ABA content in shoots and roots of pea mutants af and tl as related to their growth and morphogenesis

The comparative study of shoot and root growth was carried out, and the level of ABA therein determined in the mutant af and tl and wild-type isogenic lines of pea. The recessive af mutation transformed the leaflets into tendrils, and the tl mutation transformed the tendrils into leaflets. These mutations did not affect the length and number of internodes. In all plants, the level of ABA in the leaves was 3–10 times greater than in the roots, and in the course of vegetative growth it rose in both organs. An increase in the shoot area of tl mutant did not change the dry weight of underground and above-ground parts; therefore, the ratio shoot/root in the mutant was identical to that in the wild-type plants. The maintenance of shoot dry weight in the tl mutant at the level of wild-type plant while its area considerably increased was accounted for by a decrease in the thickness of the leaflet and stipule blades. The level of ABA in the stipules of mutant plants was greater than in the wild-type plants. A decrease in the shoot area in the af mutant brought about a decline in its dry weight; however, the ratio root/shoot was maintained at the wild-type level due to a reduced accumulation of dry weight by the root. The level of ABA in the roots of the af mutant was twice greater than in the leafy forms. ABA was assumed to participate in the control over the root growth exerted by the shoot. The absence of leaflets in the af plants was partially compensated for by expanding stipules. The level of ABA therein was three times higher than in the plants of wild type and comparable with the level in the leaflets of the tl mutant and in the wild-type plants. The role of ABA in the growth and final size of leaf blades is discussed.     

The rates of shoot and root growth in intact plants of pea mutants in leaf morphology

Isogenic lines of pea (Pisum sativum L.) with the genetically determined changes in leaf morphology, afila (af) and tendril-less (tl), were used to study the relationship between shoot and root growth rates. The time-course of shoot and root growth was followed during the pre-floral period in the intact plants grown under similar conditions. The af mutation produced afila leaves without leaflets, whereas in the case of the tl mutations, tendrils were substituted with leaflets, and acacia-like leaves were developed. Due to the changes in leaf morphology caused by these mutations, pea genotypes differed in leaf area: starting from day 7, the leaf area was lower in the af plants and larger in the tl plants as compared to the wild-type plants. Such divergence was amplified in the course of plant development and reached its maximum immediately before the transition to flowering. Plants of isogenic lines did not notably differ in stem surface areas. In spite of significant difference in total leaf area, the wild type and tl plants did not differ in leaf dry weight. Starting from leaf 9, the af plants lagged behind two leaflet-bearing genotypes (wild type and tl) in leaf dry weight, whereas stem dry weight was similar in the wild type and tl forms and slightly lower in the af plants. Root dry weights were practically similar in the wild type and tl plants until flowering. The reduction of leaf area in the af plants drastically reduced root dry weight. In other words, the latter index was related to the total weight and total area of leaves and stems. The correlation analysis demonstrated an extremely low relationship between leaf and stem area and dry weight and those of roots early in plant development (when plants develop five to seven leaves). Later, immediately before flowering (nine to eleven leaves), root weight was positively related to leaf weight and area; however, stem area and root weight did not correlate. Thus, in three genotypes (wild type, af, and tl), at the end of their vegetative growth phase, leaf and root biomass accumulated in proportion, independently of leaf area expansion.     

The effects of three genes which modify leaves and stipules in the pea plant

Summary The effects of three genes (af, st and tl) which modify leaf and stipule form and size in peas were investigated in families generated by crossing eight near-isogenic lines in all combinations but excluding the reciprocals. The eight parents and their equivalent phenotypes differed significantly for all characters due to the direct effects of all three genes, with their combined effects being especially influential in some instances. The effects of the recessive allele at any one of the three loci in homozygotes was to reduce plant productivity with stst having the most pronounced effect. The response of characters tended to be similar in direction, if not in magnitude, to any of the three genes. Partial dominance was frequently associated with the tl locus, and was especially obvious in afaf Tltl genotypes.     

