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1.
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   

2.
赖欣  戴俊  郑会明  钟增涛  朱军 《土壤学报》2008,45(4):705-709
在许多种类的细菌中均发现了luxR/luxI类型的群体感应调控系统,其中luxR是调节基因,对luxI合成自体诱导物(AI,autoinducer)起着正调控作用。通过建库筛选后,测序比对发现天山根瘤菌中存在mrtR/mrtI系统。通过基因敲除和转录水平基因表达对mrtR进行研究,发现mrtR严格调控着mrtI的表达,同时mrtR的表达也受到AI影响,这表明mrtR表达的调控是一种自调控,同时根毛吸附实验显示该基因对根瘤菌与宿主间相互作用密切相关。  相似文献   

3.
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   

4.
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   

5.
戴俊 《土壤》2008,40(4):658-661
通过PCR扩增出中慢生型天山根瘤菌中群体感应调控蛋白MrtR的基因片段,克隆至表达载体pALEX中,从而构建了原核表达质粒pALEX-mrtR.将pALEX-mrtR转化至大肠杆菌BL2/DE3菌株感受态细胞中,经IPTG诱导,表达出58 kD的GST-MnR融合蛋白.用纯化好的GST-MrtR制备多克隆抗体.经Westem blot检测,该血清可与目的蛋白发生特异性反应,证明其具有良好的免疫原性.MrtR融合蛋白的成功表达及其多克隆抗体的制备为进一步研究MrtR蛋白的功能奠定了基础.  相似文献   

6.
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   

7.
摘要: 根据苜蓿根瘤菌(Sinorhizobium meliloti ) Rm1021基因组中与Ralstonia eutropha phaZ基因同源部分序列设计1对引物,从S. meliloti基因组中用PCR扩增出835 bp phbD 基因片段并克隆到载体pGEM?襆-T Easy上;通过在phbD基因内插入ΩSmSp和基因置换构建了phbD突变体。该突变体可积累比野生型菌株多1.0~2.6倍的聚羟丁酶(PHB)。在YMA和TY平板上形成非粘液型菌落,而在以乙酰乙酸或3-羟丁酸为唯一碳源的M9基本培养基(M9-AA,M9-HB)上形成粘液型菌落。碱性磷酸酶测定结果表明,通过接合引入phbD突变体菌株中的exoF-phoA融合基因在YMB培养基中低量表达,而在M9-AA和M9-HB中高量表达。  相似文献   

8.
试验采用三亲本杂交法将cfp基因导入一株有效性较高的菜豆根瘤菌SC18,获得了一株带cfp基因标记的菌株SC18-cfp。对SC18-cfp进行性状、标记基因的遗传稳定性检测,结果表明,cfp基因不仅能有效表达,而且性状稳定。随机挑取SC18-cfp的10个转移结合子在无氮砂培条件下进行标记菌株与亲本菌株在3个菜豆品种上的竞争结瘤试验。结果证实,10个转移结合子在供试植物根系上的占瘤率均接近50%,其有效性和竞争结瘤能力与亲本菌株间的差异不显著,品种间的差异性不显著;在控制田间试验中SC18-cfp比土著根瘤菌的竞争结瘤能力强,在品种A、B、C上的占瘤率分别达到了57.8%、60.0%、55.6%。接种SC18的植株干重、全氮量、叶绿素含量和豆荚产量均显著高于对照。  相似文献   

9.
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   

10.
通过助移质粒P~(PK2013)的推动,实现了携带豌豆根瘤菌(Rhizobium Legumi-nosarum)nif D::Tn5的质粒pRK2009由大肠杆菌(E.coli)向棕色固氮菌Az-otobacter vinelandii的转移,转移频率为每受体7×10~(-5),进一步将与pRK2509不相容的质粒pPHLJI转入这个转移接合子,筛选得到了两株相同的棕色固氮菌的突变菌株。由表型特征分析和活性互补实验证明该突变菌株在染色体中含有豌豆根瘤茵nif D::Tn5,是两不同种固氮微生物的固氮基因即豌豆根瘤菌nif D与棕色固氮菌nif D之间发生同源性重组的结果  相似文献   

