外源趋化剂对气单胞菌134的趋化性及其在水稻根际定殖的影响

本研究从镉污染水稻根际土壤中分离、纯化出一株耐镉菌134,并对该菌株的生理生态特征、16S r DNA等进行了系列分析,结果表明该菌株为气单胞菌(Aeromonas sp.),革兰氏阴性,对镉离子的耐受质量浓度为75 mg/L。选取水稻根系的常见分泌物氨基酸和糖类作为外源趋化剂考察气单胞菌134对各组分的趋化性,结果表明,菌株134对组氨酸、丙氨酸、亮氨酸、葡萄糖及果糖均具有较强的正趋化性。类毛细血管定量试验进一步表明,菌株134对不同浓度趋化剂的响应不同,在亮氨酸80μmol/L、组氨酸40μmol/L、丙氨酸60μmol/L、葡萄糖60μmol/L及果糖100μmol/L时,菌株134的趋化性最强。综合考虑滴定分析试验和类毛细血管试验结果,本文设计了水培试验来研究外源添加适宜浓度的组氨酸、亮氨酸、葡萄糖及D-果糖对菌株134在水稻根际定殖的影响。试验结果表明,这4种外源趋化剂均能促进菌株134在水稻根际的定殖,尤其是葡萄糖和D-果糖的处理,其根际定殖细菌数量与对照相比差异极显著(P0.01),分别为对照组的3.48和3.87倍。     

小麦根际联合固氮菌(固氮螺菌,Azospirillum brasilense)田间接种效果的研究

据黑龙江省1990年7个中试点试验证明,小麦根际联合固氮菌田间接种效果在5.2—20.6％,平均为9.8％;每田可增产小麦19.1公斤。小麦根际联合固氮菌能促进小麦本长发育,在生育期间可使株高、鲜重增加,叶色变缘。秋收考种证明,对产量构成因子的平方米穗数、穗粒数和千粒重均有增加。     

生防菌AS818抗药性标记株在大豆根际定殖

通过₆₀Co 射线 (1 .0× 1 0 ⁴ rad)对生防菌株链霉菌AS81 8孢子悬液 (1 0 6/mL)进行诱变 ,得到抗链霉素 (>5 0μg/mL)突变株 5株。经诱导 ,RL 4抗链霉素水平达到 1 0 0μg/mL。标记株RL 4在 1 %水琼脂 (WA)和无菌土中培养的大豆根际定殖趋势有所不同。RL 4在1 %WA培养的大豆根表的定殖量呈逐步上升趋势。无菌土中种植的大豆根际和根表的检测表明 :RL 4可在无菌土中大豆根际短期定殖 ,在根际第 1     

硅酸盐细菌NBT菌株在小麦根际定殖的初步研究

对硅酸盐细菌NBT菌株进行了耐药性标记,得到稳定的链霉素抗性标记菌NBT菌株,采用选择性培养基分离计数,通过琼脂平板和盆栽试验,研究了标记菌NBT在小麦根际的定殖动态及影响因素。结果表明,在灭菌土盆栽中,播种后9d左右NBT菌株在小麦根际的定殖水平达最高(1．4×10^8cfu·g^-1根土),播种后54d左右趋向稳定,NBT菌株细胞数量为2．4×10^3cfu·g^-1根土;未灭菌土盆栽中,播种后9d左右NBT菌株的定殖水平达最高(3．8×10^8cfu·g^-1根土),60d左右趋向稳定,菌数为3．1×10^3cfu·g^-1根土,牛物和非牛物因素对NBT菌株定殖小麦根系有影响。     

淡紫拟青霉在大豆根际的定殖及对根际微生物的影响

温室实验证明淡紫拟青霉M－14菌株发酵液对大豆种子包衣,无菌土中种植1周后有1／10的生防菌能够在大豆根际定殖,其中内根际（根表与根内）可检测到菌剂包衣量的1／1000．第2周时内根际菌量提高10倍以上,4周后有所下降。2％NaClO表面消毒后,检测出植物根内也有少量淡紫拟青霉存在。大豆胞囊线虫感病土中,该菌3周后开始大量增殖。病土中引入菌剂,1周后根际其它各种微生物数量略有减少,真菌、细菌、放线菌分别减少16％、27％和27％,4周时均达到平衡,与对照一致。     

北京地区水稻根际联合固氮菌的分离鉴定及其固氮酶活性的研究

从北京地区16个水稻品种的根际分离筛选出两株固氮酶活性较高的菌株D-12和D-25。对这两个菌株进行了分类鉴定和固氮酶活性的测定。根据形态特征和理化特性的测定,菌株D-12属于肺炎克雷伯氏菌(Klebsiella pneumoniae),菌株D-25类似于克雷伯氏菌但又有区别,因此暂放在胁杆菌科(Enterobacteriaceae)。两个菌株均在厌氧条件下固氮酶活性最高。在Hill's无氮蔗糖培养基中30℃条件下,其固氮酶活性的高峰出现在第16至第18小时。     

