Identification of common genomic regions controlling resistance to <Emphasis Type="Italic">Mycosphaerella pinodes</Emphasis>, earliness and architectural traits in different pea genetic backgrounds

In a previous study a genetic map had been developed using a RIL population derived from a cross between the Pisum sativum ssp. syriacum accession P665 and the P. sativum ssp. sativum cv. Messire. This population segregated for several agricultural important traits and was successfully used to identify QTLs (Quantitative Trait Loci) controlling resistance to Mycosphaerella pinodes and Orobanche crenata, earliness, root length and aerial biomass. However, this map contained only a few markers in common with the international pea consensus map, hampering comparison with other pea maps. The objective of this study was to incorporate a set of common transferable and reproducible markers into the P665 × Messire map to favour comparative mapping and QTL validation. Seventy-eight out of the 248 SSRs assayed resulted polymorphic in the parental lines. Thirty-eight of them, uniformly distributed all over the genome, were genotyped in the whole population and included in the map. This SSR enriched map allowed identification of six new QTLs (three for resistance to M. pinodes, two for resistance to broomrape and one for root length). Inclusion of the SSRs confirmed the homology of some of the QTLs identified in the population P665 × Messire with other QTLs associated with related traits in different pea genetic backgrounds.     

Field screening ofPisum accessions to evaluate their susceptibility to the pea weevil (Coleoptera: Bruchidae)

Summary Seventeen unreplicated field trials over nine sites and four years were used to classifyPisum germplasm (P. sativum L. &P. fulvum Sibth. & Sm) as potential sources of resistance to the pea weevil,Bruchus pisorum (L.). The emergence of adult weevils from <10% of harvested seed was used as the selection criterion to indicate possible resistance. A total of 1900Pisum accessions were assessed using the field trials and 1754 of theP. sativum accessions were eliminated. However in the 18P. fulvum accessions screened, the level of infestation by pea weevil was always below the arbitrary resistance threshold selected. This suggests thatP. fulvum accessions could be a valuable source of resistance to the pea weevil.     

Identification and characterization of sources of resistance to Erysiphe pisi Syd. in Pisum spp.

A collection of 67 accessions of Pisum species originating from different countries was screened in a glasshouse test for resistance to Erysiphe pisi. All Pisum fulvum accessions were completely resistant. Incomplete resistance was identified in some accessions of P. sativum subsp. sativum var. arvense and P. sativum subsp. elatius and abyssinicum. Microscopy revealed several distinct cellular mechanisms governing resistance. In P. fulvum, it was mainly due to a high frequency of cell death that occurred both as a rapid response to attempted infection and a delayed response that followed colony establishment. Cell death following colony establishment was also key to the incomplete resistance in some accessions of P. sativum subsp. sativum var. arvense. In addition, impaired spore germination, and to a lesser extent appressorium formation, contributed to pre‐penetration resistance in some accessions. In some cases, resistance also retarded colony growth, possibly through effects on haustorial development or function in cells that survived the attack. Thus, these wild pea accessions offer diverse resistances that could be introduced to cultivated peas to increase the efficacy of powdery mildew resistance.     

Diversity analysis and genotyping in Pisum with sequence tagged microsatellite site (STMS) primers

Pisum sativum specific sequence tagged microsatellite site primers were used to amplify genomic profiles from 15accessions of P. sativum L. that represented the genetic base of the Australian field pea-breeding program and five accessions of the wild related species P. fulvum. The STMS primers were used to assess genetic relationships among the Pisum accessions in two ways. Firstly, to produce RAPD-like multiple banding marker profiles using an adapted RAMS method, for intra- and interspecific diversity analysis. From the 14 flanking primer pairs assessed, 133 markers were obtained. Conservation and reproducibility of markers among individuals within accessions was demonstrated. The largest distance observed among P. sativumaccessions was 22% and among P.fulvum accessions was 40%, similar to that revealed with other PCR-based methods. The maximum distance between P.sativum and P. fulvum accessions was 46%. Phylogenetic clustering of P. sativum accessions, using the neighbour joining method and based on simple matching distances, was distinct and distant to P. fulvum. Secondly, PCR with a higher annealing temperature and fluorescent labeling identified simple and allelic loci markers useful for creating agenotype/fingerprint database for P. sativum cultivars. This is the first report to demonstrate the use of Pisum specific STMS sequences for both diversity analysis and genotype identification. This revised version was published online in August 2006 with corrections to the Cover Date.     

