The Experts below are selected from a list of 87942 Experts worldwide ranked by ideXlab platform
P S Testillano - One of the best experts on this subject based on the ideXlab platform.
-
endogenous auxin accumulation localization during zygotic and somatic Embryogenesis of capsicum chinense jacq
Journal of Plant Physiology, 2021Co-Authors: Jacobo Perezpastrana, P S Testillano, Ivett Barany, Adriana Cantoflick, Dulce Alvarezlopez, Gema Pijeirafernandez, Susana A Avilesvinas, Laura P Penayam, Liliana S Munozramirez, Sara NahuatdzibAbstract:Zygotic and somatic Embryogenesis in plants is a fascinating event that is finely regulated through the expression of a specific group of genes and dynamic levels of plant hormones whose concerted action determines the fate that specific cells follow towards zygotic or somatic embryo development. This work studied different stages of Capsicum chinense Jacq. zygotic and somatic Embryogenesis. HPLC quantification determined that the levels of indole-3-acetic acid (IAA) increase as the zygotic or somatic Embryogenesis progresses, being higher at maturity, thus supporting a positive correlation between embryo cell differentiation and IAA increase. A monoclonal anti-IAA-antibody was used to detect IAA levels. Findings revealed a dynamic pattern of auxin distribution along the different embryogenic embryonic stages. In the early stages of zygotic embryos, the IAA gradient was observed in the basal cells of the suspensor and the hypostases, suggesting that they are the initial source of the IAA hormone. As Embryogenesis proceeds, the dynamic of the IAA gradient is displaced to the embryo and endosperm cells. In the case of induced somatic Embryogenesis, the IAA gradient was detected in the dividing cells of the endodermis, from where pre-embryogenic cells emerge. However, the analysis of somatic embryos revealed that IAA was homogeneously distributed. This study shows evidence supporting a correlation between IAA levels during zygotic or somatic Embryogenesis in Capsicum chinense species.
-
stress induced microspore Embryogenesis requires endogenous auxin synthesis and polar transport in barley
Frontiers in Plant Science, 2019Co-Authors: Yolanda Perezperez, Mariateresa Solis, Mariacarmen Risueno, Ahmedabdalla Eltantawy, P S TestillanoAbstract:Stress-induced microspore Embryogenesis is a model in vitro system of cell reprogramming, totipotency acquisition, and embryo development. After induction, responsive microspores abandon their developmental program to follow an embryogenic pathway, leading to in vitro embryo formation. This process is widely used to produce doubled-haploid lines, essential players to create new materials in modern breeding programs, particularly in cereals, although its efficiency is still low in many crop species, because the regulating mechanisms are still elusive. Stress signaling and endogenous hormones, mainly auxin, have been proposed as determinant factors of microspore Embryogenesis induction in some eudicot species; however, much less information is available in monocot plants. In this study, we have analyzed the dynamics and possible role of endogenous auxin during stress-induced microspore Embryogenesis in the monocot Hordeum vulgare, barley. The results showed auxin accumulation in early proembryo cells, from Embryogenesis initiation and a further increase with embryo development and differentiation, correlating with the induction and expression pattern of the auxin biosynthesis gene HvTAR2-like. Pharmacological treatments with kynurenine, inhibitor of auxin biosynthesis, and α-(p-chlorophenoxy)-isobutyric acid (PCIB), auxin antagonist, impaired Embryogenesis initiation and development, indicating that de novo auxin synthesis and its activity were required for the process. Efflux carrier gene HvPIN1-like was also induced with Embryogenesis initiation and progression; auxin transport inhibition by N-1-naphthylphthalamic acid significantly reduced embryo development at early and advanced stages. The results indicate activation of auxin biosynthesis with microspore Embryogenesis initiation and progression, in parallel with the activation of polar auxin transport, and reveal a central role of auxin in the process in a monocot species. The findings give new insights into the complex regulation of stress-induced microspore Embryogenesis, particularly in monocot plants for which information is still scarce, and suggest that manipulation of endogenous auxin content could be a target to improve in vitro embryo production.
