Collection and evaluation of grasspea (Lathyrus sativus L.) germplasm of central Italy

Characterization of 16 grasspea (Lathyrus sativus L.) accessions collected in central Italy (Marche region) was performed for morphological and agronomic traits and for 3‐(‐N‐oxalyl)‐L‐2,3 diaminoprop‐ionic acid (ODAP) content. Field trials, carried out in two locations, revealed high among‐population genetic variation. In particular, an evident differentiation between commercial populations, cultivated by market‐oriented farms, and household populations (not market‐oriented) was found. Genotype × environment interaction was negligible. Medium‐high values of ODAP content were observed following colorimetric and capillary electrophoresis analyses. A high positive correlation between the two methods was found (r = 0.83**), but the colorimetric values showed, on average, significant 14% lower ODAP values. This research represents a pre‐breeding activity aimed at developing a breeding programme for the preservation and utilization of Italian grasspea germplasm within sustainable or organic agricultural systems.     

The Gene Pools of the Grasspea (Lathyrus sativus L.)

Interspecific hybridization between the grasspea, Lathyrus sativus, and 15 wild species in Section Lathyrus is reported. Only two species, L. amphicarpos and L. cicera produced viable F¹ hybrids with low fertility when crossed with L. sativus as male parent. Crosses with six other species produced pods following pollination when L. sativus was the male, but seedlings were inviable, seeds did not germinate or pods were empty or had totally shrivelled seeds. When L. sativus was the female parent, only one cross with L. gorgoni produced an F¹, but the seedling was inviable. The germplasm resources of the grasspea are identified, with L. amphicarpos and L. cicera placed in the secondary gene pool and the other species in the tertiary gene pool. The definition of these germplasm resources is discussed in terms of grasspea improvement through plant breeding.     

Inheritance of neurotoxin (ODAP) content, flower and seed coat colour in grass pea (Lathyrus sativus L.)

Summary A strong epidemiological association is known to exist between the consumption of grass pea and lathyrism. A neurotoxin, -N-Oxalyl-L-, -diaminopropanoic acid (ODAP) has been identified as the causative principle. This study was undertaken to investigate the mode of inheritance of the neurotoxin ODAP, flower and seed coat colour in grass pea. Five grass pea lines with low to high ODAP concentration were inter-crossed in all possible combinations to study the inheritance of the neurotoxin. Parents, F₁ and F₂ progenies were evaluated under field condition and ODAP analyzed by an ortho-phthalaldehyde spectrophotometric method. Many of the progenies of low x low ODAP crosses were found to be low in ODAP concentration indicating the low ODAP lines shared some genes in common for seed ODAP content. The F₁ progenies of the low ODAP x high ODAP crosses were intermediate in ODAP concentration and the F₂ progenies segregated covering the entire parental range. This continuous variation, together with very close to normal distribution of the F₂ population both of low x low and low x high ODAP crosses indicated that ODAP content was quantitatively inherited. Reciprocal crosses, in some cases, produced different results indicating a maternal effect on ODAP concentration. Blue and white flower coloured lines of grass pea were inter-crossed to study the inheritance of flower colour. Blue flower colour was dominant over the white. The F₂ progenies segregated in a 13:3 ratio indicating involvement of two genes with inhibiting gene interactions. The gene symbol LB for blue flower colour and LW for white flower colour is proposed.     

Genotype-environment interaction for seed yield and ODAP concentration of Lathyrus sativus L. and L. cicera L. in Mediterranean-type environments

In 1994, 407 L. sativus and 96 L. cicera lines were evaluated for phenology, seed yield and seed ODAP (a neurotoxin) concentration in the Mediterranean-type climate of south-western Australia. A selected number of lines from this study were grown at three sites in south-western Australia in 1995 and 1996, to examine genotype-environment interactions on seed yield and ODAP concentration in the seed of the two Lathyrus spp. Principal components analysis showed that L. sativus lines grown in 1995 and 1996 could be divided into two geographical origins: Indian sub-continent and Mediterranean/European. Those lines of Mediterranean/European origin were consistently higher yielding (both in 1994 and 1995/96), with much larger seeds and later phenology. In L. cicera seed yield was closely associated with greater mean seed weight and to a lesser extent with early phenology. The genotype-environment study indicated that higher yielding lines of both species generally responded to favourable environments; in the case of L. sativus the Mediterranean/European lines and for L. cicera the larger seeded, earlier phenology lines. For both species, genotype was the most important determinant of ODAP concentration and environment had less influence. Genotype-environment interactions had no effect on seed ODAP concentrations. In terms of seed yield, both species showed substantial potential in the environments tested. Further improvements in adaptation, seed yield and reduction in ODAP are possible in both species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Molecular marker analysis of genetic variation characterizing a grasspea (Lathyrus sativus) collection from central Italy

