固氮基因nifHDK启动子的功能在大肠杆菌中的验证

近年来,在对肺炎克氏杆菌(Klebsiella pneumoniae)nifA产物研究的基础上已建立了固氮基因调控的模型。固氮基因(nif)的表达受氮调节系统ntr和nifAL操纵子的两个系统的调控。本实验将固氮基因nifHDK启动子、aphA-6基因和rbcL3’构建aphA-6基因表达盒,克隆到pSK.KmR载体,将其转入到大肠杆菌中验证nifHDK启动子的表达活性。结果表明,只有在转入外源nifA基因时,nifHDK启动子才具有转录活性,aphA-6基因才能表达。验证了NifA蛋白和σ54因子是固氮基因的正向调节蛋白,为下一步固氮生物工程研究提供了理论基础。     

樟子松固沙林经营技术的研究

通过合理的经营技术，可培育产量高，质量好，防风效能持久的樟子松固沙林。     

水稻根面争论产碱菌1AG4的分离、鉴定及其固氮和对病原菌的拮抗作用

从水稻(Oryza sativa L.)越光根面分离到一株固氮菌株 1AG4,经生理生化特性鉴定,归类为争论产碱菌(Alcaligenes paradoxus)。1AG4 的固氮酶乙炔还原活性(ARA)为13.06 C2 H4 nm ol/m l·h,固定15N2 活性(N2 FA)为 2.052 15N a.e.% ,表明 1AG4 具有较强的固氮能力。平皿拮抗试验证明 1AG4可拮抗包括水稻三大病害病原菌在内的 14 种植物病原菌(Xanthom onas oryzae pv. oryzae ZHE173、Thanatephorus cucum eris、Piricularia oryzae Cavara、Alternaria longipes(Ellis etEverhart) Tisdale etW adkins、Cochliobolus sativus、Colletotrichum agaves Cavara、Colletotrichum lagenarium (Passerini)Ellis et Halsted、Colletotrichum panacicola Uyeda et Takim oto、Fusarium oxysp orum f. niveum 、Helm inthosporium turcicum Passerini、Phytophthora parasitica Destur、Verticillium albo-atrum Reinkeet Berthold、Phytophthora m elonis Katsura、Pseudom onas solanacearum Sm ith)的生长     

Nodule phosphorus requirement and O2 uptake in common bean genotypes under phosphorus deficiency

Abstract

The effect of P deficiency on nodulation, nodule P content, nodule O₂ permeability and N fixation rates in Phaseolus vulgaris–rhizobia symbiosis was studied under glasshouse conditions. Four recombinant inbred lines (L34, L83, L115 and L147) and one variety cultivated in Morocco (Concesa) were inoculated with Rhizobium tropici CIAT 899 in hydroaeroponic culture. Two P levels i.e. 75 (deficient level) and 250 µmol plant^?1 week^?1 P (sufficient level) were applied and the trial was assessed 42 days after transplanting that coincide with plant flowering stage. Under P-deficiency, decrease of plant growth (18%) and nodule biomass (19%) was detected and significantly pronounced in the sensitive line L147 compared with the remaining genotypes. Additionally, under P-deficiency, the efficiency in use of rhizobial symbiosis, estimated by the slope of the regression model of shoot biomass as a function of nodule biomass, was significantly increased in the four lines. This constraint did not significantly influence nodule P content in Concesa, but it was 24 and 41% lower in the tolerant and in the sensitive lines, respectively. Nodule P content was positively correlated to nodule biomass, r=0.75, and shoot N, r=0.92. These genotypic variations were associated with variability in nodule O₂ permeability that was significantly affected by the P level-bean genotype interaction. Under P-deficiency, nodule O₂ permeability was significantly reduced in the tested genotypes and accompanied with a decrease in shoot N content, especially in the sensitive lines (35%). Moreover, the ratios plant N fixed: nodule P content and plant N fixed:nodule dry weight were affected under P-deficiency in four lines with an exception observed in Concesa. Depending on the observed data we concluded that N₂ fixation efficiency could be influenced by nodulation and level of nodule P requirement which depend on both bean genotypes and P level.     

