Yield and biological nitrogen fixation of common bean (Phaseolus vulgaris L.) in Peru

Two field experiments were performed to evaluate the nitrogen fixation potential of twenty common bean cultivars and breeding lines during summer and winter seasons of 1986 and 1988, respectively. The ¹⁵N isotope dilution method was used to quantify N₂ fixation. The cultivars and breeding lines were variable in terms of their N₂ fixation. The cv. Caballero was very efficient, with more than 50% N derived from the atmosphere and 60–80 kg N ha^–1 fixed in both seasons. Other cultivars were less efficient, since the poorest ones derived less than 30% of their nitrogen from the atmosphere and fixed less than 20 kg N ha^–1. After additional testing the best cultivars may be used directly by the farmers for cultivation. The experiments have provided information about which genotypes may be used to breed for enhanced fixation in common bean.     

Response to inoculation and N fertilization for increased yield and biological nitrogen fixation of common bean (Phaseolus vulgaris L.)

Common bean (Phaseolus vulgaris L.) is able to fix 20–60 kg N ha^–1 under tropical environments in Brazil, but these amounts are inadequate to meet the N requirement for economically attractive seed yields. When the plant is supplemented with N fertilizer, N₂ fixation by Rhizobium can be suppressed even at low rates of N. Using the ¹⁵N enriched method, two field experiments were conducted to compare the effect of foliar and soil applications of N-urea on N₂ fixation traits and seed yield. All treatments received a similar fertilization including 10 kg N ha^–1 at sowing. Increasing rates of N (10, 30 and 50 kg N ha^–1) were applied for both methods. Foliar application significantly enhanced nodulation, N₂ fixation (acetylene reduction activity) and yield at low N level (10 kg N ha^–1). Foliar nitrogen was less suppressive to nodulation, even at higher N levels, than soil N treatments. In the site where established Rhizobium was in low numbers, inoculation contributed substantially to increased N₂ fixation traits and yield. Both foliar and soil methods inhibited nodulation at high N rates and did not significantly increase bean yield, when comparing low (10 kg N ha^–1) and high (50 kg N ha^–1) rates applied after emergence. In both experiments, up to 30 kg N ha^–1 of biologically fixed N₂ were obtained when low rates of N were applied onto the leaves.     

Growth, nitrogen fixation and ammonium assimilation in common bean (Phaseolus vulgaris L) subjected to iron deficiency

A greenhouse experiment was carried out aiming to study the effect of iron deficiency on nitrogen fixation and ammonium assimilation in common bean nodules. Host-plant and nodule growth, symbiotic nitrogen fixation, glutamine synthetase (GS) and glutamate dehydrogenase (GDH) were analyzed in two common bean varieties subjected to iron deficiency. Results showed that host-plant and nodules growth, nitrogen fixation and GS activity decreased when under Fe-deficiency against an important increase of ammonium accumulation and GDH activity. Tolerant variety Flamingo is clearly less affected by iron deficiency than the sensitive one, Coco blanc. The allocation of iron to nodules and Fe use-efficiency for nodule growth and symbiotic nitrogen fixation were on the basis of the symbiotic performance of Flamingo under iron deprivation. Under Fe-deficiency, GDH take over GS the ammonium assimilation activity, particularly in the tolerant variety.     

Effect of phosphorus deficiency on acid phosphatase and phytase activities in common bean (Phaseolus vulgaris L.) under symbiotic nitrogen fixation

Changes in growth, symbiotic nitrogen fixation (SNF), acid phosphatase (ACP), and phytase activities to phosphorus availability (15 and 60 μmol KH₂PO₄ plant⁻¹ week⁻¹) were compared in two recombinant lines (115 and 147) of common bean. Plant growth, nodulation and SNF were genotype and P level-dependent. 147 was more affected by P shortage (15 μmol P) than 115. Four ACP types were revealed in the nodules of both lines, ACP1 exhibiting a higher specific activity under P shortage as compared to the 60 μmol P treatment, especially in 115. A single phytase was revealed for the nodules of both lines and was significantly enhanced by P deficiency. Three ACP types were found in roots and leaves, showing increasing activity under P deficiency, especially in 115. Regardless of P supply, leaf ACP specific activity was higher than that of nodules and roots in the both lines. Interestingly, phosphorus use efficiency for N₂ fixation significantly correlated to nodule ACP activity under P shortage in the both lines. The relatively better performance of 115 as compared to 147 under P deficiency could be partly ascribed to the ability of 115 to maintain higher ACP activity. This enzyme might be involved in the remobilization of the plant Pi and its utilization for SNF.     

