Genetic diversity of indigenous common bean (Phaseolus vulgaris) rhizobia in two Brazilian ecosystems

Although rhizobia for common bean (Phaseolus vulgaris L.) are established in most Brazilian soils, understanding of their genetic diversity is very poor. This study characterized bean strains from two contrasting ecosystems in Brazil, the Northeast Region, with a semi-arid climate and neutral soils and the South Region, with a humid subtropical climate and acid soils. Seedlings of the cultivars Negro Argel and Aporé were used to trap 243 rhizobial isolates from 12 out of 14 sites. An analysis of ERIC-PCR products revealed enormous variability, with 81% of the isolates representing unique strains considering a level of 70% of similarity. In general, there was no effect of either the bean cultivar, or the ecosystem on rhizobial diversity. One-hundred and one strains showing genetic relatedness (ERIC-PCR) less than 70% were further analyzed using restriction fragment length polymorphism (RFLP) of the 16 S rDNA cleaved with five restriction enzymes. Twenty-five different profile combinations were obtained. Rhizobium etli was the predominant species, with 73 strains showing similar RFLP profiles, while 12 other strains differed only by the profile with one restriction enzyme. Fifty strains were submitted to sequencing of a 16 S rDNA fragment, and 34 clustered with R. etli, including strains with RFLP-PCR profiles similar to those species or differing by one restriction enzyme. However, other strains differing by one or two enzymes were genetically distant from R. etli and two strains with identical profiles showed higher similarity to Sinorhizobium fredii. Other strains showed higher similarity of bases with R. tropici, R. leguminosarum and Mesorhizobium plurifarium, but some strains were quite dissimilar and may represent new species. Great variability was also verified among the sequenced strains in relation to the ability to grow in YMA at 40 °C, in LB, to synthesize melanin in vitro, as well as in symbiotic performance, including differences in relation to the described species, e.g. many R. etli strains were able to grow in LB and in YMA at 40 °C, and not all R. tropici were able to nodulate Leucaena.     

Polyphasic characterization of Brazilian Rhizobium tropici strains effective in fixing N2 with common bean (Phaseolus vulgaris L.)

Common bean (Phaseolus vulgaris) is native to the Americas, and Rhizobium etli is the dominant microsymbiont in both the Mesoamerican and the Andean centers of genetic diversification. Wild common beans are not found in Brazil, although the legume has been cropped in the country throughout time and all but one of the rhizobial species that nodulate it (Rhizobium gallicum) have been broadly detected in Brazilian soils. However, the majority of the effective rhizobial strains isolated so far from field-grown plants belong to R. tropici. This study describes the analysis of symbiotic and non-symbiotic genes of 15 effective R. tropici strains, isolated from four geographically distant regions in Brazil. With RFLP-PCR of the 16S and 23S rRNA genes and sequence analysis of 16S rRNA, two clusters were observed, one related to R. tropici type A and another to type B strains. Diversity in ribosomal genes was high, indicating that type A strains might represent a new species. High intraspecies diversity was also observed in the rep-PCR analysis with BOX, ERIC and REP primers. However, in the RFLP-PCR analysis of nifH and nodC genes, all R. tropici showed unique combinations of profiles, which might reflect an evolutionary strategy to maximize N₂ fixation.     

New insights into the origins and evolution of rhizobia that nodulate common bean (Phaseolus vulgaris) in Brazil

It is generally accepted that there are two major centers of genetic diversification of common beans (Phaseolus vulgaris L.): the Mesoamerican (Mexico, Colombia, Ecuador and north of Peru, probably the primary center), and the Andean (southern Peru to north of Argentina) centers. Wild common bean is not found in Brazil, but it has been grown in the country throughout recorded history. Common bean establishes symbiotic associations with a wide range of rhizobial strains and Rhizobium etli is the dominant microsymbiont at both centers of genetic diversification. In contrast, R. tropici, originally recovered from common bean in Colombia, has been found to be the dominant species nodulating field-grown common-bean plants in Brazil. However, a recent study using soil dilutions as inocula has shown surprisingly high counts of R. etli in two Brazilian ecosystems. In the present study, RFLP-PCR analyses of nodABC and nifH genes of 43 of those Brazilian R. etli strains revealed unexpected homogeneity in their banding patterns. The Brazilian R. etli strains were closely similar in 16S rRNA sequences and in nodABC and nifH RFLP-PCR profiles to the Mexican strain CFN 42^T, and were quite distinct from R. etli and R. leguminosarum strains of European origin, supporting the hypothesis that Brazilian common bean and their rhizobia are of Mesoamerican origin, and could have arrived in Brazil in pre-colonial times. R. tropici may have been introduced to Brazilian soils later, or it may be a symbiont of other indigenous legume species and, due to its tolerance to acidic soils and high temperature conditions became the predominant microsymbiont of common bean.     

