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1.
郑会明  毛怡玲  凌军  钟增涛 《土壤》2014,46(6):1109-1114
根瘤菌可以和豆科植物共生固氮,在很多根瘤菌中群体感应系统参与了结瘤固氮等相关生理过程的调节。本研究从野外不同生境紫云英宿主根部瘤体中分离了26株根瘤菌,体外定量检测了各菌株产生自体诱导物(AI)信号分子的能力,以及不同菌株AI激活中慢生华癸根瘤菌Mh菌株AHLs合成酶基因mrh I表达的情况,发现紫云英根瘤菌种内不同菌株间存在群体感应交互性。通过导入外源Att M水解酶观察菌株AI活性的变化,应用薄层层析法定性分析不同菌株的AI结构,初步揭示了不同紫云英根瘤菌产生AI特性的差异和多样性。  相似文献   

2.
细菌在高细胞密度下可以产生化学信号分子调控细菌相关基因的表达,这种信号分子被称为自体诱导物(Autoinducer,AI)。采用检测菌株JZA1在中慢生型天山根瘤菌CCBAU060A(Mesorhizobium tianshanenseCCBAU060A)培养上清液中检测到了高活性的自体诱导物;利用转座子随机突变的方法获得了AI缺失突变株,并克隆得到了相关的自体诱导物合酶基因;将自体诱导物合酶基因在大肠杆菌中进行表达,对大肠杆菌重组菌株进行自体诱导物检测发现,该合酶基因在大肠杆菌中能够合成四种自体诱导物分子。  相似文献   

3.
用质粒pKGL 3上的转座子随机插入失活的方法得到两个中慢生型天山根瘤菌胞外多糖缺失菌株,并通过随机引物PCR的方法确定了转座子插入的基因位点,该基因与豌豆根瘤菌的UDP-葡萄糖脱氢酶基因有高度的同源性.根毛吸附实验发现该基因缺失,导致胞外多糖缺失突变菌株在其宿主植物甘草上的根毛吸附量大大降低,同时生物膜的形成实验发现中慢生型天山根瘤菌的exo5-菌株不能像野生型菌株一样形成丰富的生物膜,也从侧面证明了根毛吸附实验的结果,初步推测胞外多糖可能通过影响中慢生型天山根瘤菌的根毛吸附过程来影响其与豆科植物的共生固氮.  相似文献   

4.
土壤和大豆品种是影响根瘤菌遗传多样性的主要因素。本研究通过土壤捕捉试验,分别从冬小麦茬口和玉米茬口土壤中种植的4个大豆品种根瘤中分离得到149株快生根瘤菌和49株慢生根瘤菌。对这些菌株进行16S rDNA限制性酶切分析(ARDRA)、16S-23S基因间隔(IGS)以及共生基因(nod C)的限制性片段长度多态性分析(RFLP),考察土壤茬口和大豆品种对大豆根瘤菌遗传多样性的影响。ARDRA分析结果表明,快生根瘤菌全部属于中华根瘤菌属(Sinorhizobium),慢生根瘤菌全部属于慢生根瘤菌属(Bradyrhizobium)。IGS酶切分型将所有菌株分为3种图谱类型,其中型Ⅰ与辽宁慢生根瘤菌(B.liaoningense)的模式菌株酶切图谱类型完全一致,型Ⅱ和型Ⅲ与费氏中华根瘤菌(S.fredii)的模式菌株酶切图谱类型完全一致。综合以上结果,本研究分离到的菌株分属于以上2个种。在两种茬口土壤中快生根瘤菌费氏中华根瘤菌均为优势种群,在冬小麦茬口土壤中的比例(平均95.18%)远高于玉米茬口土壤(平均53.78%)。‘冀豆12’大豆品种的费氏中华根瘤菌在两种茬口土壤中所占比重均高于其他品种。对菌株的IGS基因型与宿主大豆品种的相关性分析表明,大豆品种与根瘤菌IGS基因型之间具有一定相关性。nod C-RFLP酶切分型结果表明,土壤茬口和大豆品种对菌株的共生基因无明显影响。本研究表明根瘤菌、土壤茬口和大豆品种间存在一定的相关性。土壤茬口对根瘤菌的种群结构影响较大,大豆品种对根瘤菌的基因型具有一定的选择性。  相似文献   