-
autophagy is activated and involved in cell death with participation of cathepsins during stress induced microspore Embryogenesis in barley
Journal of Experimental Botany, 2018Co-Authors: Ivett Barany, Mariateresa Solis, Mariacarmen Risueno, Eduardo Berenguer, Yolanda Perezperez, Estrella M Santamaria, Jose L Crespo, Isabel Diaz, P S TestillanoAbstract:Microspores are reprogrammed towards Embryogenesis by stress. Many microspores die after this stress, limiting the efficiency of microspore Embryogenesis. Autophagy is a degradation pathway that plays critical roles in stress response and cell death. In animals, cathepsins have an integral role in autophagy by degrading autophagic material; less is known in plants. Plant cathepsins are papain-like C1A cysteine proteases involved in many physiological processes, including programmed cell death. We have analysed the involvement of autophagy in cell death, in relation to cathepsin activation, during stress-induced microspore Embryogenesis in Hordeum vulgare. After stress, reactive oxygen species (ROS) and cell death increased and autophagy was activated, including HvATG5 and HvATG6 up-regulation and increase of ATG5, ATG8, and autophagosomes. Concomitantly, cathepsin L/F-, B-, and H-like activities were induced, cathepsin-like genes HvPap-1 and HvPap-6 were up-regulated, and HvPap-1, HvPap-6, and HvPap-19 proteins increased and localized in the cytoplasm, resembling autophagy structures. Inhibitors of autophagy and cysteine proteases reduced cell death and promoted Embryogenesis. The findings reveal a role for autophagy in stress-induced cell death during microspore Embryogenesis, and the participation of cathepsins. Similar patterns of activation, expression, and localization suggest a possible connection between cathepsins and autophagy. The results open up new possibilities to enhance microspore Embryogenesis efficiency with autophagy and/or cysteine protease modulators.
-
early markers are present in both Embryogenesis pathways from microspores and immature zygotic embryos in cork oak quercus suber l
BMC Plant Biology, 2014Co-Authors: Hector Rodriguezsanz, Mariateresa Solis, Mariacarmen Risueno, J A Manzanera, Aranzazu Gomezgaray, Beatriz Pintos, P S TestillanoAbstract:In Quercus suber, cork oak, a Mediterranean forest tree of economic and social interest, rapid production of isogenic lines and clonal propagation of elite genotypes have been achieved by developing in vitro Embryogenesis from microspores and zygotic embryos respectively. Despite its high potential in tree breeding strategies, due to their recalcitrancy, the efficiency of Embryogenesis in vitro systems in many woody species is still very low since factors responsible for Embryogenesis initiation and embryo development are still largely unknown. The search for molecular and cellular markers during early stages of in vitro Embryogenesis constitutes an important goal to distinguish, after induction, responsive from non-responsive cells, and to elucidate the mechanisms involved in Embryogenesis initiation for their efficient manipulation. In this work, we have performed a comparative analysis of two Embryogenesis pathways derived from microspores and immature zygotic embryos in cork oak in order to characterize early markers of reprogrammed cells in both pathways. Rearrangements of the cell structural organization, changes in epigenetic marks, cell wall polymers modifications and endogenous auxin changes were analyzed at early Embryogenesis stages of the two in vitro systems by a multidisciplinary approach. Results showed that early embryo cells exhibited defined changes of cell components which were similar in both Embryogenesis in vitro systems, cellular features that were not found in non-embryogenic cells. DNA methylation level and nuclear pattern, proportion of esterified pectins in cell walls, and endogenous auxin levels were different in embryo cells in comparison with microspores and immature zygotic embryo cells from which embryos originated, constituting early Embryogenesis markers. These findings suggest that DNA hypomethylation, cell wall remodeling by pectin esterification and auxin increase are involved in early in vitro Embryogenesis in woody species, providing new evidences of the developmental pattern similarity between both Embryogenesis pathways, from microspores and immature zygotic embryos, in woody species.
-
a new microspore Embryogenesis system under low temperature which mimics zygotic Embryogenesis initials expresses auxin and efficiently regenerates doubled haploid plants in brassica napus
BMC Plant Biology, 2012Co-Authors: Deepak Prem, Mariateresa Solis, Mariacarmen Risueno, Ivett Barany, Hector Rodriguezsanz, P S TestillanoAbstract:Background Microspore Embryogenesis represents a unique system of single cell reprogramming in plants wherein a highly specialized cell, the microspore, by specific stress treatment, switches its fate towards an Embryogenesis pathway. In Brassica napus, a model species for this phenomenon, incubation of isolated microspores at 32°C is considered to be a pre-requisite for Embryogenesis induction.
Mariateresa Solis - One of the best experts on this subject based on the ideXlab platform.
