The Experts below are selected from a list of 3072 Experts worldwide ranked by ideXlab platform

Yoshihito Takahata - One of the best experts on this subject based on the ideXlab platform.

  • embryogenesis and doubled haploid production from Anther Culture in gentian gentiana triflora
    Plant Cell Tissue and Organ Culture, 2010
    Co-Authors: Ryo Takahashi, Takashi Hikage, Yoshihito Takahata
    Abstract:

    The overall goal of this study is to develop an Anther Culture system to produce doubled haploid (DH) lines of gentian (Gentiana triflora), an ornamental flowering plant, for use in an F1 hybrid breeding program. Embryogenesis was induced from Anther Cultures incubated on half-strength modified Lichter (NLN) medium containing a high concentration of sucrose (130 g/l) and subjected to heat shock treatment. Among the various parameters investigated, Anthers collected from buds 9–12 mm in length induced the highest frequency of androgenesis. Moreover, among three genotypes tested, cvs. Ashiro-no-Aki and Ashiro-no-Natsu produced 21.3 and 3.7 embryos per 100 Anthers, respectively, whereas, cv. Lovely-Ashiro failed to produce embryos. Among a total of 427 embryos transferred to a regeneration medium consisting of Murashige and Skoog (MS) medium, 138 plants were regenerated. The ploidy levels of regenerants were determined by flow cytometry and chromosome counts, revealing the presence of 5% haploids, 25% diploids, and 70% triploids. Inter simple sequence repeat (ISSR) analysis using the 6PS line obtained following self-pollination of the diploid plant obtained from Anther Culture confirmed that the diploid plant was indeed a DH.

  • embryogenesis and doubled haploid production from Anther Culture in gentian gentiana triflora
    Plant Cell Tissue and Organ Culture, 2010
    Co-Authors: Ryo Takahashi, Takashi Hikage, Yoshihito Takahata
    Abstract:

    The overall goal of this study is to develop an Anther Culture system to produce doubled haploid (DH) lines of gentian (Gentiana triflora), an ornamental flowering plant, for use in an F1 hybrid breeding program. Embryogenesis was induced from Anther Cultures incubated on half-strength modified Lichter (NLN) medium containing a high concentration of sucrose (130 g/l) and subjected to heat shock treatment. Among the various parameters investigated, Anthers collected from buds 9–12 mm in length induced the highest frequency of androgenesis. Moreover, among three genotypes tested, cvs. Ashiro-no-Aki and Ashiro-no-Natsu produced 21.3 and 3.7 embryos per 100 Anthers, respectively, whereas, cv. Lovely-Ashiro failed to produce embryos. Among a total of 427 embryos transferred to a regeneration medium consisting of Murashige and Skoog (MS) medium, 138 plants were regenerated. The ploidy levels of regenerants were determined by flow cytometry and chromosome counts, revealing the presence of 5% haploids, 25% diploids, and 70% triploids. Inter simple sequence repeat (ISSR) analysis using the 6PS line obtained following self-pollination of the diploid plant obtained from Anther Culture confirmed that the diploid plant was indeed a DH.

Maria Germana - One of the best experts on this subject based on the ideXlab platform.

  • microspore embryogenesis through Anther Culture in citrus clementina hort ex tan
    Methods of Molecular Biology, 2016
    Co-Authors: Benedetta Chiancone, Maria Germana
    Abstract:

    Anther Culture is a biotechnological method that allows to obtain, in one step, homozygous plants, very important to plant breeding, due to their numerous applications in mutation research, selection, genome sequencing, genetic analysis, and transformation. To induce the microspores, i.e., the immature male gametes, to switch from the normal gametophytic pathway to the sporophytic one, it is necessary to submit them to a type of stress, such as high or low temperature, starvation, or magnetic field. Stress can be applied to the donor plants and/or the floral buds or the Anthers or the isolated microspores, before or during the Culture. In this chapter, the protocol to induce gametic embryogenesis from Anther Culture of several cultivars of Citrus clementina Hort. ex Tan. is reported.