Genetic control of leaf-blade morphogenesis by the <Emphasis Type="Italic">INSECATUS</Emphasis> gene in <Emphasis Type="Italic">Pisum sativum</Emphasis>

To understand the role of INSECATUS (INS) gene in pea, the leaf blades of wild-type, ins mutant and seven other genotypes, constructed by recombining ins with uni-tac, af, tl and mfp gene mutations, were quantitatively compared. The ins was inherited as a recessive mutant allele and expressed its phenotype in proximal leaflets of full size leaf blades. In ins leaflets, the midvein development was arrested in distal domain and a cleft was formed in lamina above this point. There was change in the identity of ins leaflets such that the intercalary interrupted midvein bore a leaf blade. Such adventitious blades in ins, ins tl and ins tl mfp were like the distal segment of respective main leaf blade. The ins phenotype was not seen in ins af and ins af uni-tac genotypes. There was epistasis of uni-tac over ins. The ins, tl and mfp mutations interacted synergistically to produce highly pronounced ins phenotype in the ins tl mfp triple mutant. The role(s) of INS in leaf-blade organogenesis are: positive regulation of vascular patterning in leaflets, repression of UNI activity in leaflet primordia for ectopic growth and in leaf-blade primordium for indeterminate growth of rachis, delimitation of proximal leaflet domain and together with TL and MFP homeostasis for meristematic activity in leaflet primordia. The variant apically bifid shape of the affected ins leaflets demonstrated that the leaflet shape is dependent on the venation pattern.     

Regulation of stipule development by <Emphasis Type="Italic">COCHLEATA</Emphasis> and <Emphasis Type="Italic">STIPULE</Emphasis>-<Emphasis Type="Italic">REDUCED</Emphasis> genes in pea <Emphasis Type="Italic">Pisum sativum</Emphasis>

Pisum sativum L., the garden pea crop plant, is serving as the unique model for genetic analyses of morphogenetic development of stipule, the lateral organ formed on either side of the junction of leafblade petiole and stem at nodes. The stipule reduced (st) and cochleata (coch) stipule mutations and afila (af), tendril-less (tl), multifoliate-pinna (mfp) and unifoliata-tendrilled acacia (uni-tac) leafblade mutations were variously combined and the recombinant genotypes were quantitatively phenotyped for stipule morphology at both vegetative and reproductive nodes. The observations suggest a role of master regulator to COCH in stipule development. COCH is essential for initiation, growth and development of stipule, represses the UNI-TAC, AF, TL and MFP led leafblade-like morphogenetic pathway for compound stipule and together with ST mediates the developmental pathway for peltate-shaped simple wild-type stipule. It is also shown that stipule is an autonomous lateral organ, like a leafblade and secondary inflorescence.     

An Analysis of Seed Development in Pisum sativum II. The Effect of the r-Locus on the Growth and Development of the Seed

The r-locus is one of the few genetic loci known to affect thestorage product composition and the morphology of pea (Pisumsativum) seeds. Lines which are near-isogenic except for ther-locus have been developed to study the effects of this locuson seed development. The plant phenotypes of these lines werevery similar except for those characteristics previously attributedto the r-locus. The seed development of the two lines followedsimilar patterns until the endosperm was absorbed by the embryo.The fresh weight of the rr line then increased more rapidlydue to the overall effect of a higher rate of water uptake anda lower rate of dry weight increase of the rr embryos comparedwith embryos homozygous for the R-allele. The effect of the r-locus on the relationship between embryofresh weight and dry weight suggests that the alleles may beaffecting the osmotic regulation of the developing embryo. Pisum sativum, pea, seed development, r-locus, genetic variation, growth analysis     