-
stress induced microspore Embryogenesis requires endogenous auxin synthesis and polar transport in barley
Frontiers in Plant Science, 2019Co-Authors: Yolanda Perezperez, Mariateresa Solis, Mariacarmen Risueno, Ahmedabdalla Eltantawy, P S TestillanoAbstract:Stress-induced microspore Embryogenesis is a model in vitro system of cell reprogramming, totipotency acquisition, and embryo development. After induction, responsive microspores abandon their developmental program to follow an embryogenic pathway, leading to in vitro embryo formation. This process is widely used to produce doubled-haploid lines, essential players to create new materials in modern breeding programs, particularly in cereals, although its efficiency is still low in many crop species, because the regulating mechanisms are still elusive. Stress signaling and endogenous hormones, mainly auxin, have been proposed as determinant factors of microspore Embryogenesis induction in some eudicot species; however, much less information is available in monocot plants. In this study, we have analyzed the dynamics and possible role of endogenous auxin during stress-induced microspore Embryogenesis in the monocot Hordeum vulgare, barley. The results showed auxin accumulation in early proembryo cells, from Embryogenesis initiation and a further increase with embryo development and differentiation, correlating with the induction and expression pattern of the auxin biosynthesis gene HvTAR2-like. Pharmacological treatments with kynurenine, inhibitor of auxin biosynthesis, and α-(p-chlorophenoxy)-isobutyric acid (PCIB), auxin antagonist, impaired Embryogenesis initiation and development, indicating that de novo auxin synthesis and its activity were required for the process. Efflux carrier gene HvPIN1-like was also induced with Embryogenesis initiation and progression; auxin transport inhibition by N-1-naphthylphthalamic acid significantly reduced embryo development at early and advanced stages. The results indicate activation of auxin biosynthesis with microspore Embryogenesis initiation and progression, in parallel with the activation of polar auxin transport, and reveal a central role of auxin in the process in a monocot species. The findings give new insights into the complex regulation of stress-induced microspore Embryogenesis, particularly in monocot plants for which information is still scarce, and suggest that manipulation of endogenous auxin content could be a target to improve in vitro embryo production.
-
autophagy is activated and involved in cell death with participation of cathepsins during stress induced microspore Embryogenesis in barley
Journal of Experimental Botany, 2018Co-Authors: Ivett Barany, Mariateresa Solis, Mariacarmen Risueno, Eduardo Berenguer, Yolanda Perezperez, Estrella M Santamaria, Jose L Crespo, Isabel Diaz, P S TestillanoAbstract:Microspores are reprogrammed towards Embryogenesis by stress. Many microspores die after this stress, limiting the efficiency of microspore Embryogenesis. Autophagy is a degradation pathway that plays critical roles in stress response and cell death. In animals, cathepsins have an integral role in autophagy by degrading autophagic material; less is known in plants. Plant cathepsins are papain-like C1A cysteine proteases involved in many physiological processes, including programmed cell death. We have analysed the involvement of autophagy in cell death, in relation to cathepsin activation, during stress-induced microspore Embryogenesis in Hordeum vulgare. After stress, reactive oxygen species (ROS) and cell death increased and autophagy was activated, including HvATG5 and HvATG6 up-regulation and increase of ATG5, ATG8, and autophagosomes. Concomitantly, cathepsin L/F-, B-, and H-like activities were induced, cathepsin-like genes HvPap-1 and HvPap-6 were up-regulated, and HvPap-1, HvPap-6, and HvPap-19 proteins increased and localized in the cytoplasm, resembling autophagy structures. Inhibitors of autophagy and cysteine proteases reduced cell death and promoted Embryogenesis. The findings reveal a role for autophagy in stress-induced cell death during microspore Embryogenesis, and the participation of cathepsins. Similar patterns of activation, expression, and localization suggest a possible connection between cathepsins and autophagy. The results open up new possibilities to enhance microspore Embryogenesis efficiency with autophagy and/or cysteine protease modulators.