毛竹根际分离的地衣芽孢杆菌固氮特性研究

从毛竹根际分离到36株固氮菌，经初步镜检其中17株菌为芽孢杆菌，进一步鉴定确定其主要为多粘芽孢杆菌（Bacilluspolymyxa）和地衣芽孢杆菌（Bacilluslicheniformis），本文主要就其中菌号为G－8、G－12、G－14、G－17四株地衣芽孢杆菌的鉴定及固氮特性进行研究。结果显示：四株菌均为杆状，具芽孢孢孢囊稍膨大，它们的生理生化特性相同，但菌落形态有一些差异。G－8和G－17的最佳固氮温度为35℃左右，G－12和G－14的最佳固氮温度为30℃左右；G－12、G－8和G－14的最适固氮pH为6．0左右，G－17的最适固氮pH为6.0～7.5；四株菌都可利用碳源固氮，但均以葡萄糖为碳源时的固氮活性最高，依次为64．76、437．23、65．42、90．91nmolC2H4h-1，瓶-1。G－12在培养48h，G－8、G－14在培养96h，G－17在培养60h的固氮活性最高分别为138．50、122．01、170．87、109．81nmolC2H4．h-1.瓶-1。     

Genetic Effects on Host × Strain Interaction in the Symbiosis of Vicia faba and Rhizobium leguminosarum

50 Vicia faba lines were inoculated with 4 strains of Rhizobium leguminosarum in all possible combinations under greenhouse conditions in 5 replications. A root-sterile method was used for the cultures. Characters such as plant height, leaf colour, number and colour of nodules, the dry weights of shoot, root and nodules, and the nitrogen content were recorded, after 10 weeks. It was demonstrated by analysis of variance that, for example, for the dry matter of the shoot only 1.9 % of the total phenotypic variance was caused by interaction between Vicia and Rhizobium genotypes, but 24.7 % Was conditioned by the faba bean genotypes and 73.4 % by the Rhizobium strains. Differences of Vicia faba genotypes in their reaction to different Rhizobium strains were detected by joint regression analysis.     

Transfer of Grain Legume Nitrogen within a Crop Rotation Containing Winter Wheat and Winter Barley

In a crop rotation trial, conducted from 1985 to 1988 at TU-Munich's research station in Roggenstein, the transfer of grain legume nitrogen was evaluated in crop rotations containing fababeans and dry peas as well as oats (reference crop) and winter wheat and winter barley as following crops. The results obtained can be summarized as follows: Dinitrogen fixation by fababeans ranged from 165 to 240 kg N ha¹, whereas N₂-fixation by peas amounted from 215 to 246 kg N ha^?1. In all seasons the calculated N-balance where only grain was removed was positive, with a net gain being on average 106 (peas) and 84 (fababeans) kg N ha^?1. After the harvest of peas 202 kg N ha^?1 remained on the field on average over seasons (158 kg N ha^?1 in the above ground biomass and 44 kg N ha^?1 as NO₃-N in 0–90 cm depth). As compared to peas, fababeans left 41 kg N ha^?1 less due to smaller amounts of nitrogen in the straw. After oats very small amounts of residual nitrogen (33 kg N ha^?1) were detected. After the harvest of grain legumes always a very high nitrogen mineralization was observed during autumn especially after peas due to a close C/N-relationship and higher amounts of nitrogen in the straw as compared to fababeans. In comparison with fababeans, N-mineralization after the cultivation of oats remained lower by more than 50%. During winter, seepage water regularly led to a considerable decrease of soil NO₃-N content. The N-leaching losses were especially high after cultivation of peas (80 kg N ha ^?1) and considerably lower after fababeans (50 kg N ha^?1) and oats (20 kg N ha^?1). As compared to oats, a higher NO₃-N content in soil was determined at the beginning of the growing period after preceding grain legumes. Therefore, winter wheat yielded highest after preceding peas (68 dt ha^?1) and fababeans (60 dt ha^?1) and lowest after preceding oats (42 dt ha^?1). The cultivation of grain legumes had no measurable effect on yield formation of the third crop winter barley in either of the growing seasons.     