5.
对分离自西北部分地区鸡眼草(Kummerowia striata)的53株根瘤菌,进行唯一碳源和氮源利用、抗生素、染料和苯酚抗性、耐盐性等98项生理生化性状测定,并结合16S rDNA PCR-RFLP分析,对供试菌株进行了遗传多样性研究.结果表明,不同地理来源,甚至来源于同一地区的不同菌株在碳、氮源利用、耐盐性、对苯酚抗性程度等方面存在着一定差异,12.5%的菌株能耐受300 μg/mL氯霉素,58.9%的菌株能耐受2%NaCI,38%的菌株能耐受700 mg/L苯酚.通过16S rDNA PCR-RFLP分析,53株供试菌株分别属于慢生根瘤菌属(Bradyrhizobium)、中慢生根瘤菌属(Mesorhizobium)、根瘤菌属(Rhizobium)、中华根瘤菌属(Sinorhizobium)和土壤杆菌属(Agrobacterium),说明分离自鸡眼草的根瘤菌在种及属的水平上具有非常丰富的遗传多样性.  相似文献   

6.
克隆了中慢生华癸根瘤菌(Mesorhizobium huakuii)的mrh I和mrh R群体感应基因的启动子区域,至Lac Z翻译融合表达质粒载体p RA302中,经β-半乳糖苷酶活性检测发现,mrh I和mrh R的表达均受mrh I基因产物(自体诱导物)的正向调控。两种PTS糖类:葡萄糖和果糖的外源添加可以显著增加细菌的生长量,但却都同时抑制了mrh I和mrh R基因的表达,暗示在中慢生华癸根瘤菌中群体感应基因的表达可能受细菌中普遍存在的碳源代谢物阻遏现象的调节。  相似文献   

7.
以缺钼酸性紫色土为供试土壤做盆栽试验,选用celB标记的三株慢生型花生根瘤菌celB3-5、celB3-7、celB4-5接种天府9号花生。通过标记根瘤菌形成的根瘤能与检测试剂产生颜色反应的特征,检测施钼及施不同浓度的钼对花生-根瘤菌有效性和竞争性的影响。结果表明,缺钼酸性紫色土上单施钼、单接种、接种配合施钼均能促进花生与根瘤菌的共生固氮效应和竞争结瘤能力,但接种配合施钼的效果最好,单接种的效果次之,单施钼的效果差。单施钼时,0.4%的钼酸铵拌种效果好,接种根瘤菌时,0.2%的效果好。供试菌株中celB4-5的有效性和竞争性最强,celB3-7次之,celB3-5差。  相似文献   

8.
李友国  周俊初 《土壤学报》2003,40(4):613-617
研究了发光酶基因 (luxAB)标记的重组费氏中华根瘤菌HN0 1DL、大豆慢生根瘤菌TA113QD与其出发菌株HN0 1、TA11在灭菌和未灭菌土缩影中的存活动态。结果表明 :HN0 1DL和TA113QD在灭菌土缩影和未灭菌土缩影中的存活动态特征显著不同 :标记菌株在灭菌土缩影中的早期存活数量一般有所下降后而保持相对稳定的较高存活水平 ,而在未灭菌土缩影中其存活数量在整个跟踪过程中持续下降至约 4 5logcfug- 1 土。在本实验中未发现供试标记重组菌株和出发菌株在土壤缩影中的存活性存在显著差异  相似文献   

9.
nfeC基因是从慢生型大豆根瘤菌中克隆到的,与竞争结瘤有关的基因。本研究从慢生型大豆根瘤菌菌株GX201的pLAFR3为载体的基因文库中,筛选出与nfe同源基因克隆。以转座子Tn5gusA5诱变获得了gus基因表达的Tn5gusA5插入突变质粒。  相似文献   