A GENETIC SYNDROME AFFECTING LEAF DEVELOPMENT IN PISUM

A recessive foliage mutant of Pisum, designated ‘sinuate leaf’ (sil), was found to have two distinct forms of expression, depending on the background genotype. In an af/af background—wherein leaflets are converted to tendrils—sil/sil plants had adventitious tendrils arising from clefts in the distal portion of the stipule. These adventitious tendrils were morphologically modified, just as were the true tendrils on the same plant, by different allelic combinations at the tl locus. In the standard Af background, sil/sil plants had neither incised stipules nor adventitious tendrils, although they did have undulated and somewhat distorted leaflets and stipules. Because mutant expression was variable in an Af background, classification of segregating populations was uncertain. This uncertainty was removed by taking advantange of pleiotropic effects exerted by sil in the presence of one of the wax mutants, wlo, wb, or wsp. Homozygous wlo plants ordinarily have uniformly waxy stipule surfaces, but when the plants were also homozygous for sil the stipule tips were waxless. Conversely, wb/wb and wsp/wsp plants ordinarily have uniformly waxless stipules, but when wb/wb or wsp/wsp plants were also homozygous recessive for sil the stipule tips were waxy. However, sil had no observable effects of any kind on the stipule tips of plants with stipules reduced in size by the action of st/st. By their individual and combined effects, the foliage mutants used in this study revealed developmental relationships among leaf parts not otherwise evident in non-mutant plants.     

A new pea mutation in the <Emphasis Type="Italic">Tl</Emphasis> locus

A new mutant of a pea with a repeatedly odd-pinnate leaf type was obtained. The compound leaf of the mutant is different from the earlier known forms with an odd-pinnate type of leaves by very narrow and elongated leaflets that possess a tendency of entanglement. It was found that the occurrence of the new type of repeatedly odd-pinnate leaves is mediated by a mutation at the Tl locus (chromosome 7). This mutation is not analogous to White’s acacia tl ^w mutation or to Lamm’s acacia tl ^pet mutation. The recessive type of inheritance of the new mutant gene was established relative to the tl ^w allele. It was proposed to designate the new mutation as tl ^UL . The plants with the new type of acacia-like leaves (tl ^UL tl ^UL Af-) have been obtained.     

Recessive monogenic mutation in grain pea (pisum sativum) that causes pyridoxine requirement for growth and seed production

A stable pyridoxine-deficient pea mutant was obtained by screening the M2 progeny of azide-treatedPisum sativum cv Pusa Harbhajan. The mutation is visible lethal. The isolation of pyridoxine-deficient mutant demonstrates directly that pea plants synthesize their own pyridoxine and that pyridoxine is an essential growth factor for pea plants. The mutant character is determined by homozygous recessive alleles, designatedpdx-1, at a single locus. Pyridoxine-deficient plants are fertile and indistinguishable from the wild type if supplied exogenously with 2 mg of pyridoxine.     

The Photosynthetic and Respiratory Potential of the Fruit in Relation to Seed Yield of Leafless and 'Semi-leafless Mutants of Pisum sativum L.

The photosynthetic and respiratory net CO₂ exchange potentialof the fruit of Pisum sativum was evaluated in one normal andthree mutant genotypes differing widely with respect to foliagearea and morphology: AfAf.StSt, conventional; AfAf.StSt, vestigialstipules; afaf.St.St, all leaflets replaced by tendrils butstipules normal (here termed ‘semi-leafless’) andafaf.stst, tendrils and vestigial stipules (‘leafless’).Agronomically the latter phenotype offered improvement in resistanceto lodging, crop drying, and harvester through-put. On a dry weight basis, fruits of the leafless mutant were consistentlymore active photosynthetically in terms of CO² uptake from theatmosphere during the initial 18 days post-anthesis than werethe corresponding fruits of the other three phenotypes. Duringthe subsequent 16-day period of seed filling there was no markeddifference between phenotypes and the fruit continuously lostCO² to the atmosphere, but significantly less CO² was lost inthe light (40 k lux) than in the dark. That the fruit of theleafless mutant may therefore benefit from the increase in lightavailable to the fruit within a sward canopy comprised of tendrilsin place of leaflets is discussed. However, there was stilla significant reduction in seed yield associated with the leaflessmutant despite a normal complement of ovule initials. The growthcurves and mean dry weights per seed were not significantlydifferent between phenotypes. In fruit of the conventional phenotypethe attainment of maximal pod wall weight coincided with theinitiation of seed fillingonday 15 after anthesis. In a seriesof isogenic lines the gene af delayed maximal wall developmentby 6 days whilst the gene st markedly lowered the maximum wallweight attainable. Possible causes of yield reduction in theleafless mutant are discussed in relation to the observed actionof these genes in the homozygous double recessive condition.     

