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Androgenesis

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Ewa Dubas – One of the best experts on this subject based on the ideXlab platform.

  • β 1 3 glucanases and chitinases participate in the stress related defence mechanisms that are possibly connected with modulation of arabinogalactan proteins agp required for the Androgenesis initiation in rye secale cereale l
    Plant Science, 2021
    Co-Authors: Kamil Zielinski, Ewa Dubas, Monika Krzewska, Anna Nowicka, Zuzana Gersi, Agnieszka Janas, Ildiko Matusikova, Iwona żur
    Abstract:

    Abstract This work presents the biochemical, cytochemical and molecular studies on two groups of PR proteins, β-1,3-glucanases and chitinases, and the arabinogalactan proteins (AGP) during the early stages of Androgenesis induction in two breeding lines of rye (Secale cereale L.) with different androgenic potential. The process of Androgenesis was initiated by tillers pre-treatments with low temperature, mannitol and/or reduced glutathione and resulted in microspores reprogramming and formation of androgenic structures what was associated with high activity of β-1,3-glucanases and chitinases. Some isoforms of β-1,3-glucanases, namely several acidic isoforms of about 26 kDa; appeared to be anther specific. Chitinases were well represented but were less variable. RT-qPCR revealed that the cold-responsive chitinase genes Chit1 and Chit2 were expressed at a lower level in the microspores and whole anthers while the cold-responsive Glu2 and Glu3 were not active. The stress pre-treatments modifications promoted the AGP accumulation. An apparent dominance of some AGP epitopes (LM2, JIM4 and JIM14) was detected in the Androgenesis-responsive rye line. An abundant JIM13 epitopes in the vesicles and inner cell walls of the microspores and in the cell walls of the anther cell layers appeared to be the most specific for embryogenesis.

  • the effect of glutathione and mannitol on Androgenesis in anther and isolated microspore cultures of rye secale cereale l
    Plant Cell Tissue and Organ Culture, 2020
    Co-Authors: Kamil Zielinski, Ewa Dubas, Monika Krzewska, Iwona żur, Katarzyna Juzon, Przemyslaw Kopec, Anna Nowicka, Jana Moravcikova, Edyta Skrzypek
    Abstract:

    Androgenic responsiveness in anther (AC) and isolated microspore cultures (MC) was analysed using 15 lines of Polish winter rye (Secale cereale L.). The effect of low temperature (LT) alone or in combination with osmotic stress induced by mannitol treatment (MAN) and/or with reduced glutathione (GSH) on the effectiveness of the process was studied. Interestingly, each treatment had a different effect on microspore (mcs) vitality and capability to divide symmetrically. The first criterion for successful embryogenesis was to exceed the threshold number of at least 25% dividing microspores, which determined ‘embryogenic suspension culture’. In some configurations a spectacular effect was achieved, especially in lines highly recalcitrant to Androgenesis induction. Relatively high effectiveness of Androgenesis induction (up to 4.58 AS per 105 mcs per spike in MC and 21.29 AS per spike in AC) showed that the developed protocol with GSH and/or MAN tiller pre-treatments overcomes the genotypic barrier for Androgenesis initiation in rye. Moreover, relatively high, spontaneous genome diploidization (55%) of regenerated plants demonstrated that the described protocols could be effectively integrated into conventional rye breeding programmes. Glutathione and mannitol break the barrier of Androgenesis initiation by increasing the dividing and vital microspores ratio. This marker is useful for screening material in breeding programs utilizing DH technology.

  • The effect of glutathione and mannitol on Androgenesis in anther and isolated microspore cultures of rye (Secale cereale L.)
    Plant Cell Tissue and Organ Culture (PCTOC), 2020
    Co-Authors: Kamil Zielinski, Iwona Żur, Monika Krzewska, Katarzyna Juzon, Anna Nowicka, Jana Moravcikova, Edyta Skrzypek, Przemysław Kopeć, Ewa Dubas
    Abstract:

    AbstractAndrogenic responsiveness in anther (AC) and isolated microspore cultures (MC) was analysed using 15 lines of Polish winter rye (Secale cereale L.). The effect of low temperature (LT) alone or in combination with osmotic stress induced by mannitol treatment (MAN) and/or with reduced glutathione (GSH) on the effectiveness of the process was studied. Interestingly, each treatment had a different effect on microspore (mcs) vitality and capability to divide symmetrically. The first criterion for successful embryogenesis was to exceed the threshold number of at least 25% dividing microspores, which determined ‘embryogenic suspension culture’. In some configurations a spectacular effect was achieved, especially in lines highly recalcitrant to Androgenesis induction. Relatively high effectiveness of Androgenesis induction (up to 4.58 AS per 105 mcs per spike in MC and 21.29 AS per spike in AC) showed that the developed protocol with GSH and/or MAN tiller pre-treatments overcomes the genotypic barrier for Androgenesis initiation in rye. Moreover, relatively high, spontaneous genome diploidization (55%) of regenerated plants demonstrated that the described protocols could be effectively integrated into conventional rye breeding programmes.