10.
徐翔  郑会明  杜寒春  汪洋  钟增涛  朱军 《土壤》2009,41(1):67-71
通过将天山根瘤菌中的自体诱导物合成酶基因mrtI在转录水平融合肠lacZ构建了5'mrtI-lacZ融合表达菌株HMZ1.利用多株不同属的根瘤菌上清液对该菌株进行诱导,发现有3株豌豆根瘤菌和1株三叶草根瘤菌上清提取物能诱导mrtI基因的表达,且外源上清量越多,mrtI被诱导的程度越高,TLC的结果则显示它们可能有着相同的AI.该结果显示在相同功能的细菌间存在信号交流,为进一步研究群体感应调节在共生固氮上的作用提供了理论及实践依据.  相似文献   

11.
The aims of our study were to enhance growth, yield and disease control of chickpea by various combinations of microbial strains (Mesorhizobium, Azotobacter chroococcum, Pseudomonas aeruginosa and Trichoderma harzianum). Pseudomonas and Trichoderma showed positive IAA (indole-3-acetic acid) production, phosphate solubilisation and antagonistic activities against Fusarium oxysporum and Rhizoctonia solani as compared to other strains. In two year investigations, tetra-inoculants have shown significant growth attributes, yield and phytopathogen growth inhibition followed by tri-inoculants than control. Therefore, tetra-inoculants (Mesorhizobium-Azotobacter-Pseudomonas-Trichoderma) may be used as efficient biofertilizer and bio-control agent for chickpea production (Cicer arietinum L.) in eastern Uttar Pradesh.  相似文献   

12.
The present study was designed with the objective of improving the nodulation and growth of chickpea (Cicer arietinum L.) by integrating co-inoculation of Rhizobium sp. (Mesorhizobium ciceri) and plant growth promoting rhizobacteria (PGPR) carrying ACC (1-aminocyclopropane-1-carboxylate) deaminase activity with P-enriched compost (PEC) under irrigated and rainfed farming systems. PEC was prepared from fruit and vegetable waste and enriched with single super phosphate. The results demonstrated that co-inoculation significantly (P?<?0.05) increased the number of nodules per plant, nodule dry weight, pods per plant, grain yield, protein content, and total chlorophyll content under irrigated and rainfed conditions compared to inoculation with rhizobium alone. Integrating PEC with co-inoculation showed an additive effect on the nodulation and growth of chickpea under both farming systems. Analysis of leaves showed a significantly (P?<?0.05) higher photosynthetic rate and transpiration rate in comparison with inoculation with Rhizobium. Compared to irrigated farming system, co-inoculation with PEC under rainfed conditions was more beneficial in improving growth and nodulation of chickpea. Post-harvest soil analysis revealed that the integrated use of bioresources and compost enhanced microbial biomass C, available N content, dehydrogenase, and phosphomonoesterase activities.  相似文献   

13.
应用酵母双杂交系统筛选与CASTOR相互作用的蛋白   总被引:1,自引:0,他引:1  
CASTOR编码百脉根中的离子通道蛋白,它是共生途径上的功能基因,作用于结瘤因子诱导产生的钙离子激增(calcium spiking)信号转导途径的上游,突变后不能形成根瘤。为了进一步揭示CASTOR调控共生信号途径的分子机制,本文通过酵母双杂交系统在百脉根的cDNA文库中筛选可能与CASTOR相互作用的蛋白,以酵母配合的方法初筛获得111个阳性克隆,经β- Gal检测进一步确认有99个蓝色克隆子,测序以及NCBI BLAST比对分析鉴定出JAB1、CLPA、钙调素结合的翻译延伸因子等7种可能与CASTOR相互作用的蛋白,利用RT-PCR方法检测了这些基因在接种以及不接种根瘤菌的百脉根中的表达水平,分析推测CASTOR可能与上述蛋白形成复合物参与共生信号的转导。  相似文献   

