Gene flow between cultivated sunflower and <Emphasis Type="Italic">Helianthus petiolaris</Emphasis> (Asteraceae)

Helianthus petiolaris (Asteraceae) native to North America has naturalized in Argentina. The extensive overlapping with sunflower crop regions, the coincidence of life cycles and the common pollinators facilitate interspecific crosses with sunflower, H. annuus var. macrocarpus. To estimate the occurrence of crop-to-wild and wild-to-crop gene flow, off-type plants in progenies of pure stands of both species flowering in coincidence and the presence of crop alleles in H. petiolaris populations were investigated in 26 wild populations and nine crop fields. Morphological traits and RAPD markers were used to attempt hybrid characterization. Off-type individuals were found in frequencies of 0.5 and 0.3% among crop progenies and wild populations, respectively. Off-type plants showed intermediate values for metric morphological traits and low fertility traits. Some off-type plants proved to carry crop alleles based on molecular analysis. The average frequency of cultivar-marker introgression across the wild populations was very low (0.02). Although observed hybridization rates seem to be low, the extension of crop-wild overlapping in Argentina make hybrid formation a noticeable process. Therefore, the new sunflower varieties and eventually GM varieties would transfer their traits through pollen flow and they would persist in H. petiolaris populations.     

Genetic control of isozymes in eggplant and its wild relatives

Summary This study was conducted to elucidate the inheritance and linkage relationships of isozymes in aspartate aminotransferase (AAT, EC 2.6.1.1), alcohol dehydrogenase (ADH, EC 1.1.1.1), phosphogluconate dehydrogenase (PGD, EC 1.1.1.43), phosphoglucomutase (PGM, EC 2.7.5.1) and shikimate dehydrogenase (SKDH, EC 1.1.1.25) in eggplant and its wild relatives. Segregating populations were generated by backcrossing of hybrids among the species. Evidence of Mendelian inheritance was obtained for seven loci: Aat-1, Adh-1, Adh-2, Pgd-1, Pgm-1, Pgm-2 and Skdh-1. Twenty-one pairs of loci were tested for independent assortment, suggesting three linked pairs, Aat-2 with Pgd-2 (R=0.35±0.07), Adh-2 with Pgm-1 (R=0.33±0.07) and Pgd-2 with Pgm-2 (R=0.32±0.06).Abbreviations AAT aspartate aminotransferase - ADH alcohol dehydrogenase - PGD phosphogluconate dehydrogenase - PGM phosphoglucomutase - SKDH shikimate dehydrogenase     

Isozyme variation in cultivated and wild pineapple

Summary Isozyme variation was studied in 161 accessions of pineapple including four species of Ananas and one of Pseudananas. Six enzyme systems (ADH, GPI, PGM, SKDH, TPI, UGPP) involving seven putative loci revealed 35 electromorphs. Considerable variation exists within and between species of Ananas. Sixty-six distinct zymotypes were identified. Multivariate analyses of isozyme variation indicated that A. comosus contains five genetically diverse groups that do not match perfectly with the traditional varietal groups. Isozyme evidence also suggests that A. erectifolius is a conspecific variant of A. comosus, and that among other wild species, A. ananassoides is more closely related to A. comosus than A. bracteatus. Pseudananas is genetically distinct from all species of Ananas. It is evident from our study that differentiation among the species of Ananas may be due to ecological isolation rather than genetic divergence with breeding barriers and therefore may represent a species complex.This is journal series No. 3956 of the Hawaii Institute of Tropical Agriculture and Human Resources.     

