1株具解磷特性大豆根瘤菌的分离筛选与回接鉴定

从沈阳农业大学的科研基地分离得到20株根瘤菌,进一步利用乙炔还原法和溶磷圈法得到1株兼具有结瘤固氮和解磷作用的根瘤菌#6菌株。#6的固氮酶活性为4.94μmolC2H4.g-1.min-1,与参照菌株USDA110的固氮酶活性相当;用溶磷圈法定性试验#6解磷性,其可以溶解有机磷,对照菌株USDA110无解磷现象;进一步测定#6的解磷量,其解磷量分别在第7天为35.50μg/mL,第14天为21.05μg/mL;对根瘤菌#6生理生化和生长特性进行试验,以利于实际应用。     

豆科植物—根瘤菌共生固氮的基因调控和转移

豆科植物-根瘤菌共生固氮是由寄主与细菌双方基因共同参与完成的,在相互基因调控中形成特导的结构—根瘤。参与根瘤菌固氮(nif基因)和结瘤(nod基因)的基因已被克隆和分析。     

新疆干旱地区根瘤菌资源研究I．根瘤菌种类及其共生固氮作用

从新疆不同生态条件下生长的31属109种豆科植物根瘤分离获得373株根瘤菌,其中88株是从未报道过结瘤情况的39种豆科植物根瘤分离获得。它们与豆科寄主植物共生结瘤,95％以上为有效根瘤。不同种的根瘤固氮活性差异较大,黄芪属根瘤固氮活性较高,最高者为大豆根瘤固氮活性的42倍。 20属37种豆科植物根瘤中97％有吸氢活性。根瘤固氮活性、吸氢活性均与寄主植物生长发育期有相关性。     

根瘤菌nod fix和nif基因及其功能的信息汇编

nod基因 nodABC nodABC的产物是根毛弯曲和皮层细胞分裂所必需的。NodABC很可能合成一种似植物激素的物质由R.meliloti产生的这种物质称NodRm-1最近已被鉴定是末端硫酸化的,脂酰-3-N-乙基-1,4-D-葡萄糖胺还原态四聚糖。     

大豆基因工程根瘤菌田间结瘤和共生固氮效应

利用三亲本接合转移方法,将快生型大豆根病菌B_(52)的基因文库克隆转移到受体菌慢生型大豆根瘤菌2210中,获得4株大豆基因工程根瘤菌(32.43、B—2—6、B—3—8)。田间接种试验表明:在大豆分枝期(V2),43和32结瘤效分别比不接种对照增加117.9％和100.4％;在初花期(R_1),32和B—2—6结瘤数分别比对照增加122.5％和91.6％,根瘤固氮酶活性增加233.3％和195.6％;在结荚始期(R3),B—2—6和32结瘤数比对照增加78.5％和70.1％,根瘤固氟酶活性43和B—2—6分别比对照增加53.4％和49.3％。产量统计表明:32、B—2—6和43比对照分别增产16.8％、14.3％和12.2％,亩增收大豆分别为28.2、24.1和20.6公斤。以上三个菌株均比受体菌株2210增产率高。     

1株高效花生根瘤菌的筛选与鉴定

从辽宁多地花生种植土壤及其鲜根瘤初步筛选到30株花生根瘤菌,进一步通过回接盆栽花生,测定花生根瘤数、根瘤干重,以及鲜根瘤固氮酶活性、植株全氮量等,筛选出1株结瘤固氮能力较强花生根瘤菌wz-6,通过16S rDNA序列分析,鉴定为圆明慢生根瘤菌(Bradyrhizobium yuanmingense)。为开发优质花生根瘤菌菌剂奠定基础。     

豆科植物与根瘤菌共生固氮的宿主专一性

根瘤菌和豆科植物的共生固氮作用是当前最重要的植物与微生物相互作用的模式之一.本文将综述:1.根瘤菌在感染植物前发生的变化;2.根瘤形成早期的生理和发育过程;3.根瘤菌的结瘤基因的结构和功能;4.植物产生的黄酮类化合物的结构及     

我国豆科植物根瘤菌资源多样性及应用基础研究

北京农业大学菌种保藏中心(CCBAU)现已保藏根瘤菌5000余株,是全世界最大的根瘤菌资源数据库。通过对其中的2000余株根瘤菌作多相分类研究,确定根瘤菌新属2个、新种11个。结合根瘤菌宿主及其生态环境的关系,提出根瘤菌与豆科植物共生关系的新认识;并得出豆科植物接种根瘤菌的新见解,这对于西部大开发中新区种植豆科植物接种适宜的根瘤菌具有重要指导意义。     

Metabolism and translocation of fixed nitrogen in the nodulated legume

Summary Nitrogen (N₂) fixed by Rhizobium bacteroids in the legume nodule is excreted as ammonia to the surrounding host cell where it is efficiently assimilated into the amide group of glutamine. Generally glutamine is a minor exported solute of nitrogen, being further metabolised to asparagine in temperate species and to the ureides, allantoin and allantoic acid in tropical species. These solutes serve as the principal translocated forms of nitrogen in xylem. Compartmentalisation of the pathways of nitrogen metabolism and the role of ammonia in regulation of their activity is examined in nodules of both asparagine-forming (Lupinus albus L.) and ureide-forming (Vigna unguiculata L. Walp) symbioses.     