The diversity of rhizobia nodulating chickpea (Cicer arietinum) under water deficiency as a source of more efficient inoculants

Chickpea is an important pulse crop grown mainly in the arid and semi-arid regions. The effect of water deficiency on nodulation, biomass production, and competition for nodule occupancy was evaluated in three different soils with two chickpea cultivars, Amdoun I and Chetoui. Two watering regimes were considered; a control that was irrigated three times per week and a water-deficient treatment that was irrigated only one time a week. Results showed that water deficiency significantly decreased the nodule number and the shoot dry weight for both cultivars. Root-nodule bacteria were isolated and characterized by PCR-RFLP of 16S rDNA and nifD-K intergenic spacer. The results show that water deficiency affects the diversity of nodulating rhizobia. The nodulation by Mesorhizobium mediterraneum was reduced while inefficient nodulation by Ensifer meliloti was favoured. In both treatments, chickpea was preferentially nodulated by nifD-K type N6. Analysis for NaCl tolerance showed that most of rhizobia nodulating chickpea under water deficiency are NaCl tolerant. Inoculation with the selected salt-tolerant strain of M. mediterraneum LILM10 increased significantly nodule number and grain yield in the field.     

Alleviation of drought stress in the common bean (Phaseolus vulgaris L.) by co-inoculation with Paenibacillus polymyxa and Rhizobium tropici

A greenhouse experiment was performed to evaluate the influence of Rhizobium when co-inoculated with each of two Paenibacillus polymyxa strains, singly and in mixture on growth, nitrogen content, phytohormone levels and nodulation of the common bean (Phaseolus vulgaris L.) under three levels of drought stress. Stress was applied continuously by the control of matric potential (ψ_m) through a porous cup. Bean plants cv. Tenderlake were grown in pots with Fluvic Neosol eutrophic soil under three different ψ_m (S₁ −7.0; S₂ −70.0 and S₃ < −85 kPa). The seeds were inoculated with Rhizobium tropici (CIAT 899) and each of P. polymyxa (DSM 36) and P. polymyxa Loutit (L) singly and in mixture (CIAT 899 + DSM36 + Loutit). Co-inoculation of bean with Rhizobium and both Paenibacillus strains resulted increased plant growth, nitrogen content and nodulation compared to inoculation with Rhizobium alone. This was particularly evident at the most negative ψ_m (S₃ < −85 kPa) we used. Drought stress triggered a change in phytohormonal balance, including an increase in leaf abscisic acid (ABA) content, a small decline in indole acetic acid (IAA) and gibberellic acid (GA₃) and a sharp fall in zeatin content in bean leaves. The content of endogenous Cks decreased under water stress, possibly amplifying the response of shoots to increasing ABA content. We hypothesize that co-inoculation of bean with R. tropici (CIAT 899) and P. polymyxa strains (DSM 36) and Loutit (L) mitigates some of the negative effects of drought stress on bean.     