Genetics of isozyme variants in Populus tremula,P. tremuloides and their hybrids

Summary Eight Populus tremula and six P. tremuloides clones as well as 49 full-sib families were studied in GOT, LAP, 6-PGDH, and SKDH by horizontal starch gel electrophoresis. For GOT one polymorphic zone was found and segregation of seven full-sib families suggests Mendelian inheritance. For LAP and 6-PGDH two zones each were clearly scored. For LAP two polymorphic loci were proposed based on the phenotypic segregation of isozyme variants in six and 34 full-sib families, respectively. In 24 full-sibs families the presence of null alleles was inferred for both loci. The genetic control of the upper zone of 6-PGDH was demonstrated by a segregation analysis of 17 full-sib families. SKDH also demonstrated a Mendelian inheritance pattern in 12 of the full-sib families analysed. The electrophoretic patterns of pollen were similar to those of buds, but migration rates of the supposed corresponding isozymes were slightly modified (Lap-B, Skdh, 6Pgdh-A). Lap-A was not present in pollen extracts and hybrid bands were not found when gels were stained for dimeric enzymes (6-PGDH, GOT).     

Genetic analysis and nomenclature for seven isozyme systems in Brassica nigra, B. oleracea and B. campestris

General criteria for the assignment of names to enzyme systems, regions of activity, isozyme loci and allozymes have been lacking in crucifer species. This paper proposes a standard nomenclature for seven isozyme systems in the three diploid species of U's triangle: Brassica nigra, B. oleracea and B. campestris. Gel/electrode buffers, which provided the best resolution for seven isozyme systems, acid phosphatase (APS), aconitase (ACO), leucine aminopeptidase (LAP), 6-phosphogluconate dehydrogenase (6 PGD), phosphoglucoisomerase (PGI), phosphoglucomutase (PGM), and triosephosphate isomerase (TPI), were proposed as standards. Isozyme genetic analysis was determined for B. oleracea and B. campestris from previous studies and by segregation of selfed progenies of heterozygous B. nigra plants. Several populations were studied and 148 allozymes at the 18 loci observed were described for the three species. Their relative mobility was studied using a pure line of oilseed rape as reference. The comparison of the different alleles within and between the species is discussed.     

Characterization of isozymes in Salvia columbariae

Summary Salvia columbariae is a herbaceous annual species which has potential for domestication as a new source of industrial oil. Isozyme markers provide a mean by which this process may be facilitated. Isozyme survey of field grown Salvia columbariae plants showed variation for malate dehydrogenase (MDH), phosphoglucose isomerase (PGI), and phosphogllucomutase (PGM). Selfed seed was obtained from the field and was grown in the greenhouse for segregation analyses. Electrophoretic results indicated that MDH was variable at zone 1, showing presence or absence of a band. The observed segregation ratio was not significantly different from expected ratio for Pgi-4 and Pgm-3 isozymes, indicating monogenic control of the two loci. Pgi-4 locus was heterozygous for a null allele. Cross dimerization between the allozyme Pgi-3 and Pgi-4 loci resulted in an intergenic band for this isozyme system.Abbreviations ACO aconitase - ADH alcohol dehydrogenase - DTT-DL dithiothreitol - FDH formate dehydrogenase - GOT glutamate-oxalacetate transaminase - IDH isocitric dehydrogenase - MDH malate dehydrogenase - MNR menadione reductase - 6PGD 6-phosphogluconate dehydrogenase - PGI phosphoglucose isomerase - PGM phosphoglucomutase - PVP-40 polyvinylpyrrolidone - SKDH shikimate dehydrogenase - TPI triosephosphate isomerase     

A new source of cytoplasmic male sterility in sunflower

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) cv. Saturn into the cytoplasm of H. petiolaris (2n=34) by successive backcrossing, resulted in progenies with indehiscent anthers containing white, rather than normal yellow, pollen. Further backcrossing by cv. Saturn failed to restore pollen shed, suggesting that the male sterility was cytoplasmic. In vivo germination tests of pollen from 23 such plants from eight BC₅ lines, indicated complete pollen sterility for 14 plants, but normal seed set, suggesting that female fertility was not affected. Meiosis in all plants examined was normal.Crosses between seven sources of pollen-fertility restorer, one collection of wild H. annuus, and an existing source of cytoplasmic male sterility, resulted in a high frequency of plants with normal pollen shed in all F₁ progenies. However, no normal pollen shed was evident in F₁ progenies for similar crosses between BC₅ male-steriles and three of the seven restorer sources, nor for the single wild H. annuus evaluated. The foregoing suggests that the backcross substitution lines are a new source of cytoplasmic male sterility. The inheritance of restoration of pollen shed was complex and not fully elucidated. Some data suggested that two independent, complementary, dominant genes were required, but others indicated two to three independent, dominant genes.     

