Micropropagation ofAsparagus cooperi as affected by growth regulators (original) (raw)
Related papers
2009
For the conservation of high value but neglected medicinal plant (Asparagus racemosus) of Nepal an effort has been made to multiply its number rapidly using tissue culture technique. The plant has been successfully multiplied. For the multiplication, all possible explants including callus have been used and the callus and shoot explants (shoot tips and nodes) played significant role. Mainly four parameters: callus, buds, shoots and roots inductions were studied. NAA singly played a good role in all parameters except bud induction. Similarly, BAP played good roles in shoot and bud inductions, whereas combinations of NAA and BAP at various levels were found to be effective in almost all cases. The multiplied adventitious shoots were successfully rooted (in vitro and in vivo) using NAA, acclimatized and transferred to natural conditions. Abbreviations: BAP: 6-benzylaminopurine; MS: Murashige and Skoog (1962); NAA: a naphthaleneacetic acid; IAA: indole-3-acetic acid; IBA: indole-3-butyric acid; 2,4-D: 2, 4 Dichlorophenoxyacetic acid and Kin: Kinetin.
Plant Cell, Tissue and Organ Culture (PCTOC), 2020
In this study, we developed a repeatable in vitro micropropagation protocol for the medicinal plant Asparagus cochinchinensis, based on indirect organogenesis using leaf segments cut from seedlings of in vitro-germinated seeds. We obtained 85% callus induction from the leaf segments within 4-5 weeks when grown on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP, 1.0 mg/L) and 1-naphthaleneacetic acid (NAA, 0.5 mg/L). We found that MS media supplemented with Kn (1.0 mg/L) in combination with NAA (1.0 mg/L) and BAP (1.0 mg/L) combined with NAA (0.5 mg/L) were the most effective in promoting shoot regeneration, yielding plantlets with 6.72 and 6.48 shoots per culture, respectively. When cultured on PGR-free half-strength MS medium, regenerated plants developed root systems with an average 11.0 roots per shoot cluster and an average length of 36.14 mm at 9 weeks. During acclimatization, regenerated plantlets showed 96.4% survival and exhibited normal growth characteristics and morphology. We also made an attempt to directly regenerate A. cochinchinensis from shoot apices but it was futile. Histological analyses revealed the presence of crystal idioblasts in young leaves from the early stages of leaf differentiation. The leaf-based plant regeneration technique developed herein could be employed for large-scale propagation of the plants over a short time period, thereby substantially contributing to the germplasm preservation and rapid propagation of A. cochinchinensis. Key message A repeatable in vitro micropropagation protocol for Asparagus cochinchinensis was developed based on indirectorganogenesis. Histological analysis revealed crystal idioblasts for the first time in leaf primordia of this species.
Regeneration of Asparagus racemosus by shoot apex and nodal explants
In vitro propagation of Asparagus racemosus was carried out using shoot apex and nodal explants in three stages. (a) Initiation of multiple shoots on MS-medium supplemented with 6-Benzyl Adenine (8.9 µM) + Naphthalene Acetic Acid (0.27 µM) at 25 0 C having 16 h photoperiod (with 60 µ mol m-2 s-1 light intensity) and 8 h dark period. (b) Elongation of shoots on MS medium supplemented with 15% coconut milk + 2-iso Pentenyl Adenine (19.6 µM), the light intensity was increased to 80 µ mol m-2 s-1. (c) Rooting of elongated shoots by giving it a preculture treatment with MS medium augmented with Indole Butyric Acid (7.35 µM) for 48 h and then transfer to MS medium with 15% coconut milk. The rooted healthy plantlets were best hardened in a mixture of 1:1 sand and soil in a moist saturated chamber having 60-80 % humidity. Plants were transferred to fields, after 5 wks of hardening.
Micropropagation of a recalcitrant male asparagus clone (MD 22-8)
Plant Cell Tissue and Organ Culture, 1992
The Asparagus officinalis male asparagus clone MD 22-8 (Howard Scott) is highly prized by asparagus breeders but has been very difficult to micropropagate due to its slow growth rate and reluctance to initiate roots. Shoot tips cultured on indole-3-acetic acid or indole-3-propionic acid, at concentrations of 1.53 and 1.44 IxM respectively, resulted in levels of root initiation and elongation equal to or better than those of a female asparagus clone. These culture conditions also improved the rates of shoot initiation and elongation. Root initiation and elongation were further increased by culture conditions that provided enhanced gas exchange.
Plant regeneration after long-term callus culture in clones of Asparagus officinalis L
Biocell : official journal of the Sociedades Latinoamericanas de Microscopía Electronica ... et. al, 2005
Callus growth and plant regeneration from long-term callus cultures were studied in two elite clones of Asparagus officinalis cv. Argenteuil, to establish a suitable protocol for a prospective in vitro selection program. Callus initiation and growth was evaluated on MS medium with 3% sucrose, 0.9% agar, 1 mg x l(-1) kinetin, and three levels of 2,4-D. The highest callus relative growth was obtained on medium with 1.5 mg x l(-1) 2,4-D and 1 mg x l(-1) kinetin. Shoot primordia (SP) induction from > 18-months-old calluses was evaluated on several media; the highest percentage of SP induction (89%) and average number of SP per callus (8.6) were obtained with clone "265" on MS medium with 5 mg x l(-1) 2iP, 1 mg x l(-1) IAA, 3% sucrose and 0.9% agar. The highest percentage of root induction (100%) was achieved with clone '265' on MS medium with 0.1 mg x l(-1) kinetin, 0.1 mg x l(-1) NAA, 1.32 mg x l(-1) ancymidol, 7% glucose and 0.8% agar. Important medium x genotype ...
