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Indirect shoot organogenesis from leaf explants of Adhatoda vasica Nees.

Mandal J, Laxminarayana U - Springerplus (2014)

Bottom Line: Elongation of regenerated shoot was obtained on MS basal medium supplemented with 0.25 mg l(-1) TDZ.All regenerated shoots developed adventitious roots within 4 weeks when transferred to rooting medium containing SH medium supplemented with 0.5 mg l(-1) IBA.Total nine rooted plantlets were transferred from in vitro to in vivo conditions and eight plants survived and successfully acclimatized in the shaded greenhouse 12 weeks after transplanting.

View Article: PubMed Central - PubMed

Affiliation: Department of Education in Science and Mathematics, Regional Institute of Education, National Council of Educational Research and Training, Shyamla Hills, Bhopal, 462013 India.

ABSTRACT
A novel protocol for indirect shoot organogenesis of Adhatoda vasica was developed using petiole explants derived from mature shrubby plants. Media with concentrations of cytokinins in combination with auxins were used to induce callus formation in two explants types: petiole and leaf segment. The frequency of callus formation from petiole and leaf segment explants on Murashige and Skoog (MS) basal medium supplemented with 0.25 mg l(-1) thidiazuron (TDZ) and 0.25 mg l(-1) α-naphthaleneacetic acid (NAA) was 100 ± 0.0 and 83.70 ± 0.52% respectively, while on this medium supplemented with 0.25 mg l(-1) 6-(γ-γ, dimethylallyamino purine) (2iP) and 0.25 mg l(-1) NAA, the callus frequency was 100 ± 0.0 and 96.70 ± 0.67% respectively. The highest shoot regeneration (90.60 ± 0.52%) response and the maximum shoots (8.10 ± 0.28) per callus were achieved from petiole explants on MS medium containing 0.25 mg l(-1) TDZ and 0.25 mg l(-1) NAA. On the contrary, on Schenk & Hildebrandt (SH) basal medium supplemented with 0.25 mg l(-1) TDZ and 0.25 mg l(-1) NAA, the frequency of callus formation from petiole and leaf segment explants was 100 ± 0.0 and 90.50 ± 0.89% respectively while the callus frequency on this medium containing 0.25 mg l(-1) 2iP and 0.25 mg l(-1) NAA was 100 ± 0.0 and 89.90 ± 0.72% respectively. The shoot regeneration frequency for petiole explants was 89.90 ± 0.46% producing 6.00 ± 0.21 shoots per callus on SH basal medium supplemented with 0.25 mg l(-1) TDZ and 0.25 mg l(-1) NAA. Whereas petiole explants could induce 83.70 ± 0.50% shoot regeneration and 7.3 ± 1.05 shoots per callus on SH medium containing 0.25 mg l(-1) indole-3-butyric acid (IBA), 0.5 mg l(-1) 6-benzyladenine (BA) and 0.5 mg l(-1) 2iP. Elongation of regenerated shoot was obtained on MS basal medium supplemented with 0.25 mg l(-1) TDZ. All regenerated shoots developed adventitious roots within 4 weeks when transferred to rooting medium containing SH medium supplemented with 0.5 mg l(-1) IBA. Total nine rooted plantlets were transferred from in vitro to in vivo conditions and eight plants survived and successfully acclimatized in the shaded greenhouse 12 weeks after transplanting.

No MeSH data available.


Related in: MedlinePlus

Mean % of callus induction of leaf segment and petiole explants ofAdhatoda vasicaon MS medium containing plant growth regulators after 4 weeks. Different letter(s) indicate a significant difference between treatments at P ≤0.05 according to Tukey test.
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Fig1: Mean % of callus induction of leaf segment and petiole explants ofAdhatoda vasicaon MS medium containing plant growth regulators after 4 weeks. Different letter(s) indicate a significant difference between treatments at P ≤0.05 according to Tukey test.