Variability in nitrogen fixation of common bean (Phaseolus vulgaris L.) intercropped with maize

Thirty one selected bean lines were evaluated in the field for ability to support N₂ fixation when intercropped with maize which received 0, 30 and 60 kg N ha^–1 as ammonium sulphate. The amount of fixed N₂ was estimated using the natural variation of ¹⁵N and wheat as the standard non-fixing crop. Nitrogen as low as 15 kg N ha^–1 at sowing suppressed nodule weight and activity (acetylene reduction activity) but not nodule number, suggesting that the main effect of mineral N was on nodule development and function. ¹⁵N data revealed a high potential of the bean genotypes to fix N₂, with the most promising ones averaging between 50–60% of seed N coming from fixation. Bean lines CNF-480, Puebla-152, Mexico-309, Negro Argel, CNF-178, Venezuela-350 and WBR22-3, WBR22-50 and WBR22-55 were ranked as good fixers.     

Comparison of common bean (Phaseolus vulgaris L.) genotypes for nitrogen fixation tolerance to soil drying

Aims

Common bean is a major source of protein for many people worldwide. However, the crop is often subjected to drought conditions and its advantage in undertaking symbiotic nitrogen fixation can be severely decreased. The primary objective of this study was to compare the resistance of nitrogen fixation of 12 selected genotypes to soil drying.

Methods

Twelve common bean genotypes of diverse genetic background were compared. Plants were grown in pots and subjected to soil drying over about 2 weeks. Nitrogen fixation was measured daily using a flow-through acetylene reduction technique. The plants were exposed to acetylene for only a short time period allowing repeated measures. The acetylene reduction rate of plants on drying soil was normalized against the rates measured for well-watered plants.

Results

Substantial variability was identified among genotypes in the threshold soil water content at which nitrogen fixation was observed to decrease. Genotypes SER 16, SXB 412, NCB 226, and Calima were found to have the greatest delay in their decrease in nitrogen fixation rates based on soil water content. These four genotypes expressed substantial tolerance of nitrogen fixation to soil drying. These experiments also resulted in data on the threshold soil water contents at which transpiration rates decreased. A decrease in transpiration rates at high soil water contents is potentially advantageous since it allows soil water conservation for use as the severity of the drought increases. There was a general trend of those genotypes with sustained nitrogen fixation rates to low soil water contents also expressing decreased transpiration rates at high soil water contents.

Conclusions

This study identified genetic variation among common bean genotypes in their response of nitrogen fixation and transpiration to soil drying. Five genotypes (SER 16, SXB 412, NCB 226, Calima, and SEA 5) expressed the desired traits for water-limited conditions, which might be exploited in breeding efforts.     

Plant growth-promoting rhizobacteria for improving nodulation and nitrogen fixation in the common bean (Phaseolus vulgaris L.)

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.     

Races of common bean (Phaseolus vulgaris,Fabaceae)

Evidence for genetic diversity in cultivated common bean (Phaseolus vulgaris) is reviewed. Multivariate statistical analyses of morphological, agronomic, and molecular data, as well as other available information on Latin American landraces representing various geographical and ecological regions of their primary centers of domestications in the Americas, reveal the existence of two major groups of germplasm: Middle American and Andean South American, which could be further divided into six races. Three races originated in Middle America (races Durango, Jalisco, and Mesoamerica) and three in Andean South America (races Chile, Nueva Granada, and Peru). Their distinctive characteristics and their relationships with previously reported gene pools are discussed.     