  • in vitro Anther Culture of sweet orange citrus sinensis l osbeck genotypes and of a c clementina c sinensis hamlin hybrid
    Plant Cell Tissue and Organ Culture, 2014
    Co-Authors: Maria Germana, Jean Carlos Cardoso, Adriana Pinheiro Martinelli, Rodrigo Rocha Latado
    Abstract:

    Citrus, and particularly sweet oranges, are very recalcitrant to Anther Culture. In this paper it was evaluated for the first time the response of 27 genotypes of Citrus sinensis and of one hybrid C. clementina × C. sinensis, to in vitro Anther Culture. Ten genotypes of sweet oranges showed embryogenic callus induction, mostly blood sweet oranges genotypes, such as Tarocco, Moro and Sanguinelli. In vitro microspore developmental switches from the gamethophytic to the sporophytic pathway were shown by DAPI staining in microspores of these responsive genotypes, after 10 months in Culture. However, microsatellite marker analyses showed that these calli were heterozygous. The flow-cytometric analysis of these embryogenic calli showed the presence of two peaks, corresponding to haploid (n) and diploid (2n) genotypes. Differently, Anther Cultures of the hybrid C. clementina × C. sinensis produced tri-haploid (3n) embryogenic calli and the embryos obtained were homozygous when analyzed by molecular markers (sample sequence repeats), confirming the more responsive characteristic of clementine to microspore embryogenesis through Anther Culture.

  • effect of polyamines on in vitro Anther Culture of citrus clementina hort ex tan
    Plant Cell Tissue and Organ Culture, 2006
    Co-Authors: Benedetta Chiancone, Nello Bagni, Annalisa Tassoni, Maria Germana
    Abstract:

    The improvement of the induction rate in Citrus Anther Culture is important for taking practical advantage of the haploid potential in breeding. The influence of polyamines on Anther Culture of Citrus clementina, cv Nules, with particular attention to the free, soluble and insoluble-conjugated polyamine levels, has been investigated. Putrescine, spermidine and putrescine plus spermidine, were added to the standard induction medium. Before Culture, spermidine was the most abundant among the free polyamines detected in Anthers. The exogenous supply of either putrescine or spermidine, either independently or combined, effected greater uptake and accumulation of polyamines. The addition of 2 mM spermidine to the medium stimulated gametic embryogenesis in clementine Nules, whereas putrescine did not influence embryo production. Regenerants were mostly tri-haploids; a few doubled-haploids and no haploid plants were obtained.

  • correlation of sequential floral and male gametophyte development and preliminary results on Anther Culture in opuntia ficus indica
    Acta Physiologiae Plantarum, 2005
    Co-Authors: Pablo Gonzalezmelendi, Maria Germana, Nathalie Levy Guarda, Benedetta Chiancone, Maria Carmen Risueno
    Abstract:

    Before approaching Anther Culture as a tool to trigger an androgenic response in a new species, it is advisable to characterize and correlate flower and male gametophyte development to enable reproducible identification of the appropriate starting material. Buds and flowers of Opuntia ficus-indica cv. Gialla were classified in eight stages according to their total length at the earlier stages and the length of the corolla in flowers with emerging sepals. Due to the low condensation of chromatin in the microspore nucleus as well as in the vegetative nucleus of the bi- and tricellular pollen along with the high autofluorescence of the intricate exine, DAPI staining turned out not to be feasible in this species. Therefore an approach based on light-microscopy observation of semithin sections was used. These sections were stained with toluidine blue for general structure recognition and I2KI to study starch deposition. Correlations were made between the sequential floral and male gametophyte development. Using this approach we determined the timing of pollen formation and observed that pollen development is impaired in plants producing seedless fruits. Furthermore, Anther Culture was carried out with Anthers collected from flower buds at stages 2 and 3. Most of the Anthers produced callus, however no regeneration was obtained.

  • Anther Culture in citrus clementina a way to regenerate tri haploids
    Crop & Pasture Science, 2005
    Co-Authors: Maria Germana, Benedetta Chiancone, O Lain, R Testolin
    Abstract:

    Abstract. Regenerants from Anther Culture of Citrus clementina Hort. ex Tan. cvv. Nules, SRA 63, and Monreal were obtained in different experiments from 1994 to 2002. Genetic analysis of 37 such regenerants was carried out using 4 microsatellite markers that were heterozygous in the parental genotypes. The results showed that in all cases but one the regenerants carried only one or the other allele of the parental genotype, and were therefore homozygous and produced through a process of gametophytic embryogenesis. Ploidy analysis by flow cytometry of 94 regenerants showed that as many as 82% of them were tri-haploids, rather than haploids or doubled-haploids as expected, with other ploidy levels such as n, 2n, 4n, and 6n occurring at very low percentages (2, 10, 3, and 3%, respectively). Regeneration from Anther Culture is therefore proposed as a rapid, and attractive method of obtaining new triploid varieties in clementine, which could be of great interest for the fresh fruit market that now requires fruit to be seedless.

