Genetic relationship of diploid wheat (<Emphasis Type="Italic">Triticum</Emphasis> spp.) species assessed by SSR markers

Genetic diversity of 139 accessions of diploid Triticum species including Triticum urartu, Triticum boeoticum and Triticum monococcum was studied using 11 SSR (simple sequence repeats) markers. A total of 111 alleles with an average of 10 alleles per locus were detected. The polymorphism information content (PIC) of each SSR marker ranged from 0.30 to 0.90 with an average value of 0.62. Among the three Triticum species T. urartu had the highest number of total alleles (Na?=?81), private alleles (Npa?=?15) and showed higher genetic diversity (Hex?=?0.58; PIC?=?0.54). The genotypes from Turkey exhibited the highest genetic diversity (PIC?=?0.6), while the least diversity was observed among 4 Georgian accessions (PIC?=?0.11). Cluster analysis was able to distinguish 139 wheat accessions at the species level. The highest genetic similarity (GS) was noted between T. boeticum and T. monococcum (GS?=?0.84), and the lowest between T. urartu and T. monococcum (GS?=?0.46). The grouping pattern of the PCoA analysis corresponded with cluster analysis. No significant differences were found in clustering of T. urartu and T. monococcum accessions with respect to their geographic regions, while within T. boeoticum species, accessions from Iran were somewhat associated with their geographical origin and clustered as a close and separate group. The results from our study demonstrated that SSR markers were good enough for further genetic diversity analysis in einkorn wheat species.     

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.     

Production and cytogenetics of trigeneric hybrid involving <Emphasis Type="Italic">Triticum</Emphasis>, <Emphasis Type="Italic">Psathyrostachys</Emphasis> and <Emphasis Type="Italic">Secale</Emphasis>

Trigeneric hybrids may help establish evolutionary relationships among different genomes present in the same cellular-genetic background, and also offers the possibility to transfer different alien characters into cultivated wheat. In this study, a new trigeneric hybrid involving species from the Triticum, Psathyrostachys and Secale was synthesized by crossing wheat-P. huashanica amphiploid (PHW-SA) with wheat-S. cereale amphiploid (Zhongsi 828). The crossability of F₁ hybrid was high with 35.13%, and the fertility was 41.95%. The morphological characteristics of F₁ plants resembled the parent Zhongsi 828. The trigeneric hybrids pollen mother cells (PMCs) regularly revealed averagely 19.88 univalents, 9.63 ring bivalents, 3.97 rod bivalents, 0.60 trivalents and 0.03 tetravalents per cell. Multivalents consisted of trivalents and tetravalents can be observed in 52.7% of cells. A variation of abnormal lagging chromosome, micronuclei and chromosome bridge were formed at anaphase I and telophase II. The mean chromosomes number of F₂ progenies was 2n = 46.13, and the distribution range was 42–53. GISH results revealed that most F₂ plants had 6–12 S. cereale chromosomes, and only 0–2 P. huashanica chromosomes were detected. The results indicated that S. cereale chromosomes can be preferentially transmitted in the F₂ progenies of trigeneric hybrid than P. huashanica chromosomes. A survey of disease resistances revealed that the stripe rust resistance from the PHW-SA were completely expressed in the F₁ and some F₂ plants. The trigeneric hybrid could be a useful bridge for the transference of P. huashanica and S. cereale chromatins to common wheat.     

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.     

<Emphasis Type="Italic">Achromobacter insolitus</Emphasis> and <Emphasis Type="Italic">Zoogloea ramigera</Emphasis> associated with wheat plants (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

This study reports for the first time the presence of diazotrophic bacteria belonging to the genera Achromobacter and Zoogloea associated with wheat plants. These bacterial strains were identified by the analysis of 16S rDNA sequences. The bacterium IAC-AT-8 was identified as Azospirillum brasiliense, whereas isolates IAC-HT-11 and IAC-HT-12 were identified as Achromobacter insolitus and Zoogloea ramigera, respectively. A greenhouse experiment involving a non-sterilized soil was carried out with the aim to study the endophytic feature of these strains. After 40 days from inoculation, all the strains were in the inner of roots, but they were not detected in soil. In order to assess the location inside wheat plants, an experiment was conducted under axenic conditions. Fifteen days after inoculation, preparations of inoculated plants were observed by the scanning electron microscope, using the cryofracture technique, and by the transmission electron microscope. It was observed that all isolates were present on the external part of the roots and in the inner part at the elongation region, in cortex cells, but not in the endodermis or in the vascular bundle region. No colonizing bacterial cells were observed in wheat leaves.     

