Genetic diversity and population structure of an Italian landrace of runner bean (Phaseolus coccineus L.): inferences for its safeguard and on-farm conservation.

The landraces are considered important sources of valuable germplasm for breeding activities to face climatic changes as well as to satisfy the requirement of new varieties for marginal areas. Runner bean (Phaseolus coccineus L.) is one of the most cultivated Phaseolus species worldwide, but few studies have been addressed to assess the genetic diversity and structure within and among landrace populations. In the present study, 20 different populations of a runner bean landrace from Central Italy named "Fagiolone," together with 41 accessions from Italy and Mesoamerica, were evaluated by using 14 nuclear SSRs to establish its genetic structure and distinctiveness. Results indicated that "Fagiolone" landrace can be considered as a dynamic evolving open-pollinated population that shows a significant level of genetic variation, mostly detected within populations, and the presence of two main genetic groups, of which one distinguished from other Italian runner bean landraces. Results highlighted also a relevant importance of farmers' management practices able to influence the genetic structure of this landrace, in particular the seed exchanges and selection, and the past introduction in cultivation of landraces/cultivars similar to seed morphology, but genetically rather far from "Fagiolone." The most suitable on-farm strategies for seed collection, conservation and multiplication will be defined based on our results, as a model for threatened populations of other allogamous crop species. STRUCTURE and phylogenetic analyses indicated that Mesoamerican accessions and Italian landraces belong to two distinct gene pools confirming the hypothesis that Europe could be considered a secondary diversification center for P. coccineus.

Phloem cytochemical modification and gene expression following the recovery of apple plants from apple proliferation disease.

Recovery of apple trees from apple proliferation was studied by combining ultrastructural, cytochemical, and gene expression analyses to possibly reveal changes linked to recovery-associated resistance. When compared with either healthy or visibly diseased plants, recovered apple trees showed abnormal callose and phloem-protein accumulation in their leaf phloem. Although cytochemical localization detected Ca(2+) ions in the phloem of all the three plant groups, Ca(2+) concentration was remarkably higher in the phloem cytosol of recovered trees. The expression patterns of five genes encoding callose synthase and of four genes encoding phloem proteins were analyzed by quantitative real-time reverse transcription-polymerase chain reaction. In comparison to both healthy and diseased plants, four of the above nine genes were remarkably up-regulated in recovered trees. As in infected apple trees, phytoplasma disappear from the crown during winter, but persist in the roots, and it is suggested that callose synthesis/deposition and phloem-protein plugging of the sieve tubes would form physical barriers preventing the recolonization of the crown during the following spring. Since callose deposition and phloem-protein aggregation are both Ca(2+)-dependent processes, the present results suggest that an inward flux of Ca(2+) across the phloem plasma membrane could act as a signal for activating defense reactions leading to recovery in phytoplasma-infected apple trees.

The inhibitory effects of coumarin on the germination of durum wheat (Triticum turgidum ssp. durum, cv. Simeto) seeds.

The event chain leading to germination blockage in durum wheat (Triticum turgidum ssp. durum Desf.) seeds exposed to the allelochemical coumarin (2H-chromen-2-one) was studied. The physiological and biochemical aspects thought to be critical for a successful seed germination were measured. At concentrations above 200 microM: , coumarin inhibited seed germination in a concentration-dependent manner. Inhibition occurred early during seed imbibition (phase I), was rapid, and irreversible. During phase I, coumarin inhibited water uptake, electrolyte retention capacity, and O(2) consumption. Later on, coumarin delayed the reactivation of peroxidases, enhanced the activity of superoxide dismutase, decreased the activities of selected marker enzymes for metabolic resumption, and repressed the transcription of molecular chaperones involved in secretory pathways. Insufficient and/or late seed rehydration caused by coumarin could have delayed membrane stabilization or decreased respiratory O(2) consumption, both of which are conducive to an overproduction of reactive O(2) species. Being unbalanced by an adequate upsurge of antioxidant defense systems, the resulting oxidative stress might have ultimately interfered with the germination program.

Cloning and characterization of wheat PDI (protein disulfide isomerase) homoeologous genes and promoter sequences.

The genomic and cDNA sequences of three PDI homoeologous genes located on chromosomes 4A, 4B and 4D of bread wheat and their promoters were cloned and sequenced. The three sequences showed a very high conservation of the coding region and of the exon/intron structure, which consisted of ten exons. The comparison of wheat sequences with those of rice and Arabidopsis showed a significant conservation of the exon/intron structure across the three species. The expression of each gene was analysed by RT-PCR in different plant tissues (roots, coleoptiles, spikelets, leaves and developing caryopses). All the genes showed a higher expression in developing caryopses than in other analysed tissues, wherein some differences were detected. The promoter sequences of the three genes possessed some regulatory motifs typical of endosperm specific expression.

Molecular characterization of gene sequences coding for protein disulfide isomerase (PDI) in durum wheat (Triticum turgidum ssp. durum).

The organisation of the durum wheat genomic sequence (3.5 kb) coding for protein disulfide isomerase (PDI), deduced by comparison between genomic fragments and cDNA sequences (1.5 kb) isolated from immature caryopses, is described. The gene structure consists of ten exons and nine introns. The presence of consensus sequences involved in splicing, such as intron-exon junctions and branchpoint, has been observed and discussed. Although the deduced wheat PDI amino acid sequence exhibited an overall identity of only 31% to that of human PDI, their modular architecture in terms of number, size, location and secondary structure-propensities of the constituent domains are remarkably similar. The comparison of the amino acid sequences with the eight available plant PDI-like sequences showed a high identity with four of them and low with the remaining ones. Analyses of transcription levels showed that the PDI mRNA was present in all analysed tissues, with much higher expression in immature caryopses.

