A sensitive and specific PCR-based discrimination of split red vetch and lentil seeds

A PCR-based marker technique was developed to discriminate between morphologically similar split seed of vetch (Vicia sativa) and lentil (Lens culinaris subsp.culinaris). Sequence tagged microsatellite site (STMS) markers were more discriminatory than markers produced from the nontranscribed spacer (NTS) region of the 5S ribosomal RNA gene. A sequence characterized amplified region (SCAR) marker, developed from the 5S rRNA NTS region, was sensitive when resolved on agarose. However, the fluorescent-labeled 5S rRNA SCAR marker was unable to discriminate between vetch and lentil, probably because of the low copy number of the marker, and was not visualized on agarose. An STMS primer-pair (PSMPSAD123), developed from field pea, was able to discriminate split red cotyledon vetch from split red cotyledon lentil because it produced specific markers at 563 bp for lentil and 353 and 474 bp for vetch. The vetch-specific STMS marker was conserved among all species of theVicia genus used in this study and was sensitive enough to discriminate both on agarose gels and on polyacrylamide gel-based fluorescent systems. The fluorescent-tagged STMS analysis revealed peaks for vetch and lentil at the expected sizes in admixtures of milled vetch and lentil seeds, and it was sensitive enough to detect one vetch seed in 1999 lentil seeds. The development of PCR-based tests for detecting the level of vetch seed contamination in lentil export seed may provide a method for quality assurance of export lentil seed.     

Seed dispersal in relation to the domestication of Middle East Legumes

Modification of seed dispersal was perhaps one of the most important steps towards domestication of seed crops. Among the legumes of the Middle East, four patterns of such modification can be distinguished in the process of domestication. The initial stage of domestication of lentil, pea and grass pea was apparently due to a single mutation in a major gene that prevented pod dehiscente. In chick pea the domesticated type was formed by accumulation of several mutations in minor genes that reduced the amount of pod dropping and shattering. From a seed dispersal point of view, fenugreek was preadapted to cultivation since the wild species do not shatter their seeds. In the bitter vetch and common vetch partial seed shattering apparently was tolerable and desirable under cultivation since the seed served merely for sowing the next year crop.     

The Development of Desiccation-tolerance and Maximum Seed Quality During Seed Maturation in Six Grain Legumes

‘Physiological maturity’, i.e. the time when seedsreach their maximum dry weight during development, occurredwhen maturation drying on the parent plant in the field hadreduced seed moisture content to approximately 60 per cent infaba bean (Vicia faba L.), lentil (Lens culinaris Medic.), chickpea(Cicer arietinum L.), white lupin (Lupinus albus L.), soya bean(Glycine max [L.] Merr.) and pea (Pisum sativum L.) The onsetof desiccation-tolerance, i.e. the ability of seeds to germinatefollowing harvest and rapid artificial drying, coincided withphysiological maturity, except in pea where it occurred a littleearlier at about 70 per cent moisture content. Maximum seedquality as determined by maximum viability, minimum seedlingabnormalities and maximum seedling size occurred in pea, chickpeaand lupin when seeds were harvested for rapid drying at physiologicalmaturity; but for maximum seed quality in the other speciesmaturation drying had to proceed further - to about 45 per centmoisture content in soya bean and to about 30 per cent moisturecontent in lentil and faba bean seed crops. Much of this variationamongst the six species, however, was due to differences inthe variation in maturity within each seed crop. Results forindividual pods showed that peak maturity, i.e. maximum seedquality following harvest and rapid artificial drying, was achievedin all six species once maturation drying had reduced the moisturecontent of the seeds to 45–50 per cent. In pea, faba beanand soya bean there was a substantial decline in viability andan increase in seedling abnormalities when harvest was delayedbeyond the optimal moisture content for harvest.     