Mentions: Incubation of petiole and leaf segment explants of Adhatoda vasica either on MS or SH medium supplemented with different concentrations of NAA, IBA or 2,4-D (0.25 or 1.0 mg l−1) in combination with TDZ, BA or 2iP (0.25 or 1.0 mg l−1) produced callus within 2 weeks (Figures 1 and 2). Callus primarily originated in petiole explants whereas callus development was observed on cut off surface of leaf segment explants. The explants cultured on the control medium (MS or SH medium) exhibited no callusing response. MS medium produced a significantly higher callusing from both petiole and leaf segment explants than on SH medium irrespective of concentrations and combinations of hormones used (Figures 1 and 2). The callus initiation response (100 ± 0%) from petiole explants was observed on MS medium supplemented with NAA, or IBA or 2,4-D (0.25 or 1.0 mg l−1) in combination with TDZ or BA or 2iP (0.25 or 1.0 mg l−1). On the other hand, the leaf segment explants induced the maximum callus formation (100 ± 0%) when NAA, or IBA (0.25 mg l−1) was added to the MS medium containing 2iP (0.25 mg l−1) after 4 weeks of culture. The petiole explant was found to be better explant source for callus induction than leaf segment explants, as the former produced higher percentage of callus than the latter (Figures 1 and 2). MS medium containing either 0.25 or 1.0 mg l−1 NAA in combination with 0.25 or 1.0 mg l−1 TDZ or BA or 2iP produced green and compact callus from both petiole and leaf segment explants whereas friable callus was induced on MS medium containing either IBA or 2,4-D in combination with TDZ or BA or 2iP at the same level of concentrations (Figures 1 and 3a, b). In contrast, among the different concentrations and combinations of plant growth regulators tested, the best callus induction from petiole explants (100 ± 0%) was achieved when the SH medium was supplemented with 0.25 mg l−1 NAA in combination with TDZ or BA or 2iP while leaf segment explants produced the highest callus (88.50 ± 0.52 to 90.50 ± 0.89%) at the same level of concentrations after 4 weeks of culture (Figure 2). Petiole explants showed best callus induction (100 ± 0%) on SH medium containing either 0.25 mg l−1 IBA in combination with 0.25 mg l−1 BA or 2iP and 0.25 mg l−1 2,4-D in combination with 0.25 mg l−1 TDZ or BA. Leaf segment explants, in comparison, showed best callus induction in the range of 85.20 ± 0.44 to 91.50 ± 0.62% and 48.30 ± 0.30 to 88.90 ± 0.53% at the same level of concentrations (Figure 2).Figure 1


Indirect shoot organogenesis from leaf explants of Adhatoda vasica Nees.

Mandal J, Laxminarayana U - Springerplus (2014)

Mean % of callus induction of leaf segment and petiole explants ofAdhatoda vasicaon MS medium containing plant growth regulators after 4 weeks. Different letter(s) indicate a significant difference between treatments at P ≤0.05 according to Tukey test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230685&req=5

Fig1: Mean % of callus induction of leaf segment and petiole explants ofAdhatoda vasicaon MS medium containing plant growth regulators after 4 weeks. Different letter(s) indicate a significant difference between treatments at P ≤0.05 according to Tukey test.
Mentions: Incubation of petiole and leaf segment explants of Adhatoda vasica either on MS or SH medium supplemented with different concentrations of NAA, IBA or 2,4-D (0.25 or 1.0 mg l−1) in combination with TDZ, BA or 2iP (0.25 or 1.0 mg l−1) produced callus within 2 weeks (Figures 1 and 2). Callus primarily originated in petiole explants whereas callus development was observed on cut off surface of leaf segment explants. The explants cultured on the control medium (MS or SH medium) exhibited no callusing response. MS medium produced a significantly higher callusing from both petiole and leaf segment explants than on SH medium irrespective of concentrations and combinations of hormones used (Figures 1 and 2). The callus initiation response (100 ± 0%) from petiole explants was observed on MS medium supplemented with NAA, or IBA or 2,4-D (0.25 or 1.0 mg l−1) in combination with TDZ or BA or 2iP (0.25 or 1.0 mg l−1). On the other hand, the leaf segment explants induced the maximum callus formation (100 ± 0%) when NAA, or IBA (0.25 mg l−1) was added to the MS medium containing 2iP (0.25 mg l−1) after 4 weeks of culture. The petiole explant was found to be better explant source for callus induction than leaf segment explants, as the former produced higher percentage of callus than the latter (Figures 1 and 2). MS medium containing either 0.25 or 1.0 mg l−1 NAA in combination with 0.25 or 1.0 mg l−1 TDZ or BA or 2iP produced green and compact callus from both petiole and leaf segment explants whereas friable callus was induced on MS medium containing either IBA or 2,4-D in combination with TDZ or BA or 2iP at the same level of concentrations (Figures 1 and 3a, b). In contrast, among the different concentrations and combinations of plant growth regulators tested, the best callus induction from petiole explants (100 ± 0%) was achieved when the SH medium was supplemented with 0.25 mg l−1 NAA in combination with TDZ or BA or 2iP while leaf segment explants produced the highest callus (88.50 ± 0.52 to 90.50 ± 0.89%) at the same level of concentrations after 4 weeks of culture (Figure 2). Petiole explants showed best callus induction (100 ± 0%) on SH medium containing either 0.25 mg l−1 IBA in combination with 0.25 mg l−1 BA or 2iP and 0.25 mg l−1 2,4-D in combination with 0.25 mg l−1 TDZ or BA. Leaf segment explants, in comparison, showed best callus induction in the range of 85.20 ± 0.44 to 91.50 ± 0.62% and 48.30 ± 0.30 to 88.90 ± 0.53% at the same level of concentrations (Figure 2).Figure 1

