;G2的生理生化性状和16S rDNA 与Paenibacillus polymyxa 接近,但在基于16S rDNA的系统发育树中却与P. durus 聚在最小的分支内;T7可能是Paenibacillus属中的一个新种。  相似文献   

8.
固氮芽孢杆菌N3的筛选鉴定及其对二月兰的促生效果   总被引:1,自引:1,他引:0  
从山东泰安农田土壤中筛选获得1株固氮能力强的菌株N3,通过形态观察、生理生化特征以及16S rDNA基因序列分析,确定为巨大芽孢杆菌属(Bacillus megaterium),该菌株固氮酶活性达C2H428.33 nmol/(h·ml).温室条件下进行二月兰盆栽试验,设置不接菌对照(CK)、接种巨大芽孢杆菌N3、接种...  相似文献   

9.
中国十三省市土壤中非共生固氮微生物菌种资源研究   总被引:11,自引:2,他引:9  
从全国13个省市自治区的70份土样中分离、采集到了非共生固氮微生物资源181份。形态特征、生理生化特征和16S rDNA序列分析表明,采集到的菌种资源在科学分类上属于24属66种,大约占到已报道非共生固氮微生物属的一半,具备一定的多样性和代表性。资源在分类学上的特点是分类地位相对集中,有65株菌属于类芽孢杆菌属,占总量的36%;52株菌属于芽孢杆菌属,占总量的29%;19株菌属于节杆菌属,占总量的11%;这3个属菌株合计占采集资源总量的76%。随地域和作物种类分布的特点是芽孢杆菌和类芽孢杆菌两个属的菌种资源具有很强的地域广泛性和作物广泛性,即从采自全国各地、各种作物的土壤样品几乎都可以分离到这两类菌种。这个研究结果对微生物肥料菌种选育和生产应用具有指导意义。  相似文献   

10.
对4株红螺菌(荚膜红假单胞菌PSB-2、球形红假单胞菌PSB-3、胶质红假单胞菌H1和沼泽红假单胞菌H10)的去细胞上清液(CFS)抑制水产养殖病原菌的作用进行研究,并探讨养殖水体环境对红螺菌拮抗作用的影响.结果表明,4株红螺菌在细胞生长稳定期后能有效分泌拮抗物质,其去细胞上清液(CFS)对20多株水产养殖病原菌均有抑制作用.从抑菌活性看,荚膜红假单胞菌的抑菌活性强于其他菌株:荚膜红假单胞菌对水产养殖病原菌的最低抑菌浓度(MIC)为8~64 AU/mL,而其他红螺菌株多为4~16 AU/mL.在养殖水体的生态因子中,水体pH、NH4 -N与NO2--N对红螺菌代谢产物的抑菌活性影响较大,pH7~9,NH4 -N低于5 mg/L、NO2--N浓度低于7mg/L时抑菌活性较强,而养殖水体温度与溶氧变化对红螺菌的抑菌活性基本无影响作用.  相似文献   

11.
Potent bacterial strains effective against the whitefly, Bemisia tabaci, nymphs (second instar), were isolated from tomato cultivated fields at Fayoum governorate, Giza, Egypt. Of 72 isolates, 12 with the most morphologically distinct-looking bacterial colonies were selected and named A1, A2, A3, A6, A7, A9, A12, A13, A107, B37, B45 and B100. All isolates were preliminarily identified as members of the genus Bacillus based on morphological, physiological and biochemical characteristics. When tested for their pathogenicity against Bemisia tabaci, the 12 isolates revealed varying efficiencies with isolates A1 and A9 being superior, exhibiting maximum mortality of 92.2 and 90.8% on day 10, respectively. Isolate A7 recorded the lowest percentage at 18.3%. Further genetic characterization of the 12 isolates was performed using inter simple sequence repeat (ISSR), randomly amplified polymorphic DNA (RAPD) and 16S rDNA gene sequencing analysis. RAPD and ISSR results confirmed each other. The combined ISSR and RAPD phylogenetic tree showed two major clusters. With 16S rRNA gene analysis, isolate A1 and A12 sequences recorded 100% identity with Bacillus thuringiensis, while isolates A7 and B100 showed 95.7% and 95.6% identity with Bacillus cereus and Bacillus sphaericus, respectively.  相似文献   