11.
【目的】以富士(Fuji)、 秦冠(Qinguan)嫁接在平邑甜茶(Malus hupehensis Rehd.)上的当年生盆栽苗为试验材料,采用砂培方法,研究了缺氮胁迫和干旱对富士和秦冠生长情况、 光合参数、 植株各部位氮磷钾含量及氮素利用效率的影响,分析比较了低氮干旱条件下富士和秦冠生长及氮素利用的差异,以期为果树生产高效肥水利用提供理论指导。【方法】试验共设四个处理: 正常氮正常水(ZZ)、 低氮正常水(DZ)、 正常氮干旱(ZG)、 低氮干旱(DG)。氮素和水分均设置两个水平,分别为正常氮(6 mmol/L NO-3-N)、 低氮(0.3 mmol/LNO-3-N)、 正常供水(保持盆中砂子相对含水量为饱和含水量的80%~85%)、 干旱处理(保持盆中砂子相对含水量为饱和含水量的60%~65%)。【结果】富士和秦冠的生物量(茎和叶)、 株高茎粗等生长指标以及光合速率、 气孔导度、 蒸腾速率均为正常氮正常水(ZZ)>低氮正常水(DZ)>正常氮干旱(ZG)>低氮干旱(DG),并且相对应处理下秦冠的以上指标均高于富士;正常供水下,缺氮处理使富士、 秦冠的根冠比比正常氮处理均有所增加,富士提高了2.05%,秦冠提高了22.40%。富士和秦冠的氮、 磷、 钾含量均表现出正常氮正常水(ZZ)>低氮正常水(DZ)>正常氮干旱(ZG)>低氮干旱(DG); 氮、 钾元素含量在植株各部位的分布顺序依次是叶>根>茎,磷元素则是根>叶>茎;光合氮素利用效率(PNUE)和氮素利用效率表现为秦冠处理之间差异极显著,富士处理之间差异不显著;秦冠的PNUE和NUE明显高于富士,在低氮正常水(DZ)处理下,秦冠氮肥利用率比富士高42.07%,在低氮干旱(DG)处理下高64.14%;低氮胁迫下富士和秦冠的NUE显著提高,并且秦冠提高的幅度高于富士。【结论】施用氮肥能够显著提高富士与秦冠的干物质量,同等水肥条件下,秦冠生长优于富士;水分亏缺会减少叶片对氮的吸收,干旱条件下适度增施氮肥,可提高果树的抗旱能力;低氮干旱胁迫下秦冠的生长指标、 光合指标及氮素利用效率指标均优于富士,表现出较强的抗低氮干旱胁迫的能力。  相似文献   

12.
Laser-induced breakdown spectroscopy (LIBS) is a new technique for the analysis of plant material. This study investigates the application of LIBS to pasture-based plant samples. The LIBS measurements were obtained from pelletized pasture samples (100 samples) that had been also analyzed by inductively coupled plasma–optical emission spectroscopy (ICP-OES) following microwave digestion for calibration and comparison purposes. Comparisons for elements sodium (Na), potassium (K), magnesium (Mg), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), boron (B), phosphorus (P), and sulfur (S) showed that LIBS could be used for almost all the standard profile total elements with concentrations down to low mg/kg levels (observed error of Na: 0.024 percent, K: 0.18 percent, Mg: 0.016 percent, Ca: 0.073 percent, P: 0.017 percent, Mn: 31 mg/kg, Fe: 150 mg/kg, Zn: 6.6 mg/kg, and B: 1.1 mg/kg). Elemental analysis at less than mg/kg levels was not possible using LIBS. The elements S and Cu were particularly difficult to analyze with reliability using LIBS at the concentration levels found in the plant samples. Replacing microwave digestion and subsequent ICP analysis with a direct analysis of dried plant samples using LIBS has the potential to improve the productivity and reduce the cost of testing.  相似文献   

