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.     

Novel quantitative trait loci for yield and yield related traits identified in Basmati rice (Oryza sativa)

Increasing crop productivity is one of the prime goals of crop breeding research. Rice grain yield is a complex quantitative trait governed by polygenes. Although several QTLs governing grain yield traits have been reported and limited attempts have been made to map QTLs for grain yield parameters in Basmati rice. A population from the cross Sonasal and Pusa Basmati 1121 comprising 352 RILs was generated through the single seed descent method. A total of 12 QTLs governing yield and yield-related traits were mapped on six chromosomes, namely, 1, 2, 3, 7, 8 and 9, of which five QTLs were novel. We identified a novel and robust epistatic QTL (qPH1.1 and qPL1.1) governing plant height and panicle length, flanked by the markers RM5336-RM1 on chromosome 1. The gene encoding brassinosteroid insensitive 1-associated receptor kinase 1 precursor is the putative candidate gene underlying this epistatic QTL. Another novel QTL, qNT3.1, governing tiller number was bracketed to a region of .77 Mb between the markers RM15247 and RM15281 on chromosome 3. Of the 57 annotated gene models, Os03g0437600 encoding alpha/beta-fold hydrolase, a homologous to AtKai2 is a putative candidate gene underlying the novel QTL qNT3.1. The other QTLs such as qDFF1.1 governing days to 50% flowering co-localizes with the gene Ghd7, QTL for plant height qPH1.2 co-localizes with the gene sd1, the QTLs for panicle length co-localizes with FUWA and DEP2, the QTL for tiller number co-localizes with OsRLCK57 and QTLs for thousand-grain weight co-localize with the major gene GS3. The QTLs identified in the current study can be effectively used in marker-assisted selection for developing Basmati rice varieties with a higher yield.     

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.     

Identification and validation of quantitative trait loci associated with seed yield in soybean

In soybean [Glycine max (L.) Merrill], the genetic analysis of seed yield is important to aid in the breeding of high-yielding cultivars. Seed yield is a complex trait, and the number of quantitative trait loci (QTL) involved in seed yield is high. The aims of this study were to identify QTL associated with seed yield and validate their effects on seed yield using near-isogenic lines. The QTL analysis was conducted using a recombinant inbred line population derived from a cross between Japanese cultivars ‘Toyoharuka’ and ‘Toyomusume’, and eight seed yield-associated QTL were identified. There were significant positive correlations between seed yield and the number of favorable alleles at QTL associated with seed yield in the recombinant inbred lines for three years. The effects of qSY8-1, a QTL promoting greater seed yield, was validated in the Toyoharuka background. In a two-year yield trial, the 100-seed weight and seed yield of Toyoharuka-NIL, the near-isogenic line having the Toyomusume allele at qSY8-1, were significantly greater than those of Toyoharuka (106% and 107%, respectively) without any change for days to flowering and maturity. Our results suggest that qSY8-1 was not associated with maturity genes, and contributed to the 100-seed weight.     

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.     

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.     

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     

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.     

Mapping of quantitative trait loci controlling seed longevity of rice (Oryza sativa L.) after various periods of seed storage

Seed longevity varies considerably in cultivated rice (Oryza sativa L.), but the underlying genetic mechanism of longevity has not been well elucidated. Quantitative trait loci (QTL) that control seed longevity after various periods of seed storage were sought using recombinant inbred lines derived from a combination involving ‘Milyang23’(Indica‐type) and ‘Akihikari’ (Japonica‐type). In all, 12 QTLs for germination and normal seedling growth were detected as indices of seed longevity on chromosome 7 (one region) and chromosome 9 (two regions) in treated seeds that had been stored under laboratory conditions for 1, 2 or 3 years.‘Milyang23’ alleles of all QTLs promoted germination and normal seedling growth after all durations of storage. These QTL regions were detected repeatedly in more than one seed condition. Therefore, we infer that these regions control seed longevity.     

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.     

Identification of quantitative trait loci (QTLs) for seed protein concentration in soybean and analysis for additive effects and epistatic effects of QTLs under multiple environments

Soybean protein concentration is a key trait driver of successful soybean quality. A recombination inbred lines derived from a cross between ‘Charleston’ and ‘Dongnong594’, were planted in three environments across four years in China. Then, the genetic effects were partitioned into additive main effects, epistatic main effects and their environment interaction effects by using composite interval mapping, multiple interval mapping and composite interval mapping in a mixed linear model. Forty‐three quantitative trait loci QTLs were identified on 17 of 20 soybean chromosomes excluding Ch 7, Ch 8 and Ch 17. Two QTLs showed a good stability across multiple environments, qPRO20‐1 was detected under four environments, which explained 4.4–9.95% phenotypic variances and the allele was from ‘Charleston’ among four environments. qPRO7‐5 was detected under three environments, which explained 7.2–14.5% phenotypic variances and the allele was from ‘Dongnong 594’, three pathway genes of protein biosynthesis were detected in the interval of qPRO7‐5. The additive main‐effect QTLs contributed more phenotypic variation than the epistasis and environmental interaction. This indicated that it is feasible by marker‐assisted selection to improve soybean protein concentration.     

Genetic mapping of quantitative trait loci in rye (Secale cereale L.)

