Morphological and molecular diversity within Algerian cowpea (<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.) landraces

Twenty landraces of cowpea (Vigna unguiculata (L.) Walp.) scattered throughout Algeria were compared through morphological and genetic characterization. At the morphological level, for qualitative characters there was no intra-landrace variation and for quantitative characters the variations were low except for landrace NAG2 Three different cultigroups were located in Algeria: Biflora that was dominant in the Sahara, Melanophtalmus in the North and Unguiculata including one landrace in Kabylia and two in Sahara. The AMOVA analysis indicated that the genetic variation was lower within than among agro-ecological regions. A Mantel test, revealed a correlation between the qualitative morphological data and the geographical data (R = 0.28; P < 0.01), indicating that the degree of morphological change among landraces was roughly proportional to the geographical distances separating them. Genetic diversity was analyzed by using 11 random amplified polymorphic DNA (RAPD) and 12 inter-simple sequence repeat (ISSR) markers. No intra-landrace variability was found. The eleven RAPD primers yielded 77 bands, of which 45 (58.44%) were polymorphic; the genetic similarity ranged from 66.0 to 96.7%. The twelve ISSR primers provided a total of 104 bands, of which 65 (62.5%) were polymorphic; the genetic similarity ranged from 62.8 to 97.8%. cluster analysis showed a good match between genetic background and geographical distribution, which was confirmed by the results of the Mantel test. In particular, geographical data and genetic data were found to be correlated: (R = 0.33; P < 0.01) for RAPD, (R = 0.37; P < 0.01) for ISSR, and (R = 0.33; P < 0.01) for a combined RAPD-ISSR dataset. Moreover, despite the absence of significant correlation between morphological and RAPD data (R = 0.14; P = 0.14), significant correlations between morphological data and both ISSR (R = 0.27, P < 0.05) and a combined RAPD-ISSR dataset (R = 0.22, P < 0.05) were noted. ISSR markers were better linked to morphological variation than were RAPD markers. However, despite this, genetic distances among these landraces were found to be essentially the same no matter which markers were used.     

Greenbug (<Emphasis Type="Italic">Schizaphis graminum</Emphasis>) resistance in oat (<Emphasis Type="Italic">Avena</Emphasis> spp.) landraces from Asia

Among 371 oat accessions from the world collection of the Vavilov Institute of Plant Genetic Resources (VIR) mostly represented by the landraces from Asia (Maritime Territory of Russian Federation, Mongolia, China, Japan, North Korea, India, Iran, Afghanistan, and Bhutan) ninety five were heterogeneous for resistance to the North Caucasus populations of the greenbug (Schizaphis graminum Rondani). Damage ratings of resistance in 47 accessions were high, low level of resistance was identified in 48 genotypes. Higher frequency of greenbug resistant accessions (60.5%) was found among landraces from Mongolia. Lines with high expression of the resistance were selected from seven landraces with wide range of damage ratings. North Caucasus populations of S. graminum from Krasnodar area and Dagestan significantly differ by the frequencies of virulence to host plant. Seven virulence phenotypes differentially interacting with oat genotypes were found in the Krasnodar population and 10 phenotypes were revealed in the Dagestan population. Results of the experiments with the greenbug test clones revealed that all the lines selected from the accessions VIR-2490, VIR-2539, VIR-4074, VIR-12213, VIR-12214 (Mongolia), VIR-6688 (India) and VIR-13624 (North Korea) are protected by diverse alleles of resistance genes which differ from the earlier identified gene Grb3. These lines are supposed to have aphid resistance genes nonidentical to the Grb1 and Grb2. The accessions VIR-6688, VIR-4074 and VIR-12214 possess high resistance to Krasnodar and Dagestan greenbug populations and the accession VIR-13624 is protected by the most effective gene (genes) of resistance to the both populations from North Caucasus.     