种子引发对盐胁迫下高粱种子萌发及幼苗生长的影响

以“Tx623A”ד89-363”(Sb1, 耐盐性较弱)和“黑30A”ד大粒早”(Sb2, 耐盐性较强)2 个高粱杂交组合为试验材料, 利用100 mmol·L^-1?NaCl 溶液对种子进行引发处理。采用营养液沙培试验, 设4 个NaCl 浓度(0、50 mmol·L^-1、100 mmol·L^-1和150 mmol·L^-1)模拟盐胁迫, 研究种子引发处理对盐胁迫下高粱种子萌发及幼苗生长的影响。结果表明: 随着盐胁迫强度增加, 无论经引发处理还是未经引发处理2 个高粱杂交组合种子的出苗时间均明显延迟, 出苗率和成苗率下降, 幼苗生长受到抑制, 物质积累减少。同等盐胁迫强度下,引发处理与未引发处理相比, 种子出苗时间缩短, 出苗率和成苗率提高, 幼苗地上部分干、鲜重和地下部分干、鲜重增加, 光合色素含量升高, Na⁺/K⁺值显著降低。由此得出, 种子引发处理可以不同程度地促进盐胁迫下高粱种子萌发, 减轻盐胁迫对幼苗的伤害, 促进幼苗生长, 提高耐盐性; 耐盐性不同的品种引发效果存在差异, 对耐盐性较弱的品种引发效果更好。     

Nodulation of African yam bean (Sphenostylis stenocarpa) by Bradyrhizobium sp. isolated from Erythrina brucei

African yam bean (Sphenostylis stenocarpa), which is widely cultivated in Africa because of its growth capability on marginal soils, was nodulated by an endosymbiont (characterized and designed Bradyrhizobium sp. AUEB20) isolated from the Ethiopian tree Erythrina brucei with the formation of a small number of large, indeterminate N₂-fixing nodules. In contrast, 24 other isolates from Ethiopian woody legumes were ineffective. Strain AUEB20 promiscuously nodulated a number of tropical legumes, but none out of five European crop plants tested. Received: 17 September 1996     

Elevation of atmospheric CO2 and N-nutritional status modify nodulation, nodule-carbon supply, and root exudation of Phaseolus vulgaris L.

Increased root exudation and a related stimulation of rhizosphere-microbial growth have been hypothesised as possible explanations for a lower nitrogen- (N-) nutritional status of plants grown under elevated atmospheric CO₂ concentrations, due to enhanced plant-microbial N competition in the rhizosphere. Leguminous plants may be able to counterbalance the enhanced N requirement by increased symbiotic N₂ fixation. Only limited information is available about the factors determining the stimulation of symbiotic N₂ fixation in response to elevated CO₂.In this study, short-term effects of elevated CO₂ on quality and quantity of root exudation, and on carbon supply to the nodules were assessed in Phaseolus vulgaris, grown in soil culture with limited (30 mg N kg⁻¹ soil) and sufficient N supply (200 mg N kg⁻¹ soil), at ambient (400 μmol mol⁻¹) and elevated (800 μmol mol⁻¹) atmospheric CO₂ concentrations.Elevated CO₂ reduced N tissue concentrations in both N treatments, accelerated the expression of N deficiency symptoms in the N-limited variant, but did not affect plant biomass production. ¹⁴CO₂ pulse-chase labelling revealed no indication for a general increase in root exudation with subsequent stimulation of rhizosphere microbial growth, resulting in increased N-competition in the rhizosphere at elevated CO₂. However, a CO₂-induced stimulation in root exudation of sugars and malate as a chemo-attractant for rhizobia was detected in 0.5-1.5 cm apical root zones as potential infection sites. Particularly in nodules, elevated CO₂ increased the accumulation of malate as a major carbon source for the microsymbiont and of malonate with essential functions for nodule development. Nodule number, biomass and the proportion of leghaemoglobin-producing nodules were also enhanced. The release of nod-gene-inducing flavonoids (genistein, daidzein and coumestrol) was stimulated under elevated CO₂, independent of the N supply, and was already detectable at early stages of seedling development at 6 days after sowing.     