Evaluation and utilization of <Emphasis Type="Italic">Aegilops</Emphasis> and wild <Emphasis Type="Italic">Triticum</Emphasis> species for enhancing iron and zinc content in wheat

Grains of 80 accessions of nine species of wild Triticum and Aegilops along with 15 semi-dwarf cultivars of bread and durum wheat grown over 2 years at Indian Institute of Technology, Roorkee, were analyzed for grain iron and zinc content. The bread and durum cultivars had very low content and little variability for both of these micronutrients. The related non-progenitor wild species with S, U and M genomes showed up to 3–4 folds higher iron and zinc content in their grains as compared to bread and durum wheat. For confirmation, two Ae. kotschyi Boiss. accessions were analyzed after ashing and were found to have more than 30% higher grain ash content than the wheat cultivars containing more than 75% higher iron and 60% higher zinc than that of wheat. There were highly significant differences for iron and zinc contents among various cultivars and wild relatives over both the years with very high broad sense heritability. There was a significantly high positive correlation between flag leaf iron and grain iron (r = 0.82) and flag leaf zinc and grain zinc (r = 0.92) content of the selected donors suggesting that the leaf analysis could be used for early selection for high iron and zinc content. ‘Chinese Spring’ (Ph ^I ) was used for inducing homoeologous chromosome pairing between Aegilops and wheat genomes and transferring these useful traits from the wild species to the elite wheat cultivars. A majority of the interspecific hybrids had higher leaf iron and zinc content than their wheat parents and equivalent or higher content than their Aegilops parents suggesting that the parental Aegilops donors possess a more efficient system for uptake and translocation of the micronutrients which could ultimately be utilized for wheat grain biofortification. Partially fertile to sterile BC₁ derivatives with variable chromosomes of Aegilops species had also higher leaf iron and zinc content confirming the possibility of transfer of required variability. Some of the fertile BC₁F₃ and BC₂F₂ derivatives had as high grain ash and grain ash iron and zinc content as that of the donor Aegilops parent. Further work on backcrossing, selfing, selection of fertile derivatives, leaf and grain analyses for iron and zinc for developing biofortified bread and durum wheat cultivars is in progress. Nidhi Rawat, Vijay K. Tiwari, and Neelam Singh have contributed equally to the work.     

Gliadins of Bulgarian durum wheat (<Emphasis Type="Italic">Triticum durum</Emphasis> Desf.) landraces: genetic diversity and geographical distribution

The polymorphism of gliadins was studied in 98 Bulgarian durum wheat (Triticum durum Desf.) landraces and classified according to the existing catalogues of blocks of gliadin components. In total, 31 alleles, including 12 new ones, were revealed for five gliadin-coding loci. Nine allele families, which included several alleles coding similar blocks differed only by minor components, were found. The gliadin loci had a high genetic diversity (H = 0.70), and Gli-A2 ^d was the most polymorphic locus. Significant differences in allele distribution were observed through the Bulgarian region under study. The results made it possible to explain the distribution by historical factors. Presumably, the genetic material flew into the country via two different ways and different durum wheat subgroups contributed to the formation of Bulgarian landraces. The landraces were a result of long-term selection and, probably, had a close association with the history of the human populations of the region.     

Variation in Wheat (<Emphasis Type="Italic">Triticum</Emphasis> spp.) Landraces from Different Altitudes of Three Regions of Turkey

Wheat landrace populations, collected from different altitudes of three regions of Turkey, were evaluated for variation within and among populations. Experimental material consisted of 380 accessions, from nine provinces, in Central Anatolian Plateau and North Transitions. The material was grown at Haymana-Ankara Research Farm of Central Research Institute for Field Crops in 1999–2000 planting season, in a three-meter, two-row-plot trial with three bread and three durum wheat checks. Populations were evaluated for plant height, tillers number, spike length, grains per spike, awn length, thousand kernels weight (TKW), winter survival, and days to flowering. Observations and measurements were performed on five randomly selected plants in each plots. Mean, coefficient of variation (CV), and range were computed for populations from five altitude intervals of 0–399, 400–799, 800–1199, 1200–1599, and 1600–1999 m, and geographical regions. Correlation, principal component (CPC) and cluster analyses were performed later. The highest variation was recorded for awn length and the lowest for number of days to flowering. First three principal components (PCs) accounted for 60.69% of the total variation. Cluster analysis for bread wheat, durum wheat, cultivated einkorn and cultivated emmer grouped the species meaningfully.     

