Worldwide genotyping of castor bean germplasm (<Emphasis Type="Italic">Ricinus communis</Emphasis> L.) using AFLPs and SSRs

Worldwide genetic diversity in 200 individuals comprising 41 castor bean accessions was assessed using amplified fragment polymorphisms (AFLPs) and simple sequence repeats (SSRs). We found that, despite surveying five continents and 35 countries, genetic diversity in castor bean germplasm is relatively low (overall H _e = 0.126 for AFLPs and 0.188 for SSRs) compared to estimates of genetic diversity in other plant species. Our data also show no geographic structuring of genotypes across continents or countries within continents. An assessment of the congruence between AFLP and SSRs indicates a low correlation (R ² = 0.19) between the two data sets, but each marker class nonetheless shows similar patterns of low-genetic diversity and a lack of geographic structure. Our data do suggest that SSRs yield a higher percentage of polymorphic loci, higher heterozyosity and a greater range of genetic distances, and are therefore more informative than are AFLPs on a locus-by-locus basis. Based on comparisons with numerous other plant species, we suggest that the lower genetic variation in this worldwide collection may be due to one or more factors including: sampling strategies that have not captured the full extent of genetic variation in the species; artifactual variation due to long-term germplasm storage and seed regeneration; or intense selection followed by domestic cultivation of a limited number of castor bean genotypes, which are widely propagated for their horticultural and agro-economic value.     

The influence of N concentration and NO3/NH4 ratio on the growth of lima and snap bean and southern field pea seedlings

Abstract

Lima and snap beans and southern field peas were cultured in a modified Hoagland's solution for 14 days with N supplied as Ca(NO₃)₂ and/or (NH₄)₂SO₄ at three N concentrations and five ratios of NO₃ to NH₄. The ratio of NO₃/NH₄, rather than the N concentrations influenced seedling growth of these vegetable legumes. Maximum seedling growth of lima bean was generally obtained with all combinations of NO₃ and NH₄. A preference for 75% NO₃ and 25% NH₄ was observed for snap bean. Southern field pea growth was reduced only when all of the N was supplied as 100% NH₄. Ammonium toxicity symptoms, lesions and severe wilting, developed with snap bean and southern field pea within 14 days when cultured with 100% NH₄. Lima bean, though reduced in growth, exhibited a tolerance to the 100% NH₄ treatment.     

Extensive introgression of Middle American germplasm into Chilean common bean cultivars

Summary The genetic diversity of 95. representative Chilean common bean (Phaseolus vulgaris L.) landraces was analyzed using phaseolin seed protein and eight isozyme systems as genetic markers. Four types of phaseolin were found, “C”, “T”, “S” and “H”, in decreasing order of frequency. Each type had a different distribution between the Northern and Southern regions of the country. Nei’s genetic distance based on isozyme diversity indicated that a high percentage of the total variation found in this sample occurred between landraces and only a small percentage of the variation was detected within populations. Cluster analysis based on Nei’s genetic distance and a principal component analysis of isozyme frequencies did not detect a clear association between the geographic distribution of the landraces and their isozyme constitution. However, Nei’s genetic distance analysis clustered the bean landraces into two major groups which had a specific isozyme pattern, seed color, and seed size. The genetic analysis also detected a rare polymorphism for theMdh-2 locus, a null allele at theDiap-2 locus, and polymorphism for theAco-2 locus. The principal component analysis of isozyme frequencies showed that only 30% of the genotypes analyzed were similar to the Andean check and 5% of the samples were similar to Middle American check. This finding suggests a high frequency of hybridization between the Middle America and Andean gene pools in cultivated common bean from Chile.     