Resistance to bacterial blight (Pseudomonas syringae pv. pisi) in Spanish pea (Pisum sativum) landraces

Pea bacterial blight (Pseudomonas syringae pv. pisi) has long been known to be present in pea growing areas of Spain and to cause serious crop losses, although there is no published record of its occurrence. A collection of 16 isolates from a winter pea trial in Valladolid in 1991 which were shown in this study to be P.s. pv. pisi races 4 and 6 would appear to be the first published record of the disease in Spain. This occurrence of races 4 and 6 is the same as reported for winter-sown peas in the South of France. Ten Pisum sativum landraces from different geographical areas of Spain and considered to be representative of the traditional pea crop, were tested for resistance to seven races of P.s. pv. pisi. Seedlings of each landrace were stem inoculated with the type strain of each race in a glasshouse. Resistance exhibited by the different landraces mainly conformed to those previously described in pea cultivars indicating various combinations of the main resistance genes: R3, R2+4, R3+4 and R2+3+4. R3 was the most frequent R gene, being present in all landraces. R4 was present in four and R2 in three of the landraces tested. Variation for resistance within landraces was limited except for landrace accessions ZP-0102, ZP-0109 and ZP-0137 which also showed variation for morphological traits. The resistance responses of landrace ZP-0109 were difficult to interpret, but suggested a genetic mixture with some evidence of less well documented R genes, R5 and/or R6, and possibly some unknown resistance to race 6. This revised version was published online in July 2006 with corrections to the Cover Date.     

Screening of pea germplasm for resistance to powdery mildew

Powdery mildew caused by Erysiphe pisi DC results heavy losses in the yield and quality of pods and seeds of pea crop. Germplasm comprising 701 accessions of garden and field pea originating from 60 countries were screened for powdery mildew resistance under natural epiphytotic conditions and 64 accessions found resistant in field screening for 2?years at one location were further screened both in field at two locations and artificially in laboratory to four isolates. The information was also obtained on the amount of genetic diversity and agronomic superiority in resistant accessions. Fifty-seven accessions showed resistant reaction for 3 consecutive years in field screening but only 14 accessions originating from 10 countries showed resistant reaction in laboratory screening against the four most prevalent isolates of E. pisi collected from different places in the area of experiment. Germplasm lines showed both complete and incomplete levels of resistance and variable reactions to different isolates. There was sufficient genetic diversity and agronomic superiority in the resistant accessions e.g. EC598655, EC598878, EC598704, IC278261, and IC218988, which may serve as useful genetic material to plant breeders for breeding pea varieties for powdery mildew resistance and high yield.     

Recurrent breeding method enhances the level of blackspot (Didymella pinodes (Berk. & Blox.) Vestergr.) resistance in field pea (Pisum sativum L.) in southern Australia

Blackspot, caused by Didymella pinodes (Berk. & Blox.)Vestergr., is one of the most important diseases of field pea, causing significant reduction in seed yield and quality in southern Australia and in other parts of the world. Development of resistant germplasm has been slow because of the low level of resistance found in the available germplasm, poor reliability of screening methods and the polygenic nature of inheritance. Crosses were made between agronomically suitable lines and resistant germplasm from different sources. Their progeny were advanced through the single seed descent method and single plants were selected at F₄/F₅. The F₄/F₅ derived lines were screened against blackspot in the field under disease pressure and evaluated for grain yield at multilocations over 2 years. Despite the low level of resistance in the parental germplasm, the level of resistance has increased significantly in the new germplasm. Many of the resistant lines were late and low yielding, but lines with higher resistance and early flowering and high yield potential were also identified indicating that the disease resistance, adaptation and yield potential can be combined. However, the resistance identified in this study is only partial and suitable agronomic practices may need to be supplemented to minimise the yield loss and enhance the benefits of this partial resistance.     