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.     

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.     

Development of new sources of tetraploid <Emphasis Type="Italic">Arachis</Emphasis> to broaden the genetic base of cultivated groundnut (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.)

Groundnut, an important crop of many countries of the world, is susceptible to a range of diseases and pests. High levels of resistances are not available in the cultivated gene pool as the crop is said to have a narrow genetic base. Narrow genetic base is attributed to the evolution of the crop which took place by the combination of A and B genome species, and later doubling their chromosome number, giving rise to tetraploid cultivated groundnut. Direct utilization of cross-compatible wild relatives, which are diploids, to broaden the genetic base and introduction of useful traits, is not a straight-forward process due to ploidy differences between the cultivated species and wild relatives. Hence amphiploids and autotetraploids were created by not only combining the putative genomes, but many other A and B genome species, thus producing a highly variable population of tetraploid groundnuts also called new sources of Arachis hypogaea. This study describes the development and characterization of newly generated tetraploid groundnuts and the level of molecular diversity as assessed by DArT markers.     

Complete chloroplast DNA sequences of Georgian indigenous polyploid wheats (<Emphasis Type="Italic">Triticum</Emphasis> spp.) and B plasmon evolution

Three types of plasmon (A, B and G) are present for genus Triticum. Plasmon B is detected in polyploid species - Triticum turgidum L. and Triticum aestivum L. By now, 21 complete sequences of chloroplast DNA of the genus Triticum is published by different authors. Many inaccuracies can be detected in the sequenced chloroplast DNAs. Therefore, we found it necessary to study of plasmon B evolution to use only those sequences obtained by our method in our laboratory. Complete nucleotide sequences of chloroplast DNA of 11 representatives of Georgian wheat polyploid species were determined. Chloroplast DNA sequencing was performed on an Illumina MiSeq platform. Chloroplast DNA molecules were assembled using the SOAPdenovo computer program. Using T. aestivum L. subsp. macha var. palaeocolchicum as a reference, 5 SNPs were identified in chloroplast DNA of Georgian indigenous polyploid wheats. 38 and 56 bp inversions were observed in paleocolchicum subspecies. The phylogeny tree shows that subspecies macha, durum, carthlicum and palaeocolchicum occupy different positions. According the simplified scheme based on SNP and indel data the ancestral, female parent of all studied polyploid wheats is an unknown X predecesor, from which four lines were formed.     

Untapped amaranth (<Emphasis Type="Italic">Amaranthus </Emphasis>spp.) genetic diversity with potential for nutritional enhancement

Significant genetic diversity was observed in 218 out of a total of 1309 accessions of amaranth (Amaranthus hypochondriacus L.) and its seven wild relatives, A. spinosus L., A. dubius Mart. ex Thell., A. hybridus L., A. tricolor L., A. cruentus L., A. caudatus L., A. retroflexus L. for 24 nutritional parameters including total oil content, fatty acid profile, total protein content and amino acid profile. Diversity for total oil content (6.42–12.53%), linoleic acid (25.68–54.34%), oleic acid (21.97–42.01%) of the total fatty acids, total protein content (7.84–18.01%), among important essential amino acids; lysine content (0.66–11.12 g/16 g N), methionine (0.35–4.80 g/16 g N) and half cystine and (0.12–8.32 g/16 g N) was reported. The un-weighted pair-group method using arithmetic average cluster analysis based on pair wise Euclidean genetic distance grouped the accessions into seven major clusters. Histidine, half cystine, tyrosine, essential amino acids, total oil content, linoleic acid and oleic acid content were the major parameters contributing significantly to genetic diversity. Present findings indicate that significant diversity exists for nutritional parameters in amaranth germplasm. The promising accessions with higher multiple nutritive traits; protein content (>16%), oil content (>11%), lysine content (>7.5 g/16 g N) and EAA higher than the FAO reported values, were identified. This is the first report on detailed nutritional analysis of diversity collected worldwide. These could be used as potential breeding material for nutritional enhancement through genetic improvement. This will help in overcoming the “triple burden” of malnourishment, hidden hunger, and obesity.     