Combined inoculation with Glomus intraradices and Rhizobium tropici CIAT899 increases phosphorus use efficiency for symbiotic nitrogen fixation in common bean (Phaseolus vulgaris L.)

This study compared the response of common bean (Phaseolus vulgaris L.) to arbuscular mycorrhizal fungi (AMF) and rhizobia strain inoculation. Two common bean genotypes i.e. CocoT and Flamingo varying in their effectiveness for nitrogen fixation were inoculated with Glomus intraradices and Rhizobium tropici CIAT899, and grown for 50 days in soil–sand substrate in glasshouse conditions. Inoculation of common bean plants with the AM fungi resulted in a significant increase in nodulation compared to plants without inoculation. The combined inoculation of AM fungi and rhizobia significantly increased various plant growth parameters compared to simple inoculated plants. In addition, the combined inoculation of AM fungi and rhizobia resulted in significantly higher nitrogen and phosphorus accumulation in the shoots of common bean plants and improved phosphorus use efficiency compared with their controls, which were not dually inoculated. It is concluded that inoculation with rhizobia and arbuscular mycorrhizal fungi could improve the efficiency in phosphorus use for symbiotic nitrogen fixation especially under phosphorus deficiency.     

Time course of N2 fixation in common bean (Phaseolus vulgaris L.)

Field and greenhouse experiments were conducted to assess the nitrogen fixation rates of four cultivars of common bean (Phaseolus vulgaris L.) at different growth stages. The ¹⁵N isotope dilution technique was used to quantify biological nitrogen fixation. In the greenhouse, cultivars M4403 and Kallmet accumulated 301 and 189 mg N plant^–1, respectively, up to 63 days after planting (DAP) of which 57 and 43% was derived from atmosphere. Under field conditions, cultivars Bayocel and Flor de Mayo RMC accumulated in 77 DAP, 147 and 135 kg N ha^–1, respectively, of which approximately one-half was derived from the atmosphere. The rates of N₂ fixation determined at different growth stages increased as the plants developed, and reached a maximum during the reproductive stage both under field and greenhouse conditions. Differences in translocation of N were observed between the cultivars tested, particularly under field conditions. Thus, the fixed N harvest index was 93 and 60 for cultivars Flor de Mayo and Bayocel, respectively. In early stages of growth, the total content of ureides in the plants correlated with the N fixation rates. The findings reported in the present paper can be used to build a strategy for enhancing biological N₂ fixation in common bean.     

Soil moisture and potassium affect the performance of symbiotic nitrogen fixation in faba bean and common bean

Potassium (K) is reported to improve plant's resistance against environmental stress. A frequently experienced stress for plants in the tropics is water shortage. It is not known if sufficient K supply would help plants to partially overcome the effects of water stress, especially that of symbiotic nitrogen fixation which is often rather low in the tropics when compared to that of temperate regions. Thus, the impact of three levels of fertilizer potassium (0.1, 0.8 and 3.0 mM K) on symbiotic nitrogen fixation was evaluated with two legumes under high (field capacity to 25% depletion) and low (less than 50% of field capacity) water regimes. Plants were grown in single pots in silica sand under controlled conditions with 1.5 mM N (¹⁵N enriched NH₄NO₃). The species were faba bean (Vicia faba L.), a temperate, amide producing legume and common bean (Phaseolus vulgaris L.), a tropical, ureide producing species. In both species, 0.1 mM K was insufficient for nodulation at both moisture regimes, although plant growth was observed. The supply of 0.8 or 3.0 mM K allowed nodulation and subsequent nitrogen fixation which appeared to be adequate for respective plant growth. High potassium supply had a positive effect on nitrogen fixation, on shoot and root growth and on water potential in both water regimes. Where nodulation occurred, variations caused by either K or water supply had no consequences on the percentage of nitrogen derived from the symbiosis. The present data indicate that K can apparently alleviate water shortage to a certain extent. Moreover it is shown that the symbiotic system in both faba bean and common bean is less tolerant to limiting K supply than plants themselves. However, as long as nodulation occurs, N assimilation from the symbiotic source is not selectively affected by K as opposed to N assimilation from fertilizer.     

