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1.
Common bean (Phaseolus vulgaris L.) crops hold the potential to obtain higher yields by enhancing their biological nitrogen fixation (BNF) with Rhizobium. However in contrast to other legumes, common bean has shown a lack of positive response to inoculation with Rhizobium in many cases. This has led to a limited use of rhizobial inoculants in this crop, especially in Europe. The adaptation of bacterial strains to the rhizosphere is a key factor in the success of any inoculant, especially in a promiscuous legume such as common bean. This research aimed at increasing common bean yields via inoculation with effective indigenous Rhizobium leguminosarum strains. Three highly effective strains (LCS0306, LBM1123 and ZBM1008) which were selected according to their effectiveness at BNF in hydroponic conditions were separately inoculated onto common bean in a field experiment. The experiment was carried out under three environments and three tillage systems: conventional-tillage (CONVT), no-tillage (NT) and a cover-crop (CC). The grain yield observed with seed inoculation was significantly higher than the yield obtained with uninoculated seed under CONVT and CC. However, under NT inoculation had no effect. Furthermore, under CONVT and CC, inoculation with R. leguminosarum LCS0306 produced even higher yields than those obtained in nitrogen-fertilised or control plots. This is the first attempt to explain the inoculation performance of common bean under different tillage systems in Europe. A gene–based hypothesis has been used to explain the effectiveness of indigenous common bean rhizobia as nitrogen fixers in this crop.  相似文献   

2.
Kucey  R. M. N.  Snitwongse  P.  Chaiwanakupt  P.  Wadisirisuk  P.  Siripaibool  C.  Arayangkool  T.  Boonkerd  N.  Rennie  R. J. 《Plant and Soil》1988,108(1):33-41
Controlled environment and field studies were conducted to determine relationships between various measurements of N2 fixation using soybeans and to use these measures to evaluate a number ofBradyrhizobium japonicum strains for effectiveness in N2 fixation in Thai soils.15N dilution measurements of N2 fixation showed levels of fixation ranging from 32 to 161 kg N ha−1 depending on bacterial strain, host cultivar and location. Midseason measures of N2 fixation were correlated with each other, but not related measures taken at maturity. Ranking ofB. japonicum strains based on performance under controlled conditions in N-free media were highly correlated with rankings based on soybean seed yields and N2 fixation under field conditions. This study showed that inoculation of soybeans with effectiveB. japonicum strains can result in significant increases in yield and uptake of N through fixation. The most effective strains tested for use in Thai conditions were those isolated from Thai soils; however, effective strains from other locations were also of benefit.  相似文献   

3.
The nitrogen fixation response to copper nutrition in faba bean, yellow lupin and soybean was studied. Copper nutrition significantly increased the pod yields of all tested grain legumes but faba bean gave the greatest Cu-use efficiency for pod and grain production. The accumulation of dry matter in vegetative parts, nodules, N and leghemoglobin concentration in nodules and nitrogen accumulation in the whole plants were increased by copper supply in faba bean and yellow lupin in contrast with soybean. Cu nutrition significantly increased the Cu concentrations in nodules of all cultivated plants. The differential sensitivity of N2 fixation in tested grain legume species to copper nutrition could be connected with the level of phenols in nodules and depended on both the host plants and strains of rhizobia, which differ in their ability to produce catechol-like siderophores. Copper requirements by symbiotic N2 fixation could also depend on the nature of phenols in nodules (presence of o-dihydroxyphenols or number of hydroxyls in molecule).  相似文献   

