Spatial genetic structure in wild cardoon, the ancestor of cultivated globe artichoke: Limited gene flow, fragmentation and population history.

Nuclear and chloroplast markers and phenotypic characters were integrated to analyse the population genetic structure of wild cardoon, Cynara cardunculus var. sylvestris, the ancestor of cultivated globe artichoke, Cynara cardunculus var. scolymus on the island of Sardinia, Italy. The spatial scale ranged from a few metres to ∼200km. Wild cardoon appears to be genetically fragmented, with significant genetic divergence at various scales, indicating that gene flow is insufficient to counterbalance the effects of genetic drift or founder effects. Divergence between populations was higher for chloroplast (40%) than for nuclear markers (15%), suggesting that gene flow via seed was lower than via pollen. Two main genetic groups were detected; these correlated with differences in flowering time, capitula size, glossiness, and anthocyanin pigmentation. A complex population structure of wild cardoon emerged over small spatial scales, likely resulting from the interplay between gene dispersal, colonisation history and selective forces. Indeed, Sardinia appears to be a 'hybrid zone' of different gene pools. The island has unique diverse germplasm that has originated from hybridisation among different gene pools. The sampling of seeds from a few plants but from many sites is suggested as the best strategy to harvest the genetic diversity of wild cardoon.

The effect of slope on repeated sprint ability in young soccer players.

This study aimed to describe a gradient repeated sprint ability (RSA) test in comparison with a standard level one by investigating performance, metabolic demand and muscular jumping performance as a proxy for running mechanics. Eighteen athletes performed two level RSA tests (40 m × 6) - for reliability evaluation - and one ±5% gradient RSA test, second leg downhill (RSAgrad). Rating of perceived exertion (RPE), blood lactate concentration (BLa) concentration, vertical jump heights were assessed as well. Level test measures resulted highly reliable (Intra-class correlation coefficient (ICC) ≥0.96). RSAgrad worsened only first sprints' performance (-2%) but not overall test performance (~45 s). RSAgrad resulted to be less deteriorating in terms of fatigue index (FI) (-36%), BLa (-23%), RPE (-11%), jumping performance (RSAgrad post-/pre-squat jump, countermovement jump heights (CMJh): -3%, -6%, respectively). RSAgrad could be used to diversify common training protocol without stressing excessively athletes' current metabolic-anaerobic capacity. Such physical conditioning procedures could improve acceleration/braking capability.

The Impact of Jumping during Recovery on Repeated Sprint Ability in Young Soccer Players.

This study compared the effect of counter-movement-jump (CMJ)-based recovery on repeated-sprint-ability (RSA). Eighteen male footballers (16 ± 0 years, 65 ± 10 kg, 1.74 ± 0.10 m) performed three RSA-tests. RSA-1/-3 were performed according to standard procedures, while three CMJs (over 10″) - as a potential fatigue-determinant and/or running mechanics interference--were administered during RSA-2 recoveries. RSA performance, exercise effort (fatigue index [FI], rating of perceived exertion [RPE], blood lactate concentration [BLa]), simple kinematics (steps number), vertical-jump characteristics (stretch-shortening-cycle-efficiency [SSCE] assessed before/after RSA) were investigated. ANOVA showed no differences between RSA-1,-3. During RSA-2, performance was lower than RSA-1/-3, while steps number did not change. During RSA-2, FI, BLa, RPE were higher than RSA-1/-3 (FI +21.10/+20.43%, P<0.05; BLa +16.25/+13.34%, P<0.05; RPE +12.50/+9.57%, P<0.05). During RSA-2, SSCE, as the CMJ/squat-jump-height-ratio, was not significantly different from RSA-1/-3. Passive recovery RSA allows better performance. Yet, RSA CMJ-based recovery is effective in increasing training load (FI, BLa, RPE) without perturbing running mechanics (simple kinematics, SSCE).

Improving neuromuscular performance in young basketball players: plyometric vs. technique training.