Ryo Takahashi - One of the best experts on this subject based on the ideXlab platform.

  • embryogenesis and doubled haploid production from Anther Culture in gentian gentiana triflora
    Plant Cell Tissue and Organ Culture, 2010
    Co-Authors: Ryo Takahashi, Takashi Hikage, Yoshihito Takahata
    Abstract:

    The overall goal of this study is to develop an Anther Culture system to produce doubled haploid (DH) lines of gentian (Gentiana triflora), an ornamental flowering plant, for use in an F1 hybrid breeding program. Embryogenesis was induced from Anther Cultures incubated on half-strength modified Lichter (NLN) medium containing a high concentration of sucrose (130 g/l) and subjected to heat shock treatment. Among the various parameters investigated, Anthers collected from buds 9–12 mm in length induced the highest frequency of androgenesis. Moreover, among three genotypes tested, cvs. Ashiro-no-Aki and Ashiro-no-Natsu produced 21.3 and 3.7 embryos per 100 Anthers, respectively, whereas, cv. Lovely-Ashiro failed to produce embryos. Among a total of 427 embryos transferred to a regeneration medium consisting of Murashige and Skoog (MS) medium, 138 plants were regenerated. The ploidy levels of regenerants were determined by flow cytometry and chromosome counts, revealing the presence of 5% haploids, 25% diploids, and 70% triploids. Inter simple sequence repeat (ISSR) analysis using the 6PS line obtained following self-pollination of the diploid plant obtained from Anther Culture confirmed that the diploid plant was indeed a DH.

  • embryogenesis and doubled haploid production from Anther Culture in gentian gentiana triflora
    Plant Cell Tissue and Organ Culture, 2010
    Co-Authors: Ryo Takahashi, Takashi Hikage, Yoshihito Takahata
    Abstract:

    The overall goal of this study is to develop an Anther Culture system to produce doubled haploid (DH) lines of gentian (Gentiana triflora), an ornamental flowering plant, for use in an F1 hybrid breeding program. Embryogenesis was induced from Anther Cultures incubated on half-strength modified Lichter (NLN) medium containing a high concentration of sucrose (130 g/l) and subjected to heat shock treatment. Among the various parameters investigated, Anthers collected from buds 9–12 mm in length induced the highest frequency of androgenesis. Moreover, among three genotypes tested, cvs. Ashiro-no-Aki and Ashiro-no-Natsu produced 21.3 and 3.7 embryos per 100 Anthers, respectively, whereas, cv. Lovely-Ashiro failed to produce embryos. Among a total of 427 embryos transferred to a regeneration medium consisting of Murashige and Skoog (MS) medium, 138 plants were regenerated. The ploidy levels of regenerants were determined by flow cytometry and chromosome counts, revealing the presence of 5% haploids, 25% diploids, and 70% triploids. Inter simple sequence repeat (ISSR) analysis using the 6PS line obtained following self-pollination of the diploid plant obtained from Anther Culture confirmed that the diploid plant was indeed a DH.

J E Schmid - One of the best experts on this subject based on the ideXlab platform.

  • improved production of doubled haploids by colchicine application to wheat triticum aestivum l Anther Culture
    Plant Cell Reports, 1998
    Co-Authors: Amina Redha, S Saisingtong, Peter Stamp, B Buter, T Attia, J E Schmid
    Abstract:

    The aim of this study was to optimize the in vitro chromosome-doubling procedure in wheat Anther Culture. Colchicine, at concentrations of 100–5000 mg/l, was added to the induction medium for 1–5 days. Beneficial effects were obtained with concentrations of 100 and 1000 mg/l colchicine. With time, significant reductions in embryo–like structures as well as higher doubling indices were found. Similar results were obtained with the high- and low-responding genotypes. Colchicine (100 mg/l), added 5 and 20 days after inoculation for 1 and 3 days increased the induction response, but this value was reduced when colchicine was added 10 or 15 days after inoculation. The doubling effect was similar to the control, except for a significant increase with the 3-day application 20 days after inoculation. The highest success index was reached when colchicine was added to the Culture medium after 20 days.