4.
A field experiment was carried out to quantify biological nitrogen fixation (BNF) using the 15N isotope natural abundance method in maize (Zea mays L.)/faba bean (Vicia faba L.) and wheat (Triticum aestivum L.)/faba bean intercropping systems. Faba bean was yielding more in the maize/faba bean intercropping, but not in the wheat/faba bean intercropping. Biomass, grain yield and N acquisition of faba bean were significantly increased when intercropped with maize, and decreased significantly with wheat, irrespective of N-fertilizer application, indicating that the legume could gain or lose productivity in an intercropping situation. There was yield advantage of maize/faba bean intercropping, but no in wheat/faba bean intercropping. The grain yield of the faba bean intercropped with maize was greater than that of faba bean monoculture due to increases of the stems per plant and the pods per stem of faba bean. N fertilization inhibited N fixation of faba bean in maize/faba bean and wheat/faba bean intercropping and faba bean monoculture. The responses of different cropping systems to N-fertilizer application, however, were not identical, with competitive intercropping (wheat/faba bean) being more sensitive than facilitative intercropping (maize/faba bean). Intercropping increased the percentage of N derived from air (%Ndfa) of the wheat/faba bean system, but not that of the maize/faba bean system when no N fertilizer was applied. When receiving 120 kg N/ha, however, intercropping did not significantly increase %Ndfa either in the wheat/faba bean system or in the maize/faba bean system in comparison with faba bean in monoculture. The amount of shoot N derived from air (Ndfa), however, increased significantly when intercropped with maize, irrespective of N-fertilizer application. Ndfa decreased when intercropped with wheat, albeit not significantly at 120 kg N/ha. Ndfa was correlated more closely with dry matter yield, grain yield and competitive ratio, than with %Ndfa. This indicates that that total dry matter yield (sink strength), not %Ndfa, was more critical for the legume to increase Ndfa. The results suggested that N fixation could be improved by yield maximization in an intercropping system.  相似文献   

5.
Field experiments were performed in Austria, Brazil, Chile, Colombia, Guatemala, Mexico and Peru as part of an FAO/IAEA Co-ordinated Research Programme to investigate the nitrogen fixing potential of cultivars and breeding lines of common bean (Phaseolus vulgaris L.). Each experiment included approximately 20 bean genotypes which were compared using the 15N isotope dilution method. Great differences in nitrogen fixation were observed between and within experiments, with average values of 35% N derived from atmosphere (% Ndfa) and highest values of 70% Ndfa being observed. These values which were larger than had been reported previously for common bean, were observed only when environmental factors were favorable. Therefore, common bean lines are available, which can support high biological nitrogen fixation. These can be used either directly as cultivars for production or in breeding programmes to enhance nitrogen fixation in other cultivars.  相似文献   

6.
Moawad  H.  Badr El-Din  S.M.S.  Abdel-Aziz  R.A. 《Plant and Soil》1998,204(1):95-106
The diversity of rhizobia nodulating common bean ( Phaseolus vulgaris), berseem clover (Trifolium alexanderinum) and lentil (Lens culinaris) was assessed using several characterization techniques, including nitrogen fixation efficiency, intrinsic antibiotic-resistance patterns (IAR), plasmid profiles, serological markers and rep-PCR fingerprinting. Wide diversity among indigenous rhizobial populations of the isolates from lentil, bean and clover was found. Strikingly, a large percentage of the indigenous rhizobial population was extremely poor at fixing nitrogen. This emphasizes the need to increase the balance of highly efficient strains within the rhizobial population. Use of high-quality inocula strains that survive and compete with other less-desired and less-efficient N2-fixing rhizobia represents the best approach to increase biological nitrogen fixation of the target legume. In field-grown lentils, the inoculant strains were not able to outcompete the indigenous rhizobia and the native lentil rhizobia occupied 76–88% of the total nodules formed on inoculated plants. Nitrogen fixation by lentils, estimated using the 15N isotope dilution technique, ranged between 127 to 139 kg ha-1 in both inoculated and un-inoculated plants. With berseem clover, the inoculant strains were highly competitive against indigenous rhizobia and occupied 52–79% of all nodules. Inoculation with selected inocula improved N2 fixation by clover from 162 to 205 kg ha-1 in the three cuts as compared with 118 kg ha-1 in the un-inoculated treatment. The results also indicated the potential for improvement of N2 fixation by beans through the application of efficient N2-fixing rhizobia.  相似文献   