Resistance to Spodoptera litura (Fabr.) in Helianthus species and backcross derived inbred lines from crosses involving diploid species

Spodoptera litura (Fabr.), the tobacco caterpillar, is a major defoliator on sunflower in the tropics. Genetic variability for resistance to S. litura is limited in the cultivar germplasm of sunflower. In the present investigation, 43 accessions of 17 wild Helianthus species of the annual and perennial habit groups were evaluated along with cultivated sunflower under field and no-choice conditions in the laboratory for resistance to this pest. Under field conditions, H. occidentalis and H. argophyllus were found to be immune with no leaf damage and few accessions of the species belonging to section Divaricati were found to be resistant. Laboratory bioassays against neonate, two and 4-day-old larvae confirmed resistance both in terms of high larval mortality and low larval weight gain in eight species viz., H. occidentalis, H. argophyllus, H. tuberosus, H. maximiliani, H. mollis, H. simulans, H. divaricatus and H. hirsutus. Intra-accessional variability was observed and accessions of few species showed varied reaction (resistant, partially resistant to susceptible) to the target pest. Field evaluation of 224 backcross derived inbred lines from five cross combinations involving diploid species under high natural pest incidence revealed low damage in plants derived from crosses involving H. argophyllus and H. petiolaris.     

Inheritance of isozyme variants and their linkage relationships in Chinese fir (Cunninghamia lanceolata Hook.)

Summary 49 single-clone seed samples of Chinese fir (Cunninghamia lanceolata) were used to investigate genetic variation for nine enzyme systems. LAP, PGM, and SOD were invariant, whereas GDH, GOT, IDH, MDH, 6PDH, and SKDH were highly variable. Inheritance patterns of the variable enzyme systems were mostly in accordance with Mendelian expectations. Linkage was studied by analysing 43 two-locus combinations with the following two-locus combinations significantly linked in at least 50% of the respective double-heterozygous clones: Gdh1: Got3; Gdh1: Idh1; Got2: 6Pdh2; Idh2: Skdh1; Mdh1: 6Pdh1. Estimates of recombination frequencies for each locus pair varied considerably among clones.Supported by the Carl Duisberg Gesellschaft.     

Interspecific hybrids between helianthus PetiolarisNutt. and H. Annuus L.: Effect of backcrossing on meiosis

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) into the cytoplasm of H. petiolaris (2n=34) were obtained by successive backcrossing using cultivated sunflower, H. annuus, as the recurrent pollen parent.Meiosis in the F₁ was characterized by multivalents, suggesting that 10 of the 34 chromosomes were heterozygous for chromosomal interchanges. An additional pair of chromosomes also contained a paracentric inversion. Continued backcrossing resulted in rapid elimination of the meiotic aberrations evident in the F₁. In the BC₁, 1 of 11 plants had normal meiosis and by the BC₂, only 13 of 54 plants had meiotic aberrations similar to those of the F₁. However, trisomic progeny (2n=35) were found in 3 of the 11 BC₁ plants and 20 of the 54 BC₂ plants. No meiotic aberrations were observed in BC₃ or BC₄ plants. Plants with indehiscent anthers, and considered to be male sterile (M.S.), first occurred in the BC₁ and, by the BC₂, 51 of 54 plants were M.S. All 19 BC₃ and 16 BC₄ plants were M.S. Preliminary investigations suggest that the pollen from such plants is sterile and that the sterility is cytoplasmic rather than genetic.Disc-flower measurements were a useful technique for selecting samples at the correct stage of microsporogenesis, but could not be used to distinguish between successive backcrosses.     