The HFC/HCFC breakdown product trifluoroacetic acid (TFA) and its effects on the symbiosis between Bradyrhizobium japonicum and soybean (Glycine max)

The environmental impact of hydrofluorocarbon (HFC) and hydrochlorofluorocarbon (HCFC) are under intense study due to the resistance of the breakdown product, trifluoroacetic acid (TFA), to further degradation. TFA has come under scrutiny due to its rapid and complete partitioning into aqueous phases of the environment, eventually allowing for deposition of TFA into soil via precipitation. Evidence exists that TFA may be toxic to soil microbes and plants, with little or no degradation occurring in soils. Uptake by plants and microorganisms and its similarity to acetate, implies its potential to effect the symbiotic nitrogen-fixing partners Bradyrhizobium japonicum and soybean (Glycine max). A preliminary study was performed in accordance with the Alternative Fluorocarbon Environmental Acceptability Study (AFEAS). Those results are presented here in addition to the findings of further experimentation on the initial interaction of B. japonicum with soybean. We used three levels of TFA (0.67, 6.74 and 67.40 μl TFA kg⁻¹ soil; 0.003, 0.031 and 0.314 μl TFA l⁻¹) for soil and hydroponics conditions and three levels (10, 100 μM and 1 mM) in bacterial culture. The results demonstrate that TFA affects growth of B. japonicum significantly, but does not affect PHB accumulation. Also no F⁻ was found in cultures grown on TFA. Attachment of B. japonicum to soybean roots was enhanced with the lowest level of acetate or TFA and was significantly reduced with 1 mM acetate or TFA. Cultures grown on acetate or acetate with TFA do not attach well, with those grown with 1 mM TFA the least. Both effects may be attributed to pH. Soybean seedlings had significantly retarded development with levels of TFA at or above 6.74 μl TFA kg⁻¹ soil and 0.031 μl TFA l⁻¹ nutrient solution. No nodules formed on those plants treated with these levels of TFA except in the hydroponics trials. Nodule location was not affected regardless of the TFA level. At the lowest level used we found no effects on soybean or symbiotic nitrogen fixation. In some cases, nodulation was enhanced, but nodule weight reduced. Anaerobically isolated bacteroids had normal levels of acetylene reduction activity regardless of the level of TFA used. In summary, soybean is much more sensitive to low levels of TFA than its symbiotic counterpart B. japonicum. No detrimental effects on symbiotic nitrogen fixation in soybean should be expected unless large bioaccumulation of TFA occurs in agricultural areas.     

Ascophyllum nodosum Extract and Its Organic Fractions Stimulate Rhizobium Root Nodulation and Growth of Medicago sativa (Alfalfa)

The effects of the seaweed Ascophyllum nodosum extracts (ANE) on nitrogen (N)–fixing nodules and growth of alfalfa plants were studied under greenhouse conditions. The treatment of alfalfa roots increased the number of total nodules per plant with ANE (69%) and organic sub-fractions methanol (20%) and chloroform (35%) at 1 g L^?1 concentration. The number of functional nodules was greater per plant in ANE (36%) and its organic sub-fraction chloroform (105%). Maximum increase in shoot length was observed in ANE-treated plants (42%) and chloroform-treated plants (42%). Root length was longer in the chloroform fraction (15%), whereas the shoot dry-weight accumulation was greater in plants treated with ANE (118%), methanol (85%), and chloroform (85%) than the control. Root dry-weight accumulation increased in plants treated with ANE (118%) and chloroform (69%) compared to the control. Further studies are under way to identify the chemical components in ANE and organic fractions.     

The excretion of ammonium by enchytraeids (Cognettia sphagnetorum)

Enchytraeids are involved both directly and indirectly in decomposition processes and nitrogen mineralization in soil. Their influence is especially important in nitrogen poor ecosystems such as heathland where the enchytraeid species, Cognettia sphagnetorum, is often abundant and playing a significant role in the N-cycling. The objective of this study was to quantify NH₄⁺-N excretion of C. sphagnetorum at different temperatures. The results were combined with investigations of population dynamics during one year to estimate annual NH₄⁺-N excretion of the population of C. sphagnetorum in a dry Danish heath soil. C. sphagnetorum significantly increased its NH₄⁺-N excretion rate with increasing temperature. At 5 °C about 0.5 μg NH₄⁺-N mg dry weight⁻¹ day⁻¹ was excreted increasing to about 3.3 μg NH₄⁺-N mg dry weight⁻¹ day⁻¹ at 20 °C. Average enchytraeid biomass in the field was 2.5-3.5 g dry weight m⁻² during cool and wet periods. Dry and warm conditions in May and June, 2008, had a drastic and long-term negative impact on the enchytraeid community. The excretion of NH₄⁺-N by enchytraeids was therefore highest during the cool and moist months despite low temperatures (October 2007-May 2008) and amounted to about 2 mg NH₄⁺-N m⁻² day⁻¹ during this period. The estimated annual NH₄⁺-N excretion of the enchytraeid community was approximately 0.3 g N m⁻² year⁻¹. The results of the present study and the method described for estimation of N-excretion can increase our understanding of enchytraeids’ role in nitrogen mineralization.     

