Gene expression profiles of seed coats and biochemical properties of seed coats and cotyledons of two field pea (Pisum sativum) cultivars contrasting in green cotyledon bleaching resistance

Visual quality is one of the major factors determining the market value of field pea (Pisum sativum L.). Breeding for improved visual quality of pea seeds is currently a challenging task, mainly because of the complexity and the lack of sound genetic knowledge of the traits. The objectives of this research were to characterize post-harvest cotyledon bleaching resistance in green pea at the biochemical and gene expression levels. Seed coats and cotyledons of two pea cultivars, CDC Striker (bleaching resistant) and Orb (bleaching susceptible) at three developmental stages [14, 21 and 28 days after flowering (DAF)] and following exposure to accelerated bleaching conditions after harvest (0, 3, 6, and 13 days after bleaching (DAB) were evaluated. CDC Striker had a slower rate of chlorophyll degradation in cotyledons, and a higher total carotenoids to chlorophyll ratio in seed coats, than Orb when seed samples were exposed to high intensity light. An oligo-nucleotide microarray (Ps6kOLI1) revealed that gene expression profiles of the CDC Striker and Orb seed coats were significantly different during seed developmental stages. A significant up regulation of genes involved in the production and accumulation of secondary metabolites responsible for antioxidant properties including epiafzelechin, epicatechin, epigallocatechin, kaempferide, kaempferol 3-O-β-d-sophorotrioside, O-quercetin and rutin, in the seed coats of CDC Striker were observed. Thus, bleaching resistance in field pea could be due to the accumulation of specific carotenoids and phenolic compounds which quench excess light or scavenge free radical singlet oxygen molecules. The candidate genes identified in this project will need to be validated using qPCR to further support their involvement with bleaching resistance before being used for the development of gene specific markers.     

Identification of quantitative trait loci for grain yield, seed protein concentration and maturity in field pea (Pisum sativum L.)

Breeding efforts to improve grain yield, seed protein concentration and early maturity in pea (Pisum sativum L.) have proven to be difficult. The use of molecular markers will improve our understanding of the genetic factors conditioning these traits and is expected to assist in selection of superior genotypes. This study was conducted to identify genetic loci associated with grain yield, seed protein concentration and early maturity in pea. A population of 88 recombinant inbred lines (RILs) that was developed from a cross between 'Carneval' and 'MP1401' was evaluated at 13 environments across the provinces of Alberta, Manitoba and Saskatchewan, Canada in 1998, 1999 and 2000. A linkage map consisting of 193 AFLPs (amplified fragment length polymorphism), 13 RAPDs (random amplified polymorphic DNA) and one STS (sequence tagged site) marker was used to identify putative quantitative trait loci (QTL) for grain yield, seed protein concentration and early maturity. Four QTL were identified each for grain yield and days to maturity, and three QTL were identified for seed protein concentration. A multiple QTL model for each trait showed that these genomic regions accounted for 39%, 45% and 35% of the total phenotypic variation for grain yield, seed protein concentration and days to maturity, respectively. The consistency of these QTL across environments and their potential for marker-assisted selection are discussed in this report.     

Total phenolics and condensed tannins in field pea (Pisum sativum L.) and grass pea (Lathyrus sativus L.)

Seed samples of seventeen field pea cultivars grown at five locations, and nine grass pea lines grown at two locations, in western Canada during 1993 and 1994 were analysed for total phenolics and condensed tannins. Each location in each year was considered as one environment. Total phenolics in field pea differed significantly among cultivars, ranging from 162 mg/kg DM (dry matter) (CE, catechin equivalents) for AC Tamor to 325 mg/kg DM (CE) for Richmond. Field pea had barely detectable levels of condensed tannins. Total phenolics in grass pea ranged from 868 mg/kg DM (CE) for L880388 to 2059 mg/kg DM (CE) for LS89110. Condensed tannins in grass pea ranged from 0.89 g/kg DM (CE) for L880388 to 5.18 g/kg DM (CE) for LS89125. Cultivar had a larger relative contribution to total phenolic levels in field pea and to total phenolic and condensed tannin levels in grass pea than environment. Total phenolic and condensed tannin levels were not correlated with seed yield and seed protein content in field pea or grass pea. Levels of total phenolics and condensed tannins were positively correlated in grass pea. Grass pea seeds with darker seed coat colour contained higher levels of condensed tannins. This revised version was published online in July 2006 with corrections to the Cover Date.     

