Induction of multiple shoots from stem explants of Chardonny grape.
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In order to induce multiple shoots from Chardonny grape(Vitis vinifera)explants,the effects of explants,media,hormone concentrations and dark treatment on the induction of multiple shoots were studied.The shoots were induced only from the stem explants,the induction frequency reached to 17.5%.There was no difference in the average number of shoots a-mong 3 media of MS,NN1969 and B5.The shoots were induced from the stems on the MS medium supplemented with BA and IBA,and there was significant difference in the frequency of shoot induction among different concentration treatments of BA and IBA.However,there was no significant difference in the average number of shoots and most of shoots induced were single shoot,indicated that there was no significant effect of the combination of BA with IBA on the induction of multiple shoots in this study.The significant effect of the combination of BA with NAA on both the induction frequency of shoots and the number of multiple shoots was observed.The highest induction frequency(9.7%)of shoots and the highest average number of shoots(10.1)were induced from the stems on the MS medium supplemented with 3.0 mg·L-1 BA and 0.1 mg·L-1 NAA,the highest number from one stem reached up to 27.The roots were induced from the shoots on the 1/2MS medium supplemented with 0.05 mg·L-1 IAA.Cite
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<p style="text-align: justify;"><strong>Aim</strong>: To optimize the concentrations of growth regulators in the media for the proficient micropropagation of grapevine (<em>Vitis vinifera </em>L.) cv. King’s Ruby.</p><p style="text-align: justify;"><strong>Methods and results</strong>: Apical meristems of the grape cultivar were used to establish <em>in vitro</em> shoot cultures. Nodal explants, each containing an axillary bud, taken from <em>in vitro</em> grown shoots were inoculated in shoot proliferation medium, i.e., half strength Murashige and Skoog (MS) medium supplemented with benzyl aminopurine (BAP), kinetin, glycine and gibberellic acid (GA<sub>3</sub>). A higher number of shoots (5.33) with greater shoot length (2.75 cm) was produced in the medium supplemented with 1.0 mg L<sup>-1</sup> BAP and 0.1 mg L<sup>-1</sup> GA<sub>3</sub>. Calluses were induced from leaf explants taken from <em>in vitro</em> grown shoots. Callus induction was greater (73.00%) on the medium containing 2.0 mg L<sup>-1</sup> 2,4-dichlorophenoxyacetic acid (2,4-D), 0.3 mg L<sup>-1</sup> BAP and 0.2 mg L<sup>-1</sup> α-naphthaleneacetic acid (NAA). The maximum frequency of shoot regeneration (53.33%) was achieved on the medium supplemented with 1.5 mg L<sup>-1</sup> BAP and 0.5 mg L<sup>-1</sup> NAA, and the regenerated shoots successfully formed roots on growth regulator-free half strength MS medium.</p><p style="text-align: justify;"><strong>Conclusion</strong>: Optimizing the concentration of BAP and GA<sub>3</sub> and omitting the glycine and kinetin in the culture medium increased the number and length of shoots. Similarly, for inducing the callus of the leaf explants, taken from <em>in vitro</em> grown shoots, it is recommended to adjust the medium with the higher concentration of 2,4-D and lower concentrations of BAP. Moreover, the maximum number of shoots was regenerated on a medium supplemented with relatively high levels of both BAP and NAA (1.5 and 0.5 mg L<sup>-1</sup>, respectively). Finally, we suggest the half strength MS medium that is free from growth regulators for the root formation of the regenerated shoots.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: Optimizing the concentration of growth regulators is crucial for the efficient micropropagation of a grape cultivar. Knowing the specific balance between the growth regulators is necessary to establish <em>in vitro</em> shoot cultures, callus induction and shoot regeneration and, hence, to propagate disease-free true to type grape cultivars in a short time.</p>
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Effects of N6-benzyladenine (BA) or thidiazuron (TDZ) on adventitious shoot regeneration and axillary shoot multiplication of Sedum sarmentosum was investigated. Leaf and shoot tip explants obtained from in vitro-grown shoots of S. sarmentosum were cultured on Murashige and Skoog (MS) medium supplemented with 0, 2.0, 4.0 or 8.0 µM BA or TDZ. Of the two cytokinins studied, BA was found to be more responsive as compared to TDZ with respect to number of shoots produced per explant. High frequency of shoot regeneration (92.2%) with a mean of 12.3 shoots was produced when the leaf explants were cultured on MS medium supplemented with 8.0 µM BA. The highest number of shoots (25.4) was obtained when shoot tip explants were cultured on MS medium devoid of cytokinins after 35 days of culture. For root induction, regenerated shoots were cultured on MS medium supplemented with 0, 2.0, 4.0 or 8.0 µM indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA). The highest number of (17.6) roots per shoot was obtained on MS medium supplemented with 2.0 µM IBA after 30 days of culture. Regenerated plantlets were successfully acclimatized in the greenhouse with 100% survival rate.
