Band composition of electrophoretic spectra of storage proteins in interspecific pea hybrids

Electrophoretic spectra of storage proteins in parental plants and interspecific F1 and F2 hybrids Pisum sativum x Pisum fulvum have been studied. Correspondence between the polymorphism levels of protein components among the species and within the species P. sativum was established. Accessions of P. fulvum 1609881 and 1609885 manifested low polymorphism. Storage proteins of both parents were observed in spectra of F1 hybrids. F2 hybrids segregated at a limited set of bands. Accession 1609881 of P. fulvum is characterized by unique band 7, which was inherited in F1.     

Genome size in wild Pisum species

Genome size was measured in 75 samples of the wild pea species Pisum abyssinicum, P. elatius, P. fulvum and P. humile by ethidium-bromide (EB) flow cytometry (internal standard: Triticum monococcum) and Feulgen densitometry (internal standard: Pisum sativum Kleine Rheinländerin). Total variation of EB-DNA between samples covered 97.7% to 114.9% of the P. sativum value, and Feulgen DNA values were strongly correlated with EB-DNA values (r=0.9317, P < 0.001). Only P. fulvum was homogeneous in genome size (108.9% of P. sativum). Wide variation was observed between samples in P. abyssinicum (100.9–109.7%), P. elatius (97.7–114.9%) and P. humile (98.3–111.1% of P. sativum). In view of the world-wide genome size constancy in P. sativum, the present data are interpreted to show that the pea taxa with variable genome size are genetically inhomogeneous and that the current classification is not sufficient to describe the biological species groups adequately.     

High-Throughput Development of SSR Markers from Pea (Pisum sativum L.) Based on Next Generation Sequencing of a Purified Chinese Commercial Variety

Pea (Pisum sativum L.) is an important food legume globally, and is the plant species that J.G. Mendel used to lay the foundation of modern genetics. However, genomics resources of pea are limited comparing to other crop species. Application of marker assisted selection (MAS) in pea breeding has lagged behind many other crops. Development of a large number of novel and reliable SSR (simple sequence repeat) or microsatellite markers will help both basic and applied genomics research of this crop. The Illumina HiSeq 2500 System was used to uncover 8,899 putative SSR containing sequences, and 3,275 non-redundant primers were designed to amplify these SSRs. Among the 1,644 SSRs that were randomly selected for primer validation, 841 yielded reliable amplifications of detectable polymorphisms among 24 genotypes of cultivated pea (Pisum sativum L.) and wild relatives (P. fulvum Sm.) originated from diverse geographical locations. The dataset indicated that the allele number per locus ranged from 2 to 10, and that the polymorphism information content (PIC) ranged from 0.08 to 0.82 with an average of 0.38. These 1,644 novel SSR markers were also tested for polymorphism between genotypes G0003973 and G0005527. Finally, 33 polymorphic SSR markers were anchored on the genetic linkage map of G0003973 × G0005527 F₂ population.     

Phosphorylation of chloroplast membrane proteins partially protects against photoinhibition

Thylakoids isolated from peas (Pisum sativum cv. Kelvedon Wonder) and phosphorylated by incubation with ATP have been compared with non-phosphorylated thylakoids in their sensitivity to photoinhibition by exposure to illumination in vitro. Assays of the kinetics of fluorescence induction at 20° C and the fluorescence emission spectra at-196° C indicate a proportionally larger decrease in fluorescence as a result of photoinhibitory treatment of non-phosphorylated compared with phosphorylated thylakoids. It is concluded that protein phosphorylation can afford partial protection to thylakoids exposed to photoinhibitory conditions.Abbreviations and symbols DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F ₀ Level of chlorophyll fluorescence when photosystem 2 traps are open - F _m Level of chlorphyll fluorescence when photosystem 2 traps are closed - P Maximum level of fluorescence reached in the absence of DCMU - PSI (II) photosystem I(II)     

Flow cytometric and Feulgen densitometric analysis of genome size variation in Pisum