Using the marker information of 275 F₂ plants quantitative traits determining morphological and yield characters were studied analyzing F₃progenies grown in four different experiments at three sites. The map constructed contains 113 markers including the major dwarfing gene Ddw1 with an average distance of about 10 cM between adjacent markers. Of the 21 QTLs detected ten were found to map on chromosome 5RL in the region of Ddw1. Beside the expected effects on plant height and peduncle length that are most probably due to the presence of the major dwarfing gene, additional effects on yield characters and flowering time were discovered in that region which may be caused by pleiotropic effects of Ddw1. An additional supposed gene cluster consisting of four QTLs controlling flowering time and yield components was discovered in the centromere region of chromosome 2R. Further loci are distributed on chromosomes 1R (1), 4R (1) 6R (3) and 7R (1). The map positions of the quantitative trait loci detected in rye are discussed in relation to major genes or QTLs determining agronomically important traits in other cereals. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Tagging quantitative trait loci associated with grain yield and root morphological traits in rice (Oryza sativa L.) under contrasting moisture regimes

Plant breeding for drought-prone habitats envisages a favorable combination of grain yield and drought resistance. Though several components enhancing drought resistance have been identified in rice,their association with grain yield, under low-moisture stress, has been established in very few instances. We attempt to study the associations between rice grain yield and root system parameters both at phenotypic and genotypic levels. The doubled haploid population of IR64/Azucena was evaluated for root related traits at peak vegetative stage and grain yield related traits under both low-moisture stress and non stress conditions. ‘Mean environment’ was computed for yield related traits. Correlation and QTL mapping was attempted to find out the associations. The correlation between maximum root length and grain yield was positive under stress and negative in non stress. Genotypes with thicker and deeper roots, manifested higher biomass and grain yield under stress. Only one QTL found to increase days to flowering in non stress was also found to influence root volume and dry weight negatively under stress. The study suggests that loci enhancing grain yield and related traits were not pleiotropic with loci for desirable root morphological traits studied under low-moisture stress at vegetative stage, in the genetic material used in the study. It is thus possible to combine higher grain yield and desirable root morphological traits, favorably, to enhance productivity of rice under low-moisture stress. In rainfed ecologies, where deep roots contribute to enhanced drought resistance in rice, the results indicate the possibility of combining drought resistance with higher levels of grain yield. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetics of tolerance to high concentrations of soil boron in peas (Pisum sativum L.)

Summary The inheritance of tolerance to high concentrations of soil boron in pea (Pisum sativum L.) was studied in five cross combinations and their reciprocals. Segregation patterns for boron response in F₂ populations and F₃ derived families were established by visual assessment of leaf damage. The segregation ratios were explained in terms of two major gene loci interacting in an additive manner with incomplete dominance at each locus. Evaluation of selected tolerant and susceptible families indicated that tolerant families contained a significantly lower concentration of boron in shoots than susceptible families.     

Identification and confirmation of quantitative trait loci for stachyose content in soybean seed

Stachyose is an unfavorable sugar in soybean meal that causes flatulence for non‐ruminant animals. Understanding the genetic control of stachyose in soybean will facilitate the modification of stachyose content at the molecular level. The objective of this study was to identify quantitative trait loci (QTL) associated with seed stachyose content using simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers. A normal stachyose cultivar, ‘Osage’, was crossed with a low stachyose line, V99‐5089, to develop a QTL mapping population. Two parents were screened with 33 SSR and 37 SNP markers randomly distributed on chromosome 10, and 20 SSR and 19 SNP markers surrounding a previously reported stachyose QTL region on chromosome 11. Of these, 5 SSR and 16 SNP markers were used to screen the F_3:4 lines derived from ‘Osage’ x V99‐5089. Seed samples from F_3:5 and F_3:6 lines were analyzed for stachyose content using high‐performance liquid chromatography (HPLC). Composite interval mapping analysis indicated that two stachyose QTL were mapped to chromosome 10 and 11, explaining 11% and 79% of phenotypic variation for stachyose content, respectively. The SSR/SNP markers linked to stachyose QTL could be used in breeding soybean lines with desired stachyose contents. Chi‐square tests further indicated that these two QTL probably represent two independent genes for stachyose content. Therefore, a major QTL was confirmed on chromosome 11 and a novel QTL was found on chromosome 10 for stachyose content.     

Detection of quantitative trait loci in Pinus sylvestris L. across years

In an earlier study, we reported the characterisation of quantitative trait loci (QTLs) for economically important traits related to wood production for a specific year of measurement. However, validating the detected QTLs across years is important for any strategy for marker-assisted selection (MAS). Therefore, we evaluated the consistency of the QTLs across four years in 9 to 12 year-old trees. Data related to tree height were analysed in two ways. The total height was used to characterise QTLs likely to be useful in MAS, and the increment values to evaluate growth regulation. Among the 11 QTLs detected, 4 were found in both the 1996 and 1997 evaluations. QTL numbers and locations related to total height were found to be highly stable. However, the QTL effects progressively changed in successive years, following either an increasing, decreasing or curvilinear trend. Depending on their specific trends, some QTLs may not be expressed at all later in maturity, or they may be expressed at a higher level. This will have direct consequences on the applicability of molecular markers in early screening programs, since a study at the mature stage may also be required. More variation in QTL number and effect was observed relating to height increment than to total height, indicating significant differences in gene expression during the growth periods involved. This revised version was published online in August 2006 with corrections to the Cover Date.