Response to Mycosphaerella pinodes in a germplasm collection of Pisum spp

Little resistance against Mycosphaerella pinodes is available in pea. In this work 78 accessions of Pisum were screened for resistance to M. pinodes. Fourteen accessions showed a good level of resistance in seedlings under controlled conditions, and in mature plants in the field. The highest levels of resistance were found in P. fuivum, followed by P. sativum ssp. eiatius and P. sativum ssp. syriacum. Resistance of five selected accessions was effective against different isolates of M. pinodes originating from different countries. Resistant accessions reported in this paper have been successfully hybridized with field pea 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.     

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.     

Current status and future strategy in breeding pea to improve resistance to biotic and abiotic stresses

The economic importance and current progress made in studies of the host-parasite relationship and identification of sources of resistance and breeding strategies of some important biotic diseases of pea are reviewed in this paper. The root rot complex caused by Rhizoctonia solani, Fusarium solani, Aphanomyces euteiches, Pythium ultimum and Fusarium oxysporum f. sp. pisi, race 1 and 2 has been reported from all commercial pea growing areas of the world. Adequate sources of resistance have been identified and there has been impressive success in the control of the Fusarium wilt pathogen following the introduction of wilt-resistant cultivars. Leaf and stem diseases of pea caused by the Ascochyta complex, Peronospora viciae and Erysiphe pisi are prevalent in most temperate pea growing regions of the world. Several sources of resistance are available, some of which are surprisingly durable. The biochemical genetic parameters of phenolic content used for assaying resistance to Erysiphe pisi offers an alternative method of evaluating breeding material. Wild relatives of pea (Pisum fulvum and P. humile) are valuable additional sources of genetic variation and provide good sources of resistance to pests and diseases. In temperate rainfed pea growing areas of southern Australia, pea seed yield is more closely related to dry matter production than harvest index. Tall and leafy cultivars proved more productive than afila types.     

Genetic analysis of adult plant resistance to powdery mildew in pea (Pisum sativum L.)

Summary An analysis of adult plant resistance of powdery mildew in 15 F₁, F₂ and F₃ populations of pea derived from crossing 15 diverse and susceptible lines with one resistant line revealed that resistance to powdery mildew is controlled by duplicate recessive genes. The genes were designated as er₁ and er₂.Disease reaction showed independent segregation with three known markers in the resistant parent, namely, af (afila, chromosome 1), st (stipule reduced, chromosome 3) and tl (clavicula, chromosome 7).Contribution form the Department of Genetics and Plant Breeding Banaras Hindu University, Varanasi-221005, India.     

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.     

Genetics of powdery mildew resistance in pea

The genetics of powdery mildew (Erysiphe pisi) resistance in pea (Pisum sativum) was studied using five isolates of E. pisi in a controlled environment. Resistance was controlled by a single recessive gene in all the pea cvs. included in the study. The same recessive gene controlled resistance to all the five isolates of E. pisi. Tests for allelism showed that resistance in cvs. HPPC-63, HPPC-95, DPP-26, DPP-54, Mexique-4, SVP-950, Wisconsin-7104 and P-6588 is conferred by the same recessive gene. This revised version was published online in July 2006 with corrections to the Cover Date.     

Somatic embryogenesis in pea (Pisum sativum L. and Pisum arvense L.): Diallel analysis and genetic control

Summary Six lines of Pisum were tested in vitro for their ability to produce somatic embryos from apices. Significant quantitative variation was observed. Inheritance of the ability to form somatic embryos was studied using a diallel cross among six different lines. About 80% of the observed genotypic variation was due to additive effects. There is a tendency for the favourable genes to be recessive. It appears that there are two genetic systems involved. Analysis of the distribution of F₃ families means from a cross among two extreme lines seems to indicate the presence of a few major genes in the control of somatic embryogenesis of pea.     

The response of Pisum sativum L. germplasm to high concentrations of soil boron

Summary Tolerance to high levels of boron in the soil is an important aspect of the adaptation of crop varieties to southern Australian conditions. This paper reports investigations aimed at exploring the extent of genetic variation in Pisum sativum and at defining appropriate selection criteria for selection for boron tolerance in breeding programs.A collection of 617 accessions of Pisum was screened in controlled conditions and visually assessed for symptoms of boron toxicity. A high proportion of accessions were sensitive with only 3.5% being more tolerant than any of the Australian varieties. Relatively high proportions of tolerant and moderately tolerant accessions originated from Asia and South America.In a second experiment the responses of selected tolerant accessions were evaluated with respect to different parameters. The objectives were to confirm the performance of the putative boron tolerant accessions and identify appropriate parameters for selecting boron tolerant genotypes. In addition to the visual assessment of boron toxicity, measurements at the time of harvest included dry matter yield and concentration of boron in tissues. Symptom expression was highly correlated with dry matter yield and concentration of boron in tissues under high boron conditions and so could be used as a non-destructive selection criteria. A low degree of symptom expression by tolerant accessions could usually be attributed to low levels of boron in the vegetative tissues. The results of this study indicate that considerable genetic variation exists among exotic accessions of Pisum sativum and tolerance to boron could be transferred to sensitive varieties.     