<Emphasis Type="Italic">Ziziphus </Emphasis><Emphasis Type="Italic">spina-christi</Emphasis> (L.) Willd.: a multipurpose fruit tree

Neglected and underutilized species often play a vital role in securing food and livestock feed, income generation and energy needs of rural populations. In spite of their great potential little attention has been given to these species. This increases the possibility of genetic erosion which would further restrict the survival strategies of people in rural areas. Ziziphus spina-christi is a plant species that has edible fruits and a number of other beneficial applications that include the use of leaves as fodder, branches for fencing, wood as fuel, for construction and furniture making, and the utilization of different parts e.g. Fruits, leaves, roots and bark in folk medicine. Moreover, the plant is adapted to dry and hot climates which make it suitable for cultivation in an environment characterized by increasing degradation of land and water resources. Lack of research in Z. spina-christi hinders its successful improvement and promotion. Therefore, studies are needed to fully exploit this species. This article aims at summarizing information on different aspects of Z. spina-christi to stimulate interest in this crop which is of importance in Sudan and other countries of the semi-arid tropics.

Agro-morphological characterization of chili pepper landraces (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) cultivated in Northern Benin

Variability study of agro-morphological characteristics is important for varietal improvement and plant breeding program. For this, 148 chili pepper landraces (Capsicum annuum L.) collected from Northern Benin were characterized using 39 (21 qualitative and 18 quantitative) morphological characters. The trial was conducted in a Fisher block design with 3 replicates at the experimental station of the Faculty of Science and Technology (FAST) of Dassa based in Central Benin. The results revealed considerable morphological variability between landraces. Twenty nine (16 quantitative and 13 qualitative) variables among the 39 examined were discriminatory and group the landraces into 4 classes when considering the qualitative variables. With quantitative variables, 9 groups were obtained. The correlations between the studied quantitative variables show that the yield parameters are correlated with the variables related to the vegetative part of the plant. This study will help breeders to better select their parents for crossing in order to produce desirable and high yielding varieties.     

Comparison of the genetic diversity between <Emphasis Type="Italic">Triticum aestivum</Emphasis> ssp. <Emphasis Type="Italic">tibetanum</Emphasis> Shao and Tibetan wheat landraces (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) by using intron-splice junction primers

In order to evaluate and compare the germplasm resources of wheat in Tibet, we analyzed the genetic diversity of 136 Triticum aestivum ssp. tibetanum Shao and 119 Tibetan wheat landraces (Triticum aestivum L.) by using Intron-Splice Junction (ISJ) primers. The results showed that polymorphism of PCR products were obtained by 33 primer combinations, which accounted for 11% of the 300 primer combinations produced by 26 ISJ primers. A total of 333 stable bands can be amplified from the T. aestivum ssp. tibetanum Shao and 243 bands were polymorphic, which accounted for 72.9% of the total bands. Tibetan wheat Landraces produced 316 stable bands, of which 197 bands were polymorphic. The polymorphic bands accounted for 62.34% of the total bands produced from Tibetan wheat landraces. The genetic diversity of T. aestivum ssp. tibetanum Shao was higher than that of Tibetan wheat landraces in Tibet, suggesting that T. aestivum ssp. tibetanum Shao can be used as important genetic resource for the breeding and genetic improvement of wheat in Tibet. Matrix (1, 0) was generated according to the presence or absence of the bands produced from a particular wheat accession. Clustering and principle coordinates analysis showed that T. aestivum ssp. tibetanum Shao and Tibetan wheat landraces were divided into two groups. We conclude that high polymorphisms produced by ISJ primers can reflect the genetic diversity between T. aestivum ssp. tibetanum Shao and Tibetan wheat landraces.     

Joint toxicity of three plant protection products to <Emphasis Type="Italic">Triticum aestivum</Emphasis> (L.) and <Emphasis Type="Italic">Brassica rapa</Emphasis> (L.)