-
initiation of leaf somatic Embryogenesis involves high pectin esterification auxin accumulation and dna demethylation in quercus alba
Journal of Plant Physiology, 2017Co-Authors: Elena Corredoira, Mariateresa Solis, Mariacarmen Risueno, Vanesa Cano, Ivett Barany, Hector Rodriguez, Ana M Vieitez, P S TestillanoAbstract:Abstract Somatic Embryogenesis is considered a convenient tool for investigating the regulating mechanisms of embryo formation; it is also a feasible system for in vitro regeneration procedures, with many advantages in woody species. Nevertheless, trees have shown recalcitrance to somatic Embryogenesis, and its efficiency remains very low in many cases. Consequently, despite the clear potential of somatic Embryogenesis in tree breeding programs, its application is limited since factors responsible for Embryogenesis initiation have not yet been completely elucidated. In the present work, we investigated key cellular factors involved in the change of developmental program during leaf somatic Embryogenesis initiation of white oak ( Quercus alba ), aiming to identify early markers of the process. The results revealed that pectin esterification, auxin accumulation and DNA demethylation were induced during Embryogenesis initiation and differentially found in embryogenic cells, while they were not present in leaf cells before induction or in non-embryogenic cells after Embryogenesis initiation. These three factors constitute early markers of leaf Embryogenesis and represent processes that could be interconnected and involved in the regulation of cell reprogramming and Embryogenesis initiation. These findings provide new insights into the mechanisms underlying plant cell reprogramming, totipotency and embryogenic competence acquisition, especially in tree species for which information is scarce, thus opening up the possibility of efficient manipulation of somatic Embryogenesis induction.
-
auxin biosynthesis accumulation action and transport are involved in stress induced microspore Embryogenesis initiation and progression in brassica napus
Plant and Cell Physiology, 2015Co-Authors: Hector Rodriguezsanz, Mariateresa Solis, Mariacarmen Risueno, Mariafernanda Lopez, Aurelio Gomezcadenas, P S TestillanoAbstract:Isolated microspores are reprogrammed in vitro by stress, becoming totipotent cells and producing embryos and plants via a process known as microspore Embryogenesis. Despite the abundance of data on auxin involvement in plant development and Embryogenesis, no data are available regarding the dynamics of auxin concentration, cellular localization and the expression of biosynthesis genes during microspore Embryogenesis. This work involved the analysis of auxin concentration and cellular accumulation; expression of TAA1 and NIT2 encoding enzymes of two auxin biosynthetic pathways; expression of the PIN1-like efflux carrier; and the effects of inhibition of auxin transport and action by N-1-naphthylphthalamic acid (NPA) and α-(p-chlorophenoxy) isobutyric acid (PCIB) during Brassica napus microspore Embryogenesis. The results indicated de novo auxin synthesis after stress-induced microspore reprogramming and Embryogenesis initiation, accompanying the first cell divisions. The progressive increase of auxin concentration during progression of Embryogenesis correlated with the expression patterns of TAA1 and NIT2 genes of auxin biosynthetic pathways. Auxin was evenly distributed in early embryos, whereas in heart/torpedo embryos auxin was accumulated in apical and basal embryo regions. Auxin efflux carrier PIN1-like gene expression was induced in early multicellular embryos and increased at the globular/torpedo embryo stages. Inhibition of polar auxin transport (PAT) and action, by NPA and PCIB, impaired embryo development, indicating that PAT and auxin action are required for microspore embryo progression. NPA also modified auxin embryo accumulation patterns. These findings indicate that endogenous auxin biosynthesis, action and polar transport are required in stress-induced microspore reprogramming, Embryogenesis initiation and progression.
-
early markers are present in both Embryogenesis pathways from microspores and immature zygotic embryos in cork oak quercus suber l
BMC Plant Biology, 2014Co-Authors: Hector Rodriguezsanz, Mariateresa Solis, Mariacarmen Risueno, J A Manzanera, Aranzazu Gomezgaray, Beatriz Pintos, P S TestillanoAbstract:In Quercus suber, cork oak, a Mediterranean forest tree of economic and social interest, rapid production of isogenic lines and clonal propagation of elite genotypes have been achieved by developing in vitro Embryogenesis from microspores and zygotic embryos respectively. Despite its high potential in tree breeding strategies, due to their recalcitrancy, the efficiency of Embryogenesis in vitro systems in many woody species is still very low since factors responsible for Embryogenesis initiation and embryo development are still largely unknown. The search for molecular and cellular markers during early stages of in vitro Embryogenesis constitutes an important goal to distinguish, after induction, responsive from non-responsive cells, and to elucidate the mechanisms involved in Embryogenesis initiation for their efficient manipulation. In this work, we have performed a comparative analysis of two Embryogenesis pathways derived from microspores and immature zygotic embryos in cork oak in order to characterize early markers of reprogrammed cells in both pathways. Rearrangements of the cell structural organization, changes in epigenetic marks, cell wall polymers modifications and endogenous auxin changes were analyzed at early Embryogenesis stages of the two in vitro systems by a multidisciplinary approach. Results showed that early embryo cells exhibited defined changes of cell components which were similar in both Embryogenesis in vitro systems, cellular features that were not found in non-embryogenic cells. DNA methylation level and nuclear pattern, proportion of esterified pectins in cell walls, and endogenous auxin levels were different in embryo cells in comparison with microspores and immature zygotic embryo cells from which embryos originated, constituting early Embryogenesis markers. These findings suggest that DNA hypomethylation, cell wall remodeling by pectin esterification and auxin increase are involved in early in vitro Embryogenesis in woody species, providing new evidences of the developmental pattern similarity between both Embryogenesis pathways, from microspores and immature zygotic embryos, in woody species.