Nodulation pattern and acetylene reduction (nitrogen fixation) activity of some highland and lowland Acacia species of Ethiopia

 Endosymbionts from the Ethiopian highland acacia species Acacia abyssinica, A. negrii and A. etbaica, and the lowland species A. nilotica, A. prasinata, A.senegal, A. seyal, A. tortilis and Faidherbia (Acacia) albida were isolated and characterized. Seven tree species were found to be nodulated by species of both Rhizobium and Bradyrhizobium. F. (Acacia) albida and A. senegal were nodulated by only Bradyrhizobium or Rhizobium, respectively. In A. abyssinica, both genera were isolated from the same nodule, whereas in A. nilotica and A. tortilis, both strains were isolated from different nodules of the same plant. The nitrogen fixation (acetylene reduction) activities varied considerably and showed no correlation with the nitrogen content of the plant. Highland species were as effective as lowland plants, thus demonstrating good potential for soil reclamation. The endosymbionts isolated proved rather promiscuous, efficiently nodulating other Acacia spp. and some tropical grain legumes, but did not nodulate temperate legumes. Received: 7 August 1997     

Arbuscular mycorrhizae affect the N and C economy of nodulated Phaseolus vulgaris (L.) during NH4 nutrition

In the symbiosis between nodulated legume roots and arbuscular mycorrhizal (AM) fungi, the C and N economy can be influenced by the source of N-supply from either AM-derived NH₄⁺ uptake or nodule-derived biological nitrogen fixation (BNF). This relationship was investigated in terms of NH₄⁺ supply and BNF by the two symbionts. Nodulated Phaseolus vulgaris seedlings with and without AM, were hydroponically grown with either 0 N or 1 mM NH₄⁺ supply. Plants were harvested at 30 days after emergence and measurements were taken for biomass, N₂ fixation, photosynthesis, CO₂ and O₂ root respiration, calculated C and N economy. AM roots had higher NH₄⁺ uptake and this was associated with the suppression of BNF and nodule growth. The higher NH₄⁺ uptake in AM roots occurred with lower root maintenance respiration, compared to when N was derived from BNF. There was also an increase in the below-ground sink strength of NH₄⁺ fed AM roots compared to NH₄⁺ fed non-AM roots, as evidenced by the increases in root CO₂ and O₂ respiration and photosynthetic stimulation. These results indicate that although the AM root had higher total below-ground respiratory costs during NH₄⁺ nutrition, there were lower respiratory C costs associated with N derived from AM symbionts in comparison to N from BNF.     

Monitoring stomatal conductance of Jatropha curcas seedlings under different levels of water shortage with infrared thermography

Model simulations and experimental measurements were used to investigate the applicability of infrared thermography for the estimation of stomatal conductance and drought stress under sub-optimal meteorological conditions. The study focused on the stomatal conductance index I_g, calculated from the leaf temperature and the temperature of a dry and wet reference leaf. The simulations revealed that I_g is influenced by leaf dimension, wind speed and air temperature and not or hardly by leaf angle, albedo, relative humidity or insolation. In addition, I_g was found to be very sensitive to differences in wind speed, air temperature, insolation, leaf dimension and leaf angle between the measured and the reference leaves. In the experimental part, we evaluated if infrared thermography can be used to improve the knowledge on the water use of Jatropha curcas L., a tropical biofuel crop. Thermal images from Jatropha seedlings grown under three different drought treatments were made on a day with very variable insolation and a day with very low insolation. Smaller newly formed leaves and the active control of the leaf angle proved efficient ways of Jatropha to protect leaves under drought stress from overheating. I_g, assessed in four different ways, and four simplified drought stress indices were derived and related to the measured stomatal conductance (g_s) of the seedlings. The strongest correlation with g_s and the highest discriminative power between the different water treatments were achieved when I_g was calculated by taking the average leaf temperature per plant and the temperature of the dry and wet reference leaves of this plant, rather than the average temperature of several reference leaves. Using the difference between the dry reference and the measured leaf (T_dry − T_l) as a simplified index gave similar results, although correlations were weaker. Other simplified thermal indices were not well correlated with leaf stomatal conductance or with water treatment. Recommendations were formulated for the measurement of I_g and (T_dry − T_l).     