7.
Rhizobium etli, as well as some other rhizobia, presents nitrogenase reductase (nifH) gene reiterations. Several R. etli strains studied in this laboratory showed a unique organization and contained two complete nifHDK operons (copies a and b) and a truncated nifHD operon (copy c). Expression analysis of lacZ fusion demonstrated that copies a and b in strain CFN42 are transcribed at lower levels than copy c, although this copy has no discernible role during nitrogen fixation. To increase nitrogenase production, we constructed a chimeric nifHDK operon regulated by the strong nifHc promoter sequence and expressed it in symbiosis with the common bean plant (Phaseolus vulgaris), either cloned on a stably inherited plasmid or incorporated into the symbiotic plasmid (pSym). Compared with the wild-type strain, strains with the nitrogenase overexpression construction assayed in greenhouse experiments had, increased nitrogenase activity (58% on average), increased plant weight (32% on average), increased nitrogen content in plants (15% at 32 days postinoculation), and most importantly, higher seed yield (36% on average), higher nitrogen content (25%), and higher nitrogen yield (72% on average) in seeds. Additionally, expression of the chimeric nifHDK operon in a poly-beta-hydroxybutyrate-negative R. etli strain produced an additive effect in enhancing symbiosis. To our knowledge, this is the first report of increased seed yield and nutritional content in the common bean obtained by using only the genetic material already present in Rhizobium.  相似文献   

8.
Biological nitrogen fixation of leguminous crops is becoming increasingly important in attempts to develop sustainable agricultural production. However, these crops are quite variable in their effectiveness in fixing nitrogen. By the use of the 15N isotope dilution method some species have been found to fix large proportions of their nitrogen, while others like common bean have been considered rather inefficient. Methods for increasing N2 fixation are therefore of great importance in any legume work. Attempts to enhance nitrogen fixation of grain legumes has been mainly the domain of microbiologists who have selected rhizobial strains with superior effectiveness or competitive ability. Few projects have focused on the plant symbiont with the objective of improving N2 fixation as done in the FAO/IAEA Co-ordinated Research Programme which is being reported in this volume. The objective of the present paper is to discuss some possibilities available for scientists interested in enhancing symbiotic nitrogen fixation in grain legumes. Examples will be presented on work performed using agronomic methods, as well as work on the plant and microbial symbionts. There are several methods available to scientists working on enhancement of N2 fixation. No one approach is better than the others; rather work on the legume/Rhizobium symbiosis combining experience from various disciplines in inter-disciplinary research programmes should be pursued.  相似文献   

9.
Nitrogen fixation in nine common bean (Phaseolus vulgaris L.) lines was estimated using the 15N isotope dilution method at two locations in two seasons. In the first season at one location no N2 fixation was detected while in the second season up to 51 kg N ha–1 were estimated. There were significant differences between lines and correlations between trials were significant for the amounts of N2 fixed, but not for total shoot nitrogen. The plants that fixed the most nitrogen nodulated rapidly after germination. Differences in maximum nodule mass, but not specific nodule activity, were detected also.  相似文献   

10.
One of the most cultivated and consumed vegetables in Brazil is the common bean, Phaseolus vulgaris L. The symbiosis of this plant species with nitrogen-fixing bacteria that are adapted to the stresses commonly found in tropical soils can increase production. The aim of this study was to evaluate the symbiotic effectiveness of bacterial strains from soils under different land uses in the Amazon region. Further, rhizobia tolerance to acidity and aluminium and the involvement of some possible physiological mechanisms of such tolerance were also investigated. In assessing the efficiency of biological nitrogen fixation, inoculation with strains UFLA04-195, UFLA04-173 and UFLA04-202, belonging to the genus Rhizobium, resulted in greater plant growth, higher shoot nitrogen content and good nodulation compared to the inoculation with the strain CIAT 899 (R. tropici), and to the mineral nitrogen control or Burkholderia fungorum strains that nodulated or not bean plants. These efficient strains grew better at pH 5.0 than at pH 6.0 or pH 6.9; they also tolerated up to 1 mmol l−1 of Al3+ and showed an increased production of exopolysaccharides where the growing rates were less (pH 6.0 and pH 6.9). With respect to aluminium, the highest production of EPS produced greater tolerance to this element. Taken together, these results indicate that the strains evaluated in this study were tolerant to acidity and aluminium; they appeared to have developed resistance mechanisms such as EPS production and a resistant cell outer membrane (indicated by resistance to polymyxin and methyl violet). As these strains also gave increased yields of the host species, further studies on whether to recommend these strains as inoculants are already underway.  相似文献   