14.
杜寒春  高轶静  郑会明  钟增涛  朱军 《土壤》2013,45(3):529-532
通过二亲接合的方式,将含有sacB基因和nodD基因部分片段的质粒与中慢生华癸根瘤菌Mesorhizobium huakuii)93(Mh93)菌株进行同源重组,在含10%蔗糖的TY平板上成功地筛选到了nodD基因突变株Mh93-27.利用温室盆栽试验研究了紫云英单作及紫云英-小麦间作体系Mh93-27对紫云英的促生作用,结果表明,接种Mh93-27后,紫云英的地上部生物量和全氮含量以及根部生长得到了不同程度的提高,但只有在小麦孕穗期和收获期时,根瘤菌才表现出显著的促生作用.  相似文献   

15.
袁全  杨梦华  郑会明  钟增涛  朱军 《土壤》2009,41(3):459-463
细菌在高细胞密度下通过产生一类化学信号分子(autoinducer,自体诱导物)感知细胞群体密度变化,从而对基因表达进行调控,这一现象被称为群体感应(quorum sensing).以中慢生型华癸根瘤菌 Mesorhizobium huakuii AS9 为出发菌株,利用转座子随机突变的方法获得了自体诱导物缺失突变株 YQ1,并克隆得到了其自体诱导物合成酶基因,命名为 mrhI;构建PmrhI-lacZ 转录融合表达质粒 pYQ1,通过同源重组的方法得到在华癸根瘤菌中 mrhI-lacZ 融合表达而且 mrhI 基因突变菌株YQ3.突变株 YQ3 的培养液上清不含有自体诱导物分子,通过 lacZ 报告基因的表达发现 mrhI 的表达受自体诱导物的调控,说明 mrhI 基因是 M. huakuii AS9 的群体感应体系中的自体诱导物合成酶基因.  相似文献   

16.
The purpose of this study was to evaluate the responses of lentil (Lens culinariscv. ‘Ziba’) to co-inoculation with arbuscular mycorrhizal (AM) fungi and some indigenous rhizobial strains varying in phosphorus (P)-solubilizing ability in a calcareous soil with high pH and low amounts of available P and nitrogen (N). A factorial experiment with completely randomized block design was conducted under controlled greenhouse conditions. The treatments consisted of (1) three inoculants of Rhizobium leguminosarum bv. viciae strains and a mixed rhizobial inoculant with an effective P-solubilizer strain of Mesorhizobium ciceri, (2) two AM fungal species, Glomus mosseae and Glomus intraradices, (3) two P sources, superphosphate and phosphate rock. Four replications were prepared for each treatment and a related control. After the growth period of three months, the dry matter of shoots plus seeds, their P and N contents, and percent of root colonized by AM fungus were measured. The results showed that the effects of AM fungi, rhizobial strains, and P fertilizers were highly significant (p < 0.01) for all the characteristics studied. The rhizobial strain with P-solubilizing ability showed a more beneficial effect on plant growth and nutrient uptake than the strain without this ability, although both strains had similar effectiveness for N2-fixation in symbiosis with lentil. Synergistic relationships were observed between AM fungi and some rhizobial strains that related to the compatible pairing of these two microsymbionts. The P-uptake efficiency was increased when P fertilizers were applied along with AM fungi and/or P-solubilizer rhizobial strains.  相似文献   

17.
《Journal of plant nutrition》2013,36(9):1367-1375
A study was conducted to compare the responses to acid pH of Medicago sativa and Lotus glaber, two forage legumes with different environmental requirements, either supplied with inorganic nitrogen (N) or inoculated with different strains of their nodule bacteria. Medicago sativa showed, in both treatments, a significant reduction in total dry weight at pH below 6.0. In contrast, Lotus glaber grew equally well at all the pHs assayed in the presence of adequate N. Under inoculated conditions, in the absence of N supply, plant growth was dependent on the bacterial strain used. When the ability of each strain to multiply in culture medium was examined, it was observed that Sinorhizobium meliloti strains showed a pH-sensitive response that inhibited growth at pH 4.0, whereas Mesorhizobium loti strains showed normal growth at this pH. These results suggest that for the effectiveness of Mesorhizobium loti–Lotus glaber symbiosis in acid soils the major factor to be considered is the tolerance of the bacterial strain to acid conditions, while the limiting factors for the Sinorhizobium meliloti–Medicago sativa symbiosis are the sensitivity to low pH of both the plant and its bacteria.  相似文献   