An Analysis of Seed Development in Pisum sativum XVII. The Effect of the rb Locus Alone and in Combination with r on the Growth and Development of the Seed

Near-isogenic lines have been produced in pea to examine theeffects of genes at the r and rb loci on the growth and developmentof the seed. The rb alleles were found to have an effect onthe growth of the testa as well as on that of the embryo incontrast to those at the r locus which only affect the latter.The relationship between the fresh and dry weights of the embryosof rb isoline and the double mutant differed from that of theround-seeded wild-type line. Furthermore, for this relationship,the double mutant line very closely resembled the rr isoline.Copyright1994, 1999 Academic Press Embryo, growth, pea, Pisum sativum L., rugosus loci, seed development, testa, wrinkled-seeded     

Molecular mapping around the centromere of tomato chromosome 6 using irradiation-induced deletions

 Irradiation-induced deletion mapping was exploited to construct a detailed locus-order map around the centromere of tomato chromosome 6 (CEN  6). An F₁ hybrid heterozygous for the marker loci thiamineless (tl), yellow virescent (yv) and potato leaf (c), and homozygous recessive for the nematode resistance gene mi, was pollinated with γ-irradiated pollen from cultivar VFNT Cherry carrying the wild-type alleles at the corresponding loci. A dose of 100 Gy was found optimal for inducing mutants. By screening for pseudo-dominant plants showing the marker phenotypes and/or nematode susceptibility, 30 deletions encompassing one or more of the four loci were detected in the M₁ generation. Molecular-marker analysis revealed that 29 of these mutants included the tl and mi loci on the short arm and originated from terminal deletions of different sizes. Remarkably, the breakpoints of these deletions were not randomly distributed along the short arm but located within the centromeric heterochromatin. Only one yv interstitial deletion and no c mutations on the long arm of the chromosome were detected. Mapping of the various chromosomal breakpoints in the isolated mutants permitted the resolution of a cluster of molecular markers from the centromeric heterochromatin that was hitherto unresolvable by genetic linkage analysis. The usefulness of such a deletion-mapping approach for whole-genome mapping is discussed. Received: 4 March 1997 / Accepted: 2 June 1997     

An analysis of seed development in Pisum sativum XIX. Effect of mutant alleles at the r and rb loci on starch grain size and on the content and composition of starch in developing pea seeds

The effects of mutant alleles at the r and rb loci on starchgrain size and the levels of starch and amylose in developingpea (Pisum sativum L.) seeds have been examined. Four lines,near-isogenic except for genes at these loci, have been usedto show that both mutations reduce levels of starch throughoutembryo development and reduce levels still further when combinedin the ‘double mutant’. The reduction in starchcontent was due, at least in part, to a reduction in starchgranule size. Although the proportion of starch in mature embryoswas similar in the rrRbRb and RRrbrb lines, the starch contentdiffered between these two lines during development, as a percentageof embryo dry weight. This difference was due to a reductionin the absolute growth of the embryo caused by the rb mutation.Lines homozygous for the mutant r allele with either wild-type(RbRb) or mutant (rbrb) alleles at the rb locus contained increasedproportions of amylose in their starch throughout development,due to a reduced production of amylopectin. The presence ofthe rb mutation, however, also reduced the amount of amylosein relation to the reduction in total starch levels. Mutantalleles at both loci also reduced starch levels in the testaduring development, the reduction due to rb being more extreme.Reciprocal crosses showed a maternal effect of the rb mutationon final seed size and on the absolute amount of starch in theembryo. Key words: Pisum sativum L., seed, starch, development, mutant     

A mutation, tl2, in pea (Pisum sativum L.) affects leaf development only in the heterozygous state

After gamma irradiation of pea seeds, a mutation designated as tendril-less2 (tl2) was induced. In the heterozygous state, it transforms tendrils into very narrow leaflets that resemble the heterozygote phenotype of the classic tl mutation. The tendrils of the double heterozygote tl2/+, tl/+ are converted into oval leaflets. Unlike tl, the novel mutation in the homozygous state does not affect tendrils. The leaf phenotype of homozygotes tl2/tl2 and Tl2/Tl2 do not differ in the tl/+ background. However, the anthocyanin pigmentation is strongly suppressed in petals of tl2/tl2 plants. Some hypotheses to explain the unusual phenotypic manifestation of tl2 are suggested.     