11.
Pot experiments were carried out to investigate the effect of inoculation with pure and mixed cultures of nitrogen fixers Azospirillum lipoferum 137, Arthrobacter mysorens 7 and the phosphate-solubilizing strain Agrobacterium radiobacter 10 on growth and mineral nutrition of two barley cultivars. A significant positive effect on grain yield both of the studied barley cultivars was obtained after inoculation with mixtures of A. lipoferum 137 + A. radiobacter 10 and A. lipoferum 137 + A. mysorens 7 only. The acetylene reduction activity on roots or in batch culture was significantly higher when A. lipoferum 137 and A. radiobacter 10 were combined. Using 15N isotope dilution technique it was established that these mixed cultures significantly increased the accumulation of nitrogen fertilizer in the plants. The strain A. radiobacter 10 promoted a better accumulation of phosphorus fertilizer by plants and A. mysorens 7 increased the total phosphorus content in plant tissues. The maximum positive effect of joint inoculation on plant development was observed when the combined nitrogen in soil was in short supply. It was concluded that inoculation with bacterial mixtures provided a more balanced nutrition for the plants and the improvement in root uptake of nitrogen and phosphorus was the major mechanism of interaction between plants and bacteria. The introduced bacteria were able to colonize actively the rhizoplane of barley. No interspecific competition or antagonism were established between components of the bacterial mixtures in the rhizoplane. The strains A. mysorens 7 and A. radiobacter 10 improved viability of A. lipoferum 137 when the plants were grown in acid soil. Field experiments carried out on 3 barley cultivars confirmed the assertion that inoculation with mixed cultures significantly increases the grain yield and nitrogenous nutrition of plants as compared with single cultures.  相似文献   

12.
Summary Three field experiments were conducted on ten cultivars of winterwheat and four cultivars of springwheat to estimate the growth promoting effect ofAzospirillum brasilense under varying nitrogen doses. Independent of cultivar selection or nitrogen dose a highly significant yield increase could be observed in winterwheat: strains S631 and SpBr14 increased the average grain yield with 9.14% and 14.82% respectively. When the yield components were studied a coinciding increase in ear density could be demonstrated of resp. 10.57% and 13.55%. Less significant results were obtained with springwheat although in one experiment strain SpBr14 significantly increased grain yield. As with winterwheat tillering of the plant was markedly affected by inoculation with both strains. In a companion greenhouse experiment it was found that inoculation with Azospirillum can cause a decrease in the root mass of wheatplants except when strain SpBr14 is used. Therefore it is suggested that the presence of a higher tillering together with an undisturbed nutrient uptake capacity can result in yield increases after inoculation withAzospirillum brasilense.  相似文献   

13.
Summary Two experiments were performed to examine the effects of inoculation of field grown wheat with various Azospirillum strains. In the first experiment the soil was sterilized with methyl bromide to reduce the Azospirillum population and15N labelled fertilizer was added to all treatments. Two strains ofAzospirillum brasilense isolated from surface sterilized wheat roots and theA. brasilense type strain Sp7 all produced similar increases in grain yield and N content. From the15N and acetylene reduction data it was apparent that these increases were not due to N2 fixation. In the second experiment performed in the same (unsterilized) soil, twoA. brasilense strains (Sp245, Sp246) and oneA. amazonense strain (Am YTr), all isolated from wheat roots, produced responses of dry matter and N content while the response to the strain Sp7 was much smaller. These data confirm earlier results which indicate that if natural Azospirillum populations in the soil are high (the normal situation under Brazilian conditions), strains which are isolated from wheat roots are better able to produce inoculation responses than strains isolated from other sources. The inoculation of a nitrate reductase negative mutant of the strain Sp245 produced only a very small inoculation response in wheat. This suggests that the much greater inoculation response of the original strain was not due to N2 fixation but to an increased nitrate assimilation due to the nitrate reductase activity of the bacteria in the roots. Consultant Inter-American Institute for Cooperation in Agriculture IICA/EMBRAPA World Bank Project.  相似文献   