18.
王平  冯新梅  李阜棣 《土壤学报》2001,38(2):265-270
在土壤一盒栽、盆栽微宇宙系统(Microcosm)中和田间条件下,研究了发光酶基因(luxAB)标记的华癸根瘤菌JS5A16L在紫云英根圈的定殖动态、分布范围及结瘤情况。在盒栽系统中,JS5A16L的定殖密度在紫云英出苗2天后达到最高水平(7.88 lpg cfu/克根)(cfu为colony forming unit的缩写),然后开始下降,并保持在一个相对稳定的水平;58天后又有所回升,且能散布至种子下方22cm处的根段部位。盆栽条件下的定殖动态与盒栽系统中的相似,紫云英播种3天后,JS5A16L可达最高定殖水平(6.92 log cfu/株根系),随后开始缓慢下降,直至32天时仍维持在一个相对稳定水平。JS5A16L在田间条件的定殖动态则不同,播种后30天时定殖密度达到最大值(7 .03 log cfu/克根),90天后降到最低值(5. 24 log cfu/克根),然后又开始上升,160天时为7.87 log cfu/克根,甚至在地上部植株收割后20天仍可维持在7.89 log cfu/克根。接种该菌株可显著提高紫云英的生物学产量。  相似文献   

19.
Abstract

We studied the effects of the application of organic matter (OM) and chemical fertilizer (CF) on soil alkaline phosphatase (ALP) activity and ALP-harboring bacterial communities in the rhizosphere and bulk soil in an experimental lettuce field in Hokkaido, Japan. The ALP activity was higher in soils with OM than in soils with CF, and activity was higher in the rhizosphere for OM than in the bulk soil. Biomass P and available P in the soil were positively related to the ALP activity of the soil. As a result, the P concentration of lettuce was higher in OM soil than in CF soil. We analyzed the ALP-harboring bacterial communities using polymerase chain reaction based denaturing gradient gel electrophoresis (DGGE) on the ALP genes. Numerous ALP genes were detected in the DGGE profile, regardless of sampling time, fertilizer treatment or sampled soil area, which indicated a large diversity in ALP-harboring bacteria in the soil. Several ALP gene fragments were closely related to the ALP genes of Mesorhizobium loti and Pseudomonas fluorescens. The community structures of the ALP-harboring bacteria were assessed using principal component analysis of the DGGE profiles. Fertilizer treatment and sampled soil area significantly affected the community structures of ALP-harboring bacteria. As the DGGE bands contributing to the principal component were different from sampling time, it is suggested that the major bacteria harboring the ALP gene shifted. Furthermore, there was, in part, a significant correlation between ALP activity and the community structure of the ALP-harboring bacteria. These results raise the possibility that different ALP-harboring bacteria release different amounts and/or activity of ALP, and that the structure of ALP-harboring bacterial communities may play a major role in determining overall soil ALP activity.  相似文献   

20.
Chickpea is an important pulse crop grown mainly in the arid and semi-arid regions. The effect of water deficiency on nodulation, biomass production, and competition for nodule occupancy was evaluated in three different soils with two chickpea cultivars, Amdoun I and Chetoui. Two watering regimes were considered; a control that was irrigated three times per week and a water-deficient treatment that was irrigated only one time a week. Results showed that water deficiency significantly decreased the nodule number and the shoot dry weight for both cultivars. Root-nodule bacteria were isolated and characterized by PCR-RFLP of 16S rDNA and nifD-K intergenic spacer. The results show that water deficiency affects the diversity of nodulating rhizobia. The nodulation by Mesorhizobium mediterraneum was reduced while inefficient nodulation by Ensifer meliloti was favoured. In both treatments, chickpea was preferentially nodulated by nifD-K type N6. Analysis for NaCl tolerance showed that most of rhizobia nodulating chickpea under water deficiency are NaCl tolerant. Inoculation with the selected salt-tolerant strain of M. mediterraneum LILM10 increased significantly nodule number and grain yield in the field.  相似文献   

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