Mariacarmen Risueno - One of the best experts on this subject based on the ideXlab platform.
-
stress induced microspore Embryogenesis requires endogenous auxin synthesis and polar transport in barley
Frontiers in Plant Science, 2019Co-Authors: Yolanda Perezperez, Mariateresa Solis, Mariacarmen Risueno, Ahmedabdalla Eltantawy, P S TestillanoAbstract:Stress-induced microspore Embryogenesis is a model in vitro system of cell reprogramming, totipotency acquisition, and embryo development. After induction, responsive microspores abandon their developmental program to follow an embryogenic pathway, leading to in vitro embryo formation. This process is widely used to produce doubled-haploid lines, essential players to create new materials in modern breeding programs, particularly in cereals, although its efficiency is still low in many crop species, because the regulating mechanisms are still elusive. Stress signaling and endogenous hormones, mainly auxin, have been proposed as determinant factors of microspore Embryogenesis induction in some eudicot species; however, much less information is available in monocot plants. In this study, we have analyzed the dynamics and possible role of endogenous auxin during stress-induced microspore Embryogenesis in the monocot Hordeum vulgare, barley. The results showed auxin accumulation in early proembryo cells, from Embryogenesis initiation and a further increase with embryo development and differentiation, correlating with the induction and expression pattern of the auxin biosynthesis gene HvTAR2-like. Pharmacological treatments with kynurenine, inhibitor of auxin biosynthesis, and α-(p-chlorophenoxy)-isobutyric acid (PCIB), auxin antagonist, impaired Embryogenesis initiation and development, indicating that de novo auxin synthesis and its activity were required for the process. Efflux carrier gene HvPIN1-like was also induced with Embryogenesis initiation and progression; auxin transport inhibition by N-1-naphthylphthalamic acid significantly reduced embryo development at early and advanced stages. The results indicate activation of auxin biosynthesis with microspore Embryogenesis initiation and progression, in parallel with the activation of polar auxin transport, and reveal a central role of auxin in the process in a monocot species. The findings give new insights into the complex regulation of stress-induced microspore Embryogenesis, particularly in monocot plants for which information is still scarce, and suggest that manipulation of endogenous auxin content could be a target to improve in vitro embryo production.
-
autophagy is activated and involved in cell death with participation of cathepsins during stress induced microspore Embryogenesis in barley
Journal of Experimental Botany, 2018Co-Authors: Ivett Barany, Mariateresa Solis, Mariacarmen Risueno, Eduardo Berenguer, Yolanda Perezperez, Estrella M Santamaria, Jose L Crespo, Isabel Diaz, P S TestillanoAbstract:Microspores are reprogrammed towards Embryogenesis by stress. Many microspores die after this stress, limiting the efficiency of microspore Embryogenesis. Autophagy is a degradation pathway that plays critical roles in stress response and cell death. In animals, cathepsins have an integral role in autophagy by degrading autophagic material; less is known in plants. Plant cathepsins are papain-like C1A cysteine proteases involved in many physiological processes, including programmed cell death. We have analysed the involvement of autophagy in cell death, in relation to cathepsin activation, during stress-induced microspore Embryogenesis in Hordeum vulgare. After stress, reactive oxygen species (ROS) and cell death increased and autophagy was activated, including HvATG5 and HvATG6 up-regulation and increase of ATG5, ATG8, and autophagosomes. Concomitantly, cathepsin L/F-, B-, and H-like activities were induced, cathepsin-like genes HvPap-1 and HvPap-6 were up-regulated, and HvPap-1, HvPap-6, and HvPap-19 proteins increased and localized in the cytoplasm, resembling autophagy structures. Inhibitors of autophagy and cysteine proteases reduced cell death and promoted Embryogenesis. The findings reveal a role for autophagy in stress-induced cell death during microspore Embryogenesis, and the participation of cathepsins. Similar patterns of activation, expression, and localization suggest a possible connection between cathepsins and autophagy. The results open up new possibilities to enhance microspore Embryogenesis efficiency with autophagy and/or cysteine protease modulators.