AIM: The aim of this study was to compare the effects of plyometric training versus basketball technique training on improving neuro-muscular performance. METHODS: Thirty-six (age 14.9±0.9 years, body height 164.0±7.6 cm, body weight 54.0±8.7 kg, BMI 20.1±2.4 kg·m-2) basketball players girls were randomly allocated to 2 groups: Basketball Plyometric Training (BPT, N.=18) and Basketball Technique Training (BTT, N.=18). The players were tested by two specific tests: counter movement jump (CMJ) and squat jump (SJ) before and after 6 training weeks. RESULTS: The jump height, as dependent variable, showed a different trend as an effect of the different training protocols, in contrast with the current knowledge. Manova did not show significant interactions between the two groups for the height of jumps, while significant differences were found for interaction time × training (P<0.05) and for main effect × time (P<0.001). After training, the BPT group increased significantly CMJ performance by 11.3% (P<0.05), whereas the BTT group increased by 4.6%. Likewise, the BPT group demonstrated a significant greater improvement of jump height than BTT group (an increase of 15.4% vs. 7.5%, P<0.01; respectively). CONCLUSION: These results suggest that both training protocols proposed in this study improved vertical jump performance. However, a combination of the two protocols, plyometric training and sport-specific-exercises, could be useful to optimize performance by an easy transition from controlled a-specific to sport-specific performance requirements. In conclusion, BPT is a safe and effective method of achieving a favourable neuro-muscular performance than BTT in female basketball players.

Sprint vs. intermittent training in young female basketball players.

AIM: This study aimed at comparing the effects of intermittent and repeated sprint ability training on physiological variables. METHODS: Sixteen young female basketball players were randomly allocated to intermittent training (IT=8) or repeated sprint ability training (RST=8) groups. The following outcomes were measured at baseline and after 6 weeks of training: Yo-Yo intermittent recovery (Yo-Yo) and repeated sprint ability (RSA) tests. RESULTS: For all the variables investigated the effect of training type showed a different trend respect at current knowledge. In the RSA, best time (BT) was a significant main effect of training time (pre- vs. post-) (P<0.0001), and of the interaction training type/time (P=0.03). The RST showed a decrease in BT of 3.1% (P=0.005) while the IT showed a decrease of 6.2% (P<0.0001). In the IT there was a significant main effect of time for the total distance with an increment of 26.9%, and a significant main effect of time in the final speed with an increment of 1.23%. CONCLUSION: These findings suggest that the two training methods used in this study can be an effective training strategy for inducing anaerobic and basketball-specific training schedules. Besides, even when IT training is not done at very high speed, it can increase the maximum speed of the RSA.

The acute effect of whole body vibration on repeated shuttle-running in young soccer players.

The aim of this study was to investigate the acute effects of whole-body vibration (WBV) on Repeated Sprint Ability (RSA). Seventeen male soccer players (16.71±0.47 y) performed three RSA tests (Randomized crossover study design). The second RSA test was done with WBV (RSA2) to assess the effect of WBV. The studied variables were: best time (BT), worst time (WT), total time (TT), the fatigue index (FI) of RSA, and post-test blood lactate (BLa). ANOVA with repeated measures showed no differences between RSA1 and RSA3, while there were significant differences in all variables studied. TT= [RSA2 0.93% and 1.68% lower than RSA1 and RSA3 respectively; p<0.05], BLa= [RSA2 16.97% and 14.73% greater than RSA1 and RSA3 respectively; p<0.001], WT= [RSA2 1.90% and 2.93% lower than RSA1 and RSA3 respectively; p<0.01], and FI = [RSA2 30.64% and 40.15% lower than RSA1 and RSA3 respectively; p<0.0001]. When comparing individual sprints, WBV showed a significant effect at the 5th sprint: RSA2 2.29% and 2.95% lower than RSA1 and RSA3 respectively (p<0.005), while at the 6th sprint: RSA2 2.75% and 4.09% lower than RSA1 and RSA3 respectively; p<0.005. In conclusion, when applying WBV during the recovery periods of Repeated Sprint Ability efforts, most of the performance variables improved.