Genetic structure of Lima bean (<Emphasis Type="Italic">Phaseolus lunatus</Emphasis> L.) landraces grown in the Mayan area

Lima bean (Phaseolus lunatus L.) is an important crop in the Mayan culture. The Mayan area, considered as a main center of Mesoamerican diversity, has been divided into two subareas: the Mayan lowlands and the Mayan highlands. The Yucatan Peninsula is part of the Mayan lowlands and holds the highest number of Lima bean landraces of Mexico, but Lima beans are in high risk of genetic erosion due to intensification of the traditional Mayan agriculture. However, information on genetic diversity of Lima beans of the Mayan highlands is lacking. By using 46 landraces collected in the Mayan area (23 from each subarea) and 73 ISSR loci (inter-simple sequence repeats), we analyzed the structure, diversity and genetic relationships of Lima beans of this part of Mesoamerica. High levels of diversity (H _BAY = 0.45) and genetic structure (F _ST = 0.66) were found for the whole Mayan area. Genetic diversity in the Mayan lowlands was apparently higher than the Mayan highlands (H _BAY = 0.44 and 0.36, respectively); but differences were not statistically significant. Genetic structure between the subareas was high (AMOVA = 30% of total variation), most landraces grouping according to their geographic origin. This study shows the importance of the Mayan culture in the diversification and conservation of Lima beans. The results provide important information that should be considered when implementing strategies to collect Lima bean landraces and planning in situ and ex situ programs to conserve these landraces in the Mayan region.     

Genetic erosion and in situ conservation of Lima bean (Phaseolus lunatus L.) landraces in its Mesoamerican diversity center

Lima bean (Phaseolus lunatus L.) is an important crop in traditional Mayan agriculture of the Yucatan Peninsula, Mexico, its Mesoamerican center of diversity. Genetic erosion in this species is currently a threat in this region out of 3 of 21 landraces dominate 71.24% of the cultivated area, and 12 are rare landraces grown only in 6.29%. Using 90 ISSR loci, we estimated the diversity and genetic relationships for 21 landraces to analyzing their risk of genetic erosion, and generate data for their in situ conservation. Total genetic diversity was high (h = 0.29), however it was lower than wild gene pool reported (h = 0.69). The abundant landraces had genetic diversity values lower (h = 0.13, I = 0.17) than the common (h = 0.26, I = 0.33) and rare landraces (h = 0.24, I = 0.27). However, the rare landraces are in a higher risk of genetic erosion due to local extinction. The cluster analysis showed no groups corresponding to morpho-phenological characteristics, geographic origin or traditional classification, which resulted from high inter-landraces gene flow levels. The molecular data confirmed that the domesticated Lima bean pool of the Yucatan Peninsula has a high risk of genetic erosion. If current tendencies in landrace cultivation continue, many will no longer be planted within two to three generations, with a consequent loss of their alleles. Programs urgently need to be established for in situ conservation of Lima bean landraces in this region.     

Morphotype relationships in Lima bean (Phaseolus lunatus L.) deduced from variation of the evolutionary marker phaseolin

Summary Variation of the seed storage protein phaseolin was analysed in a collection of 100 accessions of Phaseolus lunatus L. using one-dimensional SDS/PAGE. Cultivated small-seeded genotypes belonging to Sieva and Potato morphotypes and intermediate Sieva-Potato and Sieva-Big Lima morphotypes showed the M (Mesoamerican) pattern, confirming their origin in the same gene pool. Cultivated Big Lima morphotypes showed the A (Andean) pattern, confirming that they belong to a distinct gene pool.     

Variation and genomic polymorphism of lectin-related proteins in Lima bean (Phaseolus lunatus L.) seeds

Variation of the lectin and the two lectin-related proteins, AIL (-amylase inhibitor-like) and ARL (arcelin-like) was examined in wild and cultivated accessions of Lima bean (Phaseolus lunatus L.) using electrophoresis of total seed proteins, immunoblot and RFLP analysis of lectin-related genes. Results confirm that divergence of the two major Lima bean gene pools, Andean and Mesoamerican, also apply to this protein family. All three members of the family are present in both gene pools, with differences in size, abundance and composition between gene pools, giving the possibility to distinguish Andean from Mesoamerican lectin pattern types. Both patterns show some variants, such as lack of lectin or its presence as an abundant protein. The observed variation reflects, at least in part, into genomic polymorphism. The presence of arcelin- and -amylase-related proteins in Lima bean could represent a tool to increase our knowledge in the evolution of the lectin family in Phaseolus species.     