-
initiation of leaf somatic Embryogenesis involves high pectin esterification auxin accumulation and dna demethylation in quercus alba
Journal of Plant Physiology, 2017Co-Authors: Elena Corredoira, Mariateresa Solis, Mariacarmen Risueno, Vanesa Cano, Ivett Barany, Hector Rodriguez, Ana M Vieitez, P S TestillanoAbstract:Abstract Somatic Embryogenesis is considered a convenient tool for investigating the regulating mechanisms of embryo formation; it is also a feasible system for in vitro regeneration procedures, with many advantages in woody species. Nevertheless, trees have shown recalcitrance to somatic Embryogenesis, and its efficiency remains very low in many cases. Consequently, despite the clear potential of somatic Embryogenesis in tree breeding programs, its application is limited since factors responsible for Embryogenesis initiation have not yet been completely elucidated. In the present work, we investigated key cellular factors involved in the change of developmental program during leaf somatic Embryogenesis initiation of white oak ( Quercus alba ), aiming to identify early markers of the process. The results revealed that pectin esterification, auxin accumulation and DNA demethylation were induced during Embryogenesis initiation and differentially found in embryogenic cells, while they were not present in leaf cells before induction or in non-embryogenic cells after Embryogenesis initiation. These three factors constitute early markers of leaf Embryogenesis and represent processes that could be interconnected and involved in the regulation of cell reprogramming and Embryogenesis initiation. These findings provide new insights into the mechanisms underlying plant cell reprogramming, totipotency and embryogenic competence acquisition, especially in tree species for which information is scarce, thus opening up the possibility of efficient manipulation of somatic Embryogenesis induction.
-
auxin biosynthesis accumulation action and transport are involved in stress induced microspore Embryogenesis initiation and progression in brassica napus
Plant and Cell Physiology, 2015Co-Authors: Hector Rodriguezsanz, Mariateresa Solis, Mariacarmen Risueno, Mariafernanda Lopez, Aurelio Gomezcadenas, P S TestillanoAbstract:Isolated microspores are reprogrammed in vitro by stress, becoming totipotent cells and producing embryos and plants via a process known as microspore Embryogenesis. Despite the abundance of data on auxin involvement in plant development and Embryogenesis, no data are available regarding the dynamics of auxin concentration, cellular localization and the expression of biosynthesis genes during microspore Embryogenesis. This work involved the analysis of auxin concentration and cellular accumulation; expression of TAA1 and NIT2 encoding enzymes of two auxin biosynthetic pathways; expression of the PIN1-like efflux carrier; and the effects of inhibition of auxin transport and action by N-1-naphthylphthalamic acid (NPA) and α-(p-chlorophenoxy) isobutyric acid (PCIB) during Brassica napus microspore Embryogenesis. The results indicated de novo auxin synthesis after stress-induced microspore reprogramming and Embryogenesis initiation, accompanying the first cell divisions. The progressive increase of auxin concentration during progression of Embryogenesis correlated with the expression patterns of TAA1 and NIT2 genes of auxin biosynthetic pathways. Auxin was evenly distributed in early embryos, whereas in heart/torpedo embryos auxin was accumulated in apical and basal embryo regions. Auxin efflux carrier PIN1-like gene expression was induced in early multicellular embryos and increased at the globular/torpedo embryo stages. Inhibition of polar auxin transport (PAT) and action, by NPA and PCIB, impaired embryo development, indicating that PAT and auxin action are required for microspore embryo progression. NPA also modified auxin embryo accumulation patterns. These findings indicate that endogenous auxin biosynthesis, action and polar transport are required in stress-induced microspore reprogramming, Embryogenesis initiation and progression.