土壤有益细菌在植物根际竞争定殖的影响因素

在土壤有益微生物应用于生物肥料、生物杀虫剂、植物生长刺激剂和生物处理剂的过程中,根际定殖具有重要作用。细菌在植物根际定殖是一个比较复杂的过程,影响定殖能力的因素也是复杂多样的。本文综述了参与根部竞争定殖的生物因素,包括受细菌遗传控制的某些特性如鞭毛/运动性、趋化性、多糖、位点特异重组酶/菌落阶段变异、NADH脱氢酶,植物根的分泌物和植物种类等;影响微生物根际定殖的非生物因素如土壤类型、土壤特性和土壤温度等,探讨了影响微生物根际定殖的主要研究方向。     

芽胞杆菌Q13和Q14在猕猴桃叶面的定殖及其对叶面菌群的影响

为了进一步明确芽胞杆菌Q13、Q14菌株对猕猴桃叶枯病的生防作用,分别对芽胞杆菌Q13、Q14菌株在猕猴桃叶面上的定殖作用及其对叶面其他微生物种类和数量的影响进行了考察。结果表明,在田间条件下,芽胞杆菌Q13、Q14能在猕猴桃叶面定殖,但随时间的变化呈下降趋势;在无降雨等恶劣天气影响的情况下,该菌能在猕猴桃叶面有效定殖8 d左右,8 d后叶面检测到的Q13、Q14菌株的菌量为4.5×106cfu/g成熟叶。与未喷施菌剂的猕猴桃叶面菌群对比,喷施菌剂后的猕猴桃叶面细菌种类多了芽胞杆菌属(Bacillus subtilis),少了条件致病菌不动杆菌属(Acinetobacter soli);真菌在数量上癣囊腔菌(Plectosphaerellacucumerina)、Pseudozyma flocculosa和米曲霉菌(Aspergillus oryzae)增加了,泡状莫氏黑粉菌(Moesziomyces bullatus)和尖孢镰刀菌(Fusarium oxysporum)减少了。     

茶树内生木霉种的鉴定及其在植物体内的定殖

采用分离自健康茶树叶片组织的一株长枝木霉Trichoderma longibrachiatum菌株CSN-18,研究茶树内生木霉的人工接种、再分离及其在茶树地上部组织中的内生性。该菌在PDA培养基上生长速度快,产孢量大。用CSN-18回接茶苗,接种后一个月可以从茶苗的茎和叶组织中再分离获得该真菌。接种后的茶苗没有观察到明显的病害表现;与对照相比,接种后6个月组培苗生长良好,叶色更绿;接种后1个月,实生苗生长正常。通过石蜡切片和苯胺蓝染色,在已接种的茶树组培苗的叶片内部组织中可观察到木霉的存在,从而证明了木霉能够在茶树地上部组织内定殖,是茶树的一种内生真菌。     

盐胁迫下粪产碱菌在水稻根表的定殖和异源固氮基因的表达

异源nif LacZ融合基因在粪产碱菌A15 6 1中的表达活性随盐浓度增加而升高 ,然后逐渐降低 ,nifH LacZ融合基因可以正常表达的盐浓度在 0 .1%～0 .5 %之间 ,盐浓度为 0 .0 5 %时活性最高。A15 6 1在盐浓度为 0 .0 6 %时趋化能力最强 ,随着盐浓度的提高逐渐下降 ,当盐浓度为 3 .0 %时完全丧失趋化能力。一定的盐浓度 (0 .5 % )对固氮粪产碱菌的根表定殖有促进作用 ,该条件下根表定殖的菌体数远大于对照。 3种nif LacZ融合基因在根内的表达部位有显著差异。nifH的表达部位主要分布于根的皮层薄壁组织细胞间隙 ,在条件适宜 (无铵和微量氧 )的部位或某些特殊位置如侧根伸出部位高水平表达。盐胁迫下水稻 耐盐粪产碱菌A15 6 1的联合固氮效率明显高于A15 6 1纯培养物     

Symbiosome-like intracellular colonization of cereals and other crop plants by nitrogen-fixing bacteria for reduced inputs of synthetic nitrogen fertilizers