High regeneration potential in vitro of sunflower (Helianthus annuus L.) lines derived from interspecific hybridization

Successful selection of interspecific hybrid progenies with superior ability to regenerate shoots from apical meristems was performed in sunflower which now allows for the development of lines for improved biotechnological applications. Early generations of interspecific hybrids originating from crosses between the two H. annuus CMS lines ‘HA89’ and ‘Baso’, and 9 wild species were screened for their ability to regenerate in vitro. Evaluation of 36 progenies allowed to identify seven progenies from crosses involving H. mollis, H. giganteus, H. strumosus, and H. decapetalus which showed a significantly higher regeneration potential than the commercial hybrid ‘Albena’ regarding the number of shoots per explant. Among these progenies, 47.2 to 62.4% of explants produced shoots with an average of 2.3 to 3.5 shoots per cultured explant. Regeneration in vitro was significantly determined by the genotype. More than half of the investigated interspecific hybrids performed better than the inbred ‘HA89’ demonstrating that the high regeneration potential available in the wild species can be efficiently transferred to cultivated sunflower. The seven progenies with high regeneration potential in vitro were characterised by agronomic performance in the field. Two of the interspecific hybrids derived from H. strumosus and H. decapetalus not only showed a superior regeneration potential but also proved to be competitive to commercial hybrids with regard to important agronomic traits, e.g. fat content and TGW. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chloroplast DNA diversity in eggplant (Solanum melongena) and its related species S. incanum and S. marginatum

Summary The potential of isozymes for distinguishing asparagus varieties was carried out by a survey on 21 varieties using 10 enzyme systems: GOT, SkDH, DIA, PGM, MDH, IDH, PGD, ACP, PGI, MR and ADH. Only 3 enzymes, SkDH, GOT and PGM, showed useful polymorphisms. The varieties were found heterogeneous according to their genetic structure: open pollinated varieties were more heterogeneous than clonal hybrids; the F1 hybrid and the vitroclones were homogeneous. As expected from the narrow genetic basis of the varieties, only a few alleles per isozyme locus were present. Moreover, for each enzyme, one allele or type was predominant so that the discriminating power of the method was low. However some of the varieties could be identified and different applications of the results are presented.Abbreviations D.U.S.- Distinction-Uniformity-Stability - ACO- aconitase - ACP- acid phosphatase - ADH- alcohol dehydrogenase - CAT- catalase - DIA- diaphorase - END- endopeptidase - GOT- glutamate oxaloacetate transaminase - IDH- isocitrate dehydrogenase - MDH- malate dehydrogenase - MR- menadione reductase - PGI- phosphoglucoisomerase - PGM- phosphoglucomutase - PGD- phosphoglucose dehydrogenase - POX- peroxidase - SkDH- shikimate dehydrogenase     

Interspecific hybrid of Helianthus annuus × H. simulans: Characterization and utilization in improvement of cultivated sunflower (H. annuus L.)

The first success at interspecific hybridization between cultivated sunflower(H. annuus) and a diploid perennial species, H. simulans is reported. The F₁s from both direct and reciprocal directions exhibited dominance of the wild species phenotype and were pollen sterile. Meiosis was irregular in the F₁ hybrids and both univalents and multivalents were observed. Multiplication of the interspecific hybrids was achieved through in vitro culture of nodal sections on Murashige & Skoog medium supplemented with 0.5 mg l^-1 benzyladenine. Fertility of the interspecific hybrids was improved by subjecting the in vitro proliferating shoots to 0.001% colchicine incorporated in the shoot multiplication medium. The amphiploids serve as fertile bridges and facilitate interspecific gene transfer through conventional breeding. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular mapping of the fertility restoration locus Rf1 in sunflower and development of diagnostic markers for the restorer gene