Neolithic cultivated plants from Albania

Macroremains of late Neolithic cultivated plants are reported from the archaeological site of Maliq, Korça District, south-eastern Albania. The material comprises einkorn (Triticum monococcum L.), emmer (Triticum dicoccon Schrank), barley (Hordeum vulgare L.), lentil (Lens culinaris Medik.) and bitter vetch (Vicia ervilia (L.) Willd.). These are the first remains of cultivated plants described from an archaeological site in Albania.     

Comparative Characteristics of Carbohydrate Binding by Lectins from Broad Bean,Pea, Common Vetch,and Lentil Seeds

Lectins from the seeds of broad bean (Vicia faba L.), pea (Pisum sativum L.), common vetch (V. sativa L.), and lentil (Lens culinaris Medik.) were isolated and purified by affinity chromatography. The hemagglutinating activity of lectins was most effectively inhibited by methyl--D-mannopyranoside, trehalose, and D-mannose. Other carbohydrate haptens, such as methyl--D-glucopyranoside, maltose, and alginic and D-glucuronic acids were less effective. Two lectins obtained from different lentil cultivars, unlike other lectins, had a relatively high affinity for melecitose, N-acetyl-D-glucosamine, L-sorbose, and sucrose. Furthermore, these lectins interacted with soluble starch. All the lectins examined had similar, but not identical, carbohydrate-binding properties. Because of their similar D-mannose/D-glucose specificity, these lectins interacted with lipopolysaccharides and exopolysaccharides of Rhizobium leguminosarum bv. viciae, root nodule bacteria that infect broad-bean, pea, common-vetch, and lentil plants with the formation of nitrogen-fixing symbiosis. However, owing to individual distinctions of carbohydrate-binding properties, these lectins showed a higher affinity for the polysaccharides of those microsymbionts within the R. leguminosarum bv. viciae species that were better specialized towards one or the other host plant from the cross inoculation group of legumes.     

From the cradle of agriculture a handful of lentils: History of domestication

Literature on lentil domestication is reviewed, particularly considering archeobotanical, phylogenetic, and molecular evidence. Lentils are one of the oldest crops cultivated and domesticated by man. Carbonized small lentil seeds have been found in several archaeological remains starting from the Neolithic. It is probable, however, that the most ancient remains refer to wild lentils; this is difficult to ascertain since seed size was probably selected after the establishment of a domesticated lentil. It is general opinion that cultivation occurred before domestication, but for how long is still an open question. It is now well accepted that the domestication of lentils was accomplished in the Near East, in an area called “the cradle of agriculture”. The genus Lens is very small, containing only 6 taxa. A wide range of morphological and molecular evidence supports the idea that the lentil wild progenitor is Lens culinaris ssp. orientalis. On the other hand, the most distantly related species within the genus appears to be L. nigricans, whose domestication was also attempted without success. The first characters involved in lentil domestication were pod dehiscence and seed dormancy. These traits are under a simple genetic control, and therefore mutants must have been fixed in a relatively short time. These and other morphological traits possibly involved in lentil domestication have been mapped in several linkage maps. However, generally these maps are not easily integrated since they are based on a limited number of markers. Newer maps, mainly built on different kinds of molecular markers, have been more recently produced. A consensus map is needed to fill the gap in lentil breeding and, at the same time, endow with deeper information on the genetics of lentil domestication, giving new insight into the origins of this crop, which present fragmented knowledge is unable.      

Germination and Early Growth of Plants Studied Using Neutron Radiography

Seed swelling, germination, root extension, lateral root initiationand shoot growth were studied in soils of different water contents,using non-destructive, serial neutron radiography. Seeds fromthree varieties of soya beans (Glycine max L.) and one varietyeach of maize (Zea mays L.) and vetch (Vicia sativa L.) wereused. The seeds germinated when they had increased in size bya certain amount, if germination is taken as the time when theradicle first appears. The rate at which roots and shoots extendalso depend on soil water content. Glycine max L., Vicia sativa L., Zea mays L., Soya bean vetch, maize, seed germination, root extension, lateral root initiation, neutron radiography     

Differences in accumulation of soluble α-galactosides during seed maturation of several <Emphasis Type="Italic">Vicia</Emphasis> species