Response of Determinate and Indeterminate Common Bean Genotypes to Rhizobium Inoculant in a Short Season Rainfed Production System in the Canadian Prairie

ABSTRACT

Common bean (Phaseolus vulgaris L.) has been shown to be a poor di-nitrogen (N₂) fixer and nitrogen (N) fertilizers are usually recommended in bean production. Recent research results suggest that the success of the bean/Rhizobium symbiosis may depend, in part, on the specific bean genotype. Twelve dry bean genotypes differing in growth habit, commercial class, and maturity were evaluated for N₂ fixation in field experiments. Response to inoculant application was highly influenced by environmental conditions. Genotypes differed in nodule dry mass, seed yield, seed N yield, and in amount of N₂ fixed. Growth habit alone was not adequate in classifying bean genotypes for N₂ fixation. The actual amount of N₂ fixed was low ranging from 16 kg ha^?1 to 27 kg ha^?1, suggesting that the symbiotic process alone may not provide adequate N for optimum seed yield in dry environments.     

Proton toxicity interferes with the screening of common bean (Phaseolus vulgaris L.) genotypes for aluminium resistance in nutrient solution

Common bean (Phaseolus vulgaris L.) proved to be very sensitive of low pH (4.3), with large genotypic differences in proton sensitivity. Therefore, proton toxicity did not allow the screening of common bean genotypes for aluminium (Al) resistance using the established protocol for maize (0.5 mM CaCl₂, 8 μM H₃BO₃, pH 4.3). Increasing the pH to 4.5, the Ca²⁺ concentration to 5 mM, and addition of 0.5 mM KCl fully prevented proton toxicity in 28 tested genotypes and allowed to identify differences in Al resistance using the inhibition of root elongation by 20 μM Al supply for 36 h as parameter of Al injury. As in maize, Al treatment induced callose formation in root apices of common bean. Aluminium‐induced callose formation well reflected the effect of Ca supply on Al sensitivity as revealed by root‐growth inhibition. Aluminum‐induced callose formation in root apices of 28 bean genotypes differing in Al resistance after 36 h Al treatment was positively correlated to Al‐induced inhibition of root elongation and Al contents in the root apices. However, the relationship was less close than previously reported for maize. Also, after 12 h Al treatment, callose formation and Al contents in root apices did not reflect differences in Al resistance between two contrasting genotypes, indicating a different mode of the expression of Al toxicity and regulation of Al resistance in common bean than in maize.     

Genetic diversity in the Lima bean (Phaseolus lunatus L.) as revealed by chloroplast DNA (cpDNA) variations

Genetic diversity in the Lima bean (P. lunatus L.) was assessed bymeans of two chloroplast DNA probes. Based on data obtained from 152accessions including wild forms and landraces, the two separateMesoamerican and Andean groups were confirmed and a transition poolof genetic diversity was observed in the two botanical formsconsistent with their distribution range. Three primary centres ofgenetic diversity and one secondary diversification spot are proposedfor the wild Lima bean whereas only two domestication sites areunderlined for the landraces. The importance of human intervention inwidening the distribution range and the genetic diversity inlandraces is discussed.     

Growth,P uptake in grain legumes and changes in rhizosphere soil P pools

In soils with low P availability, several legumes have been shown to mobilise less labile P pools and a greater capacity to take up P than cereals. But there is little information about the size of various soil P pools in the rhizosphere of legumes in soil fertilised with P although P fertiliser is often added to legumes to improve N₂ fixation. The aim of this study was to compare the growth, P uptake and the changes in rhizosphere soil P pools in five grain legumes in a soil with added P. Nodulated chickpea (Cicer arietinum L.), faba bean (Vicia faba L.), white lupin (Lupinus albus L.), yellow lupin (Lupinus luteus L.) and narrow-leafed lupin (Lupinus angustifolius L.) were grown in a loamy sand soil low in available P to which 80 mg P kg⁻¹ was added and harvested at flowering and maturity. At maturity, growth and P uptake decreased in the following order: faba bean > chickpea > narrow-leafed lupin > yellow lupin > white lupin. Compared to the unplanted soil, the depletion of labile P pools (resin P and NaHCO₃-P inorganic) was greatest in the rhizosphere of faba bean (54% and 39%). Of the less labile P pools, NaOH-P inorganic was depleted in the rhizosphere of faba bean while NaOH-P organic and residual P were most strongly depleted in the rhizosphere of white lupin. The results suggest that even in the presence of labile P, less labile P pools may be depleted in the rhizosphere of some legumes.     