12.
The ability of a few soil bacteria to transform unavailable forms of potassium (K) to an available form is an important feature in plant growth-promoting bacteria for increasing plant yields of high-K-demand crops. In this research, isolation, screening, and characterization of six isolates of K solubilizing bacteria (KSB) from some Iranian soils were carried out. The ability of all isolates were tested in three treatments including acid-leached soil, biotite, and muscovite by analyzing the soluble K content after 5 days of incubation at 28 ± 2°C. Identification and phylogenetic analyses were also carried out by morphological, biochemical, and 16S rDNA analyses. Among the six efficient isolates, five isolates belonged to Bacillus megaterium (JK3, JK4, JK5, JK6, and JK7), while isolate JK2 belonged to Arthrobacter sp. The soluble K contents in all isolated-treatments were significantly (< 0.01) higher than the contents in nonbacteria treatment. Herein, isolate JK2 had lower potential for K solubilization (910 mg kg?1) compared with other isolates in acid-leached soils. The six bacterial strains showed higher solubilized K in biotite treatment than other two treatments. Overall, it can be concluded that the isolates belong to B. megaterium are the most efficient KSB under in vitro condition.  相似文献   

13.
Whole-cell fatty acids methyl ester (FAME) profile and 16S rDNA sequence analysis were employed to isolate and identify the bacterial groups that actively solubilized phosphates in vitro from rhizosphere soil of various crops of Korea. Out of several hundred colonies that grew on Pikovskaya's medium 13 best isolates were selected based on the solubilization of insoluble phosphates in liquid culture and further characterized and identified. They were clustered under the genera Enterobacter, Pantoea and Klebsiella and the sequences of three representative strains were deposited in the GenBank nucleotide sequence data library under the accession numbers AY335552, AY335553, AY335554.  相似文献   

14.
ABSTRACT

Monosporascus root rot and vine decline (MRVD) of muskmelon, caused by Monosporascus cannonballus, is an economically important disease worldwide. The objectives of this study were to isolate native rhizobacterial strains and to evaluate their ability to promote plant growth and to provide protection against M. cannonballus. Thirty eight native bacterial isolates from the rhizosphere soil of muskmelon and cucumber were screened for their antagonism against M. cannonballus in a dual culture assay. Among them, five isolates viz., Bacillus amyloliquefaciens (B4), Pseudomonas mendocina (B7), Bacillus endophyticus (B10), Pseudomonas resinovorans B11 and P. aeruginosa AT3, identified based on the 16S rRNA gene sequence analysis, showed a significant level of antagonism and the inhibition zone ranged from 5.6 mm to 25.9 mm. Scanning electron microscopic observation of the hyphae of M. cannonballus at the inhibition zone revealed morphological abnormalities including shrinkage, loss of turgidity, pit formation and deformation. These bacterial isolates showed compatibility with one another and with Trichoderma viride. Greenhouse experiments revealed that P. resinovorans B11 was the most effective among the bacterial antagonists in controlling MRVD in melon. When applied as seed treatment and soil application, P. resinovorans B11 reduced the incidence of MRVD by 93.1%, relative to the infected control.  相似文献   

15.
【目的】在田间原位条件下研究丛枝菌根(Arbuscular mycorrhizal, AM)真菌根外菌丝表面有无解磷细菌定殖,并对存在的解磷细菌的种类进行鉴定,对其活化有机磷的能力进行检测,从而为更好地认识菌丝际土壤有机磷的周转和磷的生物地球化学循环过程提供依据。【方法】利用河北省曲周县中国农业大学实验站的玉米长期定位试验,采用田间埋膜方式从玉米根系周围收集AM真菌的根外菌丝,用蒙金娜有机磷固体培养基筛选菌丝表面具有矿化植酸钙能力的细菌,对筛选出的细菌进行分离、 培养,然后提取细菌DNA,通过16S rDNA测序分析来确定解磷细菌的种类。分离鉴定的菌株先用蒙金娜有机磷固体培养基通过测定菌落直径(d)及溶磷圈直径(D)初步鉴定其活化植酸钙的能力,再用无菌的蒙金娜有机磷液体培养基确定每株解磷细菌矿化植酸磷的能力,并对溶液的pH进行测定,每个菌株重复3次。最后采用两室隔网根盒将分离纯化的解磷细菌回接至AM真菌根外菌丝,鉴定回接成功率,确定分离出的解磷细菌能否成功定殖于菌丝表面。【结果】从AM真菌根外菌丝表面分离得到了29株具有活化有机磷能力的细菌,分属于芽胞杆菌、 假单胞菌、 沙雷氏菌、 葡萄球菌和肠杆菌5个不同的属。通过有机磷液体培养进一步检测这些菌株活化植酸磷的能力,发现它们对植酸磷的矿化率为1.9%~21.9%。其中假单胞菌属细菌的解磷能力相对较强,对植酸磷的矿化率达14%以上,液体培养基的pH值下降2~4个单位。将分离纯化的细菌回接至两室隔网根盒的菌丝室,培养30 d后,从菌丝表面再次检测到除假单胞菌属外的芽胞杆菌属(Bacillus)、 沙雷氏菌属(Serratia)、 葡萄球菌属(Staphylococcus)和肠杆菌属(Enterobacter)细菌,另外还检测到贪铜菌属(Cupriavidus)细菌。【结论】在田间原位条件下,与玉米共生的AM真菌的根外菌丝表面有多种解磷细菌定殖,它们活化有机磷能力存在差异,其中以假单胞菌属细菌的解磷能力相对较强。  相似文献   