Composition and levels of soluble α-galactosides: raffinose family oligosaccharides (RFOs) and galactosyl cyclitols (Gal-C) in developing seeds were measured by high resolution gas chromatography (HRGC) method. The studies were performed on maturing seeds of several wild and cultivated Vicia species: Vicia angustifolia L. (common vetch), Vicia cracca L. (bird vetch), Vicia grandiflora Scop. (large yellow vetch), Vicia hirsuta (L.) S.F.Gray (tiny vetch), Vicia sativa L. (garden vetch, spring-growing cultivar Kwarta), and Vicia villosa Roth (winter vetch). In all Vicia species similar patterns in the accumulation of RFOs were observed. Galactinol — the donor of galactosyl moieties in α-galactosides biosynthesis was present in the middle stage of seed development, before appearing measurable levels of RFOs. Accumulation of RFOs started parallel with seed desiccation process. At first accumulation of the raffinose, then few days later stachyose and finally verbascose was noticed. In the final stage of seed maturation the verbascose was the main soluble α-galactoside (up to 3% of dry weight, V. sativa). Besides the RFOs seeds of three Vicia species (V. cracca, V. hirsuta, and V. villosa) accumulated d-pinitol and its α-galactosides (Gal-C). Mono-galactosylpinitols (similar to raffinose) appeared in these species 2–4 days after galactinol, di-galactosyl pinitol A (common name: ciceritol) and di-galactosyl myo-inositol were present several days later than raffinose, and accumulation of tri-galactosyl pinitol A (TGPA) began after accumulation of stachyose. Matured seeds of V. hirsuta contained much more RFOs than Gal-C, opposite to seeds of V. villosa, and V. cracca where concentration of Gal-C was 4–8-fold higher than RFOs. In V. cracca seeds RFOs were almost replaced by Gal-C. In seeds of V. cracca and V. villosa the level of d-pinitol was significantly higher, than the level of myo-inositol. Contents of both cyclitols declined rapidly at the beginning of seed desiccation, when accumulation of RFOs and Gal-C quickly increased. We suggest that α-galactosides of d-pinitol can substitute raffinose family oligosaccharides and play similar role during seed maturation and storage.     

Plant remains from Asikli Höyük,a pre-pottery Neolithic site in central Anatolia

Cereal crop plants at Asikli Höyük included einkorn wheat (Triticum monococcum), emmer wheat (T. dicoccum), free-threshing wheat (T. cf. durum), hulled two-rowed barley (Hordeum distichum) and naked barley (H. vulgare var. nudum). As for pulses, bitter vetch (Vicia ervilia), lentil (Lens culinaris) and probably pea (Pisum sativum) were grown. Hackberry (Celtis tournefortii) fruits were gathered intensively. In addition, fruits of pistachio (Pistacia cf. atlantica) and almond (Amygdalus spec.) were collected from the wild.     

The role of Bh4 in parallel evolution of hull colour in domesticated and weedy rice

The two independent domestication events in the genus Oryza that led to African and Asian rice offer an extremely useful system for studying the genetic basis of parallel evolution. This system is also characterized by parallel de‐domestication events, with two genetically distinct weedy rice biotypes in the US derived from the Asian domesticate. One important trait that has been altered by rice domestication and de‐domestication is hull colour. The wild progenitors of the two cultivated rice species have predominantly black‐coloured hulls, as does one of the two U.S. weed biotypes; both cultivated species and one of the US weedy biotypes are characterized by straw‐coloured hulls. Using Black hull 4 (Bh4) as a hull colour candidate gene, we examined DNA sequence variation at this locus to study the parallel evolution of hull colour variation in the domesticated and weedy rice system. We find that independent Bh4‐coding mutations have arisen in African and Asian rice that are correlated with the straw hull phenotype, suggesting that the same gene is responsible for parallel trait evolution. For the U.S. weeds, Bh4 haplotype sequences support current hypotheses on the phylogenetic relationship between the two biotypes and domesticated Asian rice; straw hull weeds are most similar to indica crops, and black hull weeds are most similar to aus crops. Tests for selection indicate that Asian crops and straw hull weeds deviate from neutrality at this gene, suggesting possible selection on Bh4 during both rice domestication and de‐domestication.     