A DAPI and ethidium bromide flow cytometric and Feulgen densitometric analysis of genome size variation in Pisum was conducted. The material included 38 accessions of P. sativum of widely different geographic origin and altogether 14 samples of P. elatius, P. abyssinicum, P. humile and P. fulvum. The relative genome size values obtained with the three staining methods were strongly correlated. No evidence for genome size variation was found among P. sativum cultivars. In particular, certain Italian cultivars, for which strongly deviating C-values have been reported, proved to be invariant. The only occasion when ambiguous evidence for marginal genome size variation was found was when all 38 accessions taxonomically affiliated with P. sativum were considered. Pisum abyssinicum and P. fulvum differed from P. sativum by about 1.066-and 1.070-fold, respectively; 1 accession of P. humile differed by 1.089-fold, and 2 of P. elatius by 1.122- and 1.195-fold, respectively (ethidiumbromide comparison), while the other accessions of these taxa were not different from P. sativum. This variation may indicate taxonomic inhomogeneity and demands further investigation. Cultivated P. sativum has long been suspected of not being constant with respect to genome size. As shown here, these findings were not based on genuine differences, but rather were technical in origin.     

Genetic analysis of nuclear-cytoplasmic incompatibility in pea associated with cytoplasm of an accession of wild subspecies Pisum sativum subsp. elatius (Bieb.) Schmahl.

The genetic basis of nuclear-cytoplasmic incompatibility was examined using the wild pea (Pisum sativum subsp. elatius) accession VIR320. When this accession is used as the female parent in crosses with domesticated peas (Pisum sativum subsp. sativum) the F₁ is highly sterile and displays chlorophyll deficiency, chlorophyll variegation, reduction of leaflets and stipulae while the reciprocal cross produces hybrids that appear normal. A mapping recombinant inbred line (RIL) population was established based on a cross in a compatible direction of a tester line WL1238 with VIR320. The ability to cause nuclear-cytoplasmic conflict was analysed by crossing individual RIL plants as pollen parents with VIR320 as donor of cytoplasm and scoring each F₁ for major signs of the conflict. It is concluded that two unlinked nuclear genes are involved in the genetic control of the observed incompatibility. One of the genes, denoted as Scs1, is closely linked to the PhlC gene on linkage group III and the other, denoted as Scs2, is closely linked to the gp gene on linkage group V. Alleles of both genes in WL1238 are dominant and appear to be lethal in the homozygous condition in the VIR320 cytoplasm background. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The comparison of seed proteins of several representatives of the genusPisum with respect to their relationships An immunological comparison

Using immunochemical methods a comparison was made of a complex of water soluble (albumins) and salt soluble (globulins) seed proteins, especially vicilin and legumin, in selected species of the genusPisum, to determine the degree of their taxonomic relationship. Within the genusPisum the interspecific differences betweenP. abyssinicum, P. cinereum,P. elatius, P. fulvum, P. sativum, andP. syriacum are much smaller, and thus the taxonomic distances are shorter than is the casee.g. in the genusPhaseolus. In spite of this fact one may state thatPisum sativum, P. elatius andP. syriacum constitute a definite group, whereasP. abys-sinicum, P. cinereum and P.fulvum have longer taxonomic distances.     

RELATIONSHIP OF WOODY PARTHENIUM ARGENTATUM AND HERBACEOUS P. HISPIDUM VAR. AURICULATUM (ASTERACEAE)

Interspecific hybrids between woody Parthenium argentatum Gray (guayule), native to Mexico and Southwest Texas, and herbaceous perennial P. hispidum var. auriculatum (Britton) Rollins, native to the United States, were obtained successfully. The F₁ hybrids were intermediate for most morphological characters with the exception of the short woody stem, yellow pollen color, and the trichome morphology. Chromosome counts revealed the presence of 2n = 36 A-chromosomes in P. argentatum. The same number of A-chromosomes and four B-chromosomes were found in P. hispidum var. auriculatum. Observations of pollen mother cells showed regular meiosis in both parental species. At diakinesis, chiasmata averaged 1.12 and 1.24 per bivalent for P. argentatum and P. hispidum var. auriculatum, respectively. Meiotic behavior of the F₁ hybrids was irregular. F₁ hybrids averaged 4.43 univalents at metaphase I, 1.95 laggards at anaphase I, and 1.62 micronuclei at the tetrad stage. The low pollen stainability (5.1%) in the F₁ hybrids and the limited number of viable BC₁ seeds (4.07%) may be reflections of the irregular meiosis. Although these primary hybrids are partially fertile, they can be used to introduce desirable characteristics of P. hispidum var. auriculatum, such as herbaceous perennial habit, regrowth ability, and cold tolerance into guayule.     