Iwona Żur – One of the best experts on this subject based on the ideXlab platform.

  • The effect of glutathione and mannitol on Androgenesis in anther and isolated microspore cultures of rye (Secale cereale L.)
    Plant Cell Tissue and Organ Culture (PCTOC), 2020
    Co-Authors: Kamil Zielinski, Iwona Żur, Monika Krzewska, Katarzyna Juzon, Anna Nowicka, Jana Moravcikova, Edyta Skrzypek, Przemysław Kopeć, Ewa Dubas
    Abstract:

    AbstractAndrogenic responsiveness in anther (AC) and isolated microspore cultures (MC) was analysed using 15 lines of Polish winter rye (Secale cereale L.). The effect of low temperature (LT) alone or in combination with osmotic stress induced by mannitol treatment (MAN) and/or with reduced glutathione (GSH) on the effectiveness of the process was studied. Interestingly, each treatment had a different effect on microspore (mcs) vitality and capability to divide symmetrically. The first criterion for successful embryogenesis was to exceed the threshold number of at least 25% dividing microspores, which determined ‘embryogenic suspension culture’. In some configurations a spectacular effect was achieved, especially in lines highly recalcitrant to Androgenesis induction. Relatively high effectiveness of Androgenesis induction (up to 4.58 AS per 105 mcs per spike in MC and 21.29 AS per spike in AC) showed that the developed protocol with GSH and/or MAN tiller pre-treatments overcomes the genotypic barrier for Androgenesis initiation in rye. Moreover, relatively high, spontaneous genome diploidization (55%) of regenerated plants demonstrated that the described protocols could be effectively integrated into conventional rye breeding programmes.

  • Failure of Androgenesis in Miscanthus × giganteus in vitro culture of cytologically unbalanced microspores
    Plant reproduction, 2013
    Co-Authors: Iwona Żur, Ewa Dubas, Aneta Słomka, Franciszek Dubert, Elżbieta Kuta, Agnieszka Płażek
    Abstract:

    Miscanthus × giganteus is a popular energy crop, which due to its hybrid origin is only vegetatively reproduced. Asexual embryogenesis in anther and microspore culture leading to double haploids production could allow to regain the ability for sexual reproduction and to increase the biodiversity of the species. Therefore, the goal of this paper was to investigate the requirements of Androgenesis in Miscanthus. The standard protocols used for monocotyledonous plants were applied with many modifications regarding the developmental stage of the explants at the time of culture initiation, stress treatment applied to panicles and isolated anthers as well as various chemical and physical parameters of in vitro culture conditions. Our results indicated that the induction of Androgenesis in M. × giganteus is possible. However, the very low efficiency of the process and the lack of regeneration ability of the androgenic structures presently prevent the use of this technique.

  • Endogenous ABA concentration and cytoplasmic membrane fluidity in microspores of oilseed rape (Brassica napus L.) genotypes differing in responsiveness to Androgenesis induction
    Plant cell reports, 2013
    Co-Authors: Ewa Dubas, Franciszek Janowiak, Monika Krzewska, Tomasz Hura, Iwona Żur
    Abstract:

    Key message A better understanding of Androgenesis with a focus on the changes in plasma membrane fluifluidity and endogenous ABA content affecting embryogenesis induction in microspore suspension ofB. napus.

A. S. Grunina – One of the best experts on this subject based on the ideXlab platform.

  • Dispermic Androgenesis in sturgeons with the use of cryopreserved sperm: Production of androgenetic siberian sturgeon and androgenetic hybrids between Siberian and Russian sturgeons
    Russian Journal of Developmental Biology, 2011
    Co-Authors: A. S. Grunina, A. V. Rekoubratsky, L. I. Tsvetkova, A. E. Barmintseva, Ekaterina D. Vasil’eva, K. V. Kovalev, O. G. Poluektova
    Abstract:

    Dispermic Androgenesis was used to produce, for the first time, an androgenetic progeny of the Siberian sturgeon (Acipenser baerii) and the androgenetic nucleo-cytoplasmic hybrids (Siberian sturgeon, A. baerii × Russian sturgeon, A. gueldenstaedtii) using cryopreserved sperm. Microsatellite DNA analysis confirmed exclusively paternal inheritance in the androgenetic progeny of Siberian sturgeon. Heterozygotes for certain microsatellite loci were detected among the androgenetic hybrids, thereby confirming a dispermic nature of Androgenesis. According to the data of comparative morphological analysis, the obtained androgenetic hybrid, by the age of 15 months old, was completely identical to the paternal species. Both a female and a male were detected in the androgenetic sturgeon progenies, which is of interest for producing bisexual progenies via Androgenesis. The data of this study confirm the feasibility of dispermic Androgenesis using cryopreserved sperm to preserve and recover the gene pools of endangered sturgeon species.

  • Dispermic Androgenesis in sturgeons with the help of cryopreserved sperm: production of androgenetic hybrids between Siberian and Russian sturgeons
    Ontogenez, 2011
    Co-Authors: A. S. Grunina, L. I. Tsvetkova, A. E. Barmintseva, Ekaterina D. Vasil'eva, Kirill Kovalev, O. G. Poluektova
    Abstract:

    Dispermic Androgenesis was used to produce, for the first time, an androgenetic progeny of the Siberian sturgeon (Acipenser baerii) and the androgenetic nuclear cytoplasmic hybrids (Siberian sturgeon, A. baerii x Russian sturgeon, A. gueldenstaedtii) using cryopreserved sperm. Microsatellite DNA analysis confirmed exclusively paternal inheritance in the androgenetic progeny of Siberian sturgeon. Heterozygotes for certain microsatellite loci were detected among the androgenetic hybrids, thereby confirming a dispermic nature of Androgenesis. According to the data of comparative morphological analysis, the obtained androgenetic hybrid, by the age of 15 months old, was completely identical to the paternal species. Both a female and a male were detected in the androgenetic sturgeon progenies, which is of interest for producing bisexual progenies via Androgenesis. The data of this study confirm the feasibility of dispermic Androgenesis using cryopreserved sperm to preserve and recover the gene pools of endangered sturgeon species.

  • Investigation on dispermic Androgenesis in sturgeon fish. The first successful production of androgenetic sturgeons with cryopreserved sperm
    International Journal of Refrigeration, 2006
    Co-Authors: A. S. Grunina, L. I. Tsvetkova, A. V. Recoubratsky, V. A. Barmintsev
    Abstract:

    Induced diploid Androgenesis is considered as a valuable tool for restoration of endangered or extinct species from cryopreserved spermatozoa. The method of dispermic Androgenesis was developed in sturgeon fishes whose threatened status requires urgent conservation efforts. The method includes genetic inactivation of eggs, their insemination with concentrated sperm (to cause polyspermy), and the heat shock that facilitates the fusion of male pronuclei. Restoration of diploid state of androgenotes by fusion of two sperm nuclei allows androgenetic progeny to have a heterozygosity level as similar as in a regular crossing. Using this method, viable androgenetic progenies were obtained for the first time in the Siberian, Russian, stellate, and beluga sturgeons. Then a number of androgenetic nucleocytoplasmic hybrids was obtained. Androgenetic hybrids were shown to have nuclear DNA of paternal species and mitochondrial DNA of maternal species. The first experiments in which cryopreserved sperm was used have demonstrated that dispermic Androgenesis may be applied to restore sturgeons from their cryopreserved spermatozoa. Haploid Androgenesis was also used to examine whether cryopreservation procedures cause lesions in DNA of sturgeon spermatozoa.

Kamil Zielinski – One of the best experts on this subject based on the ideXlab platform.

  • β 1 3 glucanases and chitinases participate in the stress related defence mechanisms that are possibly connected with modulation of arabinogalactan proteins agp required for the Androgenesis initiation in rye secale cereale l
    Plant Science, 2021
    Co-Authors: Kamil Zielinski, Ewa Dubas, Monika Krzewska, Anna Nowicka, Zuzana Gersi, Agnieszka Janas, Ildiko Matusikova, Iwona żur
    Abstract:

    Abstract This work presents the biochemical, cytochemical and molecular studies on two groups of PR proteins, β-1,3-glucanases and chitinases, and the arabinogalactan proteins (AGP) during the early stages of Androgenesis induction in two breeding lines of rye (Secale cereale L.) with different androgenic potential. The process of Androgenesis was initiated by tillers pre-treatments with low temperature, mannitol and/or reduced glutathione and resulted in microspores reprogramming and formation of androgenic structures what was associated with high activity of β-1,3-glucanases and chitinases. Some isoforms of β-1,3-glucanases, namely several acidic isoforms of about 26 kDa; appeared to be anther specific. Chitinases were well represented but were less variable. RT-qPCR revealed that the cold-responsive chitinase genes Chit1 and Chit2 were expressed at a lower level in the microspores and whole anthers while the cold-responsive Glu2 and Glu3 were not active. The stress pre-treatments modifications promoted the AGP accumulation. An apparent dominance of some AGP epitopes (LM2, JIM4 and JIM14) was detected in the Androgenesis-responsive rye line. An abundant JIM13 epitopes in the vesicles and inner cell walls of the microspores and in the cell walls of the anther cell layers appeared to be the most specific for embryogenesis.

  • the effect of glutathione and mannitol on Androgenesis in anther and isolated microspore cultures of rye secale cereale l
    Plant Cell Tissue and Organ Culture, 2020
    Co-Authors: Kamil Zielinski, Ewa Dubas, Monika Krzewska, Iwona żur, Katarzyna Juzon, Przemyslaw Kopec, Anna Nowicka, Jana Moravcikova, Edyta Skrzypek
    Abstract:

    Androgenic responsiveness in anther (AC) and isolated microspore cultures (MC) was analysed using 15 lines of Polish winter rye (Secale cereale L.). The effect of low temperature (LT) alone or in combination with osmotic stress induced by mannitol treatment (MAN) and/or with reduced glutathione (GSH) on the effectiveness of the process was studied. Interestingly, each treatment had a different effect on microspore (mcs) vitality and capability to divide symmetrically. The first criterion for successful embryogenesis was to exceed the threshold number of at least 25% dividing microspores, which determined ‘embryogenic suspension culture’. In some configurations a spectacular effect was achieved, especially in lines highly recalcitrant to Androgenesis induction. Relatively high effectiveness of Androgenesis induction (up to 4.58 AS per 105 mcs per spike in MC and 21.29 AS per spike in AC) showed that the developed protocol with GSH and/or MAN tiller pre-treatments overcomes the genotypic barrier for Androgenesis initiation in rye. Moreover, relatively high, spontaneous genome diploidization (55%) of regenerated plants demonstrated that the described protocols could be effectively integrated into conventional rye breeding programmes. Glutathione and mannitol break the barrier of Androgenesis initiation by increasing the dividing and vital microspores ratio. This marker is useful for screening material in breeding programs utilizing DH technology.

  • The effect of glutathione and mannitol on Androgenesis in anther and isolated microspore cultures of rye (Secale cereale L.)
    Plant Cell Tissue and Organ Culture (PCTOC), 2020
    Co-Authors: Kamil Zielinski, Iwona Żur, Monika Krzewska, Katarzyna Juzon, Anna Nowicka, Jana Moravcikova, Edyta Skrzypek, Przemysław Kopeć, Ewa Dubas
    Abstract:

    AbstractAndrogenic responsiveness in anther (AC) and isolated microspore cultures (MC) was analysed using 15 lines of Polish winter rye (Secale cereale L.). The effect of low temperature (LT) alone or in combination with osmotic stress induced by mannitol treatment (MAN) and/or with reduced glutathione (GSH) on the effectiveness of the process was studied. Interestingly, each treatment had a different effect on microspore (mcs) vitality and capability to divide symmetrically. The first criterion for successful embryogenesis was to exceed the threshold number of at least 25% dividing microspores, which determined ‘embryogenic suspension culture’. In some configurations a spectacular effect was achieved, especially in lines highly recalcitrant to Androgenesis induction. Relatively high effectiveness of Androgenesis induction (up to 4.58 AS per 105 mcs per spike in MC and 21.29 AS per spike in AC) showed that the developed protocol with GSH and/or MAN tiller pre-treatments overcomes the genotypic barrier for Androgenesis initiation in rye. Moreover, relatively high, spontaneous genome diploidization (55%) of regenerated plants demonstrated that the described protocols could be effectively integrated into conventional rye breeding programmes.

Monika Krzewska – One of the best experts on this subject based on the ideXlab platform.