Enzymatic synthesis of (R)-cyanohydrins by a novel (R)-oxynitrilase from Vicia sativa L.

The defatted seed meal of common vetch (Vicia sativa L.) served as a source of (R)-oxynitrilase which catalyzed the enantioselective addition of HCN to aromatic, heteroaromatic, fluoro-substituted aromatic aldehydes to produce the corresponding enantiomeric pure cyanohydrins in yields of 52–100% and 88–99% ee at 12°C under micro-aqueous medium (diisopropyl ether) without addition of buffer solution.     

Malaysian weedy rice shows its true stripes: wild Oryza and elite rice cultivars shape agricultural weed evolution in Southeast Asia

Weedy rice is a close relative of domesticated rice (Oryza sativa) that competes aggressively with the crop and limits rice productivity worldwide. Most genetic studies of weedy rice have focused on populations in regions where no reproductively compatible wild Oryza species occur (North America, Europe and northern Asia). Here, we examined the population genetics of weedy rice in Malaysia, where wild rice (O. rufipogon) can be found growing in close proximity to cultivated and weedy rice. Using 375 accessions and a combined analysis of 24 neutral SSR loci and two rice domestication genes (sh4, controlling seed shattering, and Bh4, controlling hull colour), we addressed the following questions: (i) What is the relationship of Malaysian weedy rice to domesticated and wild rice, and to weedy rice strains in the USA? (ii) To what extent does the presence of O. rufipogon influence the genetic and phenotypic diversity of Malaysian weeds? (iii) What do the distributions of sh4 and Bh4 alleles and associated phenotypes reveal about the origin and contemporary evolution of Malaysian weedy rice? Our results reveal the following: independent evolutionary origins for Malaysian weeds and US strains, despite their very close phenotypic resemblance; wild‐to‐weed gene flow in Malaysian weed populations, including apparent adaptive introgression of seed‐shattering alleles; and a prominent role for modern Malaysian cultivars in the origin and recent proliferation of Malaysian weeds. These findings suggest that the genetic complexity and adaptability of weedy crop relatives can be profoundly influenced by proximity to reproductively compatible wild and domesticated populations.     

Expansion of rice enzymatic rhizosphere: temporal dynamics in response to phosphorus and cellulose application

Plant and Soil - Evaluate potential N benefit from chickpea (Cicer arietinum L.), faba bean (Vicia faba L.), lentil (Lens culinaris L.) and field pea (Pisum sativum L.). This is the first phase of...     

Urartian plant cultivation at Yoncatepe (Van), eastern Turkey

Plant remains were recovered from an Urartian settlement, Yoncatepe, situated in the Van province of eastern Turkey and dating to the Iron Age period (first millennium B.C.E.). Large quantities of hulled barley (Hordeum vulgare L.) and of bread/macaroni wheat (free-threshing wheat) Triticum aestivum L./T. durum Desf.), both mixed with small quantities of domesticated emmer wheat (T. dicoccum Schübl.), were found in the storerooms of the Yoncatepe palace, indicating the storage of agricultural surplus. Rye (Secale cereale L.) grains occur very occasionally, while pulses include lentil (Lens culinaris Medik.), chickpea (Cicer arietinum L.), and bitter vetch (Vicia ervilia [L.] Willd.). Grape seeds unearthed in a tomb at Yoncatepe provide physical evidence supporting written records of vineyards. Numerous seeds of gold of pleasure (Camelina sativa [L.] Crantz), found in a storage vessel, provide evidence of the cultivation of this plant. It is likely, that the Urartians used the seeds for oil extraction.     