Footstep manipulation during uphill running.

The present study investigated the effects of step frequency manipulation during training on slopes (2%) on biomechanical parameters at Iso-Efficiency Speed (without increasing the metabolic demand). 24 male marathon runners were randomly allocated to one of 2 training groups for 3 weeks: step frequency manipulation group (SFM, n=12) and free step frequency group (SFF, n=12). Lower limb kinematic parameters were measured before and after the 3 weeks training. The SFM group increased step length 4.30% (p<0.001), flight time 29.48% (p<0.001) and decreased contact time 14% (p<0.01). These findings coincide with characteristics of better running performances. The SFF group did not elicit such results. The results from the study could help coaches to devise training methods which could improve an athlete's performance through increasing step length. The method provided may aid faster race times for athletes.

Chloroplast Microsatellite Diversity in Phaseolus vulgaris.

Evolutionary studies that are aimed at defining the processes behind the present level and organization of crop genetic diversity represent the fundamental bases for biodiversity conservation and use. A Mesoamerican origin of the common bean Phaseolus vulgaris was recently suggested through analysis of nucleotide polymorphism at the nuclear level. Here, we have used chloroplast microsatellites to investigate the origin of the common bean, on the basis of the specific characteristics of these markers (no recombination, haploid genome, uniparental inheritance), to validate these recent findings. Indeed, comparisons of the results obtained through analysis of nuclear and cytoplasmic DNA should allow the resolution of some of the contrasting information available on the evolutionary processes. The main outcomes of the present study are: (i) confirmation at the chloroplast level of the results obtained through nuclear data, further supporting the Mesoamerican origin of P. vulgaris, with central Mexico representing the cradle of its diversity; (ii) identification of a putative ancestral plastidial genome, which is characteristic of a group of accessions distributed from central Mexico to Peru, but which have not been highlighted beforehand through analyses at the nuclear level. Finally, the present study suggests that when a single species is analyzed, there is the need to take into account the complexity of the relationships between P. vulgaris and its closely related and partially intercrossable species P. coccineus and P. dumosus. Thus, the present study stresses the importance for the investigation of the speciation processes of these taxa through comparisons of both plastidial and nuclear variability. This knowledge will be fundamental not only from an evolutionary point of view, but also to put P. coccineus and P. dumosus germplasm to better use as a source of useful diversity for P. vulgaris breeding.

Nucleotide diversity of a genomic sequence similar to SHATTERPROOF (PvSHP1) in domesticated and wild common bean (Phaseolus vulgaris L.).

Evolutionary studies in plant and animal breeding are aimed at understanding the structure and organization of genetic variations of species. We have identified and characterized a genomic sequence in Phaseolus vulgaris of 1,200 bp (PvSHP1) that is homologous to SHATTERPROOF-1 (SHP1), a gene involved in control of fruit shattering in Arabidopsis thaliana. The PvSHP1 fragment was mapped to chromosome Pv06 in P. vulgaris and is linked to the flower and seed color gene V. Amplification of the PvSHP1 sequence from the most agronomically important legume species showed a high degree of interspecies diversity in the introns within the Phaseoleae, while the coding region was conserved across distant taxa. Sequencing of the PvSHP1 sequence in a sample of 91 wild and domesticated genotypes that span the geographic distribution of this species in the centers of origin showed that PvSHP1 is highly polymorphic and, therefore, particularly useful to further investigate the origin and domestication history of P. vulgaris. Our data confirm the gene pool structure seen in P. vulgaris along with independent domestication processes in the Andes and Mesoamerica; they provide additional evidence for a single domestication event in Mesoamerica. Moreover, our results support the Mesoamerican origin of this species. Finally, we have developed three indel-spanning markers that will be very useful for bean germplasm characterization, and particularly to trace the distribution of the domesticated Andean and Mesoamerican gene pools.