Fertility restoration by dominant nuclear genes is essential for hybrid breeding based on cytoplasmic male sterility (CMS) to obtain heterotic effects and high seed yields. In sunflower, only the PET1 sterility inducing cytoplasm has been used in commercial hybrid breeding until now. This particular male sterility was derived from an interspecific hybrid Helianthus petiolaris × H. annuus. For the recent work we used the segregating population RHA325(CMS) × HA342, based on the PET1 cytoplasm. Molecular markers were mapped within 1.1 cM around the restoration locus Rf1. At the distal side, the marker OP-K13_454 mapped at a distance of 0.9 cM and E32M36-155R at 0.7 cM from Rf1. At the proximal side the markers E44M70-275A, E42M76-125A and E33M61-136R were mapped at 0.1, 0.2, and 0.3 cM from the restorer locus, respectively. These markers provide an excellent basis for a map based cloning approach and for marker-assisted sunflower breeding.     

Genetic and linkage analysis of isozyme loci in Daucus carota L.

Summary Electrophoretic polymorphisms of eight enzyme systems were studied in leaves of Daucus carota ssp. sativus in order to identify additional isozyme loci and generate first linkage groups of genetic markers. The genetic analysis of aconitase (ACO), leucin aminopeptidase (LAP), menadione reductase (MDR), phosphoglucomutase (PGM), 6-phosphogluconic dehydrogenase (6-PGD), shikimate dehydrogenase (SKD), and triose phosphate isomerase (TPI) zymograms resulted in the identification of 8 isozyme marker loci, designated as Aco-1, Lap-1, Pgm-1, Pgm-3, 6-Pgd-2, Skd-1, Tpi-1, and Tpi-2. All loci segregated with codominant alleles and encoded for monomers (ACO, LAP, PGM, SKD), and dimers (6-PGD, TPI), respectively. MDR enzymes of the variable region MDR-2 appeared to be identical with Dia-2 isozymes. Tests of joint segregation for pairwise comparisons of all 14 isozyme marker loci now available in carrots indicate that 12 loci are linked in 4 linkage groups (marked K1 to K4) in the following order: Aco-1, Pgi-1, and Dia-3 (K1), Tpi-2, Got-2, and Lap-1 (K2), Got-3 and Tpi-1 (K3) and Pgm-1, Pgm-3, 6-Pgd-2 and Skd-1 (K4). Dia-2 and Got-1 remained unlinked. The possible duplication of a PGM locus and a 6-PGD locus is discussed.     

Ploidy Manipulation and Introgression of Resistance to Alternaria Helianthi from Wild Hexaploid Helianthus Species to Cultivated Sunflower (H. annuus L.) Aided by Anther Culture

The present investigation discusses the scope for transferring of resistance to leaf spot disease incited by Alternaria helianthi from two hexaploid wild species (H. tuberosus and H. resinosus) to diploid cultivated sunflower. Interspecific hybrids produced between sunflower and these two hexaploid species were partially fertile with tetraploid chromosome status. Backcrosses of these interspecific hybrids with cultivated sunflower resulted in the formation of sterile triploid plants. To overcome the problem of sterility and facilitate backcrosses with cultivated sunflower, anther culture of the tetraploid interspecific hybrids was carried out to bring down their chromosome number to diploid status. Anthers from both interspecific hybrids were cultured on basal Murashige and Skoog media supplemented with varying concentrations of organics and the growth regulators benzyladenine and naphthaleneacetic acid. Anthers of interspecific hybrids involving H. resinosus responded well and regenerated through an embryogenic route at a frequency of 98.7%. But in interspecific hybrids with H. tuberosus, anthers formed callus and subsequently regenerated shoots through an organogenic pathway. DNA ploidy analysis of anther culture plants of interspecific hybrids derived from H. tuberosus crosses was carried out to identify plants with desired diploid status. In vitro screening of parents, interspecific hybrids and anther culture plantlets against A. helianthi showed resistance in 68.5% of the anther culture plants of interspecific hybrids from H. tuberosus and in 24.3% of the plants derived from interspecific hybrids involving H. resinosus.     