Heterosis for yield and related characters in pea

Summary To determine the levels of heterosis in F1 hybrids, four current pea (Pisum sativum L.) cultivars from southern Australia were used as female parents and crossed with 18 introduced genotypes. The 22 parents, 72 F1 hybrids and, depending on the environment, either 54 or all 72 F2 families were grown in replicated plots in four environments. Grain yield, total dry matter, harvest index, branches per plant, pods per plant, seeds per pod, hundred seed weight, plant height, onset of flowering and flowering periods were evaluated. For both the F1 and F2 generation, heterosis was determined as the superiority over the mid-parent and also over the better parent. In addition, the superiority over the best commercial cultivar was calculated. Most hybrids were higher yielding than their mid-parent but were less stable in yield across environments. Four F1 hybrids were significantly higher yielding than the best parent, by up to 26%. There were significant correlations between F1 hybrid and mid-parent value for plant height, pods per plant and hundred seed weight but not for yield. Overall, grain yield heterosis was mainly due to more pods per plant in the hybrids. The level of heterosis for yield in a poor yielding environment was higher than that in a high yielding one. Both additive and non-additive gene effects were important in the expression of all studied traits. The average level of heterosis for grain yield and total dry matter in the F2 population was half of that in F1 hybrids. The low level of inbreeding depression from the F1 to the F2 generation suggested that epistatic gene action also contributed to the expression of grain yield. Some F2 populations maintained the high yield levels of the corresponding F1 hybrids.     

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.     

Genetic male sterility in the pea (Pisum sativum L.)

Summary Genetic male sterility has been described in the pea (Pisum sativum L.), but no comprehensive effort has been made to study the phenomenon. A preceding companion paper reported the inheritance, allelism and linkage relations of thirteen male sterile mutants obtained from seed mutagen treatments. In the present study, the same male sterile mutants were investigated cytologically to determine the cause of sterility. Normal microsporogenesis and microgametogenesis was compared to that of the mutants. The ms-2, ms-3, and ms-4 mutants exhibited meiotic abnormalities similar to those described by Gottschalk and colleagues except that ms-3 had a high degree of female sterility. Chromosome clumping and spindle abnormalities leading to formation of coenocytic microspores and degeneration were characteristic of ms-2 and ms-4. The ms-2 and ms-4 mutants were previously found to be allelic, and were nearly identical cytologically in the present study. The ms-3 mutant exhibited a lack of chromosome condensation in meiosis I, and a lack of spindle formation in both meiotic divisions. Two mutations (ms-6 and ms-10) affected meiosis, with univalents at metaphase, and asynchronous divisions during meiosis II. Microspores of ms-6 completely degenerated whereas those of ms-10 showed some development. Sticky chromosomes, bridges and fragments, tripolar spindles, and lack of a second division were characteristic of ms-10. The ms-10 mutant also showed reduced female fertility. Two male steriles (ms-5 and ms-9) had abnormalities associated with premature degeneration of the tapetum. Three others (ms-7, ms-8, and ms-11) aborted pollen during meicrogametogenesis. Pollen grains of ms-11 had thinner walls than normal and lacked sculptured exine. The ms-10 mutant, and those affecting microgametogenesis (ms-5, ms-7, ms-8, ms-9, and ms-11) produced some stainable and viable pollen.     

Analysis of tissue culture-derived variation in pea (Pisum sativum L.)-preliminary results

Summary The possibility of producing agronomically-useful somaclones via organogenesis and somatic embryogenesis from callus cultures of pea (Pisum sativum L.) was studied. Organogenic calli were induced from immature leaflets on MSB medium with NAA and BAP. Embryogenic calli were derived either from immature zygotic embryos (using 2,4-D) or from shoot apices (using picloram) of aseptically-germinated seedlings.The seed progenies (T₁ to T₃-generation) of primary regenerants were grown in field conditions and their phenotypic variation was evaluated and compared with control, non-tissue culture-derived plant material. In addition, electrophoretic analyses of selected isoenzyme systems and total proteins have been done. The results do not show dramatic changes in qualitative and quantitative traits. The evaluation of at least two future generations (T₄, T₅) is planned.Abbreviations BAP 6-benzylaminopurine - IBA indole-3-butyric acid - MSB medium (mineral salts after Murashige & Skoog, 1962, vitamins after Gamborg et al., 1968) - NAA -naphthalene-acetic acid, picloram-4-amino-3,5,6-trichloro picolinic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - ORG organogenesis - SE somatic embryogenesis     

Selection for partial resistance to downy mildew in peas by means of greenhouse tests