Micro-morphology of common bean (<Emphasis Type=Italic>Phaseolus vulgaris</Emphasis> L.) nodules undergoing senescence

We studied morphological changes over time by nodules formed on the root system of the common bean (Phaseolus vulgaris L.). Two cultivars, Bayomex and Cacahuate 72 with growth habit Type I and the Rhizobium etli strain CE-3 were used. The results showed the collapse of the infected zone, degradation of the cell walls and membranes, changes in the number and distribution of the starch granules, appearance of protein granules, and disintegration of the central tissue of the nodule with ageing. Additionally, we describe the influence of time on the progress of the nodular senescence.     

Host-genetic control of nitrogen fixation in the legume-Rhizobium symbiosis: complication in the genetic analysis due to maternal effects

Summary In pea cv. Afghanistan a recessive gene sym 6 prevents the full expression of nitrogenase activity in root nodules, induced byRhizobium leguminosarum strain F 13. In contrast, nitrogenase activity is fully expressed in pea cv. Iran. A comparison of the reciprocal hybrids of these two plants showed that the size of the plant was determined by the mother plant (maternal effect). Therefore the shoot weight and the total amount of nitrogen fixed are not suitable as parameters for a genetic analysis. The % nitrogen of the shoot and the specific activity of the nodules per gram of nodules are more reliable, but for practical purposes the specific activity of the nodules expressed per gram of shoot tissue can be used.     

Non-destructive semi-quantitative determination of acetylene reduction in the field

Summary Direct injection of acetylene into soil around plant roots, followed by determination of ethylene/acetylene ratios in the soil atmosphere has been tested as a rapid, non-destructive method of estimating acetylene reducing activity. In pots of artificial media as well as in field soil, the ratios determined within 10 min. after injection were significantly correlated with the rates of acetylenedependent ethylene production in detached roots. The method may be useful in preliminary screening of large numbers of plant-bacteria combinations.     

Immunolocalization of ferritin in determinate and indeterminate legume root nodules

Summary In eukaryotic organisms ferritin is a protein involved in the storage of iron. The occurrence of ferritin and its relationship to the effectiveness of the nitrogen-fixing activity have been previously studied during the early stages of the nodule development by biochemical methods. We have used immunocytochemistry techniques to determine the precise location of ferritin and the behavior of this protein along the nodule development. The major localization was found in plastids and amyloplasts of infected and uninfected cells of the three legume nodules studied. A decrease of the immunolabelling was observed in infected cells of lupin and soybean senescing nodules and in the senescent zone of indeterminate alfalfa nodules. In the cortex of soybean and lupin nodules, ferritin increased during nodule ageing and the immunogold particles were mainly located in crystalline structures. The putative role of ferritin and plastids during nodule development is discussed.     

Mutagenesis of pea (Pisum sativum L.) and the isolation of mutants for nodulation and nitrogen fixation

Pea mutants for nodulation have been obtained by treating seeds with ethyl methane sulfonate (EMS) followed by 2 screening procedures. In one, mutants resistant to nodulation (nod⁻), or with ineffective nodules (nod⁺, fix⁻) were obtained, whilst in the other 4 hypernodulated mutants (nod⁺⁺) with 5–10 times more nodules than cv. Frisson and expressing a character of nitrate tolerant symbiosis (nts) were discovered. All mutations are under the control of single recessive genes. (nod⁻), (nod⁺, fix⁻) and (nod⁺⁺, nts) mutations result from mutation events at 6, 7 and 1 different loci respectively.

Grafting experiments showed the (nod⁻) and (nod⁺, fix⁻) phenotypes are associated with the root genotypes and that (nod⁺⁺, nts) phenotype is associated with the shoot genotype.     

Nodule Organogenesis in Lotus japonicus

Lotus japonicus. Received 25 September 2000/ Accepted in revised form 9 October 2000     

Changes in growth and pigment concentrations with leaf age in pea under modulated UV-B radiation field treatments

We assessed whether growth of garden pea (Pisum sativum mutant Argenteum) was reduced under ecologically relevant enhancements of ultraviolet-B radiation (UV-B, 280–320 nm) by employing modulated field lampbanks which simulated 0, 16 or 24% ozone depiction. In addition, we determined whether enhanced UV-B altered the concentration and distribution of chlorophyll and UV-B-absorbing compounds in leaves, and whether this was dependent on leaf age. There were no significant UV-B effects on the four whole-plant parameters we examined (height, above-ground biomass, total leaflet area or average leaflet area). Of the 12 leaf-level parameters we examined, UV-B had a significant effect (P < 0.05) on only one parameter: the ratio of UV-B-absorbing compounds to chlorophyll, which was greatest at the highest UV-B level. Total chlorophyll concentrations tended to be lower under enhanced UV-B (P= 0.11), while the proportion of UV-B-absorbing compounds in the adaxial epidermis tended to be higher (P= 0.11). Total leaf concentrations of UV-B-absorbing compounds were unaffected by UV-B level. Cooler, suboptimal growing conditions during this late summer/early autumn experiment may have masked some potential UV-B effects. In contrast to the UV-B effects, we found strong leaf-age effects on nearly all parameters that we assessed. On an area basis, concentrations of total chlorophyll and UV-B-absorbing compounds increased with leaf age, while Chlorophyll a/b) ratios decreased. One of the few parameters unaffected by leaf age was the ratio of UV-B-absorbing compounds to total chlorophyll, which remained constant within a given UV-B treatment. Pea was much less sensitive to enhanced UV-B than in previous growth-chamber and greenhouse studies, and in nearly all cases UV-B treatment effects were overshadowed by leaf-age effects. In view of the large effect leaf age had on concentrations of UV-B-absorbing compounds, as well as their distribution within leaves, researchers may need to consider leaf age in UV-B experimental designs.