It has been forecast that the challenge of meeting increased food demand and protecting environmental quality will be won or lost in maize, rice and wheat cropping systems,and that the problem of environmental nitrogen enrichment is most likely to be solved by substituting synthetic nitrogen fertilizers by the creation of cereal crops that are able to fix nitrogen symbiotically as legumes do. In legumes, rhizobia present intraceliularly in membrane-bound vesicular compartments in the cytoplasm of nodule cells fix nitrogen endosymbiotically. Within these symbiosomes, membrane-bound vesicular compartments, rhizobia are supplied with energy derived from plant photosynthates and in return supply the plant with biologically fixed nitrogen, usually as ammonia. This minimizes or eliminates the need for inputs of synthetic nitrogen fertilizers. Recently we have demonstrated, using novel inoculation conditions with very low numbers of bacteria, that cells of root meristems of maize, rice, wheat and other major non-legume crops, such as oilseed rape and tomato, can be intracellularly colonized by the non-rhizobial, non-nodulating, nitrogen fixing bacterium, Gluconacetobacter diazotrophicus that naturally occurs in sugarcane. G. diazotrophicus expressing nitrogen fixing (nifH) genes is present in symbiosome-like compartments in the cytoplasm of cells of the root meristems of the target cereals and non-legume crop species, somewhat similar to the intracellular symbiosome colonization of legume nodule cells by rhizobia. To obtain an indication of the likelihood of adequate growth and yield, of maize for example, with reduced inputs of synthetic nitrogen fertilizers,we are currently determining the extent to which nitrogen fixation, as assessed using various methods, is correlated with the extent of systemic intracellular colonization by G. diazotrophicus,with minimal or zero inputs.     

Symbiosome-like intracellular colonization of cereals and other crop plants by nitrogen-fixing bacteria for reduced inputs of synthetic nitrogen fertilizers

It has been forecast that the challenge of meeting increased food demand and protecting environmental quality will be won or lost in maize, rice and wheat cropping systems, and that the problem of environmental nitrogen enrichment is most likely to be solved by substituting synthetic nitrogen fertilizers by the creation of cereal crops that are able to fix nitrogen symbiotically as legumes do. In legumes, rhizobia present intracellularly in membrane-bound vesicular compartments in the cytoplasm of nodule cells fix nitrogen endosymbiotically. Within these symbiosomes, membrane-bound vesicular compartments, rhizobia are supplied with energy derived from plant photosynthates and in return supply the plant with biologically fixed nitrogen, usually as ammonia. This minimizes or eliminates the need for inputs of synthetic nitrogen fertilizers. Recently we have demonstrated, using novel inoculation conditions with very low numbers of bacteria, that cells of root meristems of maize, rice, wheat and other major non-legume crops, such as oilseed rape and tomato, can be intracellularly colonized by the non-rhizobial, non-nodulating, nitrogen fixing bacterium, Gluconacetobacter diazotrophicus that naturally occurs in sugarcane. G. diazotrophicus expressing nitrogen fixing (nifH) genes is present in symbiosome-like compartments in the cytoplasm of cells of the root meristems of the target cereals and non-legume crop species, somewhat similar to the intracellular symbiosome colonization of legume nodule cells by rhizobia. To obtain an indication of the likelihood of adequate growth and yield, of maize for example, with reduced inputs of synthetic nitrogen fertilizers, we are currently determining the extent to which nitrogen fixation, as assessed using various methods, is correlated with the extent of systemic intracellular colonization by G. diazotrophicus, with minimal or zero inputs.     

The survival and development of inoculant ectomycorrhizal fungi on roots of outplanted Eucalyptus globulus Labill

The survival and development of two inoculant ectomycorrhizal fungi (Hebeloma westraliense Bough. Tom. and Mal. and Setchelliogaster sp. nov.) on roots of outplanted Eucalyptus globulus Labill. was examined at two expasture field sites in the south-west of Western Australia. Site 1 was a gravelly yellow duplex soil, and Site 2 was a yellow sandy earth. Plants were grown in steamed or unsteamed soil, in root bags designed as field containers for young growing trees. Three, 6 and 12 months after outplanting, plants were removed from these bags and assessed for dry weights of shoots and ectomycorrhizal colonization of roots.The inoculant ectomycorrhizal fungi (identified on the basis of the colour and morphology of their mycorrhizas) survived on roots of E. globulus for at least 12 months after outplanting at both field sites. At Site 1, however, colonization of new fine roots by the inoculant fungi was low (less than 20% of fine root length). Inoculation had no effect on the growth of E. globulus at this site. In contrast, at Site 2 the inoculant ectomycorrhizal fungi colonized up to 30–50% of new fine root length during the first 6 months after outplanting. There was a corresponding growth response to ectomycorrhizal inoculation at this site, with a close relationship (r²=0.82^**) between plant growth at 12 months and root colonization at 3 months. Plant growth at 12 months was related less closely with root colonization at 6 or 12 months. Root colonization by resident ectomycorrhizal fungi increased with time at both field sites. At Site 2, this increase appeared to be at the expense of colonization by the inoculant fungi, which was reduced to less than 10% of fine root length at 12 months. Steaming the soil had little effect on colonization by the inoculant ectomycorrhizal fungi at either field site, but decreased colonization by the resident ectomycorrhizal fungi.     