ABSTRACT Somatic embryogenesis and plantlet formation were obtained from callus derived from the subapical region of spears of Aspara$us cooperi Baker. Callus was obtained in Murashige and Skoog's medium supplemented with l-naphthaleneacetic acid and kinetin. Increase in the concentration of potassium nitrate in subsequent subcultures resulted in the formation of embryos. Rapid multiplication of embryos was secured on transfer to a medium containing a different source of nitrogen and a low level (0.01 rag/l) of gibberellic acid. Media containing zeatin or gibberellic acid led to the formation of complete plantlets from embryos. Regenerated plants were cytologically and phenotypically stable.
In order to study the in vitro callus induction and plant regeneration in the asparagus plant (Asparagus officinalis ) and indirect organogenesis in explants, an experiment in a completely randomized design was performed in tissue culture laboratory of faculty of agriculture, Islamic Azad University of Dezfool. In this study, the nodal explants treated with different concentrations of NAA and BAP, and it was found that the presence of two plant growth regulators NAA And BAP Is necessary for seedlings regeneration and callus induction in asparagus,. Treatment that leads to achieving the maximum amount of callus buds was MS medium supplied with 0.5 mg/lit of NAA and 1 mg/lit BAP which leads to the production of 22 buds per explants. Two different levels of 2, 4-D were used for induce roots in regenerated seedlings. The best treatment was 1.25 mg/ lit 2, 4-D with an average of 52% rooting in regenerated plants.
HortScience, 1995
The effects of culture media, culture modes, and carbon sources on plating efficiencies of protoplasts of two genotypes of Asparagus officinalis L. were investigated. Protoplasts grew best in a semisolid culture system containing half-strength MS medium with 1 mg NAA/liter, 0.5 mg zeatin/liter, 0.6 M glucose, and 0.1% Gelrite. The plating efficiencies were 12.5% and 8.1% for genotypes G 203 and G 171, respectively. Embryogenic calli were produced from protoplast-derived microcalli after culturing on MS medium with 1 mg 2,4-D, 3% sucrose, and 0.2% Gelrite. The somatic embryos were initiated, matured, and then germinated to plantlets in MS medium containing 0.1 mg NAA/liter, 0.3 mg 2-iP/liter (EMM), and different levels of carbohydrates. Transfer of somatic embryos from EMM with 10% glucose to EMM containing 2% sucrose produced the highest number of bipolar embryos and plantlets. The plantlets regenerated shoots and roots in MS medium with 3% sucrose, 0.1 mg NAA/liter, 0.1 mg kinetin/...
Callus induction and axillary shoot formation in Asparagus racemosus Willd
Current Botany
An experiment was performed to establish a regeneration protocol for an important medicinal plant, Asparagus racemosus. In the present investigation, nodal and internodal explants were employed for callus induction and axillary shoot formation. Maximum callus induction frequency was found on MS medium fortified with 2,4-D (1.0 mg/L) along with NAA (1.0 mg/L) and BAP (0.5 mg/L). However, individual effects of 2,4-D or NAA with BAP showed least callus induction. The higher concentrations of 2,4-D and BAP decreased the response of explants. However, maximum axillary shoot formation was observed on MS medium adjuvanted with BAP (2.0 mg/L) and NAA (0.5 mg/L).
African Journal of Biotechnology, 2008
In vitro plant regeneration was achieved from embryogenic cell suspension culture of Astragalus chrysochlorus. When 30-day-old aseptically grown seedlings were cultured on Murashige and Skoog (MS) medium containing 0.1 mg/l α-naphthaleneacetic acid (NAA) plus 1.0 mg/l 6-benzyladenine (BA), friable callus was formed within two weeks from the mesocotyl of the seedling. After three weeks, proliferated actively growing calli were transferred to MS liquid medium containing 2,4dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA) or NAA and subcultured at two week intervals. After two weeks, induction of somatic embryos up to the torpedo stage occured at all tested concentrations of 2,4-D, IAA or NAA. Somatic embryos developed only in MS medium containing 0.5 mg/l IAA within two weeks and 2% of globular embryos were developed into the cotyledonary stage embryos. Eighty one percent of somatic embryos cultured in MS medium supplemented with 0.5 mg/l IAA were found to be diploid by flow cytometric analysis. Plantlet propagation was achieved on half strength MS liquid medium supplemented with 3% (w/v) sucrose after four weeks of culture. After a month on half strength MS medium [1.5% (w/v) sucrose and 0.8% (w/v) agar] 29 of 71 shoots developed into rooted plantlets.