14.
Summary The total amount of nitrogen derived from symbiotic nitrogen fixation in two pea and one field bean cultivar, supplied with 50 kg N ha−1 at sowing (‘starter’-N), was estimated to 165, 136, and 186 kg N ha−1, respectively (three-year means). However, estimates varied considerably between the three years. At the full bloom/flat pod growth stage from 30 to 59 per cent of total N2 fixation had taken place. The proportion of total N derived from N2 fixation at maturity was higher in seeds than in vegetative plant parts and amounted to 59.5, 51.3 and 66.3 per cent of total above-ground plant N in the two pea cultivars and field bean, respectively (three-year means). The recovery of fertilizer N was 62.2, 70.2, 52.1, and 69.5 per cent in the two pea cultivars, field bean and barley, respectively. Growth analysis indicated that barley did not meet the claims for an ideal reference crop in the15N fertilizer dilution technique for estimating N2 fixation in pea and field bean. ‘Starter’-N neither increased the seed yield nor the N content of the grain legumes.  相似文献   

15.
Rhizobium etli, as well as some other rhizobia, presents nitrogenase reductase (nifH) gene reiterations. Several R. etli strains studied in this laboratory showed a unique organization and contained two complete nifHDK operons (copies a and b) and a truncated nifHD operon (copy c). Expression analysis of lacZ fusion demonstrated that copies a and b in strain CFN42 are transcribed at lower levels than copy c, although this copy has no discernible role during nitrogen fixation. To increase nitrogenase production, we constructed a chimeric nifHDK operon regulated by the strong nifHc promoter sequence and expressed it in symbiosis with the common bean plant (Phaseolus vulgaris), either cloned on a stably inherited plasmid or incorporated into the symbiotic plasmid (pSym). Compared with the wild-type strain, strains with the nitrogenase overexpression construction assayed in greenhouse experiments had, increased nitrogenase activity (58% on average), increased plant weight (32% on average), increased nitrogen content in plants (15% at 32 days postinoculation), and most importantly, higher seed yield (36% on average), higher nitrogen content (25%), and higher nitrogen yield (72% on average) in seeds. Additionally, expression of the chimeric nifHDK operon in a poly-β-hydroxybutyrate-negative R. etli strain produced an additive effect in enhancing symbiosis. To our knowledge, this is the first report of increased seed yield and nutritional content in the common bean obtained by using only the genetic material already present in Rhizobium.  相似文献   

16.
A greenhouse experiment was performed to evaluate the effects of plant growth-promoting rhizobacteria (PGPR) on nodulation, biological nitrogen fixation (BNF) and growth of the common bean (Phaseolus vulgaris L. cv. Tenderlake). Single and dual inoculation treatments of bean with Rhizobium and/or PGPR were administered to detect possible changes in the levels of and interactions between the phytohormones IAA and cytokinin. Bean plants cv. Tenderlake were grown in pots containing Fluvic Neosol eutrophic (pH 6.5). Fourteen kilogram aliquots of soil contained in 15-l pots were autoclaved. Bean seeds were surface sterilized and inoculated with Rhizobium tropici (CIAT 899-standard strain) alone and in combination with one of the PGPR strains: Bacillus endophyticus (DSM 13796), B. pumilus (DSM 27), B. subtilis (DSM 704), Paenibacillus lautus (DSM 13411), P. macerans (DSM 24), P. polymyxa (DSM 36), P. polymyxa (Loutit L.) or Bacillus sp.(65E180). The experimental design was randomized block design with three replications. Beans co-inoculated with Rhizobium tropici (CIAT899) and Paenibacillus polymyxa (DSM 36) had higher leghemoglobin concentrations, nitrogenase activity and N2 fixation efficiency and thereby formed associations of greater symbiotic efficiency. Inoculation with Rhizobium and P. polymyxa strain Loutit (L) stimulated nodulation as well as nitrogen fixation. PGPR also stimulated specific-nodulation (number of nodules per gram of root dry weight) increases that translated into higher levels of accumulated nitrogen. The activities of phytohormones depended on their content and interactions with Rhizobium tropici and Paenibacillus and/or Bacillus (PGPR) strains which affect the cytokinin in content in the common bean.  相似文献   