Purpose  

Few studies have been conducted to evaluate the effects of mixtures of chemicals in terrestrial environment. Thus, it seems important to evaluate if the combined application of pesticides currently used in agricultural fields may pose a risk to terrestrial plants.     

<Emphasis Type="Italic">Calamintha nepeta</Emphasis> (L.) Savi and <Emphasis Type="Italic">Micromeria thymifolia</Emphasis> (Scop.) Fritsch cultivated in Italy

Calamintha nepeta and Micromeria thymifolia have been traditionally used in the Mediterranean area as condiments and medicinal plants for a long time. Whereas in parts of Italy C. nepeta (special recipes have been developed in Lazio and Tuscany) is also an established garden plant showing different evolutionary products and their interaction among each other and the wild progenitor, M. thymifolia is being developed into a new crop plant. Both plants and their uses are described with regard to Italy. There is a marked tendency to broaden the use of condiments and spices which results in new crop plants which have to be documented and elaborated in further studies. Many species of Labiatae are predisposed to use by man and new items can be found even in areas which have to be considered as well studied.     

Genetic diversity of European pear cultivars (<Emphasis Type="Italic">Pyrus communis</Emphasis> L.) and wild pear (<Emphasis Type="Italic">Pyrus pyraster</Emphasis> (L.) Burgsd.) inferred from microsatellite markers analysis

The aim of this study was to identify the group of highly polymorphic microsatellite markers for the identification of six pear cultivars (P. communis) and two individuals of wild pear (P. pyraster). From among 40 tested SSR markers, 19 were selected to profile genetic diversity in pear genotypes due to high polymorphisms. These markers showed high heterozygosity levels (0.5–1) and, on average, 6.4 alleles per marker were found. The set of microsatellite markers employed in this study demonstrated usefulness of microsatellite markers for the identification of pear genotypes. The examined wild forms were represented in this study by only two individuals of P. pyraster. It can be assumed that these forms were distinctly different from the cultivated pear cultivars.     

AFLP analysis of genetic diversity in leafy kale (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. convar. <Emphasis Type="Italic">acephala</Emphasis> (DC.) Alef.) landraces,cultivars and wild populations in Europe

AFLP markers were used to characterize diversity and asses the genetic structure among 17 accessions of kale landraces, cultivars and wild populations from Europe. The range of average gene diversity in accessions was 0.11–0.27. Several landraces showed higher levels of diversity than the wild populations and one cultivar had the lowest diversity measures. The landraces that were most genetically diverse were from areas where kales are known to be extensively grown, suggesting in situ conservation in these areas as a supplement to storage of seeds in gene banks. An analysis of molecular variance (AMOVA) showed that 62% of the total variation was found within accessions. For most accessions, genetic distance was not related to geographic distance. Similarities among accessions were probably not caused by recent gene flow since they were widely separated geographically; more likely the relationship among them is due to seed dispersal through human interactions. Our results indicate that a kale population found in a natural habitat in Denmark was probably not truly wild but most likely an escape from a cultivated Danish kale that had subsequently become naturalized.     

Genetic structure and history of Swiss maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. ssp. <Emphasis Type="Italic">mays</Emphasis>) landraces

Between 1930 and 2003 with emphasis on the 1940s maize landraces (Zea mays L. ssp. mays) from all over Switzerland were collected for maintenance and further use in a new Swiss breeding program. The genetic relationship and diversity among these accessions stored in the Swiss gene bank is largely unknown. Our hypothesis was that due to the unique geographic, climatic, and cultural diversity in Switzerland a diverse population of maize landraces had developed over the past three centuries. The aims were to characterize the genetic diversity of the Swiss landraces and their genetic relationship with accessions from neighbouring regions as well as reviewing their history, collection, and maintenance. The characterization and grouping was based on analyses with ten microsatellite markers. Geographic, cultural, and climatic conditions explained a division in two distinct groups of accessions. One group consisted of landraces collected in the southern parts of Switzerland. This group was related to the Italian Orange Flints. The other group contained accessions from northern Switzerland which were related to Northern European Flints in particular German Flints. Historic evidence was found for a frequent exchange of landraces within the country resulting in a lack of region-specific or landrace-specific genetic groups. The relatively large separation between the accessions, indicated by high F _ST (0.42), might be explained partly by a bottleneck during the collection and maintenance phase as well as by geographical and cultural separation of north and south of the country. Due to the high genetic diversity, the accessions here are a potential resource for broadening the European flint pool.     