13.
正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,  相似文献   

14.
The responses of tallgrass prairie plant communities and ecosystem processes to fire and grazing are well characterized. However, responses of invertebrate consumer groups, and particularly soil-dwelling organisms, to these disturbances are not well known. At Konza Prairie Biological Station, we sampled soil macroinvertebrates in 1994 and 1999 as part of a long-term experiment designed to examine the effects and interactions of annual fire, mowing, and fertilization (N and P) on prairie soil communities and processes. For nearly all taxa, in both years, responses were characterized by significant treatment interactions, but some general patterns were evident. Introduced European earthworms (Aporrectodea spp. and Octolasion spp.) were most abundant in plots where fire was excluded, and the proportion of the total earthworm community consisting of introduced earthworms was greater in unburned, unmowed, and fertilized plots. Nymphs of two Cicada genera were collected (Cicadetta spp. and Tibicen spp.). Cicadetta nymphs were more abundant in burned plots, but mowing reduced their abundance. Tibicen nymphs were collected almost exclusively from unburned plots. Treatment effects on herbivorous beetle larvae (Scarabaeidae, Elateridae, and Curculionidae) were variable, but nutrient additions (N or P) usually resulted in greater densities, whereas mowing usually resulted in lower densities. Our results suggest that departures from historical disturbance regimes (i.e. frequent fire and grazing) may render soils more susceptible to increased numbers of European earthworms, and that interactions between fire, aboveground biomass removal, and vegetation responses affect the structure and composition of invertebrate communities in tallgrass prairie soils.  相似文献   

15.
Abstract

A 3-year study was carried out to investigate quality parameters in 14 tree fruit and berry species grown in southern Norway. The species were blueberry, apple, aronia, sour cherry, sweet cherry, red raspberry, strawberry, blackcurrant, gooseberry, red currant and elderberry, harvested along with wild bilberry, cloudberry and lingonberry. Significant differences between species were identified for all quality parameters. The coefficient of variation between species was lowest for pH (12.5%), dry matter (18.9%) and soluble solids (25.3%), followed by titratable acids (59.3%), total phenolics (83.8%), antioxidant capacity FRAP (85.7%) and antiradical power by the DPPH-assay (97.8%), total monomeric anthocyanins (132%) and ascorbic acid (137%). Average coefficient of variation within species were lower and ranged from 4 (pH) to 62% (ascorbic acid). Only the FRAP values were significantly affected by harvesting year with lower levels in 2004 than in 2005 and 2006. There were significant interactions between species and harvesting year for dry matter, soluble solids, pH, ascorbic acid and FRAP. The results indicate generic ranges in composition within species independent upon growing location and climate, and the composition of the tree fruits and berries is not likely to deviate from these ranges. It is concluded that desirable composition of tree fruits and berries and their products should primarily be achieved by selection among species rather than searching fors broadened variation within individual species.  相似文献   

16.
Potassium (K) fixation and release in soil are important factors in the long-term sustainability of a cropping system. Changes in K concentration and characteristics of K fixation and release in rhizosphere and nonrhizosphere soils in the rapeseed (Brassica napus L.)–rice (Oryza sativa L.) rotation were investigated using a rhizobox system. The concentrations of different forms of K in both rhizosphere and nonrhizosphere soils decreased with plants compared to without plants, regardless of K fertilizer application. Potassium uptake by crops mainly came from the rhizosphere soil. In the treatment without K fertilizer (–K), the main form of K supplied by the soil to the crops was 1.0 mol L?1 nitric acid (HNO3) nonextractable K, followed by nonexchangeable K, and then exchangeable K. In the treatment with K fertilizer (+K), the main K forms supplied by the soil to the crops were exchangeable K and nonexchangeable K. The amount and rate of K fixation after one cycle of the rapeseed–rice rotation was greater in rhizosphere soil than in nonrhizosphere soil. The amount and rate of K fixation of soil in the +K treatment were significantly less than in the –K treatment. The cumulative amounts of K released with 1.0 mol L?1 ammonium acetate (NH4OAc) and 1.0 mol L?1 HNO3 extraction increased with the increasing numbers of extractions, but the K-releasing power of soil by successive extraction decreased gradually and finally became almost constant. The release of K was less in rhizosphere soil than in nonrhizosphere soil. The release of K in the +K treatment was similar to that in the –K treatment in rhizosphere soil, but the K release in nonrhizosphere soil was greater with the +K than the –K treatment. Overall, the information obtained in this study will be helpful in formulating more precise K fertilizer recommendations for certain soils.  相似文献   