17.
18.
Two Rhizobium etli strains, EBRI 2 and EBRI 26, isolated from Egypt were tested for nodulation competitiveness on beans using Rhizobium tropici CIAT 899G as the competing strain. The insertion of the gus-reporter transposon mTn5ssgusA30 did not alter the nodulation or nitrogen fixation capacity of mutant strain CIAT 899G compared to the wild type. At neutral pH, R. etli strains EBRI 2 and EBRI 26 were more competitive than CIAT 899G with the bean cultivar Saxa. These two strains gave nodule occupancies of 52.1 and 61.1% competing with equal cell numbers of CIAT 899G. Nodule occupancies from these two native strains increased with the bean cultivar Giza 6 from Egypt to 66 and 67.5%. Based on these results, cultivar Giza 6 was used to select the most competitive strains under stress of salinity or alkalinity as a major problem for a large part of Egyptian soils. Under stress of salinity (0.2% NaCl or 34.2 mM NaCl), the salt-sensitive strain EBRI 2 was more competitive than the salt-resistant strain EBRI 26. Strain EBRI 2 gave 87.4% but strain EBRI 26 gave 63.7% nodule occupancy against CIAT 899G. The same trend of results was observed under stress of alkalinity (pH 8). Strain EBRI 2 occupied 83% while Strain EBRI 26 occupied 53.2%.  相似文献   

19.
5株耐盐碱促生细菌的筛选鉴定及其对红小豆的促生作用   总被引:1,自引:3,他引:1  
【背景】黑龙江省大庆市地处松嫩平原,土地盐碱化程度较重。盐碱土中含有大量的耐盐碱菌株,其中部分具有特殊的促生功能,可以作为有益促生菌促进植物生长,提高植物对盐碱的抗性。【目的】有效解决土地盐碱化问题,提高粮食作物产量。【方法】采集大庆地区的盐碱土壤为实验材料,在NaHCO_3终浓度为50 mmol/L、pH 9.0的条件下对土壤中耐盐碱细菌进行分离,用功能性培养基进行促生功能细菌菌株的筛选,并对相应促生物质产量进行测定;挑选促生功能较全、促生效果较好的细菌菌株,采用形态学观察、生理生化法以及16S rRNA基因序列分析法进行菌种鉴定,以确定菌株的分类学地位;在pH 8.5、15 mmol/L Na HCO3胁迫条件下测试施用促生菌株对红小豆种子发芽率、根长、芽长及插条生根数和根长的影响,以验证促生效果。【结果】对筛选获得的细菌菌株进行鉴定,29号细菌为卓贝尔氏菌属(Zobellella sp.),23、34、36和41号细菌属于盐单胞菌属(Halomonas sp.)。5株细菌均具有固氮、产植物激素吲哚乙酸(Indole-3-Acetic Acid,IAA)及ACC脱氨酶功能,34号和23号细菌具有最高植物激素IAA和ACC脱氨酶产量,分别达到了17.66 mg/L和0.31 U/mg。另外,23号细菌具有产铁载体能力,铁载体产量为3.19%;29和41号细菌具有溶有机磷功能,溶磷量分别为20.05 mg/L和34.61 mg/L。植物促生试验结果显示,在盐碱胁迫条件下,红小豆种子接菌后种子的发芽率和根长分别提高了41.67%和32.63%以上;红小豆插条接菌后,幼苗的根长和生根数与盐碱胁迫对照相比分别提升了129%和160%以上。【结论】这5株耐盐碱促生细菌能够缓解盐碱胁迫对红小豆生长的抑制作用,可以较好地促进其根系生长发育,可为研制耐盐碱促生菌肥提供优良菌种。  相似文献   

20.
Indigenous rhizobial population is among the factors which influence increased crop yield through inoculation with elite strains. In this study, we compared in greenhouse conditions the competitiveness of Rhizobium strain ISRA 355 for nodulation of the common bean (Phaseolus vulgaris) cultivated in different unsterile Senegal soils in terms of pH, N and C contents. The strain ISRA 355 produced a stable GUS+ transconjugant which was used for competition with indigenous soil rhizobia in six localities. At Bayakh, the transconjugant ISRA 355gusA was less competitive than the indigenous rhizobial strains, whereas in the other localities, it was more competitive since it occupied more than 90% of the nodules. Thus the Rhizobium strain ISRA 355 should be used for successfully inoculating the common bean in Senegal soils.  相似文献   

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