Relationships between an Italian Strawberry Ecotype and its Ancestor using RAPD Markers

Random-amplified polymorphic DNA (RAPD) markers were used to evaluate genetic variability among populations of an Italian strawberry ecotype, and to determinate genetic relationships between genotypes and their putative ancestor. A total of 65 selections and one cultivar ‘Madame Moutot’ (MM), were analysed to evaluate genetic variability present in Etna mountain area and to confirm as MM was one of the cultivars that originated the ecotype. A total of 222 RAPD markers was obtained using 16 decamer primers and 6 longer primers, 90.8% of the markers obtained by selected primers resulted polymorphic at least within analysed genotypes. RAPDs were used to calculate genetic similarity coefficients and to generate dendrograms representing genetic relationships among genotypes analysed. Cluster analysis displays as RAPD polymorphisms were able to characterize the genotype variability among closely related groups. The data show as MM could be considered the ancestral genotypes introduced in that area. The results obtained confirm that RAPD markers could be used as reliable markers to perform phylogenetic studies in Fragaria×ananassa Duch. ex Rozier. Giuseppe Bertino and Piero Spada - Coauthor involved in genotype selection and field management     

Genetic differentiation among selected tepary bean collections revealed by morphological traits and simple sequence repeat markers

Understanding the genetic relationship of crop ideotypes is essential for genetic analysis and breeding. The objective of this study was to determine genetic differentiation present among 20 selected tepary bean (Phaseolus acutifolius A. Gray) genotypes using morphological traits and simple sequence repeat (SSR) markers in order to identify genetically unique parental lines to develop breeding populations. Phenotypic diversity was estimated using Shannon-Weaver diversity index (H’), principal component analysis (PCA) and cluster analysis (CA). Genotypic diversity was estimated using Jaccard's genetic distances and CA. Shannon-Weaver diversity index values for the studied morphological traits ranged from 0.89 to 0.99, with a mean of 0.95 revealing high phenotypic differences among test genotypes. PCA identified four useful principal components (PC's) which contributed to 73% of the total phenotypic variation of collections. PC1 accounted for 28% of the total variation correlated to dry shoot mass and number of pods per plant. PC2 correlated with number of seeds per pod, grain yield and harvest index and contributed to 21% of total variation. The mean observed (H_o) and expected (H_e) heterozygosity values were 0.45 and 0.51, respectively revealing moderate genetic differentiation among genotypes. The mean polymorphic information content was 0.52, suggesting efficient discriminatory power of the SSR loci useful in future tepary bean genetic diversity analysis. The current study revealed moderate genetic differentiation among the studied tepary bean genotypes. Morphological traits and SSR markers well-correlated in allocating the tepary bean genotypes. The following genotypes were genetically distinct: G40201, G40237, G40068, G40033 and G40063 which are recommended for further crosses, selection and population development.     

Competition and persistence of Rhizobium tropici and Rhizobium etli in tropical soil during successive bean (Phaseolus vulgaris L.) cultures

Strains of Rhizobium tropici IIB, CIAT899 and F98.5, both showing good N₂ fixation, and a R. etli strain W16.3SB were introduced into a field which had no history of bean culture. Plant dilution estimates showed that in the presence of its host (Phaseolus vulgaris cv. Carioca) during the cropping seasons and the subsequent fallow summer periods, the bean rhizobial populations increased from less than 30 to 10³ g^–1 dry soil after 1 year and to 10⁴ g^–1 dry soil after 2 years. In the 1st year crop, the inoculated strains occupied most of the nodules, which resulted in a higher nodulation and C₂H₂ reduction activity. Without reinoculation for the second and third crops, however, little R. tropici IIB was recovered from the nodules and the bean population consisted mainly of R. etli, R. leguminosarum bv. phaseoli, and R. tropici IIA. Reinoculation with our superior R. tropici IIB strains before the second crop resulted in R. tropici IIB occupying the main part of the nodules and a positive effect on nodulation and C₂H₂ reduction activity, but reintroduction of the inoculant strain in the third season did not have any effect.     