Six new botanical varieties of <Emphasis Type="Italic">Triticum</Emphasis> from Oman

Due to its geographic position on the northeastern tip of the Arabian Peninsula and its sea trade relationships with Asia, East Africa and the Middle East, Oman has for millennia been at the cross-roads of inter-regional exchange of cultivated plants. This is reflected in recent findings of new cultivars of banana (Musa spp.) and wheat (Triticum spp.) in remote oases of the Hajar Mountains in northern Oman. Material collected in 2003 and 2004 contained six new botanical varieties of wheat which are described here. One of them belongs to the tetraploid T. aethiopicum, the others are hexaploid.     

<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.

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 ?

     

Genetic diversity of HMW glutenin subunits in diploid,tetraploid and hexaploid <Emphasis Type="Italic">Triticum</Emphasis> species

The genetic variations of high-molecular-weight (HMW) glutenin subunits in 1051 accessions of 13 Triticum subspecies were investigated using sodium dodecyl sulfate polyacrylamide-gel electrophoresis. A total of 37 alleles were detected, resulting in 117 different allele combinations, among which 20, 68 and 29 combinations were observed in diploid, tetraploid and hexaploid wheats, respectively. Abundance and frequency of allele and combinations in tetraploid wheats were higher than these in hexaploid wheats. Allele Glu-A1c was the most frequent subunit at Glu-A1 locus in tetraploid and hexaploid wheats. Consequently, the results also suggested that the higher variations occurred at Glu-B1 locus compared to Glu-A1 and Glu-D1. Therefore, carthlicum wheat possessing the allele 1Ay could be presumed a special evolutional approach distinguished from other tetraploid species. Furthermore, this provides a convenient approach of induction of the 1Ay to common wheat through direct cross with carthlicum wheat. Alleles Glu-B1c and Glu-B1i generally absent in tetraploid wheats were also found in tetraploid wheats. Our results implied that tetraploid and hexaploid wheats were distinguished in dendrogram, whereas carthlicum and spelta wheats and however displayed the unique performance. In addition, founder effect, no-randomness of diploidization, mutation and artificial selection could cause allele distribution of HMW-GS in Triticum. All alleles of HMW-GS in Triticum could be further utilized through hybrid in the quality improvement of common wheat.     

<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.     

Exploitation of <Emphasis Type="Italic">Malus</Emphasis> EST-SSRs and the utility in evaluation of genetic diversity in <Emphasis Type="Italic">Malus</Emphasis> and <Emphasis Type="Italic">Pyrus</Emphasis>

A total of 8117 suitable SSR-contaning ESTs were acquired by screening from a Malus EST database, among which dinudeotide SSRs were the most abundant repeat motif, within which, CT/TC followed by AG/GA were predominant. Based on the suitable sequences, we developed 147 SSR primer pairs, of which 94 pairs gave amplifications within the expected size range while 65 pairs were found to be polymorphic after a preliminary test. Eighteen primer pairs selected randomly were further used to assess genetic relationship among 20 Malus species or cultivars. As a result, these primers displayed high level of polymorphism with a mean of 6.94 alleles per locus and UPGMA cluster analysis grouped twenty Malus accessions into five groups at the similarity level of 0.6800 that were largely congruent to the traditional taxonomy. Subsequently, all of the 94 primer pairs were tested on four accessions of Pyrus to evaluate the transferability of the markers, and 40 of 72 functional SSRs produced polymorphic amplicons from which 8 SSR loci selected randomly were employed to analyze genetic diversity and relationship among a collection of Pyrus. The 8 primer pairs produced expected bands with the similar size in apples with an average of 7.375 alleles per locus. The observed heterozygosity of different loci ranged from 0.29 (MES₉₆) to 0.83 (MES₁₃₈), with a mean of 0.55 which is lower than 0.63 reported in genome-derived SSR marker analysis in Pyrus. The UPGMA dendrogram was similar to the previous results obtained by using RAPD and AFLP markers. Our results showed that these EST-SSR markers displayed reliable amplification and considerable polymorphism in both Malus and Pyrus, and will contribute to the knowledge of genetic study of Malus and genetically closed genera.     

<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.