-
early markers are present in both Embryogenesis pathways from microspores and immature zygotic embryos in cork oak quercus suber l
BMC Plant Biology, 2014Co-Authors: Hector Rodriguezsanz, Mariateresa Solis, Mariacarmen Risueno, J A Manzanera, Aranzazu Gomezgaray, Beatriz Pintos, P S TestillanoAbstract:In Quercus suber, cork oak, a Mediterranean forest tree of economic and social interest, rapid production of isogenic lines and clonal propagation of elite genotypes have been achieved by developing in vitro Embryogenesis from microspores and zygotic embryos respectively. Despite its high potential in tree breeding strategies, due to their recalcitrancy, the efficiency of Embryogenesis in vitro systems in many woody species is still very low since factors responsible for Embryogenesis initiation and embryo development are still largely unknown. The search for molecular and cellular markers during early stages of in vitro Embryogenesis constitutes an important goal to distinguish, after induction, responsive from non-responsive cells, and to elucidate the mechanisms involved in Embryogenesis initiation for their efficient manipulation. In this work, we have performed a comparative analysis of two Embryogenesis pathways derived from microspores and immature zygotic embryos in cork oak in order to characterize early markers of reprogrammed cells in both pathways. Rearrangements of the cell structural organization, changes in epigenetic marks, cell wall polymers modifications and endogenous auxin changes were analyzed at early Embryogenesis stages of the two in vitro systems by a multidisciplinary approach. Results showed that early embryo cells exhibited defined changes of cell components which were similar in both Embryogenesis in vitro systems, cellular features that were not found in non-embryogenic cells. DNA methylation level and nuclear pattern, proportion of esterified pectins in cell walls, and endogenous auxin levels were different in embryo cells in comparison with microspores and immature zygotic embryo cells from which embryos originated, constituting early Embryogenesis markers. These findings suggest that DNA hypomethylation, cell wall remodeling by pectin esterification and auxin increase are involved in early in vitro Embryogenesis in woody species, providing new evidences of the developmental pattern similarity between both Embryogenesis pathways, from microspores and immature zygotic embryos, in woody species.
Sonia Goncalves - One of the best experts on this subject based on the ideXlab platform.
-
transcriptome dynamics of cork oak quercus suber somatic Embryogenesis reveals active gene players in transcription regulation and phytohormone homeostasis of embryo development
Tree Genetics & Genomes, 2019Co-Authors: Tiago Capote, Anabel Usie, Pedro Barbosa, Marcos Ramos, Leonor Moraiscecilio, Sonia GoncalvesAbstract:Cork oak (Quercus suber L.) is one of the most important Mediterranean forest tree species. The last decades have been marked by a decline in this species. Implementation of breeding programs is fundamental to revert this trend. Somatic Embryogenesis is the system of choice for clonal propagation, constituting a valuable tool for embryo production and improved genotype testing. In this study, the cork oak transcriptome during somatic Embryogenesis was characterized in four stages of development to identify relevant genes in the process and to understand the molecular and biochemical events occurring in each specific stage. A total 66,693 candidate coding regions were predicted from the generated de novo transcriptome assembly. Differential gene expression analysis identified 11,507 genes distributed in 30 clusters with distinct gene expression patterns and enriched in various biological process GO terms. Results show 1159 differentially expressed genes coding for transcription regulators, namely transcription factors (76%) with important roles in Embryogenesis, like orthologous of AINTEGUMENTA-like, PLETHORA, CYTOKININ RESPONSE FACTOR, GATA transcription factors, and AUXIN RESPONSE FACTORs genes. Results also show 250 differentially expressed phytohormone-related genes involved in important aspects of Embryogenesis as tissue specification, differentiation, and Embryogenesis competence. Finally, we identified a group of genes with functions in cellular protection and abiotic stress tolerance coding for LATE Embryogenesis ABUNDANT proteins. Cork oak Embryogenesis transcriptome characterization represents a tool for future biotechnological applications. Our results provide a molecular insight into embryo development, establishing a basis for further research towards improvement of somatic Embryogenesis in cork oak.
Hector Rodriguezsanz - One of the best experts on this subject based on the ideXlab platform.