  • colchicine mediated chromosome doubling during Anther Culture of maize zea mays l
    Theoretical and Applied Genetics, 1996
    Co-Authors: S Saisingtong, Peter Stamp, J E Schmid, B Buter
    Abstract:

    Efficient methods of chromosome doubling are critical for the production of microspore-derived, doubled-haploid (=DH) plants, especially if, as in maize Anther Culture, spontaneous chromosome doubling occurs infrequently. In the present study, colchicine (5–1000 mg/l) was added to the induction medium and maize Anthers were incubated in the colchicine-containing medium for different durations (1–7 days). In order to improve overall Anther Culture response, the Culture temperature was adjusted to 14°C during the first 7 days. Colchicine applied at low concentration, i.e. 5 mg/l (7 days), or for short duration, i.e. 1–3 days (250 mg/l), showed beneficial effects on the formation of embryolike structures (=ES) and thus led to increased plant production, but was comparatively ineffective regarding chromosome doubling. Optimal doubling effects were observed when Anthers had been exposed to Culture medium containing 250 and 1000 mg/l of colchicine (7 days); in these treatments the doubling index (=DI), defined as the quotient of the number of DH plants and the number of totally regenerated plants in a specific treatment, rose to 0.56 and 0.53, respectively, compared to 0.20 in the untreated control. However, colchicine administered at concentrations higher than 250 mg/l seemed to be detrimental to general plant production; thus, in spite of a high DI, the overall DH plant production was even lower than in the control treatment. Maximum DH plant production for three different genotypes was accomplished with Culture medium containing 250 mg/l of colchicine (7 days). With the best-responding genotype (ETH-M 36) a DH plant production of 9.9 DH plants/100 Anthers was accomplished, i.e. a 7-fold increase compared to the non-treated Anthers. This is the first report on efficient chromosome doubling in Anther Culture by subjecting Anthers to colchicinecontaining induction medium during a post-plating cold treatment. Chromosome doubling as described here becomes an integral part of the maize Anther Culture protocol and thus represents a rapid and economical way to produce DH plants.

Pauk János - One of the best experts on this subject based on the ideXlab platform.

  • In vitro Anther Culture as efficiently applied technique for doubled haploid production of wheat (Triticum aestivum L.)
    'Centre for Evaluation in Education and Science (CEON CEES)', 2021
    Co-Authors: Kanbar Osama Zuhair, Lantos Csaba, Pauk János
    Abstract:

    In vitro Anther Culture is the most commonly-used technology for doubled haploid production in wheat (Triticum aestivum L.) and various cereals. The technology application allows the genetic attainment of homozygous pure lines from heterozygous breeding material in a single generation, and has become a rapid alternative to the conventional breeding methods. This technology also contributes to more accurately assess QTL × environmental interactions and has been applied in: genetic researches for marker-trait association study, genomics and as a target for transformation, genetic engineering, gene mapping, and mapping of quantitative trait loci (QTLs). This paper reviews the most important factors that limit the efficiency of winter wheat (Triticum aestivum L.) Anther Culture, such as genotype dependency and albinism, in addition to other affecting factors including the collection time of tillers and physiological growth environments of plants. Factors that improve the efficiency of wheat Anther Culture in mitigating both genotypic dependency and albinism have been included in this review, too, for example, genetic improvements by following a strategy to realize this purpose through using responsive breeding material for crossing, application of convenient stress pre-treatments (cold pre-treatments, heat shock treatment, colchicine, hormones, and chemicals) to induce androgenesis, and improvement of the composition of Anther Culture media and Culture conditions. We have also highlighted the methods of chromosome doubling for the haploid wheat produced by Anther Culture androgenesis, such as spontaneous chromosome doubling or use of colchicine and anti-mitotic chemicals at an early stage of Anther Culture or as root immersion treatment

  • Generation of doubled haploid lines from winter wheat (Triticum aestivum L.) breeding material using in vitro Anther Culture
    'Czech Academy of Agricultural Sciences', 2020
    Co-Authors: Kanbar Osama Zuhair, Lantos Csaba, Chege Paul, Kiss Erzsébet, Pauk János
    Abstract:

    We investigated the Anther Culture (AC) efficiency of thirteen F4 combinations of winter wheat (Triticum aestivum L.). The genotype dependency was assessed during the induction of the androgenic entities, i.e. embryo-like structures (ELS), regenerated-, green-, albino-, and transplanted plantlets. The number of green plantlets per 100 Anthers (GP/100A) varied from 0.36 to 24.74 GP/100A with a mean of 8.31 GP/100A. Albino plantlets (AP) occurred in each combination, ranging from 0.20 to 22.80 AP/100A with an average value of 5.59 AP/100A. Between 25–87.76 doubled haploid (DH) plants per 100 acclimatised plantlets (DH/100ADP), depending on the combination, with a mean of 59.74% were recovered. We have found the highest DH production in the combinations Béres/Midas, Kalász/Tacitus, Béres/Pamier, and Premio/5009. This improves remarkably the choice of basic genetic material in subsequent crossing programmes. These observations emphasise the usability and efficiency of in vitro AC in producing a large number of DH lines for breeding and the applied researches of winter wheat. Although albinism was found in each combination, it was mitigated by the in vitro AC application

  • Androgenic responses of winter wheat (Triticum aestivum L.) combinations in in vitro Anther Culture
    'National Library of Serbia', 2020
    Co-Authors: Kanbar Osama Zuhair, Lantos Csaba, Kiss Erzsébet, Pauk János
    Abstract:

    The androgenic parameters were investigated by in vitro Anther Culture (AC) on nine F25 breeding combinations of winter wheat (Triticum aestivium L.). Each combination produced embryo-like structures (ELS), green plantlets, albino plantlets, transplanted plantlets and acclimatized plantlets, with respect to the number of Anthers in AC. The number of AC-derived ELS was between 11.73 and 52.76 ELS/100 Anthers with the mean of 26.22 ELS/100 Anthers, out of which the number of regenerated green plantlets varied from 3.20 to 26.40 green plantlets/100 Anthers and the mean was 9.76 green plantlets/100 Anthers, while the number of transplanted plantlets ranged from 2.16 to 21.77 transplanted plantlets/100 Anthers. Furthermore, the number of albinos/100 Anthers was mitigated and varied between 0.72 and 6.20 albinos/100 Anthers. We also studied the number of green and albino plantlets per 100 ELS. The rate of green plantlets per 100 ELS ranged between 14.81% and 64.01%, with the overall mean 33.59%, while the rate of albinos per 100 ELS ranged from 4.82% to 20.70% with the overall mean 11.93%. In our experiment, the rate of acclimatized plantlets (70.15-91.57%) depended mostly on the combination. This study asserted the importance of AC method in wheat for in vitro production of green plants. Although albinism was found in each combination, it did not hinder the production of green plantlets. The satisfying results were achieved in green plantlets production compared to the previously published data, but further improvement will be needed continuously, experiment by experiment. The generated acclimatized plantlets will be used in the wheat breeding program as doubled haploid (DH) lines

  • Utilization of in Vitro Anther Culture in Spelt Wheat Breeding
    'MDPI AG', 2019
    Co-Authors: Lantos Csaba, Purgel Szandra, Ács Katalin, Langó Bernadett, Bóna Lajos, Boda Krisztina, Békés Ferenc, Pauk János
    Abstract:

    The efficiency of in vitro Anther Culture was screened in a full diallel population of four spelt wheat genotypes and ten F1 hybrids. Genotype dependency was observed based on the data of embryo-like structures (ELS), green-, albino plantlets. In the diallel population and ten F1 hybrids, the green plantlets production ranged from 13.75 to 85.00 and from 6.30 to 51.00, respectively. The Anther Culture-derived plants of F1 hybrids were grown up in the nursery. At the harvest, 436 spontaneous doubled haploid (DH) plants were identified among the 1535 Anther Culture-derived transplanted and grown up individual plants. The mean of spontaneous rediploidization was 28.4% which ranged from 9.76% to 54.24%. In two consecutive years, the agronomic values of ‘Tonkoly.pop1’ advanced line were compared with seven DH lines of ‘Tonkoly.pop1’ in the nursery. The DH lines achieved competitive values in comparison with ‘Tonkoly.pop1’ advanced line based on the 11 measured parameters (heading date, plant height, yield, hardness, width and length of seed, TKW, hulling yield, flour yield, protein and wet gluten content). These observations presage the efficient utilization of Anther Culture in spelt wheat breeding