Levels of genetic similarity characterizing 20 grasspea (Lathyrus sativus L.) populations collected in central Italy (17 populations in the Marche region and three populations in the Abruzzo region) were analysed with amplified fragment length polymorphism (AFLP) molecular markers. Two main clusters were found: one included large‐seeded populations from farms that were not market‐oriented (named Household populations) and the second, small‐seeded populations, cultivated in market‐oriented farms (named Commercial populations). Relationships among populations collected in different regions were found, although one population of the Abruzzo region was placed between the two main clusters, suggesting a possible further genetic differentiation within this grasspea germplasm collection. Principal component analysis based on AFLP marker frequency was effective in identifying polymorphic markers showing high discriminating ability between clusters H and C. In particular, seven markers showing high positive and three markers with low negative PC1 scores showed an almost cluster‐specific distribution. These results will be useful for enhancing Italian grasspea germplasm use in plant‐breeding programmes and for extending grasspea cultivation within the sustainable agricultural systems of central Italy.     

Variation and association of morphological and biochemical characters in grass pea (Lathyrus sativus L.)

Fifty grass pea land race populations of Ethiopia that were selected based on administrative regions and different altitude classes were used in this study to determine variability in morphological characters, Oxalyl Diamino Propanoic acid(ODAP) – a causative agent of lathyrism, and protein contents and their associations. Highly significant differences were observed among the populations for most of the morphological characters. Populations collected from Gondar region and the higher altitude group (>2550 m) showed the highest mean and coefficient of variation indicating the presence of high genetic diversity in this region and altitude group. ODAP analysis from single plants showed significant variation both within a population and between populations. Four individual plants with low ODAP contents ranging from 0.149% to 0.182% (range within safe level) were identified in populations collected from different regions, indicating the diversity and variation of this trait with in one population and between regions. These low ODAP lines are considered important for further breeding. The association of ODAP with grain yield, biomass, plant height and seed size was negative, suggesting that the selection of tall and late maturing varieties with large seed size and high grain yield potential will enable the development of varieties with low ODAP content. This revised version was published online in August 2006 with corrections to the Cover Date.     

Contents of total protein and β-N-oxalyl-l-α,β-diaminopropionic acid (ODAP) in a collection of Lathyrus cicera of the Bank of Plant Germplasm of Cuenca (Spain)

One hundred and thirty-four accessions of Lathyrus cicera from the Bank of Plant Germplasm of Cuenca (BGV-Cuenca, Spain) have been analysed for total protein and β- N -oxalyl- l -α,β-diaminopropionic acid (ODAP) contents in order to identify those with a suitable nutritional quality for animal feeding. There was a large variability in both traits, the content of total protein ranged from 22.95% to 30.98% with a mean value of 27.62%, and the ODAP from 0.09% to 0.30% with a mean value of 0.17%. The accessions from Castilla y León and Castilla-La Mancha showed higher percentages of protein and ODAP as compared with those from Andalucía, Greece and Extremadura. Nevertheless, total protein and ODAP contents showed a low positive correlation ( r  = 0.396, P = 0.01). Several accessions showed a content of ODAP similar to that of the variety 'Chalus' and two accessions seemed very suitable for animal feeding because of their high content of protein and low ODAP.     

Grass pea (<Emphasis Type="Italic">Lathyrus sativus</Emphasis>): Is there a case for further crop improvement?

An ideal legume for resource-poor farmers, grass pea [Lathyrus sativus] is a drought tolerant crop that thrives with minimal external inputs. It is grown on 1 million ha throughout South Asia, mainly as a relay crop after rice. It is also grown extensively in Ethiopia, where it is an important legume for human consumption. Traditionally used for human consumption and as a source of animal feed, this protein-rich legume is favoured for its excellent flavour. There is great potential for an expansion in the utilization of grass pea in drought-prone economies, such as Ethiopia. However, L. sativus produces small quantities of a neurotoxin, β-N-oxalyl –L-α-diaminopropanoic acid (ODAP), which, when consumed alone in large quantities, may cause ‘lathyrism’, an irreversible paralysis of the legs. Lathyrism is a medical condition closely associated with poverty in rural areas. Despite efforts in some countries to discourage production of L. sativus, poor farming communities continue to rely on the crop to supplement their meagre diets. As local land races with high toxin levels generally outperform introduced varieties with lower toxin levels, farmers have little option but to continue to grow their locally-adapted varieties. While there has been some success in breeding grass pea lines with low levels of ODAP, crop improvement programmes are scarce and under-resourced. Facing rising food prices and more frequent natural disasters associated with climate change, we should not neglect the ability of grass pea to provide human and animal feed, under conditions unsuitable for economic production of other legume crops.     