Genetical analysis of resistance to Mycosphaerella pinodes in pea seedlings

Summary In studies of the inheritance of resistance, pea seedlings of seven lines in which stems and leaves were both resistant to Mycosphaerella pinodes were crossed with a line in which they were both susceptible. With seven of the crosses resistance was dominant to susceptibility. When F₂ progenies of five crosses were inoculated on either stems or leaves independently, phenotypes segregated in a ratio of 3 resistant: 1 susceptible indicating that a single dominant gene controlled resistance. F₂ progenies of one other cross gave ratios with a better fit to 9 resistant: 7 susceptible indicating that two co-dominant genes controlled resistance. The F₂ progeny of another cross segregated in complex ratios indicating multigene resistance.When resistant lines JI 97 and JI 1089 were crossed with a susceptible line and leaves and stems of each F₂ plant were inoculated, resistance phenotypes segregated independently demonstrating that leaf and stem resistance were controlled by different genes. In two experiments where the F₂ progeny of the cross JI 97×JI 1089 were tested for stem and leaf resistance separately, both characters segregated in a ratio of 15 resistant:1 susceptible indicating that these two resistant lines contain two non-allelic genes for stem resistance (designated Rmp1 and Rmp2) and two for leaf resistance (designated Rmp3 and Rmp4). Evidence that the gene for leaf resistance in JI 1089 is located in linkage group 4 of Pisum sativum is presented.     

Controlled environment assessment of partial resistance to Mycosphaerella pinodes in pea (Pisum sativum L.) seedlings

We evaluated the effect of various genotypes of pea (Pisum sativum L.) to delay the development of ascochyta blight, caused by the fungus Mycosphaerella pinodes, at the seedling stage under controlled conditions. We assessed leaflets, stipules and stems, separately. We compared genotypes on the basis of four variables assessing resistance: incubation period (IP), appearance of disease scoring 3 (DS3), disease progress rate (r_d) and area under the disease progress curve (AUDPC).For leaves, a continuous variation was observed between pea lines in disease severity at all times and disease progress curves were similar for the various genotypes. Partial resistance was defined as a delay in first symptom appearance and disease development. Resistance factors are thought to be involved in the very early stages of the interaction. Similar results were obtained for stipules and leaflets, suggesting similar mechanisms of resistance in these two organs. Stem and leaf resistances were also strongly correlated. We found that IP could discriminate genotypes but that factors associated with disease progress (DS3, AUDPC and r_d) were more informative. We suggest that further genetic analyses should be based on growth-chamber methods, with AUDPC as the variable measured. This revised version was published online in August 2006 with corrections to the Cover Date.     

Agronomical performance and breeding value of selected strains of diploid wheat,Triticum monococcum

Summary Twenty-one germplasm accessions and breeding lines of einkorn wheat (Triticum monococcum ssp. monococcum and ssp. sinskajae) were grown at two sites in Italy and evaluated for various field and seed characteristics. Grain yields of germplasm accessions were relatively high (317–3238 kg/ha), but distinctly lower than those of four modern cultivars of tetraploid (T. turgidum ssp. durum) and hexaploid wheat (T. aestivum ssp. Aestivum) included in the experiments as controls. As expected, all Einkorns-including some substantially higher yielding crossbred lines (3415–4362 kg/ha)-were defective for one or more agronomically relevant features. However, a few of the accessions examined were found to contain, as a group, practically all the genes needed to breed monococcums having the main field attributes of a modern wheat cultivar: high yielding capacity, good threshability, large kernel size, earliness, short stature and adequate lodging resistance. Still higher yielding diploid wheats, more responsive to improved growing conditions and of better seed quality, could probably be obtained from crosses with wild monococcums bearing mostly two-seeded florets and with accessions producing less slender-shaped kernels. Some of the Einkorns examined were found to carry minor genes for easy threshing which might enhance the efficacy of the major gene for soft glumes carried by T. monococcum ssp. sinskajae, a partially free-threshing diploid wheat taxon. Seed protein percentage of monococcums was markedly higher than that of durum and bread wheat cultivars even in those cases where their grain yields surpassed those of the polyploid checks. The possibilities offered by diploid wheat in the exploitation of novel endosperm mutants and F₁ hybrid vigour, as well as in the fields of celiac disease, crop diversification and resistance to agro-biological stresses are discussed. Breeding priorities and strategies are also proposed.     