Bottom Line: Elongation of regenerated shoot was obtained on MS basal medium supplemented with 0.25 mg l(-1) TDZ.All regenerated shoots developed adventitious roots within 4 weeks when transferred to rooting medium containing SH medium supplemented with 0.5 mg l(-1) IBA.Total nine rooted plantlets were transferred from in vitro to in vivo conditions and eight plants survived and successfully acclimatized in the shaded greenhouse 12 weeks after transplanting.

View Article: PubMed Central - PubMed

Affiliation: Department of Education in Science and Mathematics, Regional Institute of Education, National Council of Educational Research and Training, Shyamla Hills, Bhopal, 462013 India.

ABSTRACT
A novel protocol for indirect shoot organogenesis of Adhatoda vasica was developed using petiole explants derived from mature shrubby plants. Media with concentrations of cytokinins in combination with auxins were used to induce callus formation in two explants types: petiole and leaf segment. The frequency of callus formation from petiole and leaf segment explants on Murashige and Skoog (MS) basal medium supplemented with 0.25 mg l(-1) thidiazuron (TDZ) and 0.25 mg l(-1) α-naphthaleneacetic acid (NAA) was 100 ± 0.0 and 83.70 ± 0.52% respectively, while on this medium supplemented with 0.25 mg l(-1) 6-(γ-γ, dimethylallyamino purine) (2iP) and 0.25 mg l(-1) NAA, the callus frequency was 100 ± 0.0 and 96.70 ± 0.67% respectively. The highest shoot regeneration (90.60 ± 0.52%) response and the maximum shoots (8.10 ± 0.28) per callus were achieved from petiole explants on MS medium containing 0.25 mg l(-1) TDZ and 0.25 mg l(-1) NAA. On the contrary, on Schenk & Hildebrandt (SH) basal medium supplemented with 0.25 mg l(-1) TDZ and 0.25 mg l(-1) NAA, the frequency of callus formation from petiole and leaf segment explants was 100 ± 0.0 and 90.50 ± 0.89% respectively while the callus frequency on this medium containing 0.25 mg l(-1) 2iP and 0.25 mg l(-1) NAA was 100 ± 0.0 and 89.90 ± 0.72% respectively. The shoot regeneration frequency for petiole explants was 89.90 ± 0.46% producing 6.00 ± 0.21 shoots per callus on SH basal medium supplemented with 0.25 mg l(-1) TDZ and 0.25 mg l(-1) NAA. Whereas petiole explants could induce 83.70 ± 0.50% shoot regeneration and 7.3 ± 1.05 shoots per callus on SH medium containing 0.25 mg l(-1) indole-3-butyric acid (IBA), 0.5 mg l(-1) 6-benzyladenine (BA) and 0.5 mg l(-1) 2iP. Elongation of regenerated shoot was obtained on MS basal medium supplemented with 0.25 mg l(-1) TDZ. All regenerated shoots developed adventitious roots within 4 weeks when transferred to rooting medium containing SH medium supplemented with 0.5 mg l(-1) IBA. Total nine rooted plantlets were transferred from in vitro to in vivo conditions and eight plants survived and successfully acclimatized in the shaded greenhouse 12 weeks after transplanting.

No MeSH data available.


Related in: MedlinePlus