16.
Cellulolytic bacteria have a bio-activating role in the composting process. A study was carried out to isolate and identify cellulolytic bacteria from various sources. The isolates were cultured in the carboxymethyl cellulose (CMC) agar medium and incubated at 30°C for 3–7 days. Based on morphological characteristics of the isolates, maximum diameter of a clear zone around the colony and maximum cellulolytic activity, eight isolate were selected for further studies regarding composting experiments.

Molecular tests based on PCR amplification and sequencing of 16S rRNA gene of isolates showed the closest phylogenetic similarity with the species of Stenotrophomonas rhizophila DSM14405 (99.8%), Brevibacterium halotolerans DSM8802 (99.6%), Achromobacter marplatensis B2 (99.8%), Bacillus methylotrophicus CBMB205 (100%), Pseudomonas azotoformans IAM 1603 (99.7%), Bacillus sonorensis NBRC 101234 (99.8%), Bacillus subtilis KCTC 13429 (100%) and Ochrobactrum thiophenivorans DSM 7216 (99.3%). The study of the isolates impact on the composting of palm wastes in a randomized complete block design with 11 treatments in 3 replications showed that strain IB (B. methylotrophicus) caused a significant decrease in C:N ratio (58%). The increasing of microbial respiration compared with control after 30 days incubation at 37°C showed that the B. methylotrophicus strain IB with cellulolytic characteristics can be applied to hydrolysis of cellulosic biomass in the composting processes.  相似文献   


17.
香蕉束顶病毒海南分离物DNA组分的克隆及序列分析   总被引:7,自引:0,他引:7  
摘要:以中国海南岛地区表现束顶病毒(Banana bunchy top virus, BBTV)症状的香蕉组织总DNA为模板, 通过PCR方法克隆了BBTV海南分离物的6个DNA组分全序列,并进行了核苷酸序列分析。测序结果表明, DNA1~6序列全长分别为1104、1067、1059、1045、1014和1081 nt,均已登录到GenBank(登录号分别为AY450396、AY606084、AY494786、AY494788、AY606085和AY494787)。与世界不同地域分离物进行序列同源分析,发现DNA1序列较保守,与越南分离物的同源性达到了95.4%;而DNA2和DNA4变异较大,其中DNA2序列与广州分离物的同源性仅有83%。 根据DNA1的序列差异分析初步确定BBTV海南分离物属于亚洲亚组。  相似文献   

18.
为明确不同土壤条件对甘蔗内生固氮菌多样性的影响,对分别种植在红壤土和水稻土的4个甘蔗品种的内生固氮菌进行分离和鉴定,并利用数学生态学方法分析不同来源内生固氮菌的多样性。结果表明,从红壤土的甘蔗组织中分离筛选获得23个内生固氮菌株,它们在基于16S r DNA序列分析结果上属于9个不同的菌属;而从水稻土的甘蔗组织中获得59个内生固氮菌株,分类上属于19个不同的菌属。群落生态学分析结果显示,来源于水稻土的甘蔗内生固氮菌的群落丰富度指数(dma)、多样性指数(H')以及均匀度指数(Jsw)均明显高于红壤土,表明水稻土条件下的甘蔗内生固氮菌群落结构相对复杂、稳定和均匀;而相同土壤条件下4个甘蔗品种间的内生固氮菌丰富度指数和多样性指数差异并不明显。本研究结果反映了土壤生态条件是影响甘蔗内生固氮菌多样性的重要因子。  相似文献   

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