不同施氮量下干旱胁迫对棉花生长及种内关系的影响

土壤水分和氮肥是影响荒漠绿洲区作物生长和种内关系的主要因素。为明确不同施氮量下干旱胁迫对棉花产量以及种内关系的影响,本研究以集群栽培(1穴3株)下棉花为研究对象,设置3种施氮量[300 kg(N)·hm~(-2)、225 kg(N)·hm~(-2)、150 kg(N)·hm~(-2)]和3种水分处理(正常水分、中度干旱胁迫、重度干旱胁迫),测定了不同处理下棉花的生长指标和棉花产量,分析了不同处理下棉花种内关系的变化。结果表明:1)同一施肥条件下,2016年和2017年棉花株高在重度干旱胁迫下显著降低,而茎粗随干旱胁迫加剧显著降低,但叶面积随干旱胁迫加剧却有所增加,且在中度干旱胁迫下的叶面积最大。2)同一施肥条件下,与正常水分相比,中度和重度干旱胁迫下棉花的茎秆生物量均显著下降;叶片生物量和籽棉产量在中度干旱胁迫下最高,在重度干旱胁迫下却降低。3)在当地施氮量300kg·hm~(-2)下,随着干旱胁迫的加剧,相对邻体效应(relativeneighbor effect, RNE)数值从正值变为负值;在氮肥减少处理(225 kg·hm~(-2))下,随着干旱胁迫的加剧, RNE数值先升高后降低,而在施氮量为150 kg·hm~(-2)处理下, RNE数值随着干旱的加剧逐渐降低,且均为负值。总之,在当地施氮量和中度干旱胁迫下棉花能够获得较高的籽棉产量,且在此处理下棉花的种内关系为助长关系;其他处理均降低了棉花的产量,并使得棉花的种内关系转变为竞争关系。     

The role of potassium fertilizer in nodulation and nitrogen fixation of faba bean (Vicia faba L.) plants under drought stress

Three-week-old nodulated faba bean plants were subjected to different levels of drought stress (onehalf, one-quarter, or one-eighth field capacity) for 5 weeks. Half the stressed plants were treated with KCl at 10 mg kg^-1 soil or 150 mg kg^-1 soil at the beginning of the drought stress. Nodulation and nitrogenase activity were significantly decreased by increasing drought stress. Leghaemoglobin and protein contents of nodule cytosol were also severely inhibited by drought sttess. This decline was attributed to the induction of protease activity. However, carbohydrate contents of the nodule cytosol increased significantly. This accumulation was attributed to a sharp decline in invertase activity and low use of sugar by the bacteroids We conclude that harmful effects of water deficits can be alleviated by increasing K⁺ supplementation.     

Recovery of reproduction after drought in the soil living Folsomia candida (Collembola)

Soil dwelling (euedaphic) Collembola have evolved to live in soil pores where the atmosphere is saturated with water vapour. Here we show that reproduction in the euedaphic Folsomia candida ceases during drought where soil water potentials are lower than −7 bar (∼99.4% RH). Recovery of the moulting cycle after cessation of drought showed increasing delay with increasing previous drought pressure. However, reproduction was rapidly resumed. Only in F. candida recovering from the most severe drought (−55 bar, 96% RH) was the time lapse between drought and resumed reproduction significantly different from the control (99.4% RH). Furthermore, the final numbers of eggs were unaffected by the degree of drought exposure. Hence, these results suggest that while even very mild drought will stop the reproductive cycle, subsequent recovery is rapid and complete.     