Overcoming hybridization barriers between pea and some of its wild relatives

Pea would benefit from the plasticity and adaptability of its cross-incompatible relatives Pisum fulvum and Lathyrus sativus L., and we have tested reciprocal sexual crossings by manually cross-pollinating plants of genotypes of these three species. Studies of in situ germination of pollen grains on stigmata showed that pollen tubes were generally unable to germinate or could not reach the ovary. A few putative hybrid pods were nevertheless harvested, with one grain per pod germinated in vitro, then micropropagated for flow cytometry, isoenzyme, molecular (ribosomal ITS PCR-RFLP) and genomic in situ hybridization (GISH) studies. One such grain was recovered from an inter-generic cross of P. sativum x L. sativus and four from an inter-specific P. sativum x P. fulvumcross. A strong cross-incompatibility was shown between pea and grass pea, where the putative hybrid turned out to be pea. Conversely, with the interspecific, P. sativum x P. fulvum cross, flow cytometry and isoenzymes with leaf tissues strongly suggested hybridity, while molecular approaches and GISH confirmed the production of inter-specific hybrids, and without the need for a wild type P. sativum accession as a bridging cross.     

Beet mild yellowing virus resistance derived from wild and cultivated Beta germplasm

Sugar beet (Beta vulgaris ssp. vulgaris L.) has a relatively narrow genetic base and several programmes have sought to evaluate the potential for introducing novel traits from wild and cultivated Beta germplasm into the crop. In particular, resistance to important sugar beet diseases has been identified within individual Beta accessions. We report here the successful transfer of resistance to Beet mild yellowing virus (BMYV) from garden beet, fodder beet and leaf beet accessions to progeny populations in initial crosses with sugar beet. Twelve plant populations derived from different Beta accessions were inoculated with viruliferous aphids carrying BMYV and the virus content of individual plants subsequently quantified by an ELISA test. Seven populations were significantly more resistant than a control sugar beet cultivar (P ≤ 0.05). BMYV resistance was successfully inherited in BC₁ and BC₂ generations, suggesting that resistance could potentially be introgressed from these sources into elite sugar beet lines.     

Resistance in spelt wheat to yellow rust

Summary Seven spelt wheat accessions of different origin were hybridized with the susceptible bread wheat cultivar Taichung 29 in order to study the genetics of their resistance to yellow rust (Puccinia striiformis Westend. f. sp. tritici). One Iranian and five European accessions were found to carry Yr5 of Triticum aestivum ssp. spelta var. album, whereas a factor for resistance in the Iranian accession 415 was confirmed to be genetically distinct from Yr5. The alleles for resistance in each of the accessions studied showed a monogenic dominant mode of inheritance. Twenty-eight spelt wheat accessions, including those studied for their resistance to yellow rust, were subjected to polyacrylamide-gel-electrophoresis to study variation for gliadin storage protein patterns. Thirteen distinct patterns were revealed, implying the presence of duplicates within the studied spelt wheat collection.     