Genetic control of high stearic acid content in seed oil of two soybean mutants

 Stearic acid is one of the two saturated fatty acids found in soybean [Glycine max (L.) Merr.] oil, with its content in the seed oil of commercial cultivars averaging 4.0%. Two mutants, KK-2 and M25 with two- and six-fold higher stearic acid contents in the seed oil than cv ‘Bay’, were identified after X-ray seed irradiation. Our objective was to determine the genetic control of high stearic acid content in these mutants. Reciprocal crosses were made between each mutant and ‘Bay’, and between the two mutants. No maternal effect for stearic acid content was observed from the analysis of F₁ seeds in any of the crosses. Low stearic acid content in ‘Bay’ was partially dominant to high stearic acid content in KK-2 and M25, and high stearic acid content in KK-2 was partially dominant to high stearic acid content in M25. Cytoplasmic effects were not observed, as demonstrated by the lack of reciprocal cross differences for stearic acid content in our analysis of F₂ seeds from F₁ plants. The stearic acid content in F₂ seeds of KK-2בBay’ and M25בBay’ crosses segregated into three phenotypic classes which satisfactorily fit a 1:2:1 ratio, indicating that high stearic acid content in KK-2 and M25 was controlled by recessive alleles at a single locus. The data for stearic acid content in F₂ seeds of the KK-2×M25 cross satisfactorily fit a 3:9:1:3 phenotypic ratio. The F₂ segregation ratio and the segregation of F₃ seeds from individual F₂ plants indicated that KK-2 and M25 have different alleles at different loci for stearic acid content. The alleles in KK-2 and M25 have been designated as st ₁ and st ₂, respectively. The stearic acid content (>30.0%) found in the st ₁ st ₁ st ₂ st ₂ genotype is the highest known to date in soybean, but it was not possible to develop the line with this genotype because the irregular seeds failed to grow into plants after germination. Therefore, tissue culture methods must be developed to perpetuate this genotype. Received: 28 March 1997 / Accepted: 18 April 1997     

Association of gene-linked SSR markers to seed glucosinolate content in oilseed rape (Brassica napus ssp. napus)

Breeding of oilseed rape (Brassica napus ssp. napus) has evoked a strong bottleneck selection towards double-low (00) seed quality with zero erucic acid and low seed glucosinolate content. The resulting reduction of genetic variability in elite 00-quality oilseed rape is particularly relevant with regard to the development of genetically diverse heterotic pools for hybrid breeding. In contrast, B. napus genotypes containing high levels of erucic acid and seed glucosinolates (++ quality) represent a comparatively genetically divergent source of germplasm. Seed glucosinolate content is a complex quantitative trait, however, meaning that the introgression of novel germplasm from this gene pool requires recurrent backcrossing to avoid linkage drag for high glucosinolate content. Molecular markers for key low-glucosinolate alleles could potentially improve the selection process. The aim of this study was to identify potentially gene-linked markers for important seed glucosinolate loci via structure-based allele-trait association studies in genetically diverse B. napus genotypes. The analyses included a set of new simple-sequence repeat (SSR) markers whose orthologs in Arabidopsis thaliana are physically closely linked to promising candidate genes for glucosinolate biosynthesis. We found evidence that four genes involved in the biosynthesis of indole, aliphatic and aromatic glucosinolates might be associated with known quantitative trait loci for total seed glucosinolate content in B. napus. Markers linked to homoeologous loci of these genes in the paleopolyploid B. napus genome were found to be associated with a significant effect on the seed glucosinolate content. This example shows the potential of Arabidopsis-Brassica comparative genome analysis for synteny-based identification of gene-linked SSR markers that can potentially be used in marker-assisted selection for an important trait in oilseed rape. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

QTL analysis of yield traits in an advanced backcross population derived from a cultivated Andean × wild common bean (Phaseolus vulgaris L.) cross