Marker association study of yield attributing traits in common bean (Phaseolus vulgaris L.)

Molecular Biology Reports - Common bean is gaining acceptance as one of the most valuable major food consumed worldwide owing to innumerable nutritional and therapeutic benefits. Comparatively less...     

Multiple paternity in common bean (Phaseolus vulgaris L., Fabaceae)

We report on two field experiments that were conducted in 1991 and 1992 at the South Coast Extension and Research Center, Irvine, CA, to study the incidence of multiple paternity in the common bean (Phaseolus vulgaris L.). Hypocotyl color and shikimate dehydrogenase (Skdh) isozymes were used as genetic markers. The white-seeded cultivar ‘Ferry Morse 53’ (FM 53) was used as the female parent. This cultivar is homozygous recessive (pp) for hypocotyl color. The pollen source parents were three homozygous dominant (PP) purple-hypocotyled, black-seeded cultivars. Three cultivars, ‘ICA Pijao,’ G4459, and the maternal parent FM 53, are of Mesoamerican origin and homozygous for the fast (F) allele at the Skdh locus. The other cultivar, Black Valentine, is of Andean origin and is homozygous for the slow (S) allele at the Skdh locus. Overall, 6 125 pods were obtained from 57 and 111 plants harvested individually in 1991 and 1992, respectively. All progeny, 28938 seeds, were scored for hypocotyl color at the seedling stage. The purple-hypocotyled seedlings were genotyped for the Skdh locus to identify their pollen parents. Multiple paternity was identified in all the pods with hybrid seeds (i.e., those of intercultivar crosses) at 5.8% and 8.1% in 1991 and 1992, respectively. All multiply sired pods produced both nonhybrid and hybrid seeds. As many as three successful fathers per pod were identified, but the number of markers limited measuring higher levels of multiple paternity. Most multiply sired pods (≈70%) were filled by nonhybrid seeds plus a single hybrid seed. Ovule position effect within multiply sired pods was inferred from the nonrandom distribution of hybrid seeds within a pod. On average, hybrid seeds occurred more frequently in ovules in position 7 (most basal) and in position 1 (most stylar) than in ovules in the middle positions of the pod.     

Inheritance of partial resistance to bean rust in the common bean (Phaseolus vulgaris L.)

The inheritance of partial resistance within eight bean cultivars to a single-pustule isolate of bean rust was studied by means of a F₁ diallel test. General combining ability (GCA) and specific combining ability (SCA) were very highly significant over two seasons and in interaction with seasons. The partitioning of the sums of squares indicated the greater importance of GCA in the inheritance of the resistance. Reciprocal effects were not significant. The estimates of narrow-sense heritability in the two seasons were 0.899 ± 0.056 and 0.603 ± 0.065.     

Host plant nodule parameters associated with nitrogen fixation efficiency in French bean (Phaseolus vulgaris L) cultivars

Two cultivars of French bean (Phaseolus vulgaris L.) viz. contender and arka komal were planted in polythene bags containing sand and grown under glasshouse conditions. The nodulation status, shoot/root biomass, activities of several nodule enzymes, total soluble protein and leghaemoglobin contents were monitored over the entire growth period. Allantoinase activity in leaves was measured to monitor the ureide degrading capacity. Significant genotype difference was observed in both the cultivars. All the parameters showed a decline after flowering except uricase, which declined before flowering. Malate dehydrogenase and isocitrate dehydrogenase showed a constant decline throughout the growth period. Degree of decline varied with the genotype for all the parameters. Leghaemoglobin content, PEP carboxylase activity and ureide degrading capacity of leaves did not show an appreciable decline in contender and were significantly higher than in arka komal. These factors can be used to increase nitrogen fixation in French bean.     