Phenotypic polymorphism of six enzymes in the grasspea (Lathyrus sativus L.)

Summary The phenotype variation in six enzymes, 6-phosphogluconate dehydrogenase (6-PGD), malate dehydrogenase (MDH), peroxidase (PRX), isocitrate dehydrogenase (IDH), galactose dehydrogenase (GD) and glutamate oxaloacetate transaminase (GOT), was investigated using horizontal starch gel electrophoresis in 52 accessions of the grasspea, Lathyrus sativus. Phenotypic polymorphism was observed for all six enzymes. High phenotypic polymorphism (Pj) was observed for PRX and 6-PGD, while there was little polymorphism for GOT, with only two accessions showing variation. There was no correlation between phenotypic polymorphism and region of origin, or groupings of accessions made on the basis of flower colour. Tentative genetic interpretations of banding patterns are given for five of the enzyme systems. The level of apparent heterozygosity was higher than expected in this predominantly autogamous species. The level of variation in the grasspea is discussed in terms of its potential for exploitation through plant breeding.     

Photosynthesis is Reduced,and Seeds Fail to Set and Fill at Similar Soil Water Contents in Grass Pea (Lathyrus sativus L.) Subjected to Terminal Drought

Grass pea (Lathyrus sativus L.) is an indeterminate grain legume considered adapted to dry environments, but the mechanisms of its adaptation are not well understood. Grass pea plants were exposed to terminal drought from podding, and the development of water deficit was measured together with its effects on leaf photosynthesis, stomatal conductance, carbon remobilisation to the seeds, flower production and abortion, pod production and abortion, seed set, seed growth and the neurotoxin β‐N‐oxalyl‐L‐a, β‐diaminopropionic acid (β‐ODAP) concentration. Predawn leaf water potential (Ψ_leaf), stomatal conductance (gs), rate of leaf photosynthesis (Pn), flower production, pod production, filled pod number, seed number, seed size and yield decreased, while flower abortion, pod abortion and seed abortion increased, and the concentration of β‐ODAP was unchanged under terminal drought conditions. gs and Pn began to decrease at a higher plant‐available soil water content (PAWC) (67.2 ± 2.3 % and 62.9 ± 2.3 %) than Ψ_leaf (43.7 ± 1.1 %). Flowers and pods ceased being produced only when the PAWC decreased below 40.1 ± 4.6 % and 35.3 ± 3.0 %, respectively, but seed set and seed growth ceased when PAWC decreased below 55.5 ± 1.6 % and 58.0 ± 3.7 %, respectively. The mobilization of ¹³C labelled assimilates from the stems was greater under terminal drought than under well‐watered conditions, but the transfer to the seed was small. We conclude that seed set and seed growth decreased as the soil dried due to a reduction in current photosynthesis as a result of stomatal closure.     

The Influence of Day Length on Plant and Nodule Development of Fababeans (Vicia faba L.) and Grasspeas (Lathyrus sativus L.)

The valuation of the influence of different day length (DL) on plant and especially nodule development of fababeans ( Vicia faba L.) and grasspeas ( Lathyrus sativus L.) was the aim of these experiments. For that, experiments in growth chambers (Exp. 1) with DL of 10, 13, 16 and 19 hours and an experiment in vegetation boxes (Exp. 2) with DL of 7, 10, 13 hours and natural DL as control (CON) were conducted. The development of plants was studied by 6 and 7 harvests (HA) respectively beginning at 27 and 31 days after emergence (DAE) respectively, as well as consecutive determinations of leaf area, dry matter (DM) of the different plant organs and nodule number.
Total- and pod-DM of both species increased with higher DL (19 h, CON). Nodule-DM was positive influenced by longer assimilation duration and bigger leaf areas of the higher DL treatments. With onset of pod ripening, which occurred early under higher DL, lower nodule-DM was determined due to the beginning of nodule senescence. Under 19 h DL the fababeans established highest nodule-DM, 1,100 mg/plant, whereas the grasspea 410 mg only. Nodules under shorter DL (7 h, 10 h) showed less intensive red tissue and less nodules > 6 mm diameter. The grasspea established more bigger nodules than the fababeans. Total nodule number was not unequivocal affected by DL, however, it increased with the course of vegetation. Senescence of nodules was most evident at time of highest growth rates of the pod-DM.     