Genetic structure of a hybrid zone between two violets,Viola rossii Hemsl. and V. bissetii Maxim.: dominance of F1 individuals in a narrow contact range

The genetic composition of a hybrid zone can provide insight into the evolution of diversification in plants. We carried out morphological and amplified fragment length polymorphism analyses to investigate the genetic composition of a hybrid zone between two violets, Viola bissetii Hemsl. and Viola rossii Maxim. Our aim was to clarify the formation and maintenance of hybrids between these Viola species. We found that most hybrid individuals (V. bissetii × V. rossii) were of the F₁ generation, with a few of the F₂ generation. We found no backcrosses. The scarcity of post‐F₁ hybrids indicates that a species barrier is established between the parental species. The F₁‐dominated hybrid zone occupied only a narrow, intermediate ecotone between the parental habitats, suggesting that selection by environmental factors against hybrids may help to maintain the current conditions in this hybrid zone.     

ARTIFICIAL HYBRIDIZATION OF RUBBER-BEARING GUAYULE WITH COLD-TOLERANT PARTHENIUM LIGULATUM

ABSTRACT Caespitose and cold-tolerant plants of Parthenium ligulatum (Jones) Barneby (Asteraceae) from a native population in the Uinta Basin, Utah, were uprooted, potted, and transferred to a greenhouse in California. Approximately two years after transfer, the plants flowered and subsequently were crossed to diploid guayule (Parthenium argentatum Gray), the rubber-bearing species, native to the state of Durango, Mexico. Only female guayule × male P. ligulatum crosses produced F₁ hybrids. Only crosses involving guayule as female parent and F₁ plants as male parents produced backcross (BC,) plants. Hybrid plants were variable with respect to their growth habit, inflorescence, and leaf shape. Both parents and F₁ hybrids had 2n = 36 chromosomes. Unlike the parents, however, meiosis was irregular in the hybrids which showed a range of 0–5 and an average of 2.1 univalents at metaphase I. Hybrids averaged 0.87 laggards at anaphase I and 0.83 micronuclei at the tetrad stage. The crossability of guayule and P. ligulatum, the high degree of chromosome pairing of the F₁ hybrids, and the production of BC₁ plants indicate that the two species are related in spite of their distinct morphological and ecological differences. This study suggests that the cold-tolerance trait of P. ligulatum may be transferred to guayule through interspecific hybridization followed by backcrossing. The development of cold-tolerant guayule cultivars is expected to expand the areas of guayule production beyond that of the Chihuahuan desert and similar climates.     

Analysis of a lipoxygenase pseudogene in Pisum

 Lipoxygenase (LOX) enzymes play important roles in plant biology, and in the quality of plant-derived foods, through the production of fatty acid hydroperoxides that are metabolized either to jasmonate, or to volatile aldehydes that are part of plant defence systems and/or impart tastes and aromas to fruits and vegetables. We have identified a lipoxygenase pseudogene in peas that is composed of three elements: rearranged LOX-2 and LOX-3 genes and an unidentified tract of DNA. We present evidence that such an arrangement is normally present in the genome of Pisum sativum, but is absent from Pisum fulvum lines, including a mutant line that lacks LOX-2 polypeptides. The absence of LOX-2 polypeptides and the pseudogene co-segregate. The pseudogene therefore has utility as a molecular marker for the introgression of the LOX-2-null phenotype into commercial Pisum sativum genotypes. Received: 27 April 1998 / Accepted: 17 September 1998     

Meiotic abnormalities as expression of nuclear-cytoplasmic incompatibility in crosses of <Emphasis Type="Italic">Pisum sativum</Emphasis> subspecies

Meiosis in anthers and mitosis in somatic cells were studied in reciprocal F₁ hybrids of the accession VIR320, which belonged to wild Pisum sativum ssp. elatius (Bieb.) Schmal., and the laboratory line Sprint-1. When VIR320 was used as a maternal form, the hybrids displayed nuclear-cytoplasmic conflict, which caused chlorophyll defects and meiotic abnormalities. One or two chromosomes lagged in the equatorial region during chromosome segregation to the poles, distorting cytokinesis and yielding abnormal microspores. Chlorophyll defects were not observed, and meiotic abnormalities were far less frequent in reciprocal hybrids and in the case of an abnormal paternal inheritance of plastids from Sprint-1. Mitosis lacked overt abnormalities in all of the hybrids.     