Impact of heat stress on pod-based yield components in field pea (Pisum sativum L.)

Elevated temperatures associated with climate change result in crops being exposed to frequent spells of heat stress. Heat stress results in reduced yield in field pea (Pisum sativum L.); it is therefore important to identify cultivars with improved pod and seed retention under heat to mitigate this loss. Objectives were to investigate the effect of heat stress on phenology, yield and pod-based yield components. Sixteen pea cultivars were evaluated at normal and late (hot) seeding dates in the field in Arizona 2012 and in growth chambers with two temperature regimes (24/18°C and 35/18°C day/night temperature for 7 days) during reproductive development. We measured variation in the pattern of pod retention at four-node positions on plants, seed retention by ovule position (stylar, medial and basal) within pods and screened cultivars for pod retention, seed retention and yield. Heat stress reduced seed yield by accelerating the crop lifecycle and reducing pod number and seed size. Heat stress had the most damaging effect on younger reproductive growth (flowers and pods developed later), resulting in ovary abortion from developing flowers. Heat also accelerated seed abortion in all ovule positions within pods. Two high-yielding cultivars under control temperature, “Naparnyk” and “CDC Meadow”, maintained high yield in heat, and “MFR043” had the lowest yield. Cultivars “40-10” and “Naparnyk” retained the most ovules and seeds per pod, and “MFR043” aborted seeds when exposed to heat. In half of the cultivars, ovules at the basal peduncle end of pods were likely to abort while ovules at the medial and stylar end positions developed into seeds. For seven of the field cultivars, ovules at the medial pod position also produced mature seeds. Cultivars “40-10”, “Naparnyk” and “CDC Meadow” had greater pod and ovule retention or maintained high yield under heat stress, and were identified as heat-tolerant cultivars. Our results allow for a better understanding of pod-based yield components in field pea under heat stress and developing heat-tolerant cultivars.     

Inheritance of resistance to pea blight (Ascochyta pinodella) and induction of resistance in pea (Pisum sativum L.)

Summary Pea blight caused by Assochyta pinodella does considerable damage to the pea crop every year. To ascertain the inheritance of resistance to pea blight and incorporate resistance in the commercial cultivars, crosses were made between Kinnauri resistant to pea blight and four highly susceptible commercial pea cultivars — Bonneville, Lincoln, GC 141 and Sel. 18. Studies of the F₁'s, F₂'s, back crosses and F₃'s indicated that Kinnauri carries a dominant gene imparting resistance to pea blight.     

Maintenance breeding and multiplication of pea and faba bean cultivars. Maintenance of protein peas (Pisum sativum) and field beans (Vicia faba)

Summary The maintenance of protein pea (Pisum sativum)and field bean (Vicia faba)cultivars is a part of the breedingscheme of these pulses at CEBECO-ZADEN. As soon as the cultivar is pure breeding a large quantity ofseed is stored under conditioned circumstances.     

Factors affecting in vitro flowering and fruiting of green pea (Pisum sativum L.)

Multiple shoots were efficiently regenerated from cotyledonary node and shoot tip explants of Pisum sativum within 15 days on MS medium containing B₅ vitamins and supplelmented with 2.0 mgl^-1 6-benzylaminopurine. The elongated shoots produced on the same medium were excised and transferred to MS medium containing half strength ammonium nitrate (8.25 gml^-1) and supplemented with auxins (indole-3-butyric acid or naphthalene acetic acid) either alone or in combinations with gibberellic acid. Rooting and flowering were observed on the 7th and 15th day after their transfer to rooting medium. The flowers self-fertilised in vitro and produced mature pods within 25 days of rooting. These seeds were germinable both in vitro and in vivo. In vitro seeds sown in pots under field conditions developed into flowering plants, and subsequently produced pods with viable seeds. This revised version was published online in July 2006 with corrections to the Cover Date.     

Study of the variation in the somatic embryogenesis ability of some pea lines (Pisum sativum L. and Pisum arvense L.)

Summary Thirty lines of pea were tested in vitro to evaluate their ability to produce somatic embryos. Three distinct genotypic classes were detected (strong, medium and weak). The best responses were obtained in Pisum sativum. Abnormal somatic embryos and secondary embryogenesis seem to constitute the principal obstacle to the development of these structures.     