  • β 1 3 glucanases and chitinases participate in the stress related defence mechanisms that are possibly connected with modulation of arabinogalactan proteins agp required for the Androgenesis initiation in rye secale cereale l
    Plant Science, 2021
    Co-Authors: Kamil Zielinski, Ewa Dubas, Monika Krzewska, Anna Nowicka, Zuzana Gersi, Agnieszka Janas, Ildiko Matusikova, Iwona żur
    Abstract:

    Abstract This work presents the biochemical, cytochemical and molecular studies on two groups of PR proteins, β-1,3-glucanases and chitinases, and the arabinogalactan proteins (AGP) during the early stages of Androgenesis induction in two breeding lines of rye (Secale cereale L.) with different androgenic potential. The process of Androgenesis was initiated by tillers pre-treatments with low temperature, mannitol and/or reduced glutathione and resulted in microspores reprogramming and formation of androgenic structures what was associated with high activity of β-1,3-glucanases and chitinases. Some isoforms of β-1,3-glucanases, namely several acidic isoforms of about 26 kDa; appeared to be anther specific. Chitinases were well represented but were less variable. RT-qPCR revealed that the cold-responsive chitinase genes Chit1 and Chit2 were expressed at a lower level in the microspores and whole anthers while the cold-responsive Glu2 and Glu3 were not active. The stress pre-treatments modifications promoted the AGP accumulation. An apparent dominance of some AGP epitopes (LM2, JIM4 and JIM14) was detected in the Androgenesis-responsive rye line. An abundant JIM13 epitopes in the vesicles and inner cell walls of the microspores and in the cell walls of the anther cell layers appeared to be the most specific for embryogenesis.

  • the effect of glutathione and mannitol on Androgenesis in anther and isolated microspore cultures of rye secale cereale l
    Plant Cell Tissue and Organ Culture, 2020
    Co-Authors: Kamil Zielinski, Ewa Dubas, Monika Krzewska, Iwona żur, Katarzyna Juzon, Przemyslaw Kopec, Anna Nowicka, Jana Moravcikova, Edyta Skrzypek
    Abstract:

    Androgenic responsiveness in anther (AC) and isolated microspore cultures (MC) was analysed using 15 lines of Polish winter rye (Secale cereale L.). The effect of low temperature (LT) alone or in combination with osmotic stress induced by mannitol treatment (MAN) and/or with reduced glutathione (GSH) on the effectiveness of the process was studied. Interestingly, each treatment had a different effect on microspore (mcs) vitality and capability to divide symmetrically. The first criterion for successful embryogenesis was to exceed the threshold number of at least 25% dividing microspores, which determined ‘embryogenic suspension culture’. In some configurations a spectacular effect was achieved, especially in lines highly recalcitrant to Androgenesis induction. Relatively high effectiveness of Androgenesis induction (up to 4.58 AS per 105 mcs per spike in MC and 21.29 AS per spike in AC) showed that the developed protocol with GSH and/or MAN tiller pre-treatments overcomes the genotypic barrier for Androgenesis initiation in rye. Moreover, relatively high, spontaneous genome diploidization (55%) of regenerated plants demonstrated that the described protocols could be effectively integrated into conventional rye breeding programmes. Glutathione and mannitol break the barrier of Androgenesis initiation by increasing the dividing and vital microspores ratio. This marker is useful for screening material in breeding programs utilizing DH technology.

  • The effect of glutathione and mannitol on Androgenesis in anther and isolated microspore cultures of rye (Secale cereale L.)
    Plant Cell Tissue and Organ Culture (PCTOC), 2020
    Co-Authors: Kamil Zielinski, Iwona Żur, Monika Krzewska, Katarzyna Juzon, Anna Nowicka, Jana Moravcikova, Edyta Skrzypek, Przemysław Kopeć, Ewa Dubas
    Abstract:

    AbstractAndrogenic responsiveness in anther (AC) and isolated microspore cultures (MC) was analysed using 15 lines of Polish winter rye (Secale cereale L.). The effect of low temperature (LT) alone or in combination with osmotic stress induced by mannitol treatment (MAN) and/or with reduced glutathione (GSH) on the effectiveness of the process was studied. Interestingly, each treatment had a different effect on microspore (mcs) vitality and capability to divide symmetrically. The first criterion for successful embryogenesis was to exceed the threshold number of at least 25% dividing microspores, which determined ‘embryogenic suspension culture’. In some configurations a spectacular effect was achieved, especially in lines highly recalcitrant to Androgenesis induction. Relatively high effectiveness of Androgenesis induction (up to 4.58 AS per 105 mcs per spike in MC and 21.29 AS per spike in AC) showed that the developed protocol with GSH and/or MAN tiller pre-treatments overcomes the genotypic barrier for Androgenesis initiation in rye. Moreover, relatively high, spontaneous genome diploidization (55%) of regenerated plants demonstrated that the described protocols could be effectively integrated into conventional rye breeding programmes.