Association of dominant loci for resistance to Pseudomonas syringae pv. pisi with linkage groups II, VI and VII of Pisum sativum

Morphological characters, isoenzymes and recombinant inbred lines were employed to assign four loci for resistance to Pseudomonas syringae pv pisi to genetic linkage groups in Pisum sativum. A total of five morphological markers and 11 isoenzyme loci were screened in two independent F₂ P. sativum populations: Vinco × Hurst’s Greenshaft (V×HGS) and Partridge × Early Onward (P×EO). Mapping was also carried out in two recombinant inbred populations, unrelated to the F₂ populations. Previously reported linkage between resistance genes Ppi3 and Ppi4 was confirmed. Linkage was also detected between resistance gene Ppi2 and the isoenzyme locus Aldo (linkage group VII). The linked loci Ppi3 and Ppi4 were associated with a (linkage group II). A further resistance gene Ppi1 was associated with linkage group VI close to the hilum colour gene P1. RAPD markers tested in the cross P×EO were not well targeted; however, one marker, OPA-20_0.71, showed linkage to Ppi3. Received: 3 July 2000 / Accepted: 27 October 2000     

Analyses of Phaseolus vulgaris L. and P. coccineus Lam. hybrids by RFLP: preferential transmission of P. vulgaris alleles

Summary Restriction fragment length polymorphism (RFLP) was determined among P. vulgaris genotypes and Phaseolus species using 19 probes. The incidence of polymorphism was high (70–86%) between species, but relatively low (22–26%) between genotypes of P. vulgaris. Suitable probes were identified for the analysis of P. vulgaris and P. coccineus hybrids. The segregation pattern in F₂ populations was Mendelian for two probes (LHB and VEE20) and non-Mendelian for GS-g, CHS, and CHI. Statistical analyses indicated gametic selection with preferential transmission of the P. vulgaris alleles, which may account for the selective recovery of P. vulgaris progeny types observed earlier. The available hybrids of P. vulgaris and P. coccineus and the high degree of interspecific RFLP will facilitate the construction of a linkage map for Phaseolus.     

Comparison of seed proteins of some representatives of the genusPisum from the point of view of their relationship comparison by disc electrophoresis

Ten taxa of the genusPisum were examined by disc electrophoresis in gels according to Davis and to Reisfeldet al. For evaluation of band patterns the Jaccard Index was applied. The results in both types of gels show thatPisum abyssinicum and especiallyP. fulvum have biochemically a relatively isolated position.Pisum elatius and its subspeciescaspicum andpalestinicum form a subgroup withP. cinereum;P. sativum var.zeylanicum and cv. Jupiter form another subgroup withP. syriacum. Our results are in good agreement with the results of Przybylskaet al. (with the exception ofP. cinereum) and also with immunoelectrophoretic analyses performed by Turkováet al. (1980), with the same exception.     

Impact of Asymmetrical Hybridization Followed By Sterile F1 Hybrids on Species Replacement in Pseudorasbora

Pseudorasbora pumila, one of the endangered freshwater minnows in eastern Japan, has been largely replaced by the accidentally introduced species, P. parva, which originated from western Japan. In the contact zone, P. pumila and P. parva have hybridized intensively, producing sterile F₁ hybrids. The present study determined the maternal parent of F₁ hybrids using mtDNA haplotypes to investigate the mating system between P. parva and P. pumila in the hybrid zone. We also pursued the successive changes in the genetic structures of hybridizing populations over a 5-year period using allozymes. A total of 100 natural F₁ hybrids collected from six different populations had P. pumila mtDNA without exception, suggesting that sterile F₁ hybrids resulted from mating only between P. pumila females and P. parva males. Such asymmetrical hybridization implies that P. pumila females waste considerably greater reproductive efforts compared with P. parva males. The data suggest that the rapid replacement of P. pumila by P. parva has been promoted by asymmetrical hybridization, resulting in sterile F₁ hybrids.     