Response of pea (Pisum sativum L.) to mepiquat chloride under varying application doses and stages

Grown as a monoculture, peas (Pisum sativum L.) exhibit severe lodging after flowering and lodging causes yield reductions considerable. This study was conducted to investigate the effects of dose (untreated, 25, 50, 75 and 100 g a.i. ha^?1) and stage (late vegetative, early blooming and early pod filling) of mepiquat chloride (MC) application on the growth, lodging control, seed yield and yield parameters of pea (Pisum sativum L.) under field conditions in Erzurum, Turkey in 2002 and 2003. Application doses of 25, 50, 75 and 100 g a.i. ha^?1 significantly reduced stem height by 5.3 %, 7.2 %, 7.5 % and 6.4 % and increased stem width by 7.5 %, 12.7 %, 12.3 % and 15.7 % respectively, when compared with the untreated control, and thereby reduced the tendency of the crop to lodging. Increases of the seed yield under different application doses of MC ranged between 13.7 % and 20.1 % over the untreated control. However, in all parameters investigated, except for stem width, higher application doses of MC gave no clear advantages compared with the application dose of 25 g a.i. ha^?1. Seed yield was also significantly influenced by application stage of MC and application at early blooming stage of crop, MC significantly increased seed yield by 11.4 % and 10.2 % when compared with the late vegetative and the early pod filling stages respectively. Furthermore, the interaction of application dose and stage was significant, and spraying of pea plants with 25 g a.i. ha^?1 MC at early blooming stage has the most beneficial effects on the characters evaluated.     

Genetic variability in tolerance to cadmium and accumulation of heavy metals in pea (Pisum sativum L.)

Ninety-nine wild growing and primitive varieties of garden pea (Pisum sativum L.) were screened for tolerance to cadmium (Cd) toxicity in quartz sand culture. Cadmium tolerance was determined by the time to plant death when treated with a lethal Cd concentration (13 mg kg^-1), and by a tolerance index (TI) calculated as a ratio between biomasses of Cd-treated and untreated plants in the presence of toxic Cd concentrations (7 mg kg^-1 and 5 mg kg^-1). The Cd-tolerance index varied significantly between pea genotypes from 35% to 90% and from 54% to 100% in the presence of 7 and5 mg Cd kg^-1, respectively. Shoot Cd concentration of tolerant and sensitive genotypes grown in the presence of 5 mg Cdkg^-1 varied between 35 mg Cd kg^-1and 135 mg Cd kg^-1 (dry weight) and was negatively correlated with TI. Certain tolerant pea genotypes were characterized by a high Cd concentration in shoots. All varieties were also screened for their ability to take up heavy metals (HMs) from a slightly contaminated soil. The concentration of Cd, chromium, copper, nickel, lead, strontium and zinc in plant shoots varied between pea genotypes by a factor of 2.8, 4.9, 2.7, 3.5, 9.7, 3.9 and 4.0, respectively. The coefficients of variation between pea genotypes for HM concentration were high, varying from 23%to 39% depending on the metal. The distribution patterns for varieties based on Cd tolerance (sand culture) and HM concentrations (soil culture) were characterised by positive skewness coefficients, suggesting that the majority of pea genotypes was relatively sensitive to Cd toxicity and tended to avoid excessive accumulation of HMs in shoots. These results show that a high genetic variability exists in pea with regards to Cd tolerance and HM accumulation. Concentrations of different HMs in plants grown in soil correlated positively with each other, with the exception of chromium. There was no correlation between Cd tolerance of the varieties in sand culture, shoot concentration of HMs in soil culture, biomass production, subspecies and geographical origin of the varieties. The genetic systems controlling Cd tolerance, HM accumulation and morphological traits are therefore independent to some extent, suggesting a possibility for breeding pea cultivars characterised by high tolerance to and low concentration of HMs in shoots. This revised version was published online in August 2006 with corrections to the Cover Date.     