Advanced backcross QTL analysis was used to identify quantitative trait loci (QTL) for agronomic performance in a population of BC₂F_3:5 introgression lines created from the cross of a Colombian large red-seeded commercial cultivar, ICA Cerinza, and a wild common bean accession, G24404. A total of 157 lines were evaluated for phenological traits, plant architecture, seed weight, yield and yield components in replicated trials in three environments in Colombia and genotyped with microsatellite, SCAR, and phaseolin markers that were used to create a genetic map that covered all 11 linkage groups of the common bean genome with markers spaced at an average distance of every 10.4 cM. Segregation distortion was most significant in regions orthologous for a seed coat color locus (R-C) on linkage group b08 and two domestication syndrome genes, one on linkage group b01 at the determinacy (fin) locus and the other on linkage group b02 at the seed-shattering (st) locus. Composite interval mapping analysis identified a total of 41 significant QTL for the eight traits measured of which five for seed weight, two for days to flowering, and one for yield were consistent across two or more environments. QTL were located on every linkage group with b06 showing the greatest number of independent loci. A total of 13 QTL for plant height, yield and yield components along with a single QTL for seed size showed positive alleles from the wild parent while the remaining QTL showed positive alleles from the cultivated parent. Some QTL co-localized with regions that had previously been described to be important for these traits. Compensation was observed between greater pod and seed production and smaller seed size and may have resulted from QTL for these traits being linked or pleiotropic. Although wild beans have been used before to transfer biotic stress resistance traits, this study is the first to attempt to simultaneously obtain a higher yield potential from wild beans and to analyze this trait with single-copy markers. The wild accession was notable for being from a unique center of diversity and for contributing positive alleles for yield and other traits to the introgression lines showing the potential that advanced backcrossing has in common bean improvement.     

Carbon Dioxide Photoassimilation in Normal-leaved and Mutant Forms of Pisum sativum L.

The reproducible steady-state carbon dioxide (CO₂) photoassimilationpotentials of three mutants and a normal form of pea (Pisumsativum L.) have been compared. The three mutants studied differed markedly in foliar morphology:genotype af af Tl Tl had leaflets converted to tendrils; AfAf tl tl had tendrils converted to leaflets; af af tl tl hadrelatively minute leaflets on a branched petiole. Interest layprimarily in the phenotype with only tendrils since it provideda potential means of reducing the volume of haulm that has tobe rapidly processed in the case of vining peas, and dried inthe case of harvest peas. These mutants had been derived from relatively unimproved cultivars.Before completion of the lengthy backcrossing required to makea full assessment of the value of such mutants an interim studyusing infra red gas analysis indicated that, in terms of CO₂ photoassimilation perunit area of youngest expanded attached leaf of glasshouse-grownplants, the mutants were comparable to normal. The phenotypewith only tendrils was the least efficient of those assayedat utilizing light of an intensity below 100 J m² sec¹ (400–700nm), and on a unit dry-weight basis it was only 18 per centas efficient as a normal-leaved pea. The other mutants werecomparable to normal in this respect. Comparison of CO₂ photoassimilation of glasshouse-grown andfield-grown plants showed them to be similar though they differedin dry weight, transpiration, and dark respiration.     

Pea Compound Leaf Architecture Is Regulated by Interactions among the Genes UNIFOLIATA, COCHLEATA, AFILA, and TENDRIL-LESS

The compound leaf primordium of pea represents a marginal blastozone that initiates organ primordia, in an acropetal manner, from its growing distal region. The UNIFOLIATA (UNI) gene is important in marginal blastozone maintenance because loss or reduction of its function results in uni mutant leaves of reduced complexity. In this study, we show that UNI is expressed in the leaf blastozone over the period in which organ primordia are initiated and is downregulated at the time of leaf primordium determination. Prolonged UNI expression was associated with increased blastozone activity in the complex leaves of afila (af), cochleata (coch), and afila tendril-less (af tl) mutant plants. Our analysis suggests that UNI expression is negatively regulated by COCH in stipule primordia, by AF in proximal leaflet primordia, and by AF and TL in distal and terminal tendril primordia. We propose that the control of UNI expression by AF, TL, and COCH is important in the regulation of blastozone activity and pattern formation in the compound leaf primordium of the pea.