Independent segregation of two isozyme markers and inter-plant differences in nuclear DNA content in the interspecific backcross (Allium fistulosum L. × A. cepa L.) × A. cepa L.

Summary From backcrosses of three interspecific hybrids (A. fistulosum x A. cepa) with a. cepa 14 diploid and 2 triploid plants were recovered.In this BC1 population introgression of A. fistulosum genetic material into the A. cepa nuclear genome was studied using two isozyme markers: Got-1 and Got-2. Both loci carried two alleles. A. cepa was monomorphic for both markers. A. fistulosum was polymorphic for Got-2. Based on their Got isozyme pattern seven out of the 14 diploid BC1 plants had a recombinant genotype. The loci appeared to be unlinked. Differences were observed in nuclear DNA contents between the diploid BC1 plants, indicating that gametes produced by the interspecific hybrids contained different combinations of chromosomal material from A. cepa and A. fistulosum.Abbreviations Adh alcohol dehydrogenase - Got glutamate oxaloacetate transaminase - Idh isocitrate dehydrogenase - Pgi phosphoglucoisomerase - Pgm phosphoglucomutase     

Construction of a crop—wild hybrid population for broadening genetic diversity in cultivated sunflower and first evaluation of its combining ability: the concept of neodomestication

The genetic base of sunflower elite lines is very narrow, due to many years of selection and breeding. To broaden the genetic diversity of the cultivated sunflower, in 1995 73 wild sunflower populations were crossed with 3 cultivated lines (Testers), and 219 hybrid offspring’s were evaluated in the field. GCA and SCA effects were computed suggesting for all traits a genetic potential for improvement through selection. Study of the hybrids revealed that the wild accessions bear different genetic abilities to combine with the testers for traits of morphological architecture, phenology and yield (seed weight and seed oil). The variance due to GCA and SCA showed that gene action was additive for days to flowering, branching and plant height. Genotypes derived from the same geographic origin may have either good or poor general combing ability. The correlation between GCA and per se genotype performance was positive for all traits except for seed oil content. This was the first attempt to evaluate wild-cultivated hybrids in sunflower on a large scale and will be the starting point for the management of hybrid Helianthus annuus populations for breeding. GCA and SCA estimations will facilitate the definition of strategies to manage and exploit the natural diversity for this crop.     

Genetic variation for isozyme genes and proteins in spanish primitive cultivars and wild subspecies of Lens

Summary An analysis of the variability for genes encoding seven isozyme systems and storage proteins in a collection of cultivated and wild accessions of Lens is reported. The collection, which is part of the Spanish INIA Cenebank, contains the ssp. culinaris, orientalis, odemensis, nigricans and ervoides, and presents a high degree of genetic diversity both within and between the accessions. A total 25 loci were examined; of these, 18 were polymorphic (the 7 genes encoding storage proteins, and the following isozyme loci: Acp-1, Acp-2, Cpx-1, Cpx-2, Aat-p, Aat-m, Lap-1, Mdh-2, Mdh-3, Mdh-4 and 6pgd-p) and 7 were monomorphic (Aat-mb, Aat-c, Mdh-1, Mdh-5, 6pgd-2, Pgm-c and Pgm-p). The phylogenetic relationships between subspecies were analyzed using the allelic frequencies. The study suggests that orientalis and odemensis share more biochemical characters than the other subspecies, and that those subspecies keep an intermediate position between Lens culinaris and Lens nigricans.     

Heredity of seventeen isozyme loci in cassava (Manihot esculenta Crantz)

Summary Starch gel electrophoresis was used to assess isozyme polymorphism in two Manihot species. Crude extracts were obtained from leaves and pollen. Ten enzymes were examined for their polymorphism in a germplasm collection of 365 cultivated plus 109 wild accessions, mainly from Africa. The inheritance of these enzymes was examined using 13 intra and interspecific progenies. Seventeen polymorphic loci were found for the ten enzyme systems, with 59 alleles. All the markers showed disomic heredity and three linkage groups were identified.