Genetic diversity and structure of a worldwide collection of Phaseolus coccineus L.

Phaseolus coccineus L. is closely related to P. vulgaris and is the third most important cultivated Phaseolus species. Little is known about the patterns of its diversity. In this work, a representative collection of its worldwide diversity was initially developed. The collection includes 28 wild forms (WFs) and 52 landraces (LRs) from Mesoamerica (the crop domestication area), and 148 LRs from Europe (where the crop was introduced in the sixteenth century). The collection was studied by using 12 SSR molecular markers that were developed for the P. vulgaris genome. They were proved to be effective and reliable in P. coccineus in this work. Fourteen LRs of P. dumosus (previously identified as a subspecies of P. coccineus) were also studied. The genetic diversity, population structure and phylogenetic relationships were investigated. The results indicate that: (a) the European and Mesoamerican gene pools are clearly differentiated, (b) a certain reduction of diversity occurred with introduction into Europe, and (c) the Mesoamerican LRs (P. dumosus included) and WFs are closely related and are connected by a high gene flow. Inferences on the domestication process of P. coccineus are also presented. This study provides a picture of the genetic diversity distribution and outcomes with introduction into the Old World, which was not available before. It also underlines that the genetic diversity of both WFs and LRs is an important source for Phaseolus spp. breeding programs and deserves to be preserved in situ and ex situ.

Beans in Europe: origin and structure of the European landraces of Phaseolus vulgaris L.

This study focuses on the expansion of Phaseolus vulgaris in Europe. The pathways of distribution of beans into and across Europe were very complex, with several introductions from the New World that were combined with direct exchanges between European and other Mediterranean countries. We have analyzed here six chloroplast microsatellite (cpSSR) loci and two unlinked nuclear loci (for phaseolin types and Pv-shatterproof1). We have assessed the genetic structure and level of diversity of a large collection of European landraces of P. vulgaris (307) in comparison to 94 genotypes from the Americas that are representative of the Andean and Mesoamerican gene pools. First, we show that most of the European common bean landraces (67%) are of Andean origin, and that there are no strong differences across European regions for the proportions of the Andean and Mesoamerican gene pools. Moreover, cytoplasmic diversity is evenly distributed across European regions. Secondly, the cytoplasmic bottleneck that was due to the introduction of P. vulgaris into the Old World was very weak or nearly absent. This is in contrast to evidence from nuclear analyses that have suggested a bottleneck of greater intensity. Finally, we estimate that a relatively high proportion of the European bean germplasm (about 44%) was derived from hybridization between the Andean and Mesoamerican gene pools. Moreover, although hybrids are present everywhere in Europe, they show an uneven distribution, with high frequencies in central Europe, and low frequencies in Spain and Italy. On the basis of these data, we suggest that the entire European continent and not only some of the countries therein can be regarded as a secondary diversification center for P. vulgaris. Finally, we outline the relevance of these inter-gene pool hybrids for plant breeding.

Development and use of chloroplast microsatellites in Phaseolus spp. and other legumes.

Chloroplast microsatellites (cpSSRs) provide a powerful tool to study the genetic variation and evolution of plants. We have investigated the usefulness of 39 primer pairs tagging cpSSR loci on a set of eight different genera of Leguminosae (Papilionoideae subfamily) and five species belonging to the genus Phaseolus. Thirty-six 'universal' primer pairs were retrieved from the literature, one was re-designed and a further two were designed de novo. The cpSSR loci analysed were highly polymorphic across the individuals examined. Twenty-seven primer pairs were polymorphic in the overall sample, 18 within Phaseolus, and 16 in both P. vulgaris and P. coccineus. Analysis of the plastome sequences of four Leguminosae species (obtained from GenBank) showed that in the loci targeted by universal primer pairs: (i) the originally tagged cpSSRs can be lost; (ii) other cpSSRs can be present; and (iii) polymorphism arises not only from differences in the numbers of cpSSR repeats, but often from other insertion/deletion events. Multilocus linkage disequilibrium analysis suggests that homoplasy is not a major problem in our dataset, and principal component analysis indicates intelligible relationships among the species considered. Our study demonstrates that this set of chloroplast markers provides a useful tool to study the diversity and the evolution of several legumes, and particularly P. vulgaris and P. coccineus.