Investigation of the genetic structure of some common bean (Phaseolus vulgaris L.) commercial varieties and genotypes used as a genitor with SSR and SNP markers

Common bean is a species belonging to the Phaseolus genus of the Leguminosae family. It has economic importance due to being rich in protein, vitamin A and C, and minerals. Being one of the most cultivated species of legumes, the determination of genetic diversity in bean genotypes or populations has an important role in terms of our genetic resources. The objective of this study was to evaluate the genetic structure of 94 genotypes which were cultivated in different parts of the world and our country with SSR and SNP markers. 10 SSR loci and 73 SNP primers were used for the determination of genetic structure in commercial cultivars and breeding lines. All of the SSR and SNP loci used in the study were found to be polymorphic. A total of 89 alleles were identified for 10 SSR loci. Mean number of alleles per locus (Na?=?8.9), effective allele number (Ne?=?3.731), Shannon information index (I?=?1.468), observed heterozygosity (Ho?=?0.023), and expected heterozygosity (He?=?0.654) were calculated based on SSR analysis. According to the results of Bayesian-based STRUCTURE analysis using SSR and SNP data, 94 bean genotypes were genetically divided into three main clusters. According to genetic distance based UPGMA dendrogram obtained from SNP analysis, 94 bean genotypes were divided into 2 main clusters corresponding Mesoamerican and Andean gene pools. The obtained results provide important information about the genetic structures of the studied bean cultivars and breeding lines. With the obtained results, it will be possible to develop breeding programs to develop new cultivars by using our gene resources.

     

Genetic relationships within Vigna unguiculata (L.) Walp. based on isozyme analyses

Summary Isozyme analyses of genetic diversity in Vigna unguiculata were performed to determine genetic relationships and level of genetic diversity between wild and cultivated cowpea. Thirty-four cultivated accessions of V. unguiculata, 56 wild accessions of V. unguiculata, and six accessions representing five related wild Vigna species were analyzed. Ten enzyme systems were polymorphic within Vigna unguiculata: AAT, ACO, G6PDH, DIAP, LAP, MUE, ME MDH, PRX, and SOD. Fourteen of 24 putative loci (58%) were polymorphic within wild V. unguiculata, but only one locus (4%) was polymorphic within cultivated cowpea; when five related Vigna species were examined, 21 of the 24 bands of activity showed polymorphisms (88%) adding 33 alleles to the 48 identified within V. unguiculata. In one F₂ population of 68 plants (UCDVg 36 × UCDVg 21) a loose linkage was indicated between Diap-2 and G6pd-1 (² = 15.39; p = 0.004) with an estimated distance of 36.0 cM ± 5.02 (recombination (r) = 0.31). Also in another F₂ population of 38 plants (CB 88 × UCDVg 21) a loose linkage was indicated between Lap-1 and Prx (\gC² = 9.62; p = 0.047) with an estimated distance of 39.8 cM ± 7.0 (r = 0.33). Total genetic diversity (H_T) was 0.085 over all of the accessions including the one classified as V. nervosa. Within accession diversity (H_s) approached zero and between accession diversity (D_si) was responsible for all of the genetic diversity present. Therefore the coefficient of gene differentiation (G_ST = D_STII_T) approached 1. Absolute gene differentiation (D_m) was 0.087. Two of the nine segregations in this study were skewed. In general, results of this study concurred with the taxonomic classification within V. unguiculata and provided a strong indication that a severe genetic bottleneck occurred during the domestication process of cowpea.Abbreviations AAT aspartate amino-transferase - ACO aconitase - ALD aldolase - AUS Australia - BDI Burundi - BWA Botswana - CHN China - CMR Cameroon - DIAP diaphorase - DZA Algeria - ETH Ethiopia - G6PDH glucose-6-phosphate dehydrogenase - GDH glutamate dehydrogenase - GHA Ghana - GUY Guyana - IDH isocitrate dehydrogenase - IND India - KEN Kenya - LAO Laos - LAP leucine aminopeptidase - MDH malate dehydrogenase - ME malic enzyme - MEX Mexico - MOZ Mozambique - MUE methylumelliferyl-esterase - MWI Malawi - MYS Malaysia - NER Niger - NGA Nigeria - PRX peroxidase - RBSC ribulose-bisphosphate carboxylase - SEN Senegal - SLE Sierra Leone - SOD superoxide dismutase - TGO Togo - TZA Tanzania - USA United States of America - XDH xanthine dehydrogenase - ZAF South Africa - ZAR Zaire - ZIM Zimbabwe - ZMB Zambia     

