The regenerative potential of stolons from C. sinuosus was evaluated for the establishment of a c... more The regenerative potential of stolons from C. sinuosus was evaluated for the establishment of a clonal propagation protocol.
In vitro propagation of Petiveria alliacea L. was achieved based on the development of axillary b... more In vitro propagation of Petiveria alliacea L. was achieved based on the development of axillary buds from nodal segments cultured on Murashige and Skoog (MS0) medium. Rooting of shoots was induced on half strength MS supplemented with 0.6 μM Indole-3-acetic acid (IAA). After rooting, plants were transferred to greenhouse conditions. Morphological abnormalities were not observed among the regenerants. Leaf segments from in vitro plants were cultured on MS medium supplemented with different concentrations of Naphthaleneacetic acid (NAA) (2.7; 5.4; 10.7; 16.1 and 26.9 μM), 6- Benzylaminopurine (BAP) (2.2; 4.4; 8.9; 13.3 and 22.2 μM), Kinetin (KIN) (2.3; 4.6; 9.2; 13.8; and 23.0 μM), Picloram (PIC) (2.0; 4.0; 8.0; 12.0; and 20.0 μM) or 2,4-Diclorophenoxyacetic acid (2,4D) (2.2; 4.5; 9.0; 13.6 and 22.6 μM) to promote development of friable calluses. Highest induction rates were obtained in the presence of 13.6 and 22.6 μM 2,4D as well as of 20.0 μM PIC. Cell suspension cultures were esta...
Journal of Horticultural Science and Biotechnology
Wild species of Arachis are restricted to South America and generally occur in regions under inte... more Wild species of Arachis are restricted to South America and generally occur in regions under intensive environmental disturbance. Both in situ and ex situ conservation strategies are required in order to maintain the availability of these genotypes. This work developed in vitro regeneration systems from seed explants of 17 wild species of Arachis from six Sections (Heteranthae, Caulorrhizae, Triseminatae, Erectoides, Procumbentes and Arachis). After seed disinfection, embryonic axes, leaflets and cotyledons were excised aseptically and cultured on Murashige and Skoog (MS) medium supplemented with 8.8 μM, 22 μM or 110 μM 6-benzylaminopurine (BAP). Cultures were maintained in a growth chamber at 28° ± 2°C with a 16 h photoperiod. Regeneration patterns from seed explants were similar among species from all Sections. Embryonic axes produced plants through meristematic amplification or multiple shoot formation, while cotyledons and embryonic leaflets produced shoots at significantly lowe...
Journal of Horticultural Science and Biotechnology
In vitro conservation techniques are considered suitable strategies for ex situ conservation of w... more In vitro conservation techniques are considered suitable strategies for ex situ conservation of wild species of Arachis. However, there is a potential risk of genetic and epigenetic alterations induced by tissue culture conditions. The goal of this work was to determine the influence of explant type and regeneration pathway on sequence modifications and changes in the DNA methylation status of in vitro-grown plants of A. villosulicarpa using Amplified Fragment Length Polymorphism (AFLP) and Methylation-Sensitive Amplified Polymorphism (MSAP) analyses. Regeneration from embryo axes occurred through multiplication of pre-existing meristems. Cotyledons and embryonic leaflets displayed direct and indirect organogenesis, respectively. No polymorphic AFLP amplification fragments were detected among the regenerants. Conversely, MSAP analysis showed a decrease in the levels of DNA methylation in cotyledon- and embryonic leaflet-derived plants.
The effects of two methods of cryopreservation involving chemical vitrification and air desiccati... more The effects of two methods of cryopreservation involving chemical vitrification and air desiccation) were studied on isolated embryonic axes of A. hypogaea. Vitrification with PVS2 and desiccation in a laminar flow cabinet resulted in high levels (70-90%) of whole plant recovery after cryopreservation. A desiccation protocol based on 1h exposure of explants to the air flow was successfully applied to six wild species of section Extranervosae, resulting in recovery levels of 70-90% after liquid nitrogen treatment.
A storage protocol at cryogenic temperature was established for shoot apices from in vitro plants... more A storage protocol at cryogenic temperature was established for shoot apices from in vitro plants of the cultivated groundnut (Arachis hypogaea) and wild Arachis species (A. retusa and A. burchellii) using a basic vitrification protocol with direct immersion in liquid nitrogen (LN). The effect of pre-treatments of donor-plants with ABA as well as of different supplements in the post-thaw culture medium was studied. After rapid warming at 40 C, the explants were cultured on MS medium devoid of growth regulators (MS0) or MS supplemented with 4.4(M benzylaminopurine (BAP) and 0.5(M naphthalene acetic acid (NAA) plus 5(M silver nitrate (AgNO3), 0.25% polyvinylpyrrolidone (PVP) or 0.2% activated charcoal. Non-frozen explants from the three species formed one shoot through meristematic amplification when cultured on MS0 medium. These explants also developed callus on MS supplemented with growth regulators (4.4(M BAP and 0.5(M NAA) alone or plus PVP or AgNO3. Callus formation was suppresse...
BIOLOGIA PLANTARUM 45 (3): 353-357,2002 Germplasm preservation of wild Arachis species through cu... more BIOLOGIA PLANTARUM 45 (3): 353-357,2002 Germplasm preservation of wild Arachis species through culture of shoot apices and axillary buds from in vitro plants RF GAGLIARDI*, GPPACHECO*, JFM VALLS** and E. MANSUR*1 ... 1985, Krapovickas and Gregory 1994). ...