Genetic improvement of legumes using somatic cell and molecular techniques

Summary The merits and limitations of somatic cell techniques involving Agrobacterium-mediated transformation, direct gene transfer and protoplast fusion, are discussed in relation to the genetic improvement of forage and grain legumes. Whilst progress with legumes is limited compared to that with plants of other families such as the Solanaceae, the fact that many legumes are readily amenable to tissue culture now permits somatic cell techniques to be targetted to these species. Future development of the subject will necessitate close collaboration between molecular biologists and plant breeders to enable novel plants generated by in vitro technologies to be incorporated into conventional breeding programmes.     

Genetic linkage map of Chinese native variety faba bean (Vicia faba L.) based on simple sequence repeat markers

Simple sequence repeat (SSR) marker is a powerful tool for construction of genetic linkage map which can be applied for quantitative trait loci (QTL) and marker‐assisted selection (MAS). In this study, a genetic map of faba bean was constructed with SSR markers using a 129 F₂ individuals population derived from the cross of Chinese native variety 91825 (large seed) and K1563 (small seed). By screening 11 551 SSR primers between two parents, 149 primer pairs were detected polymorphic and used for F₂ population analysis. This SSR‐based genetic linkage map consisted of 15 linkage groups with 128 SSR. The map encompassed 1587 cM with an average genetic distance of 12.4 cM. The genetic map generated in this study will be beneficial for genetic studies of faba bean for identification of marker‐locus‐trait associations as well as comparative mapping among faba bean, pea and grasspea.     

Revised calculation of the coefficient of parentage in plant breeding

The knowledge of the genetic similarity between two individuals has been useful for animal breeders for a long time, and one of the most common measures of similarity is the Coefficient of Parentage (COP). For two individuals X and Y, it is defined as the probability that an allele taken from a particular locus in individual X is equal by descent to another allele taken from the same locus in individual Y; thus, the COP between two individuals is the same as the expected inbreeding of their offspring, and it is now a routine calculation among plant breeders as part of crop improvement programmes. Nevertheless, plant breeders deal with the genealogy between strains or groups of individuals, and here, we provide a mathematical proof that in this case the application of standard formulae replicated from animal breeding may result in overestimation of the true amount of inbreeding between two strains.     

Development and characterization of a new 1BL.1RS translocation line with resistance to stripe rust and powdery mildew of wheat

The 1BL.1RS wheat-rye translocation from Petkus rye has contributed substantially to the world wheat production. However, following the breakdown of disease resistance genes in 1RS, its importance for wheat improvement decreased. We have developed a new 1BL.1RS line, R14, by means of crossing rye inbred line L155, selected from Petkus rye to several wheat cultivars. One new gene each, for stripe rust and powdery mildew resistance, located on 1RS of the line R14, are tentatively named YrCn17 and PmCn17. YrCn17 and PmCn17 confer resistance to Puccinia striiformis f. sp. tritici pathotypes that are virulent on Yr9, and Blumeria graminis f. sp. tritici pathotypes virulent on Pm8. These two new resistances, YrCn17 and PmCn17, are now available for wheat improvement programs. The present study indicates that rye cultivars may carry yet untapped variations as potential sources of resistance.     

Genetic improvement of grain yield and associated traits in the southern China winter wheat region: 1949 to 2000

To understand the genetic gains of grain yield in the Southern China Winter Wheat Region (SCWWR), two yield potential trials, i.e., YPT 1 including 11 leading cultivars from the Middle and Low Yangtze Valley (Zone III) and YPT 2 including 15 leading cultivars from the Southwestern China Region (Zone IV) from 1949 to 2000, were conduced during the 2001–2003 cropping seasons. A completely randomized block design of three replicates was employed with controlled field environments. Molecular markers were used to detect the presence of dwarfing genes and the 1B/1R translocation. Results showed that average annual genetic gain was 0.31% (P < 0.05) or 13.96 kg/ha/year and 0.74% (P < 0.01) or 40.80 kg/ha/year in Zones III and IV, respectively. In YPT 1, changes of all other traits were not significant, but plant height was significantly reduced. In YPT 2, the genetic improvement of grain yield was primarily attributed to the increased thousand kernel weight (TKW) (0.65%, P < 0.01) and kernel weight/spike (0.87%, P < 0.01), reduced plant height and increased harvest index (HI). The dwarfing gene Rht 8 was most frequently present (46.1%), Rht-B1b was observed in three genotypes in Zone III, and Rht-D1b was present in only one genotype in Zone IV. The 1B/1R translocation was present in four genotypes. Utilization of Italian germplasm and development of landmark cultivar Fan 7 were the key factors for grain yield improvement in SCWWR. The future challenge of wheat breeding in this region is to continue improving grain yield and disease resistance, and to develop cultivars suitable for the reduced tillage of wheat/rice double cropping. Utilization of Mexican germplasm could provide opportunities for future yield improvement.     