The productivity of some mutants of the pea (Pisum sativum L.) and their hybrids a contribution to the heterosis problem in self-fertilizing species

Summary The favourable yielding capacity of a group of X-ray inducedPisum sativum mutants makes them attractive for pea breeding. The fasciated mutant 489C produced as much as 32–92% more seed than the corresponding values of the initial line during the past 4 years. Mutants 68C and 1201A cannot exploit their favourable properties. After incorporation in different genetic backgrounds the genes in question can be expected to induce better productivity. The small-grained mutant 176A having a very good seed set could be of interest to canning industry. The early flowering mutant 46 cannot be used in practical breeding because of its heterogeneous yield properties in subsequent generations.Hybrids between these mutants were made in order to combine their useful characters. The productivity of these hybrids and their parental lines was studied and compared with the initial line. Hybrids between 176A × 68C and 1201A × 68C did not show heterosis for any characters. Hybrids of all mutants with the fasciated 489C, however, showed a marked heterosis effect for increased vigour and stem length resulting in a conspiciously high seed production.Supported by the Ministry of Scientific Research of our Government and by the Association EURATOM-ITAL in Wageningen, The Netherlands.     

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.     

Lodging and Yield of Dry Peas (Pisum sativum L.) as Influenced by Various Mixing Ratios of a Conventional and a Semi-Leafless Cultivar

Lodging is a serious problem in dry pea production in North-West Europe. Therefore, lodging and its effects on yields were studied in five field-experiments with two cultivars with two different canopy structures, both as single crop and in various mixing weight ratios. In 1985, mechanically simulated lodging decreased yield by about 45% in both the conventional type cv. Rondo and the semi-leafless type cv. Solara , by reducing both the number of seeds and seed weight. Natural lodging brought about the same effect in 'Rondo' but in 'Solara' , however, effects were less because of delayed lodging. In 1986, 1987 and 1988, mixtures of the conventional cv. Finale and Solara showed delayed lodging compared to 'Finale' single and tended to produce higher yields than the single crops of both cultivars. Polynomial analysis of the yields of all mixing experiments showed maximum yields at 53% 'Solara' (quadratic analysis) or 67% 'Solara' (cubic analysis). Thus, mixing high yielding, lodging sensitive conventional cultivars with lower yielding but lodging resistant semi-leafless cultivars may improve yield in dry peas.     

水杨酸浸种对豌豆种子萌发及幼苗生长的影响

研究了水扬酸(SA)浸种对豌豆种子萌发及幼苗生长的影响。结果表明:10~40 mg/L的SA浸种,发芽势、发芽率显著提高;40 mg/L的处理,增幅分别为对照的12%和10%;鲜重和干重分别达到对照的120%和132%;20 mg/L的处理,萌动豌豆种子的呼吸速率和幼苗的根系活力均极显著高于对照,当处理浓度超过60 mg/L时,电导率为对照的128%,相关生理指标下降,幼苗生长受到一定抑制。试验证明,40 mg/L的水杨酸处理,能改善豌豆幼苗对N和K的吸收,但SA不利于幼苗对P的吸收。     

The effect of temperature during growth and flowering on the seed set of tetraploid freesia

Summary Phytotron experiments have been carried out to study the effect of different temperatures on the seed set of tetraploid freesias during the raising of the plants and at the time of flowering and pollination.The effect of a relatively low temperature during the growth period is reflected in a comperatively greater number of flowers as a result of an increase in number of ramifications of the peduncle. This sometimes promotes the seed yield notwithstanding that the lower temperature reduces the fertility of each seperate flower. It is clearly proved that the temperature during flowering and pollination affects the fertility of the flower and the ultimate seed yield of the plant. Experiments in our phytotron showed 14°C during the growth period and 20°C at the time of pollination to be the optimum combination for a good seed yield.The effect of a lower night temperature was generally favourable, particularly so when measured by the number of seeds per pollinated flower.     

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.     

Developmental abnormalities in the ovule and embryo sac of ulluco (Ullucus tuberosus) and their effect on seed set

Summary Ulluco (Ullucus tuberosus) is a vegetatively propagated crop plant that flowers abundantly but sets only few seeds. We examined if the low seed/ovule ratio can be explained by morphological abnormalities in ovules, embryo sacs or embryos. In 35 clones growing in the actual area of cultivation in Ecuador and in 5 clones grown in Finland flowers were analysed by clearing technique. About half of all analysed clones contained ovules and embryo sacs whose development was arrested, in two clones there were embryo sacs whose cell patterns were abnormal and in four clones there were double embryo sacs. There were both normal and abnormal ovules and embryo sacs in the same plant. Genetic load is suggested to be one of the causes for the low sexual fertility found in ulluco. It may be incapable of maintaining homeostasis of development of the ovule and embryo sac in a varying environment. The occurrence of developmental abnormalities should be studied in each clone intended for inclusion in traditional breeding programmes.