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

Genetic diversity of barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) landraces from the central highlands of Ethiopia: comparison between the <Emphasis Type="Italic">Belg</Emphasis> and <Emphasis Type="Italic">Meher</Emphasis> growing seasons using morphological traits

In Ethiopia, barley is generally grown in two different planting seasons per year: during the long rainy season (Meher) and the short rainy season (Belg). The aim of the present study was to assess for the first time the role of this ‘two-season system’ on the structure of the genetic diversity of the Ethiopian barley landraces. We characterised 3,170 individual genotypes from 106 landrace populations using eight morphological spike traits. The diversity within population was higher in the season where barley is more important (Belg), and in general, where its cultivation is in larger plots because of weaker ‘competition’ with others crops. This indicates that barley diversity has a complex relationship with variations in the surrounding agro-ecosystem. Overall, the divergence between the two seasons was quite small (3.4%), suggesting that seed flow does not occur independently across the years within the two seasons. This would affect the amount of mutations and historic recombination that have accumulated within these populations.     

Crossability of <Emphasis Type="Italic">Vigna rhomboidea</Emphasis> Burtt. Davy with Cowpea (<Emphasis Type="Italic">V. unguiculata</Emphasis> (L.) Walp.)

V. rhomboidea is a wild Vigna species that is a potential source of genes for pubescence which could be incorporated into cultivated cowpea for insect pests resistance. Due to the lack of reliable records on the crossability and gene pool relationships between V. rhomboidea and cowpea, crossing trials were conducted in the screenhouse to observe if V. rhomboidea is reciprocally crossable (compatible) with cowpea. Crossability of V. rhomboidea (as seed parent) with cowpea (as pollen parent) was, for the first time, successfully achieved at the rate of 5.7% from 1145 crosses. Reciprocal crosses with cowpea as seed parent and V. rhomboidea pollen parent gave an average of 22.6% pod set from 2299 crosses. It is concluded that V. rhomboidea is reciprocally compatible with cowpea. This implies that V. rhomboidea belongs to the primary gene pool of cowpea.     

Comparative Study of Common Bean (<Emphasis Type="Italic"> Phaseolus vulgaris</Emphasis>L.) Landraces Conserved <Emphasis Type="Italic">ex situ</Emphasis> in Genebanks and <Emphasis Type="Italic">in situ</Emphasis> by Farmers

Genetic diversity of populations stored ex situ or in situ can be altered due to the management practices they are subjected to. In this paper, we compare populations of two common bean (Phaseolus vulgaris L.) landraces grown on farms with material collected from the same farms and now kept in two ex situ collections (CIAT and REGEN) with the purpose to monitor any changes that have occurred due to ex situ conservation. The diversity was measured using seven bean microsatellite markers. Further phenotypic and developmental traits were registered in a field experiment. Compared with the in situ populations, the ex situ ones had a lower level of gene diversity and we suggest that this is due to the regeneration process. Most of the phenotypic traits did not differ significantly between ex situ and in situ populations, although for yield and 100-seed weight, the CIAT material showed significant lower values. We assume that these populations have gone through an adaptational change. Overall, the conservation ex situ has been successful in maintaining the majority of the adaptations found in the landraces studied, however, the probable loss of genetic diversity that we have observed, suggest that protocols for the regeneration process must be carefully worked out if the majority of alleles are to be preserved for the future. This study also highlights the complementarity of ex situ and in situ conservation methods in order to preserve landrace adaptations and to capture new, useful diversity generated in in situ populations.     