Gliadin polymorphism in wild and cultivated einkorn wheats.

To study the relationships between different species of the Einkorn group, 408 accessions of Triticum monococcum, T. boeoticum, T. boeoticum ssp. thauodar and T. urartu were analyzed electrophoretically for their protein composition at the Gli-1 and Gli-2 loci. In all the species the range of allelic variation at the loci examined is remarkable. The gliadin patterns of T. monococcum and T. boeoticum were very similar to one another but differed substantially from those of T. urartu. Several accessions of T. boeoticum and T. monococcum were shown to share the same alleles at the Gli-1 and Gli-2 loci, confirming the recent nomenclature that considers these wheats as different subspecies of the same species, T. monococcum. The gliadin composition of T. urartu resembled that of the A genome of polyploid wheats more than did T. boeoticum or T. monococcum, supporting the hypothesis that T. urartu, rather than T. boeoticum, is the donor of the A genome in cultivated wheats. Because of their high degree of polymorphism the gliadin markers may help in selecting breeding parents from diploid wheat germ plasm collections and can be used both to search for valuable genes linked to the gliadin-coding loci and to monitor the transfer of alien genes into cultivated polyploid wheats.

Apatite deposition in polymyositis subluxing arthropathy.

We describe a 47-year-old woman who developed chronic subluxing arthropathy associated with polymyositis (PM) and positive anti-Jo1 antibodies. After a followup of 3 years, PM did not recur and polyarthritis dominated her clinical picture, leading to deformities and periarticular calcifications in her hands, shoulders, and feet. Patients with myositis, anti-Jo1 antibodies, and periarticular calcifications are at risk of developing deforming arthritis unresponsive to conventional therapy.

Storage-protein variation in wild emmer wheat (Triticum turgidum ssp.dicoccoides) from Jordan and Turkey. I. Electrophoretic characterization of genotypes.

Seed storage-protein variation at theGlu-A1,Glu-B1 andGli-B1/Glu-B3 loci in the tetraploid wild progenitor of wheat,T. dicoccoides, was studied electrophoretically in 315 individuals representing nine populations from Jordan and three from Turkey. A total of 44 different HMW-glutenin patterns were identified, resulting from the combination of 15 alleles in the A genome and 19 in the B genome. Twenty-seven new allelic variants, 12 at theGlu-A1 locus and 15 at theGlu-B1 locus, were identified by comparing the mobilities of their subunits to those previously found in bread and durum wheats. The novel variants include six alleles at theGlu-A1 locus showing both x and y subunits. The genes coding for the 1Bx and 1By subunits showed no or very little (3%) inactivity, the 1Ax gene showed a moderate degree (6.3%) of inactivity whereas the gene coding for lAy showed the highest degree of inactivity (84.8%). A high level of polymorphism was also present for the omega- and gamma-gliadins and LMW-glutenin subunits encoded by genes at the linkedGli-B1 andGlu-B3 loci (19 alleles). Some Jordanian accessions were found to contain omega-gliadin 35, gamma-gliadin 45, and LMW-2 also present in cultivated durum wheats and related to good gluten viscoelasticity. The newly-discovered alleles enhance the genetic variability available for improving the technological quality of wheats. Additionally some of them may facilitate basic research on the relationship between industrial properties and the number and functionality of HMW- and LMW-glutenin subunits.

Storage-protein variation in wild emmer (Triticum turgidum ssp.dicoccoides) from Jordan and Turkey. : II. Patterns of allele distribution.

Genetic diversity in the seed storage-proteins encoded at theGlu-A1,Glu-B1 andGli-B1/Glu-B3 loci was studied electrophoretically in 315 individuals belonging to nine populations ofT. dicoccoides from Jordan and three from Turkey. The inter- and intra-population distribution of seed storage-protein alleles at the considered loci and its link with geographical factors were investigated. Population differentiation in seed storage-proteins was in some cases very high with very weak correlations with geographic distance. Greater gene differentiation was found within and between populations which were geographically very close in Jordan than between those from Jordan and Turkey. However the distribution of alleles appeared to be non random. Samples collected from populations at locations over 900 m above sea level were less polymorphic than those collected at lower altitudes (500-700 m), whereas the relative genetic differentiation between populations was greater between those collected at higher altitudes. Seed storage-protein differentiation was significantly correlated with the altitude of the collecting sites. Although it is difficult to point out the selective pressure of altitude per se, altitude can reflect an integration of several environmental parameters. The possible adaptive value of seed storage-proteins is discussed.

Trapeziometacarpal osteoarthritis: surgical technique and results of "stabilized resection-arthroplasty".

Since 1973, 212 hands of 179 patients with disabling trapeziometacarpal osteoarthritis were treated with resection-arthroplasty stabilized by capsuloplasty and intermetacarpal ligament construction; 159 patients (180 hands) could be followed up (average, 7.8 years; range, 1 to 17.3 years). Pain relief was excellent in 89%, good in 10%, and fair in 1%. Patients were "delighted" (78%), "satisfied" (18%), "indifferent" (2%), or "disappointed" (2%) with their functional results. Palmar abduction improved 25%. The tip of the thumb reached the fifth metacarpophalangeal joint in 97% of the hands. Grip strength improved 29%; key pinch, 19%; pulp pinch, 24%. First metacarpal was stable in 94% of the cases. Scaphometacarpal space averaged 5.5 mm. There were three complete failures (2%); complications were mild and infrequent. Because of its good, predictable results, "stabilized resection-arthroplasty" is our preferred surgical treatment for osteoarthritic trapeziometacarpal joints.