Resistance to late blight (Phytophthora infestans) in ten wild Solanum species

Nineteen accessions of the tuber-bearing species Solanum berthaultii, S. chacoense, S. leptophyes, S. microdontum, S. sparsipilum, S. sucrense, S. venturii, S. vernei and S. verrucosum were tested for their resistance to late blight in two years of field experiments. Plants were artifically inoculated with zoospores of race 1.2.3.4.5.7.10.11 and the development of the disease was followed. Resistance ratings, calculated as the areas under the disease progress curves (ADPC), demonstrated a high resistance in all accessions except in S. sparsipilum, S. leptophyes and their interspecific hybrid. Segregations suggest that major genes for resistance are present in S. sucrense and S. venturii, and may also play a role in S. verrucosum. It is not yet certain wether the resistance of the other accessions is comparable to the partial and durable resistance of S. tuberosum cultivars like Pimpernel, as inheritance and mechanism have yet to be established. However, segregations suggesting the presence of single major genes with complete dominance were not found in these other accessions. Tuber initiation in the field occurred in only one accession, S. tuberosum ssp. andigena, and maturity of the clones was not related to their resistance. In the other accessions maturity types could not be assessed, as the clones require short day conditions for tuber initiation.     

Characterization of resistance response of pea (Pisum spp.) against rust (Uromyces pisi)

Components of resistance to the pea rust (Uromyces pisi) were studied at the histological level in seedlings of seven pea (Pisum spp.) accessions showing partial resistance under field conditions. Resistance was characterized macroscopically by an increased latent period and a decreased infection frequency in spite of a compatible interaction (high infection type). Histological investigations revealed little differences among accessions in spore germination and appressoria formation, differences being more evident once the stomata were penetrated by the infection structures. Resistance was mainly due to a restriction of haustorium formation with significant levels of early aborted colonies in accessions IFPI3260, PI347321 and PI347347. Furthermore, a reduction in the number of haustoria per colony, hyphal tips per colony and smaller colony size were found in all genotypes studied. Resistance was not associated with early‐ or late‐acting hypersensitivity in any of the genotypes studied.     

Development of genomic simple sequence repeat markers from an enriched genomic library of grass pea (Lathyrus sativus L.)

Simple sequence repeat (SSR) or microsatellite markers are a valuable tool for several purposes such as evaluation of genetic diversity, fingerprinting, marker‐assisted selection and breeding. In this study, a SSR genomic enriched library was developed in Lathyrus sativus (grass pea) by affinity capture of restriction fragments to biotinylated microsatellite oligonucleotides. About 400 randomly selected clones were sequenced, and SSRs were present in approximately 30% of them. Clones contained 75%, 9% and 16% of simple, interrupted and compound SSRs, respectively. Of the 10 SSRs tested, 7 primer pairs produced clearly distinguishable DNA banding patterns. Successively, SSR primer pairs were successfully tested to reveal polymorphism in a set of four different grass pea germplasm accessions. The transferability of SSR markers was high among three related species of Lathyrus, namely Lathyrus cicera, Lathyrus ochrus and Lathyrus tingitanus, and the legume crop, Pisum sativum. These results indicate that the novel SSR markers are informative and will be useful and convenient for genetic analysis in grass pea and related species.     

Diversity in root architecture and response to P deficiency in seedlings of <Emphasis Type="Italic">Cucumis melo</Emphasis> L.

Breeding for more phosphorus (P)-efficient crops is one strategy to reduce the use of P fertilizers, thus mitigating the environmental and economic impacts of agriculture. Variation in root architecture and the response to P deficiency were studied in C. melo. Forty accessions representing genetic diversity within the species were screened for their root systems in normal and deficient P conditions at the seedling stage. Various parameters of P-uptake and P-use were analyzed in a subset of accessions at 40 days. Significant differences in root architecture were observed, with the taproot systems prevailing among the wild and exotic accessions, and more branched root systems in cultivated stocks. Moreover, differences in the plastic response of roots to P starvation were observed. Variation in different P-use and -uptake traits correlated with the root architecture. Within ssp. melo, the inodorus and flexuosus landraces had larger and more branched roots and more efficient P-uptake, thereby providing a close genepool for breeding. Within ssp. agrestis, conomon and momordica accessions can be sources of interest for the enhancement of variation in root architecture and P-use efficiency into cultivated melons. Therefore, the diversity observed within C. melo species could be useful in breeding P-efficient melon cultivars.     