Summary A partially downy mildew resistant pea line was back-crossed to a susceptible cultivar with more pods per node and lower seed weight. Breeding lines with different degrees of infection in a greenhouse test were selected. These lines and the two parental lines were investigated in field trials and tested in the greenhouse for four generations. Significant genetic variation among lines was found for infection of seedlings in greenhouse tests, and infection of pods, pod set and seed weight in field-trials. Infection of seedlings in the greenhouse was correlated with infection of pods in the field. In greenhouse tests, the non-genetic variance component was large in comparison with the genetic component and a significant genotype trial interaction was found. Significant repeatability was obtained for downy mildew on seedlings and pods, number of pods per node and seed weight. An unfavourable correlation between susceptibility to downy mildew and number of pods per node was found. No single breeding line showed the ideal combination of good resistance, high number of pods per node and small seeds. However, one line showing better resistance than the susceptible parent, with smaller seeds and more pods per node than the resistant parent was found. The susceptible parent also carries some resistance factor that is not present in the resistant parent.     

Molecular mapping of quantitative trait loci determining resistance to septoria tritici blotch caused by Mycosphaerella graminicola in wheat

A set of 65 recombinant inbred lines of the ‘International Triticeae Mapping Initiative’ mapping population (‘W7984’בOpata 85’) was analysed for resistance to septoria tritici blotch at the seedling and adult plant stages. The mapping population was inoculated with two Argentinean isolates (IPO 92067 and IPO 93014). At the seedling stage, three loci were discovered on the short arms of chromosomes 1D, 2D and 6B. All three loci were detected with both isolates. At the adult plant stage, two isolate-specific QTL were found. The loci specific for isolates IPO 92067 and IPO 93014 were mapped on the long arms of chromosomes 3D and 7B, respectively. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identification of RAPD markers linked to the rust (<Emphasis Type="Italic">Uromyces fabae</Emphasis>) resistance gene in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>)

Summary Two RAPD markers linked to gene for resistance (assayed as pustule number cm⁻² leaf area) to rust [Uromyces fabae (Pers.) de Bary] in pea (Pisum sativum L.) were identified using a mapping population of 31 BC₁F₁ [HUVP 1 (HUVP 1 × FC 1] plants, FC 1 being the resistant parent. The analysis of genetics of rust resistance was based on the parents, F₁, F₂, BC₁F₁ and BC₁F₂ generations. Rust resistance in pea is of non-hypersensitive type; it appeared to be governed by a single partially dominant gene for which symbol Ruf is proposed. Further, this trait seems to be affected by some polygenes in addition to the proposed oligogene Ruf. A total of 614 decamer primers were used to survey the parental polymorphism with regard to DNA amplification by polymerase chain reaction. The primers that amplified polymorphic bands present in the resistant parent (FC 1) were used for bulked segregant analysis. Those markers that amplified consistently and differentially in the resistant and susceptible bulks were separately tested with the 31 BC₁F₁ individuals. Two RAPD makers, viz., SC10-82₃₆₀ (primer, GCCGTGAAGT), and SCRI-71₁₀₀₀ (primer, GTGGCGTAGT), flanking the rust resistance gene (Ruf) with a distance of 10.8 cM (0.097 rF and LOD of 5.05) and 24.5 cM (0.194 rF and a LOD of 2.72), respectively, were identified. These RAPD markers were not close enough to Ruf to allow a dependable maker-assisted selection for rust resistance. However, if the two makers flanking Ruf were used together, the effectiveness of MAS would be improved considerably.     

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 first record of tetrasomy in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

Tetrasomic plants with two additional small chromosomes were identified, with a frequency about 2.5%, in the trisomic pea line TRUST-R, which normally contains one extra chromosome covering a sporophyte lethal in the regular chromosome set. As compared to trisomics, tetrasomics exhibited an enhanced expression of the traits resulting from extra chromosome addition: slow growth, enlarged bracts, shortened peduncles, wavy leaflets and stipulae. They were almost sterile, their pollen contained a variable proportion of empty grains and some anomalously large, small or deformed grains. In metaphase I, two extra chromosomes did not form a stable bivalent and only in some cases were situated close to each other. In anaphase I, the extra chromosomes migrated independently to either pole or retarded in the equatorial plain, the same was observed for chromatids in anaphase II. This retardation resulted in anomalous cytokinesis, so that triads, dyads and half-divided or non-divided monads appeared. The retarded extra chromosomes may form small extra nuclei either included into one of the microspores or forming a separate miniature cell; in this way tetrasomics may eliminate extra chromosomes. One of the tetrasomics analysed formed an exceptionally high proportion of microspore pentads. In the regluar TRUST-R trisomics, the sole extra chromosome retarded in the equatorial plain in anaphases I and II. The retardation in anaphase II often makes cytokinesis in trisomics (in general more regular than in tetrasomic) to proceed in two steps: at first cell wall formation separates a pollen mother cell into two dyads and then each of them into two microspores.