Kale population improvement and cultivar production

Marrow-stem kale (Brassica oleracea var. acephala L.) is grown in the United Kingdom, New Zealand and similar climates, for feeding cattle and sheep during autumn and winter. Population improvement by half-sib family selection for higher digestibility and digestible organic matter yield, lower levels of antimetabolites, and clubroot resistance is described and analysed as a selection experiment. The foundation population consisted of 120 clubroot resistant plants from the 16 most resistant marrow-stem cultivars in an assessment of 96 cultivars of Brassica oleracea. At the end of the first generation of selection adequate resistance had been achieved. After five generations, dry-matter content and digestible organic matter yield had been increased, the latter without an undesirable increase in height. Indolyl glucosinolate (source of goitrogenic thiocyanate ion) content had been reduced, but not S-methylcysteine sulphoxide (the haemolytic factor) content. Selection differentials each generation were small and heritabilities of family means were moderate to low, making improvement per generation slow. It was demonstrated that faster progress could be made by switching from half-sib to full-sib family selection, as expected from theory. Multiplication of the half-sib family with the highest digestibility (DOMD%) achieved an improvement in this trait, without a reduction in digestible organic matter yield, and was marketed as cultivar Grampian.     

Strain improvement in the cultivated mushroom Agaricus bisporus

Summary Early attempts at genetic improvement in the cultivated mushroom Agaricus bisporus (Lange) Imbach were empirical, for little was understood of its natural breeding system. The mushroom is now known to be a secondarily homothallic species with a single multiallelic mating-type factor. This better understanding makes it possible to evaluate those breeding methods previously used and to suggest alternatives.Strain selection alone based on single spores, multispores or tissue culture may give improvement in the short term but it is unlikely to be as effective as methods involving controlled crossing. Mixing fertile strains may produce hybrids but it is difficult to identify them. It is better to use non-fertile isolates because only hybrids fruit. The earlier recognition of hybrids can be achieved using markers which are expressed in culture and genetic resistances may be especially useful in this respect.There is also a possible role for other Agaricus species which may be grown commercially and are more amenable to genetic manipulation than is A. bisporus.     

Performance of Solanum habrochaites LA1777 introgression line hybrids for marketable tomato fruit yield in Asia

Wild relatives of the cultivated tomato (Solanum lycopersicum L.) are major sources of new genetic diversity for tomato improvement. Introgression lines (IL) are near-isogenic lines homozygous for one or several mapped wild DNA fragments in a common recurrent parent. A set of ILs developed by Cornell University from Solanum habrochaites accession LA1777 and recurrent parent E6203 has been made publicly available through the Tomato Genetics Resource Center of the University of California-Davis. Our objective was to identify LA1777 introgressions with potential to increase the marketable fruit yield of tomato grown in the tropics. A subset of ILs were each crossed to CLN2498E (resistant to bacterial wilt and some begomoviruses) to create IL hybrids (ILH). ILH, IL recurrent parent E6203, CLN2498E, and CLN2498E × E6203 (Hchk) were evaluated in replicated trials in Thailand, India, and Taiwan during two dry seasons. Highly significant effects for marketable fruit yield were detected in Thailand and Taiwan. ILH heterozygous for S. habrochaites segments at the bottom of chromosome 1 yielded about 20% than the Hchk at Thailand and Taiwan. Our results agree with previous results from Cornell University researchers who found a segment of S. habrochaites DNA located between TG158 and TG27 associated with increased total fruit yield in previous trials conducted in upper state New York. Yield improvement due to this S. habrochaites introgression can occur over a wide range of environments.     

Gibberellic Acid + 2,4-D Improves Seed Quality in Hordeum vulgare L. ×H. bulbosum L. Crosses

Doubled haploid production from crosses between barley (Hordeum vulgare L.) and H. bulbosum L. is a well established technique for inducing homozygosity. One problem which occurs using certain barley cultivars is premature seed degeneration. In this paper, a post-pollination spray of gibberellic acid and 2,4-dichlorophenoxyacetic acid that delays the onset of senescence is described. The technique improved the number of high quality seeds to more than 75% of the seed set. This improvement should result in increased survival of embryos for culture and subsequent plant regeneration.