Yield response of spring wheat cultivars (Triticum aestivum and T. turgidum) to inoculation with Azospirillum brasilense under field conditions

Summary Eight commercial Israeli spring wheat cultivars (six Triticum aestivum and two T. turgidum) grown with 40 and 120 kg N/ha were tested for responses to inoculation with Azospirillum brasilense. At the low level of N fertilization (40 kg/ha), five cultivars showed significant increases in plant dry weight measured at the milky ripe stage; however, by maturation only the cultivar Miriam showed a significant increase in grain yield. Two cultivars, which had shown a positive inoculation effect at the earlier stages, had a significant decrease in grain yield. No significant effect of inoculation was found at the high N level. To confirm those results, four wheat (T. aestivum) cultivars were tested separately over 4 years in 4 different locations under varying N levels. Only Miriam showed a consistently positive effect of Azospirillum inoculation on grain yield. Inoculation increased the number of roots per plant on Miriam compared with uninoculated plants. This effect was found at all N levels. Nutrient (N, P and K) accumulation and number of fertile tillers per unit area were also enhanced by Azospirillum, but these parameters were greatly affected by the level of applied N. It is suggested that the positive response of the spring wheat cultivar Miriam to Azospirillum inoculation is due to its capacity to escape water stresses at the end of the growth season.     

Effects of Azospirillum brasilense on root morphology of common bean (Phaseolus vulgaris L.) under different water regimes

 The effects of inoculation with Azospirillum brasilense Cd on root morphology and growth of common bean (Phaseolus vulgaris L.) were studied under different growth systems and water regimes. The root systems were evaluated by image analysis. In a PVC-tube growth system, inoculation with A. brasilense at 10⁷ colony forming units (CFU) ml^–1 increased root length, root projection area, specific root length (m g^–1) and specific root area (cm² g^–1), as compared with non-inoculated controls, resulting in root systems with longer and thinner roots. Water stress induced similar root responses to those observed after inoculation with A. brasilense. No increase in plant biomass was observed in inoculated plants, suggesting that under the tested growth conditions, a relatively larger amount of resources is required for the maintenance of the thinner roots. In water-stressed potted plants, the effect of A. brasilense on tap root length was inoculum-concentration dependent. At 10⁷ CFU ml^–1 this effect was significant as compared to non-inoculated controls. In a pouch system without water stress, inoculation with A. brasilense at a concentration of 10⁵–10⁷ CFU ml^–1 2 days after germination resulted initially (2 days after inoculation) in an increase in root length (95%) and root fresh weight (66%), but reduced root diameter (20%), compared to controls. At this early stage of growth the distribution of root length among the different root diameter classes changed: the thinner-root classes had the largest proportion of longer roots. Received: 3 January 2000     

氮素对NaCl胁迫下甜高粱种子萌发及芽苗生长与生理的影响

为了研究NaCl胁迫下氮肥对甜高粱种子萌发及芽苗生长和生理特性的影响,探索提高甜高粱耐盐能力的措施,室内设置不同盐分浓度、不同氮源及浓度条件下甜高粱萌芽试验。结果表明:NaCl胁迫和不同氮源对甜高粱发芽和芽苗生长的影响各有不同。NaCl浓度对甜高粱种子萌发有显著影响,在甜高粱芽苗生长阶段,通过提高保护酶活性和渗透调节物质而增强耐盐伤害能力是有限的。100 mmol.L 1NaCl胁迫下,根系POD活性最低,而叶片MDA积累量、可溶性糖含量、POD活性最高,受盐害程度最大。没有盐胁迫情况下增加不同氮源及氮量对甜高粱根叶生理特性的影响差异显著,当氮浓度在20 mmol.L 1时,细胞受伤害程度最低,生长最好。不同形态氮源对甜高粱发芽和幼苗生长的影响差异明显,NH4Cl的促进效果优于KNO3。在100mmol.L 1的NaCl胁迫下,施加铵态氮或硝态氮源均可以增强甜高粱芽苗期的POD活性,减少MDA积累,从而缓解盐胁迫带来的伤害。研究表明采取适当的氮肥调控措施可以提高甜高粱的耐盐能力。