Screening techniques and improved biological nitrogen fixation in cool season food legumes

Dinitrogen fixation and legume productivity are greatly influenced through the interactions of legume host, Rhizobium, and the above- and below-ground environment. The benefits of improving legume N₂ fixation include reduced reliance on soil N, leading to more sustainable agricultural systems and reduced requirements for fertilizer N, enhanced residual benefits to subsequent crops, and increased legume crop yields. Most of the gains in N₂ fixation to date have been derived from management of legume cropping systems and through inoculation of legume seed with competitive and symbiotically effective rhizobia. Further gains are possible by developing plant cultivars with tolerance to soil abiotic factors, increased plant yield, and a broader and more effective matching of plant host and rhizobia. Techniques for screening material for superior N₂ fixation and examples of programs to increase fixed N, with attention to the major abiotic stresses, are discussed.     

Selection for Increased Seed Nitrogen Accumulation in Common Bean: Implications for Improving Dinitrogen Fixation and Seed Yield

Estimates of N₂ fixation in segregating populations of bean plants based on ¹⁵N-isotope methods are technically demanding and expensive; therefore, indirect measures based on related traits including total seed N were used to select for improved N₂ fixation and yield. In 1985, six populations of F₂-derived F₃ families resulting from six parental lines crossed to a common tester were grown in field trials on a low-N soil. In 1986, 25 selected half-sib families and two populations of full-sib F₄ families were grown under similar conditions. Parents and a non-nodulating soybean line were included both years. Narrow sense (H_NS) heritability estimates based on parent-offspring regressions ranged from 0.57 for total seed N to 0.39 for shoot biomass in one population, but were near zero for all measured traits in a second population. Among the criteria used to identify parental lines with superior potential for producing progenies with high total seed N, testcross population means combined with estimates of realized heritability were the most reliable. Selection of the best F₃ families for total seed N resulted in F₄ families with increased total seed N and higher seed yields, while seed protein percentage was unchanged. When plants are grown on low-N soils, selection for total seed N offers a useful alternative to selection for increased N₂ fixation based on ¹⁵N-isotopic or total-N-difference method.     

Rhizobium specificity of Trifolium ambiguum M. Bieb × T. repens L. hybrid clovers

Growth of full-sib families of an F₁ interspecific hybrid between Trifolium ambiguum M.Bieb and T. repens L, and two generations of backcross hybrids (BC₁F₂ and BC₂F₁) with T. repens as the recurrent parent, were compared to their parental species. Plants were grown in a N-free medium and inoculated with Rhizobium leguminosarum biovar trifolii rhizobia effective on T. ambiguum or T. repens. Hybridisation produced progeny that nodulated with rhizobia from either T. ambiguum or T. repens, but plant growth varied. Mean weights of T. repens and hybrids, particularly F₁ and BC₁F₂, were higher when inoculated with a mixture of strains isolated from field grown T. repens than with the New Zealand inoculant strain for T. repens. When inoculated with the mix of rhizobia from T. repens, mean weights were 242, 189, 132, 125, and 100 mg/plant for T. repens, BC₂F₁, BC₁F₂, F₁ and T. ambiguum, respectively. However, although the mean weight of BC₂F₁ lines was significantly less than T. repens, there was considerable variation in individual full-sib families indicating the potential to select within BC₂F₁ hybrids for high plant growth/symbiotic nitrogen fixation. The weight of T. ambiguumplants inoculated with the New Zealand inoculant strain for hexaploid T. ambiguum was similar to T. repens inoculated with the mix of rhizobia from T. repens (253 and 242 mg/plant, respectively). Mean fresh weights of F₁ hybrid plants were similar when inoculated with rhizobia for T. ambiguum or T. repens (125 and 130 mg/plant, respectively). However, weight of T. repens, BC₁F₂ and BC₂F₁ hybrids inoculated with rhizobia for T. ambiguum were all less than 90 mg/plant. There was a significant relationship between plant fresh weight and ethylene production. The results indicate that measuring weights of inoculated plants growing in N-free media is a rapid initial method of screening a range of plant germplasm for plant growth/symbiotic nitrogen fixation rates. This revised version was published online in August 2006 with corrections to the Cover Date.