Phylogeny and evolution of mating-type genes from Pyrenophora teres, the causal agent of barley "net blotch" disease.

The main aim of this study was to test the patterns of sequence divergence and haplotype structure at the MAT locus of Pyrenophora teres, the causal agent of barley 'net blotch' disease. P. teres is a heterothallic ascomycete that co-occurs in two symptomatological forms, the net form (NF) and the spot form (SF). The mating-type genes MAT1-1-1 and MAT1-2-1 were sequenced from 22 NF isolates (12 MAT1-1-1 and 10 MAT1-2-1 sequences) and 17 SF isolates (10 MAT1-1-1 and seven MAT1-2-1 sequences) collected from Sardinian barley landrace populations and worldwide. On the basis of a parsimony network analysis, the two forms of P. teres are phylogenetically separated. More than 85% of the total nucleotide variation was found between formae speciales. The two forms do not share any polymorphisms. Six diagnostic nucleotide polymorphisms were found in the MAT1-1-1 intron (1) and in the MAT1-1-1 (3) and MAT1-2-1 (2) exons. Three diagnostic non-synonymous mutations were found, one in MAT1-1-1 and two in MAT1-2-1. For comparison with P. teres sequence data, the mating-type genes from Pyrenophora graminea were also isolated and sequenced. Divergence between P. graminea and P. teres is of a similar magnitude to that between NF and SF of P. teres. The MAT genes of P. graminea were closer to those of SF than to NF, with the MAT1-2-1 SF peptide not different from the MAT1-2-1 peptide of P. graminea. Overall, these data suggest long genetic isolation between the two forms of P. teres and that hybridization is rare or absent under field conditions, with each form having some particular niche specialization. This indicates that research on resistance to P. teres should consider the two forms separately, as different species.

Population genetic structure of Pyrenophora teres Drechs. the causal agent of net blotch in Sardinian landraces of barley (Hordeum vulgare L.).

Monoconidial cultures of Pyrenophora teres, the causal agent of barley net blotch, were isolated from leaves collected from six populations of the barley landrace "S'orgiu sardu" growing in five agro-ecological areas of Sardinia, Italy, and genotyped using AFLPs. The 150 isolates were from lesions of either the "net form" (P. teres f. sp. teres) or the "spot form" (P. teres f. sp. maculata) of the disease. Of 121 AFLP markers, 42%, were polymorphic. Cluster analysis resolved the isolates into two strongly divergent groups (F(ST) = 0.79), corresponding to the net (45% of the isolates) and the spot (55% of the isolates) forms (designated the NFR and SFR groups, respectively). The absence of intermediate genotypes and the low number of shared markers between the two groups indicated that hybridization between the two formae is rare or absent under the field condition of Sardinia. Five of the barley populations hosted both forms but in different proportions. The SFR populations were similar in overall polymorphism to the NFR populations. However, compared to the SFR form, the NFR occurred in all fields sampled and showed a higher population divergence (F(ST) = 0.43 versus F(ST) = 0.09 with all isolates; F(ST) = 0.37 versus F(ST) = 0.06 with clone corrected samples) probably due to a lower migration rate. AFLP fingerprints resolved 117 distinct genotypes among the 150 isolates sampled (78%), 87% in SFR and 68% in NFR isolates. Although the absolute numbers may be a function of the number of AFLP markers assayed, the relative difference suggests that clonality is more prevalent among the NFR isolates (with 11 of 46 haplotypes observed more than once), compared with SFR isolates (7 of 71 haplotypes). Both digenic and multilocus linkage disequilibrium analyses suggested that sexual reproduction occurs at significant levels within the NFR and SFR populations, and that the relative contribution of sexual and asexual reproduction varies among different environments.