Endophytic colonization of plant roots by nitrogen-fixing bacteria

Plant and Soil - Nitrogen-fixing bacteria are able to enter into roots from the rhizosphere, particularly at the base of emerging lateral roots, between epidermal cells and through root hairs. In...     

Increased root exudation of14C-compounds by sorghum seedlings inoculated with nitrogen-fixing bacteria

Summary Organic components leaked fromSorghum bicolor seedlings (‘root exudates’) were examined by recovering¹⁴C labelled compounds from root solutions of seedlings inoculated withAzospirillum brasilense, Azotobacter vinelandii orKlebsiella pneumoniae nif-. Up to 3.5% of the total¹⁴C recovered from shoots, roots, and nutrient solutions was found in the root solutions. Inoculation with Azospirillum and Azotobacter increased the amounts of¹⁴C and decreased the amounts of carbohydrates in the root solutions. When sucrose was added as a carbon source for the bacteria, the increase of¹⁴C in the solutions did not occur. Quantities of¹⁴C found in the root solutions were proportional to amounts of mineral nitrogen supplied to the plants. Bacterial growth also was proportional to nitrogen levels. When sorghum plants were grown in soil and labelled with¹⁴CO₂, about 15% of the total¹⁴C recovered within 48 hours exposure was found in soil leachates.     

Adhesion of fimbriated nitrogen-fixing enteric bacteria to roots of grasses and cereals

Summary The role of fimbriae in enterobacterial adhesion to roots of grasses and cereals is discussed. All nitrogen-fixing enteric bacteria isolated in Finland had fimbriae. AllEnterobacter isolates had mannose-binding type-1 fimbriae, whereas most of theKlebsiella isolates had both type-1 and type-3 fimbriae. The strains were isolated from a total of ten different grass species, and no specific association was found between grass species and bacterial fimbriation, biogroup or serogroup. Purified, radiolabeled fimbriae bound to roots ofPoa pratensis in vitro, and bacterial adhesion was inhibited by Fab fragments specific for fimbriae.Klebsiella strains carrying type-3 fimbriae adhered to roots of various grasses and cereals more efficiently than type-1- or nonfimbriated strains, and it was concluded that type-3 fimbriae are the major adhesions ofKlebsiella. Immunofluorescence studies revealed that the bacteria preferentially adhered to root hairs, and to a lesser extent, to the zone of elongation and the root cap mucilage. No strict host specificity in enterobacterial adhesion was observed.     

Effect of diazotrophic bacteria isolated from a mycelium of arbuscular mycorrhizal fungi on colonization of maize roots by Glomus fistulosum

The inoculation of mycorrhizal maize plants with three isolates of microaerophilic diazotrophic bacteria obtained from the mycelium of arbuscular mycorrhizal fungi associated with three grasses (Arrhenatherum elatius - bacterial isolate ARR, Agropyrum repens - isolate AGR and Poa annua - isolate POA) caused no increase in nitrogen content in plant biomass. The inoculation with bacterial isolate ARR resulted in the decreased plant growth. Bacterial isolate AGR decreased the percentage of the root length colonized by arbuscular mycorrhizal fungus Glomus fistulosum. The inoculation with both mycorrhizal fungus and isolate POA increased significantly the concentration of phosphorus in plant shoots compared to uninoculated control. This revised version was published online in July 2006 with corrections to the Cover Date.     

Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110.

The complete nucleotide sequence of the genome of a symbiotic bacterium Bradyrhizobium japonicum USDA110 was determined. The genome of B. japonicum was a single circular chromosome 9,105,828 bp in length with an average GC content of 64.1%. No plasmid was detected. The chromosome comprises 8317 potential protein-coding genes, one set of rRNA genes and 50 tRNA genes. Fifty-two percent of the potential protein genes showed sequence similarity to genes of known function and 30% to hypothetical genes. The remaining 18% had no apparent similarity to reported genes. Thirty-four percent of the B. japonicum genes showed significant sequence similarity to those of both Mesorhizobium loti and Sinorhizobium meliloti, while 23% were unique to this species. A presumptive symbiosis island 681 kb in length, which includes a 410-kb symbiotic region previously reported by G?ttfert et al., was identified. Six hundred fifty-five putative protein-coding genes were assigned in this region, and the functions of 301 genes, including those related to symbiotic nitrogen fixation and DNA transmission, were deduced. A total of 167 genes for transposases/104 copies of insertion sequences were identified in the genome. It was remarkable that 100 out of 167 transposase genes are located in the presumptive symbiotic island. DNA segments of 4 to 97 kb inserted into tRNA genes were found at 14 locations in the genome, which generates partial duplication of the target tRNA genes. These observations suggest plasticity of the B. japonicum genome, which is probably due to complex genome rearrangements such as horizontal transfer and insertion of various DNA elements, and to homologous recombination.