The regenerative potential of stolons from C. sinuosus was evaluated for the establishment of a c... more The regenerative potential of stolons from C. sinuosus was evaluated for the establishment of a clonal propagation protocol.
In vitro propagation of Petiveria alliacea L. was achieved based on the development of axillary b... more In vitro propagation of Petiveria alliacea L. was achieved based on the development of axillary buds from nodal segments cultured on Murashige and Skoog (MS0) medium. Rooting of shoots was induced on half strength MS supplemented with 0.6 μM Indole-3-acetic acid (IAA). After rooting, plants were transferred to greenhouse conditions. Morphological abnormalities were not observed among the regenerants. Leaf segments from in vitro plants were cultured on MS medium supplemented with different concentrations of Naphthaleneacetic acid (NAA) (2.7; 5.4; 10.7; 16.1 and 26.9 μM), 6- Benzylaminopurine (BAP) (2.2; 4.4; 8.9; 13.3 and 22.2 μM), Kinetin (KIN) (2.3; 4.6; 9.2; 13.8; and 23.0 μM), Picloram (PIC) (2.0; 4.0; 8.0; 12.0; and 20.0 μM) or 2,4-Diclorophenoxyacetic acid (2,4D) (2.2; 4.5; 9.0; 13.6 and 22.6 μM) to promote development of friable calluses. Highest induction rates were obtained in the presence of 13.6 and 22.6 μM 2,4D as well as of 20.0 μM PIC. Cell suspension cultures were esta...
Journal of Horticultural Science and Biotechnology
Wild species of Arachis are restricted to South America and generally occur in regions under inte... more Wild species of Arachis are restricted to South America and generally occur in regions under intensive environmental disturbance. Both in situ and ex situ conservation strategies are required in order to maintain the availability of these genotypes. This work developed in vitro regeneration systems from seed explants of 17 wild species of Arachis from six Sections (Heteranthae, Caulorrhizae, Triseminatae, Erectoides, Procumbentes and Arachis). After seed disinfection, embryonic axes, leaflets and cotyledons were excised aseptically and cultured on Murashige and Skoog (MS) medium supplemented with 8.8 μM, 22 μM or 110 μM 6-benzylaminopurine (BAP). Cultures were maintained in a growth chamber at 28° ± 2°C with a 16 h photoperiod. Regeneration patterns from seed explants were similar among species from all Sections. Embryonic axes produced plants through meristematic amplification or multiple shoot formation, while cotyledons and embryonic leaflets produced shoots at significantly lowe...
Journal of Horticultural Science and Biotechnology
In vitro conservation techniques are considered suitable strategies for ex situ conservation of w... more In vitro conservation techniques are considered suitable strategies for ex situ conservation of wild species of Arachis. However, there is a potential risk of genetic and epigenetic alterations induced by tissue culture conditions. The goal of this work was to determine the influence of explant type and regeneration pathway on sequence modifications and changes in the DNA methylation status of in vitro-grown plants of A. villosulicarpa using Amplified Fragment Length Polymorphism (AFLP) and Methylation-Sensitive Amplified Polymorphism (MSAP) analyses. Regeneration from embryo axes occurred through multiplication of pre-existing meristems. Cotyledons and embryonic leaflets displayed direct and indirect organogenesis, respectively. No polymorphic AFLP amplification fragments were detected among the regenerants. Conversely, MSAP analysis showed a decrease in the levels of DNA methylation in cotyledon- and embryonic leaflet-derived plants.
The effects of two methods of cryopreservation involving chemical vitrification and air desiccati... more The effects of two methods of cryopreservation involving chemical vitrification and air desiccation) were studied on isolated embryonic axes of A. hypogaea. Vitrification with PVS2 and desiccation in a laminar flow cabinet resulted in high levels (70-90%) of whole plant recovery after cryopreservation. A desiccation protocol based on 1h exposure of explants to the air flow was successfully applied to six wild species of section Extranervosae, resulting in recovery levels of 70-90% after liquid nitrogen treatment.
A storage protocol at cryogenic temperature was established for shoot apices from in vitro plants... more A storage protocol at cryogenic temperature was established for shoot apices from in vitro plants of the cultivated groundnut (Arachis hypogaea) and wild Arachis species (A. retusa and A. burchellii) using a basic vitrification protocol with direct immersion in liquid nitrogen (LN). The effect of pre-treatments of donor-plants with ABA as well as of different supplements in the post-thaw culture medium was studied. After rapid warming at 40 C, the explants were cultured on MS medium devoid of growth regulators (MS0) or MS supplemented with 4.4(M benzylaminopurine (BAP) and 0.5(M naphthalene acetic acid (NAA) plus 5(M silver nitrate (AgNO3), 0.25% polyvinylpyrrolidone (PVP) or 0.2% activated charcoal. Non-frozen explants from the three species formed one shoot through meristematic amplification when cultured on MS0 medium. These explants also developed callus on MS supplemented with growth regulators (4.4(M BAP and 0.5(M NAA) alone or plus PVP or AgNO3. Callus formation was suppresse...
BIOLOGIA PLANTARUM 45 (3): 353-357,2002 Germplasm preservation of wild Arachis species through cu... more BIOLOGIA PLANTARUM 45 (3): 353-357,2002 Germplasm preservation of wild Arachis species through culture of shoot apices and axillary buds from in vitro plants RF GAGLIARDI*, GPPACHECO*, JFM VALLS** and E. MANSUR*1 ... 1985, Krapovickas and Gregory 1994). ...
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