Biological nitrogen fixation in Azospirillum strain-maize genotype associations as evaluated by the 15N isotope dilution technique

Few studies of the inoculation of cereal crops with N₂-fixing bacteria have included more than one or two plant genotypes. In a recent study performed in Argentina using 12 different maize genotypes, it was found in 2 consecutive field experiments that several of them responded consistently, either negatively or positively, to inoculation with a mixture of strains of Azospirillum spp. The present study in post was performed to investigate the effect of inoculation of individual strains (and a mixture) of Azospirillum spp., and their nitrate reductase negative (NR-) mutants, on the growth of four of these maize genotypes. Two of these genotypes were grown in ¹⁵N-labelled soil with the aim of quantifying any contributions of biological N₂ fixation. Two genotypes (Morgan 318 and Dekalb 4D-70) produced similar increases in grain yield when they were inoculated with a mixture of Azospirillum spp. strains or fertilized with the equivalent of 100 kg N ha^-1. The other genotypes (Dekalb 2F-11 and CMS 22) showed little response to inoculation or N fertilization. The Morgan 318 and Dekalb 4D-70 genotypes showed a large increase in total N accumulation, suggesting that the response was due to increased N acquisition, but not due to bacterial nitrate reductase as the NR- mutants generally caused plant responses similar to those of the parent strains. Despite problems with the stabilization of the ¹⁵N enrichment in the soil, the ¹⁵N isotope dilution results indicated that there were very significant biological nitrogen fixation (BNF) contributions to the Dekalb 4D-70 and CMS 22 maize genotypes.Dedicated to Professor J.C.G. Ottow on the occasion of his 60th birthday     

A Preliminary Comparative Evaluation of Genetic Diversity between Chinese and Japanese Wild Soybean (<Emphasis Type="Italic">Glycine soja</Emphasis>) Germplasm Pools using SSR markers

The wild soybean, an annual self-pollinating plant, is the progenitor of soybeans and is extensively distributed in the Far East of Russia, the Korea peninsula, China and Japan. Geographically, Japan is surrounded by sea and insulated from China. We preliminarily evaluate whether the Japanese and Chinese wild soybean germplasm pools are genetically differentiated from each other using SSR markers. The results showed that the two pools have great genetic differentiation. Some loci presented obvious differences in mean genetic divergence (G_ST) value between the two pools. The G_ST among the geographic regions in China was higher than that in Japan. The average within-geographic region gene diversity values (H_S) in the two pools were completely identical and thus the genetic difference between the two pools was mostly attributed to the relatively high level of between-geographic region gene diversity (D_ST) in China. We suggest that Japanese and Chinese wild soybeans should be comparatively independently evolving in phylogeny.     

Effect of nitrogen levels and Rhizobium strains on symbiotic N2 fixation and grain yield of Phaseolus vulgaris L. genotypes in normal and saline-sodic soils

Summary Following screening, selection, characterization, and symbiotic N₂ fixation with 12,5, 25.0, and 40.0 mg N kg^–1 in normal and saline-sodic soils, only two Phaseolus vulgaris genotypes (HUR 137 and VL 63) and two Rhizobium spp. strains (ND 1 and ND 2) produced maximum nodulation, nitrogenase activity, plant N contents, and grain yields in saline-sodic soil, with 12.5 mg N kg^–1, compared with the other strains. However, interactions between strains (USDA 2689, USDA 2674, and ND 5) and genotypes (PDR 14, HUR 15, and HUR 138) were significant and resulted in more nodulation, and greater plant N contents, nitrogenase activity, and grain yields in normal soils with 12.5 mg N kg^–1 compared with salt-tolerant strains. Higher levels of N inhibited nodulation and nitrogenase activity without affecting grain yields. To achieve high crop yields from saline-sodic and normal soils in the plains area, simultaneous selection of favourably interacting symbionts is necessary for N economy, so that bean yields can be increased by the application of an active symbiotic system.