A significant proportion of indigenous rhizobia from India associated with soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.) distinctly belong to <Emphasis Type="Italic">Bradyrhizobium</Emphasis> and <Emphasis Type="Italic">Ensifer</Emphasis> genera

The diversity among 269 rhizobia isolated from naturally occurring root nodules of soybean collected from two different agro-ecological regions of India, based on RFLP and sequences of the intergenic spacer (IGS) between the 16S and 23S rRNA genes, growth rate, and indole acetic acid production, revealed their significant, site-dependent genomic diversity. Among these bacteria, nine IGS genotypes were identified with two endonucleases. They were distributed into five divergent lineages by sequence analysis of each IGS representative strain, i.e., (1) comprising IGS genotypes I, II, III, and reference Bradyrhizobium yuanmingense; (2) with genotype IV and strains of unclassified bradyrhizobia genomic species; (3) including genotypes V, VI, and Bradyrhizobium liaoningense; (4) with IGS genotype VII and Bradyrhizobium elkanii strains; and (5) comprising IGS genotypes VIII, IX, and different Ensifer genus bacteria. Host-specificity test revealed that all rhizobia-nodulated soybean and cowpea and only part of them formed nodules on Arachis hypogeae and Cajanus cajan. The great diversity of soybean nodulators observed in this study emphasises that Indian soil is an important reservoir of nitrogen-fixing rhizobia.     

Genetic diversity of Ethiopian <Emphasis Type="Italic">Xanthosoma sagittifolium</Emphasis> (L.) Schott accessions assessed with AFLPs

Xanthosoma sagittifolium (L.) Schott originated from the American tropics. Domestication may have occurred in various places as this Araceae species is an important food source. It has been cultivated for many decades. In this study, Amplified Fragments Length Polymorphism (AFLP) markers were used to analyze the genetic relationships among 78 Ethiopian X. sagittifolium accessions, for conservation purpose. Cormels were collected from Bench-Maji, Kefa, Dawuro and Wolaita zones, representing four populations. The accessions belonged to either green (G) or purple (P) colored leaf and petiole accessions. Three different AFLP primer combinations resulted in 478 scorable bands, of which 99.2% were polymorphic. The mean Nei’s gene diversity (He) within populations was 0.35 while the G accessions featured higher He (0.38) than the P ones (0.35). The Nei’s gene diversity (He) at entire collection level was 0.38. The detected high genetic diversity may indicate the X. sagittifolium plants growing in the country may derive from diverse parental genotype stock elsewhere and/or there may be multiple introductions to the country. Low levels of genetic differentiation were detected among populations (Gst?=?0.07) and between the G and P accessions (Gst?=?0.02). Insignificant genetic and geographic correlation was revealed by Mantel test. Clustering analysis grouped 91% of the accessions together. Conservation and management of X. sagittifolium in the country should concentrate on maintaining high level genetic diversity within each population as well as at entire collection level through both ex situ and in situ conservation actions.     

Identification of <Emphasis Type="Italic">Aegilops</Emphasis> L. species and <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. based on chloroplast DNA

The genus Aegilops L. is a very important genetic resource for the breeding of bread wheat Triticum aestivum. Therefore, an accurate and easy identification of Aegilops species is required. Traditionally, identification of Aegilops species has relied heavily on morphological characters. These characters, however, are either not variable enough among Aegilops species or too plastic to be used for identification at the species level. Molecular markers that are more stable within species, therefore, could be the alternative strategy towards an accurate identification. Since the chloroplast DNA has a lower level of evolution compared to the nuclear genome, an attempt was made in this study to investigate polymorphism in the chloroplast DNA among 21 Aegilops species (including Ae. mutica that is now known as Amblyopyrum muticum) and between the latter and T. aestivum to generate markers for the diagnosis of all targeted species. Cleaved amplified polymorphic sequence (CAPS) applied on 22 coding and non-coding chloroplast regions using 80 endonucleases and sequencing of two of those regions revealed little polymorphism between T. aestivum and the various Aegilops species examined and to a less extent was the variation among Aegilops species. Polymorphism observed among species analysed allowed the discrimination of T. aestivum and 12 Aegilops species.     