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Multiple shoots were induced from the nodal meristms in Coccinia indica L. in present study. Maximum number of shoots (4.7) per explant were observed on Murashige and Skoog (MS) medium supplemented with 2.5 mgl-1 6-Benzylaminopurine (BAP). The response of explants towards the use of kinetin (Kn) was not impressive (3.6 shoots per explant) as compared to BAP. Application of auxin (Indole-3 acetic acid, IAA) in the medium caused cullogenesis from the basal part of the explants. Shoots were multiplied (47.3 shoots per culture flask) on MS medium supplemented with 1.0 mgl-1 each of BAP and Kn. Long and healthy shoots (4-5 cm long) were rooted (8-9 roots per shoot) on agar-gelled half-strength MS medium supplemented with 2.0 mgl-1 Indole-3 butyric acid (IBA). None of the shoot produced roots in ex vitro experiments even after five weeks in green house. In vitro generated plantlets were hardened in the green house and transferred to the pots and finally to the field with 100% success.
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Abstract A micropropagation protocol for Maytenus senegalensis was established using in vitro germinated 6-week-old seedlings as a source of explants. Shoot proliferation was only achieved using nodal explants. The presence of cytokinins was necessary for shoot induction and growth. While benzyl-6-aminopurine (BA) generally promoted the average number of shoots produced per explant, kinetin promoted the mean length of shoots. Inclusion of auxin (IAA and IBA) in the shoot induction and growth medium did not have significant effects on either the average number of roots produced per shoot or the mean length of shoots. Low levels of BA (0.5 or 1.0 mg l − 1 ) were optimal for shoot induction and growth. As a result, shoot multiplication was done on a medium supplemented with BA (0.5 mg l − 1 ) alone. Rooting was achieved in 2 stages. In the first stage, 8-week-old shoots (2–3 cm long) were pulse-treated in the dark using ½ strength, MS [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum 15, 473–497] liquid medium containing auxins and then transferred to a solid hormone-free medium at a 12/12 h photoperiod. The shoots pulse-treated with 25 mg l − 1 IBA for 120 h produced the highest number of roots per shoot. IBA concentration and the period of pulse treatment had significant effects on the average number of roots produced per shoot. The rooted plantlets were successfully acclimatized.
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Abstract This study established, for the first time, shoot proliferation and plant regeneration protocols via shoot organogenesis from leaf explants of a medical and ornamental plant, Portulaca pilosa L. The optimal proliferation of axillary shoots was 6.2-fold within 30 days on Murashige and Skoog (MS) medium supplemented with 3.0 µM 6-benzyladenine (BA). Shoots could be induced directly from leaf explants, forming an average of 3.8 adventitious shoots per explant, on optimal MS medium supplemented with 1.0 µM thidiazuron (TDZ) and 0.1 µM α -naphthaleneacetic acid (NAA). A higher concentration of TDZ (3.0 µM), alone or in combination with 0.1 µM NAA, induced somatic embryo-like shoot buds and then developed into real shoots. Rooting was easier since roots were induced on all rooting media within one month. Half-strength MS medium free of plant growth regulators was best for rooting. Rooted plantlets were transferred to a sand: perlite (1:1, v/v) substrate, resulting in highest survival (90%). Plantlets showed more robust growth, however, on substrates of yellow mud: perlite (1:1, v/v) or peat soil: vermiculite: perlite (1:1:1, v/v).
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A high frequency of sprouting (80%) from nodal- and (70%) from shoot tip explants and shoot differentiation was observed in the primary cultures of Morus alba L. on MS medium supplemented with BAP and Kn. In vitro proliferated shoots were multiplied rapidly by culture of shoot tips and nodal explants on MS with BAP (2 mg/l) and NAA (0.2 mg/l) as supplements. This combination proved best for multiple shoot formation. Multiplication was also achieved by culture of both the kinds of explants on MS fortified with BAP (2 mg/l) + NAA (0.2 mg/l) + aspargine (25 mg/l) + glutamine (1 mg/l). This medium facilitated the elongation of shoots and sprouting of axillary buds of in vitro grown shoots. About 80% rooting was obtained from shoots cultured on the MS supplemented with NAA (1.0 mg/l). Plants with well developed roots were transferred to soil with survival frequency of 70%.
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The effect of benzyladenine (BA) and thidiazuron (TDZ) in combination with -naphthaleneacetic acid and indole-3-butyric acid (IBA) on in vitro shoot regeneration from leaf segments of several Japanese pears was investigated. A regeneration protocol was developed and regeneration was achieved from six cultivars: – ‘Kosui’, ‘Hosui’, ‘Niitaka’, ‘Wasekaso’, ‘Okusankichi’ and ‘Whangkeumbae’ pears. The highest regeneration rate was obtained in most shoots from young leaves on a medium based on MS macroelements supplemented with 0.1-1.0 mg/L TDZ and 0.1-1.0 mg/L IBA and/or NAA in each cultivar. Physiological differences with BA and TDZ treatments were compared. In the regeneration medium with BA treatments, green foci appeared on the callus surface after 8 days. Then, some adventitious buds were induced on those green foci, resulting in normal shoots. On the other hand, in the medium with TDZ, callus surface turned compact and greenish, and many adventitious buds were formed over the whole area of the callus surface. In some shoots, cultured on the medium with TDZ, there were morphologically abnormal shoots, including vitrified shoots. A-fourth strength MS medium supplemented with 0.1 mg/L NAA produced plantlets in good quality of root number and root length.
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