Reactions of Hordeum spontaneum to infection with two cultures of Puccinia hordei from Israel and United States

Summary The reactions to infection with two cultures of Puccinia hordei were determined for 292 Hordeum spontaneum (syn. H. vulgare ssp. spontaneum) accessions, collected at 16 sites which encompassed the ecological range of H. spontaneum in Israel. Culture Tel-Aviv was from Israel and culture 57.19 was from the United States. Fifty-two percent of the accessions were resistant to culture Tel-Aviv and 67% were resistant to culture 57.19. Forty-three percent of the accessions were resistant to both cultures. The average infection type (IT) of accessions within sites ranged from 2.7 to 7.5 on a 0–9 rating scale. The results showed that the presence of Ornithogalum species, the alternate hosts of P. hordei, may increase the percentage of H. spontaneoum accessions resistant to P. hordei. More accessions were resistant at sites where humidity at 1400 was higher, the annual evaporation was lower, and where the glumes were shorter. Kernel weight and annual rainfall was not correlated with resistance. A lower percentage of H. spontaneum accessions were resistant to P. hordei culture Tel-Aviv from Israel than to culture 57.19 from the United States. In a previous study a lower percentage of H. spontaneum accessions also was found to be resistant to a culture of Erysiphe graminis hordei from Israel than to cultures from other countries. Previous studies also have shown that cultures of P. hordei and E. graminis hordei from Israel have many genes for virulence on barley, and that H. spontaneum accessions from Israel have many genes for resistance to these two pathogens. Previous results and the results reported in this paper support the hypothesis of coevolution of resistant host genes and virulent pathogen genes where hosts and pathogens have coexisted for many thousand years.     

Variation in tolerance to radiant frost at reproductive stages in field pea germplasm

Radiant frost is a major abiotic stress, particularly at the reproductive stage, in field pea (Pisum sativum L.) grown in Mediterranean environments. Here, response to frost was studied for flowering stage (FS) organs (buds, flowers and set pods) and pod development stage (PDS) organs (flat, swollen and mature pods) under controlled conditions, with plants exposed to a minimum temperature of –4.8°C for 4 h. This frost treatment adversely affected seed yield through (i) abortion of buds, flowers and set pods (ii) death of pods and (iii) reduction in seed size. FS organs were more sensitive to frost than PDS organs. Genetic variation was observed among 83 accessions collected from 34 countries worldwide for survival of FS buds, flowers and set pods. In 60 of 83 accessions, no buds, flowers or set pods survived the frost treatment. Five accessions: ATC 104 (origin: United Kingdom), ATC 377 (Estonia), ATC 968 (Italy), ATC 3992 (Kazakhstan) and ATC 4204 (China), showed the highest frost tolerance of FS organs and lowest numbers of abnormal seeds. The frost tolerant accessions identified in this study may be useful as parents for breeding field pea varieties that will be less likely to suffer yield loss due to radiant frost during the reproductive stage.     

Resistance to Brevicoryne brassicae in horticultural brassicas

Summary A wide range of cultivated brassica accessions including broccoli, Brussels sprouts, Chinese cabbage, cauliflower, collard, kale and swede material was tested against the cabbage aphid, Brevicoryne brassicae, at HRI, Wellesbourne in the field and laboratory in both 1992 and 1993. In the field, B. brassicae attack was assessed as the proportion of infested plants and the numbers of aphid colonies present. In the laboratory, aphid performance was measured in terms of the pre-reproductive and reproductive period, population increase, and insect mortality. Interpretation of the data was facilitated by plotting sorted accession means against normal order statistics. This statistical approach indicated the spread of variation amongst the accessions and permitted identification of extremes. Partial levels of antixenosis resistance were discovered in red brassicas. Glossy accessions of cabbage and cauliflower possessed antixenosis and antibiosis resistance that lasted throughout the season of crop growth in the field. Other accessions were shown to withstand aphid attack and therefore possessed tolerance. The laboratory studies provided information on mechanisms of antibiosis resistance. The potential value of the different sources of resistance is discussed.