More than one way to evolve a weed: parallel evolution of US weedy rice through independent genetic mechanisms

Many different crop species were selected for a common suite of ‘domestication traits’, which facilitates their use for studies of parallel evolution. Within domesticated rice (Oryza sativa), there has also been independent evolution of weedy strains from different cultivated varieties. This makes it possible to examine the genetic basis of parallel weed evolution and the extent to which this process occurs through shared genetic mechanisms. We performed comparative QTL mapping of weediness traits using two recombinant inbred line populations derived from crosses between an indica crop variety and representatives of each of the two independently evolved weed strains found in US rice fields, strawhull (S) and blackhull awned (B). Genotyping‐by‐sequencing provided dense marker coverage for linkage map construction (average marker interval <0.25 cM), with 6016 and 13 730 SNPs mapped in F₅ lines of the S and B populations, respectively. For some weediness traits (awn length, hull pigmentation and pericarp pigmentation), QTL mapping and sequencing of underlying candidate genes confirmed that trait variation was largely attributable to individual loci. However, for more complex quantitative traits (including heading date, panicle length and seed shattering), we found multiple QTL, with little evidence of shared genetic bases between the S and B populations or across previous studies of weedy rice. Candidate gene sequencing revealed causal genetic bases for 8 of 27 total mapped QTL. Together these findings suggest that despite the genetic bottleneck that occurred during rice domestication, there is ample genetic variation in this crop to allow agricultural weed evolution through multiple genetic mechanisms.     

Transferability of sequence tagged microsatellite site (STMS) primers across four major pulses

The transferability of genome-specific sequence tagged microsatellite site (STMS) primers from field pea (P. sativum) and chickpea (C. arietinum) to other major pulses was examined. Overall, field pea STMS primers amplified products in most of the accessions in comparison to that of the chickpea STMS primers, which amplified products in relatively few accessions. The highest level of successful amplifications with a single primer was 89% for field pea and 33% for chickpea primers respectively. The potential transferability of the STMS primers among species, expressed as the total mean percentage of positive amplifications, was 53% for the field pea STMS primers and 9% for the chickpea STMS primers. The individual mean percentage of successful transferability of field pea STMS primers across lentil, vetch and chickpea/Cicer sp. accessions was 60%, 39% and 62%, respectively. Whereas, for the chickpea STMS primers successful transferability was 5%, 3% and 18% for lentil, vetch and field pea, respectively. The trnasferability of these STMS primers indicates a high level of sequence conservation in these regions across species. Together with their locus-specificity, co-dominant nature and potential to amplify multiple alleles, their transferability makes STMS markers a powerful tool for genetic mapping, diversity analysis and genotyping.     

Flight responses in a wind tunnel of the parasitoid Microplitis croceipes to three spring and three summer host plants of Heliothis spp.

Flight responses of the parasitoid, Microplitis croceipes (Cresson) (Hymenoptera: Braconidae) to two phenological stages of spring host plants (crimson clover, Trifolium incarnatum L., hairy vetch, Vicia villosa Corbiere, and cutleaf geranium, Geranium dissectum L.) and summer host plants (cotton, Gossypium hirsutum L., sorghum, Sorghum bicolor L., and soybean Glycine max (L.) Merr.) of Heliothis virescens (F.), were measured in a dual choice situation in a wind tunnel. Preflowering clover was more attractive than preflowering geranium, followed by preflowering vetch. Clover was preferred over geranium in flowering stages as well. No within species differences were detected for the two phenological stages. Preflowering and flowering cotton and soybean were preferred over preflowering and flowering sorghum. The findings could be important in pre-determining release sites for M. croceipes in the field.     