Patterns of variation in cultivated common bean (Phaseolus vulgaris,Fabaceae)

More than 18,000 accessions of common bean (Thaseolus vulgaris, Fabaceae) from the Centro Internacional de Agricultura Tropical (CIAT) germplasm bank were examined at two locations in Colombia. A large variation in cultivated dry bean was found among accessions from primary centers of domestication in Middle and South America. For some bean types, such as medium- and large-seeded white, variation was greater among germplasm from western Asia (Turkey) and Europe (Portugal, Spain, Greece, France, Italy, and Bulgaria). Based on growth habit, on seed, pod, and leaf characteristics, and on ecological regions of adaptation, dry-bean germplasm was divided into a total of six gene pools from Middle American and four gene pools from South American centers of domestication. Most of the variation in the snap or stringless bean appears to be of relatively recent origin; it was greatest among cultivars from China, Europe, and the United States. These could be grouped into two additional gene pools. A strategy for breeding and transfer of genes across gene pools is also discussed.     

The effect of Rhizobium inoculation and nitrogen fertiliser on nitrogen fixation and seed yield of dry beans (Phaseolus vulgaris)

The potential benefit to be derived from seed inoculation of Phaseolus vulgaris beans with effective strains of Rhizobium phaseoli, was investigated in field experiments over three years on a site low in soil nitrogen and lacking indigenous effective strains of R. phaseoli. Inoculation with R. phaseoli (strain RCR 3644) produced significant increases in nodulation, nitrogenase activity and plant growth in all experiments. In trials in 1978 and 1979, with cv. Seafarer, inoculation, in the absence of nitrogen fertiliser doubled seed yields. In 1978, the seed yields from inoculated beans without nitrogen fertiliser (1–6 t/ha) were not significantly different from those obtained with uninoculated beans receiving the optimum nitrogen fertiliser treatment of 120 kg N/ha (1–75 t/ha). In 1979, with lower rainfall favouring more efficient utilisation of nitrogen fertiliser, inoculation gave seed yields (1–88 t/ha) equivalent to those obtained with 60 kg N/ha (1–70 t/ha) but significantly less than with 120 kg N/ha (2–88 t/ha). More precise estimates from nitrogen response curves showed that inoculation supplied the fertiliser equivalent of 105 and 70 kg N/ha in 1978 and 1979 respectively. In both years, significant benefits were also obtained by the combination of inoculation and nitrogen fertiliser. In a separate experiment in 1979, with four R. phaseoli strains inoculated onto eight bean cultivars, three were highly effective nitrogen fixers on all cultivars. Two strains (RCR 3644 and NVRS 963A) each increased mean yields, in the absence of nitrogen fertiliser, from 1–39 t/ha uninoculated to c. 2–5 t/ha inoculated whilst strain RCR 3622 was outstanding with a mean yield of 3-0 t/ha. An analysis of the nitrogen content of seed showed that gains from nitrogen fixation were 37–57 kg N/ha/growing cycle for the combination RCR 3644 with cv. Seafarer. However, 106 kg N/ha/growing cycle was recorded for the combination RCR 3622 and cv. Aurora.     

Sphingolipids in bean leaves (Phaseolus vulgaris)

Phytoglycolipid has been isolated for the first time from plant leaves (Phaseolus vulgaris). The purified product (almost identical with the phytoglycolipid isolated from flax seed) was a ceramide attached through phosphate diester linkage to an oligosaccharide, which consisted of the usual trisaccharide unit (inositol, hexuronic acid, hexosamine) to which were attached mannose, galactose, and arabinose. The major fatty acids were the saturated 2-hydroxy C(22), C(24), and C(26) acids; the major long-chain bases were dehydrophytosphingosine (d-ribo-1,3,4-trihydroxy-2-amino-8-trans-octadecene) (53%) and phytosphingosine (d-ribo-1,3,4-trihydroxy-2-amino-octadecane) (32%). A ceramide and a cerebroside were also isolated. In the ceramide the major fatty acids and the major long-chain bases were the same as in the phytoglycolipid. In the cerebroside, the fatty acid composition was similar to that in the ceramide and phytoglycolipid, but the long-chain bases consisted of dehydrophytosphingosine and phytosphingosine (7:1) with a substantial amount of unidentified long-chain base. The sugar component was glucose.