<Emphasis Type="Italic">Rhizobium</Emphasis>,<Emphasis Type="Italic"> Bradyrhizobium</Emphasis> and<Emphasis Type="Italic"> Agrobacterium</Emphasis> strains isolated from cultivated legumes

The present study was conducted to isolate and characterize rhizobial strains from root nodules of cultivated legumes, i.e. chickpea, mungbean, pea and siratro. Preliminary characterization of these isolates was done on the basis of plant infectivity test, acetylene reduction assay, C-source utilization, phosphate solubilization, phytohormones and polysaccharide production. The plant infectivity test and acetylene reduction assay showed effective root nodule formation by all the isolates on their respective hosts, except for chickpea isolate Ca-18 that failed to infect its original host. All strains showed homology to a typical Rhizobium strain on the basis of growth pattern, C-source utilization and polysaccharide production. The strain Ca-18 was characterized by its phosphate solubilization and indole acetic acid (IAA) production. The genetic relationship of the six rhizobial strains was carried out by random amplified polymorphic DNA (RAPD) including a reference strain of Bradyrhizobium japonicum TAL-102. Analysis conducted with 60 primers discriminated between the strains of Rhizobium and Bradyrhizobium in two different clusters. One of the primers, OPB-5, yielded a unique RAPD pattern for the six strains and well discriminated the non-nodulating chickpea isolate Ca-18 from all the other nodulating rhizobial strains. Isolate Ca-18 showed the least homology of 15% and 18% with Rhizobium and Bradyrhizobium, respectively, and was probably not a (Brady)rhizobium strain. Partial 16S rRNA gene sequence analysis for MN-S, TAL-102 and Ca-18 strains showed 97% homology between MN-S and TAL-102 strains, supporting the view that they were strains of B. japonicum species. The non-infective isolate Ca-18 was 67% different from the other two strains and probably was an Agrobacterium strain.     

Putative Role of <Emphasis Type="Italic">Flavobacterium</Emphasis>, <Emphasis Type="Italic">Dokdonella</Emphasis> and <Emphasis Type="Italic">Methylophilus</Emphasis> Strains in Paracetamol Biodegradation

Paracetamol, the most widely and globally used analgesic and antipyretic, is easily accumulated in aquatic environments. In the present study, the biodegradation of paracetamol in different media (one for general growth, one specific for sulfate reducing bacteria, a mineral salts medium and municipal wastewater) inoculated with two types of sludge (from anaerobic lagoon and from oxidation ditch) under different oxygenic conditions (anoxic; moderate oxygenation in open flasks and high oxygenation by aeration) was investigated. In addition, bacteria with relative abundances increasing simultaneously with paracetamol degradation, when this drug was the only carbon source, thus with a putative role in its degradation, were identified using 16S rRNA gene sequences. The results show that aerobic microorganisms had a major role in the degradation of paracetamol, with 50 mg/L totally removed from municipal wastewater after 2 days incubation with aeration, and that the metabolites 4-aminophenol and hydroquinone plus one compound not identified in this work were produced in the process. The identification of bacteria with a role in the degradation of paracetamol revealed a strain from genus Pseudomonas with the highest final relative abundance of 21.2%, confirming previous works reporting strains of this genus as paracetamol decomposers. Besides, genera Flavobacterium, Dokdonella and Methylophilus were also in evidence, with initial relative abundances of 1.66%, 1.48 and 0.00% (not detected) in the inoculum and 6.91%, 3.80 and 3.83% after incubation, respectively. Therefore, a putative role of these genera in paracetamol biodegradation is suggested for the first time.

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Graphical Abstract ?