Host plant species and carbohydrate supplements affecting rate of multiplication of redlegged earth mite

The effects of young plants of different legume species on the rate of multiplication of Halotydeus destructor (Tucker)(Acarina: Penthaleidae) were examined in one, two or four week experiments in the laboratory. Rates of multiplication of H. destructor were substantially greater on Vicia sativa (common vetch), Phaseolus vulgaris (dwarf bean) and Pisum sativum (snow pea), than on subterranean clover, Trifolium subterraneum; cultivars Dalkeith, Geraldton and Junee. Levels of soluble carbohydrate and soluble nitrogen were greater in vetch, bean and pea, than in the subclovers. Concentrations of 20 amino acids were not associated with differences in mite performance; pea contained higher levels of glutamine and homoserine than the other plants, which may have adversely affected mite multiplication.In further experiments a carbohydrate supplement caused a fourfold increase in rate of multiplication on subclover cv Junee, but had no effect on rate of multiplication on vetch cv Blanchefleur. The carbohydrate supplement did not cause the rate of multiplication on subclover to equal that on vetch. Subclover is not an optimum host plant for H. destructor nutritionally. The nutritional value of host plants is one factor affecting mite multiplication.     

Effect of Seed Treatment with Rhizobium leguminosarum on Pythium Damping-off, Seedling Height, Root Nodulation, Root Biomass, Shoot Biomass, and Seed Yield of Pea and Lentil

Field experiments were conducted in 2004 and 2005 to determine the effects of seed treatment with Rhizobium leguminosarum bv. viceae on damping‐off, seedling height, root nodule mass, root biomass, shoot biomass and seed yield of pea and lentil in a field naturally infested with Pythium spp. Compared with the untreated controls, treatment of pea seeds with R. leguminosarum bv. viceae strains R12, R20 or R21 significantly (P < 0.05) reduced incidence of damping‐off, promoted seedling growth and increased root nodule mass, root biomass and shoot biomass. Seed treatments with R12 or R21 also resulted in a significant (P < 0.05) increase in seed yield of pea. The strain R21 was most effective among the four strains of R. leguminosarum bv. viceae tested in peas. Although, the level of disease control by strain R21 was similar to seed treatment with the fungicide Thiram^TM, R21 was more effective in enhancing root nodule production and promoting plant growth. For lentil, treatment of seeds with R. leguminosarum bv. viceae strains R12 or R21 significantly (P < 0.05) reduced incidence of damping‐off compared with the untreated control. All of the four strains of R. leguminosarum bv. viceae tested increased lentil seedling height, root nodule mass and shoot biomass, and all except R20 increased root biomass. Seed yield was higher for the treatments of R12 and R21. The strain R12 was most effective among the four strains of R. leguminosarum bv. viceae tested in lentil. Although, strain R12 was as effective as Thiram^TM for control of damping‐off of lentil, it was more effective than Thiram^TM for the production of root nodules and promotion of plant growth. The study concludes that seed treatment with R. leguminosarum bv. viceae is effective in control of Pythium damping‐off of pea and lentil and that the efficacy of control is strain specific, strain R21 for control of the disease on pea and strain R12 for control of the disease on lentil.     

An investigation of processing and consumption of pulses among prehistoric societies: archaeobotanical, experimental and ethnographic evidence from Greece

Pulses have constituted an important food source for prehistoric communities in the Old World, yet little is known as regards their processing for consumption through the archaeobotanical record. This paper provides an overview of archaeobotanical evidence for the use of pulses in prehistoric Greece based on two case studies from the north, and explores (a) their preparation for consumption, in particular their detoxification and (b) the consumption of pulses as a component of ordinary daily meals in prehistoric times, as well as those for special occasions, within a context of feasting and ritual. The paper examines charred remains of Vicia ervilia (bitter vetch) and Lathyrus sativus (grass pea) from early Bronze Age Agios Athanasios and late Neolithic Kremasti Koiladas, respectively, as the former provides a basis for a pilot exploration of pulse detoxification and the latter, due to its origin, offers a rare opportunity to discuss the context of consumption. In the pilot exploration of pulse seed preparation for consumption, the inner cotyledon morphology of modern V. ervilia seeds which were experimentally processed with water and pounding was examined macroscopically and through SEM micrographs. Preliminary observations suggest that intentional splitting of pulse seeds as part of processing for consumption as food may be recognisable in the archaeobotanical record. Processing with water may also be detected. The particular context of the Kremasti finds suggests that pulses, in this particular case L. sativus, may have constituted special foods for particular occasions, loaded with symbolic meaning.