Reproductive interactions between hybridizing irises: analyses of pollen-tube growth and fertilization success

Pollen-tube growth and seed siring ability were measured in crosses between the Louisiana iris species Iris fulva and Iris hexagona and their F₁ and F₂ hybrids. Flowers of the parental species were pollinated with self, outcross intraspecific, and interspecific pollen. Pollen-tube lengths were similar for all three pollen types in I. fulva, but in I. hexagona interspecific pollen tubes were longer than intraspecific pollen tubes. Pollen-tube lengths also differed for F₁ and F₂ flowers pollinated with I. fulva, I. hexagona, and hybrid pollen. For both hybrid classes I. fulva pollen tubes were the shortest while pollen tubes from I. hexagona and hybrids grew the furthest. Mixtures of genetically marked pollen were used to assess the seed siring ability of intra- and interspecific pollen in the parental species by varying the proportion of each pollen type in a replacement series design. For both species, the observed proportions of hybrid seeds were lower than the expected based on the frequency of each pollen type in the mixtures across all treatments. Flowers of I. fulva produced less than 10% hybrid progeny even when 75% of the pollen applied to stigmas was derived from interspecific flowers. The frequency of hybrid seed formation was somewhat greater in I. hexagona, but was still significantly lower than expected across all mixture treatments. Seed set per fruit remained constant across the mixture treatments for both species, but in I. fulva fruit set decreased with an increase in the proportion of interspecific pollen. The data indicate that both pre- and postfertilization processes contribute to discrimination against hybrid seed formation.     

Iron Deficiency Induced Changes on Electron Transport Rate in Pisum Sativum Chloroplasts

Iron deficiency induced decrease in the rate of whole electron transport chain in chloroplasts of pea (Pisum sativum L.). Such reduction was mainly due to the loss of photosystem (PS) 2 activity. The same result was obtained when the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m) was evaluated. The loss in PS 2 activity was primarily due to a loss of 33, 23 and 17 kDa polypeptides. In contrast, iron deficiency induced the synthesis of 28 and 29 kDa polypeptides.     

Nuclear DNA contents of Pisum genotypes grown in vivo and in vitro

Summary The nuclear DNA content of prophase nuclei in root tips of two cultivars and two primitive lines of Pisum sativum and of Pisum fulvum have been determined, using a scanning microdensitometer. The nuclear DNA contents differed significantly between the genotypes investigated but there was no correlation with their supposed phylogenetic positions.A loss of 73% of the DNA from cells of aseptically cultured excised pea roots has been recently reported (Abbott, 1971). In marked contrast to this claim, our measurements of the nuclear 4C DNA content of root tip meristematic cells have shown that there is no significant loss in excised roots compared with attached roots.     

THE EFFECT OF HABITAT ON PARENTAL AND HYBRID FITNESS: TRANSPLANT EXPERIMENTS WITH LOUISIANA IRISES

We performed transplant experiments with Louisiana irises to test the assumptions of three models of hybrid zone structure: the bounded hybrid superiority model, the mosaic model, and the tension zone model. Rhizomes of Iris fulva, I. hexagona, and F₁ and F₂ hybrids were planted at four sites in southeastern Louisiana in 1994. Wild irises grew at all four sites, but differed in genotypic composition among sites. The sites were characterized by (1) pure I. fulva plants; (2) I. fulva-like hybrids; (3) I. hexagona-like hybrids; and (4) pure I. hexagona plants. The sites differed significantly in light availability, soil moisture and chemical composition, and vegetation. Survival of transplants was high in all sites and did not differ significantly among plant classes. Iris hexagona produced significantly more leaf material than I. fulva at the I. hexagona and I. hexagona hybrid sites. The two species did not differ in leaf production at the I. fulva and I. fulva hybrid sites. Leaf production by both classes of hybrid was as great as, or significantly greater than, both parental classes in all sites. Iris hexagona rhizomes gained mass in the I. hexagona and I. hexagona hybrid sites, but lost mass in the I. fulva and I. fulva hybrid sites. Iris fulva rhizomes lost mass in all sites. There were no significant differences in rhizome growth among classes at the I. fulva site. At all other sites, F₁ rhizomes grew significantly more than all other classes except for I. hexagona at the I. hexagona hybrid site. There were no significant differences among classes in the production of new ramets. Overall blooming frequencies were 30% for I. fulva, 10% for F₁s, 3% for F₂s, and 0.7% for I. hexagona. Blooming frequency did not differ among sites for I. fulva, but significantly more F₁s bloomed at the I. hexagona site than at the I. fulva site. These results are inconsistent with all three models of hybrid zone structure. They suggest that once rhizomes become established, hybrids can reproduce by clonal growth as successfully as parents in all habitats, and can outperform them in some habitats. Clonal reproduction may ensure the long-term survival of early generation hybrids and allow the establishment of introgressed populations, despite the fact that F₁ hybrids are rarely produced in nature.