-
auxin biosynthesis accumulation action and transport are involved in stress induced microspore Embryogenesis initiation and progression in brassica napus
Plant and Cell Physiology, 2015Co-Authors: Hector Rodriguezsanz, Mariateresa Solis, Mariacarmen Risueno, Mariafernanda Lopez, Aurelio Gomezcadenas, P S TestillanoAbstract:Isolated microspores are reprogrammed in vitro by stress, becoming totipotent cells and producing embryos and plants via a process known as microspore Embryogenesis. Despite the abundance of data on auxin involvement in plant development and Embryogenesis, no data are available regarding the dynamics of auxin concentration, cellular localization and the expression of biosynthesis genes during microspore Embryogenesis. This work involved the analysis of auxin concentration and cellular accumulation; expression of TAA1 and NIT2 encoding enzymes of two auxin biosynthetic pathways; expression of the PIN1-like efflux carrier; and the effects of inhibition of auxin transport and action by N-1-naphthylphthalamic acid (NPA) and α-(p-chlorophenoxy) isobutyric acid (PCIB) during Brassica napus microspore Embryogenesis. The results indicated de novo auxin synthesis after stress-induced microspore reprogramming and Embryogenesis initiation, accompanying the first cell divisions. The progressive increase of auxin concentration during progression of Embryogenesis correlated with the expression patterns of TAA1 and NIT2 genes of auxin biosynthetic pathways. Auxin was evenly distributed in early embryos, whereas in heart/torpedo embryos auxin was accumulated in apical and basal embryo regions. Auxin efflux carrier PIN1-like gene expression was induced in early multicellular embryos and increased at the globular/torpedo embryo stages. Inhibition of polar auxin transport (PAT) and action, by NPA and PCIB, impaired embryo development, indicating that PAT and auxin action are required for microspore embryo progression. NPA also modified auxin embryo accumulation patterns. These findings indicate that endogenous auxin biosynthesis, action and polar transport are required in stress-induced microspore reprogramming, Embryogenesis initiation and progression.
-
early markers are present in both Embryogenesis pathways from microspores and immature zygotic embryos in cork oak quercus suber l
BMC Plant Biology, 2014Co-Authors: Hector Rodriguezsanz, Mariateresa Solis, Mariacarmen Risueno, J A Manzanera, Aranzazu Gomezgaray, Beatriz Pintos, P S TestillanoAbstract:In Quercus suber, cork oak, a Mediterranean forest tree of economic and social interest, rapid production of isogenic lines and clonal propagation of elite genotypes have been achieved by developing in vitro Embryogenesis from microspores and zygotic embryos respectively. Despite its high potential in tree breeding strategies, due to their recalcitrancy, the efficiency of Embryogenesis in vitro systems in many woody species is still very low since factors responsible for Embryogenesis initiation and embryo development are still largely unknown. The search for molecular and cellular markers during early stages of in vitro Embryogenesis constitutes an important goal to distinguish, after induction, responsive from non-responsive cells, and to elucidate the mechanisms involved in Embryogenesis initiation for their efficient manipulation. In this work, we have performed a comparative analysis of two Embryogenesis pathways derived from microspores and immature zygotic embryos in cork oak in order to characterize early markers of reprogrammed cells in both pathways. Rearrangements of the cell structural organization, changes in epigenetic marks, cell wall polymers modifications and endogenous auxin changes were analyzed at early Embryogenesis stages of the two in vitro systems by a multidisciplinary approach. Results showed that early embryo cells exhibited defined changes of cell components which were similar in both Embryogenesis in vitro systems, cellular features that were not found in non-embryogenic cells. DNA methylation level and nuclear pattern, proportion of esterified pectins in cell walls, and endogenous auxin levels were different in embryo cells in comparison with microspores and immature zygotic embryo cells from which embryos originated, constituting early Embryogenesis markers. These findings suggest that DNA hypomethylation, cell wall remodeling by pectin esterification and auxin increase are involved in early in vitro Embryogenesis in woody species, providing new evidences of the developmental pattern similarity between both Embryogenesis pathways, from microspores and immature zygotic embryos, in woody species.
-
a new microspore Embryogenesis system under low temperature which mimics zygotic Embryogenesis initials expresses auxin and efficiently regenerates doubled haploid plants in brassica napus
BMC Plant Biology, 2012Co-Authors: Deepak Prem, Mariateresa Solis, Mariacarmen Risueno, Ivett Barany, Hector Rodriguezsanz, P S TestillanoAbstract:Background Microspore Embryogenesis represents a unique system of single cell reprogramming in plants wherein a highly specialized cell, the microspore, by specific stress treatment, switches its fate towards an Embryogenesis pathway. In Brassica napus, a model species for this phenomenon, incubation of isolated microspores at 32°C is considered to be a pre-requisite for Embryogenesis induction.