17.
Inoculation of wheat seedlings with the plant growth-promoting bacterium Azospirillum brasilense Cd was immobilized in alginate microbeads and, without applying any stress, significantly increased the quantity of several photosynthetic pigments, such as chlorophyll a, chlorophyll b, and the auxiliary photoprotective pigments violaxanthin, zeaxanthin, antheroxanthin, lutein, neoxanthin, and β-carotene. This resulted in greener plants with no apparent visible stress. After monitoring the quantity of photosynthetic pigments for 4 weeks, we observed that inoculated plants had higher quantities of pigments in shoot and stem. The greatest difference in the quantity of all pigments between inoculated and noninoculated plants occurred in the first week of growth. Regardless of treatment, the quantity of pigments in stems was three to four times less than the quantity of these pigments in shoots. Application of Azospirillum, either as liquid inoculant or as alginate microbeads, did not alter the positive effect of the bacteria on pigment production or the positive response of the plants towards A. brasilense Cd inoculation.  相似文献   

18.
We examined the community composition of microbes that colonized atrazine-containing beads buried in agricultural soils that differed in atrazine treatment history. Bacterial abundance was 5-40-fold greater in atrazine-fortified beads. In beads containing 20 mg atrazine kg−1 buried in soil with a history of atrazine application (conditioned soil), the abundance of Actinobacteria increased approximately 80-fold whereas in control soil, Actinobacteria were enriched only 10-fold and the gamma-Proteobacteria and Planctomycetes increased by 60- and 25-fold, respectively. The gamma-Proteobacteria were enriched by 120- and 230-fold in beads containing 200 mg atrazine kg−1 in conditioned and control soil, respectively. The results demonstrate that BioSep® beads are a suitable matrix for recruiting a diverse subset of the bacterial community involved in atrazine degradation.  相似文献   

19.
Arbitrary oligonucleotides were used as primers to amplifygenomic DNA of 48 wild Spanish populations of Agropyroncristatum, Elymus hispanicus,E. caninus,E. repens,Thinopyrum curvifolium, Th.junceum and Th.intermedium. Genetic diversity was analysedusing nineteen primers. The number of amplified products ranged from8 to 18 per primer and a total of 240 markers were scored. Differentlevels of intraspecific genetic diversity were found, the allogamousspecies E. repens andTh. intermedium being themost variable. Jaccard's similarity coefficients for internalmeasure within and between populations were used to produce a clusterdiagram. The results demonstrate differences in the degree ofsimilarity between taxonomic units. Interpopulational variability andinterspecific genomic relationships of these species arediscussed.  相似文献   

20.
The effects of three commonly used fungicides on the colonization and sporulation by a mixture of three arbuscular mycorrhizal (AM) fungi consisting of Glomus etunicatum (Becker & Gerd.), Glomus mosseae (Nicol. & Gerd.) Gerd. & Trappe, and Gigaspora rosea (Nicol. & Schenck) in symbiosis with pea plants and the resulting response of the host-plant were examined. Benomyl, PCNB, and captan were applied as soil drenches at a rate of 20 mg active ingredient kg-1 soil 2 weeks after transplanting pea seedlings in a silty clay-loam soil containing the mixed inocula of AM fungi (AM plants). Effects of fungicides were compared to untreated plants that were inoculated with fungi (AM control). The effect of mycorrhizal inoculation on plant growth was also examined by including nonmycorrhizal, non-fungicide-treated plants (non-AM control). Fungicides or inoculation with AM fungi had only a small effect on the final shoot weights of pea plants, but had greater effects on root length and seed yield. AM control plants had higher seed yields and lower root lengths than the corresponding non-AM plants, and the fungicide-treated AM plants had intermediate yields and root lengths. Seed N and P contents were likewise highest in AM control plants, lowest in non-AM plants, and intermediate in fungicide-treated AM plants. All three fungicides depressed the proportion (%) of root length colonized by AM fungi, but these differences did not translate to reductions in the total root length that was colonized, since roots were longer in the fungicide-treated AM plants. Pea plants apparently compensated for the reduction in AM-fungal metabolism due to fungicides by increasing root growth. Fungicides affected the population of the three fungi as determined by sporulation at the final harvest. Captan significantly reduced the number, relative abundance, and relative volume of G. rosea spores in the final population relative to the controls. The relative volume of G. etunicatum spores was greater in all the fungicide-treated soils, while G. mosseae relative volumes were only greater in the captan-treated soil. These findings show that fungicides can alter the species composition of an AM-fungal community. The results also show that AM fungi can increase seed yield without enhancing the vegetative shoot growth of host plants.  相似文献   

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