scholarly journals High Frequency Plant Regeneration System of Aerides odorata Lour. Through Foliar and Shoot Tip Culture

2013 ◽  
Vol 41 (1) ◽  
pp. 169 ◽  
Author(s):  
Huidrom Sunitibala DEVI ◽  
Sanglakpam Irabati DEVI ◽  
Thingbaijam Dikash SINGH

An efficient protocol for rapid clonal propagation from different explants of Aerides odorata Lour.- an endemic orchid of Manipur has been established. Leaf base explants showed significant response in ½ strength Murashige and Skoog medium supplemented with thidiazuron (TDZ) and 6-benzylaminopurine (BAP). Callus were initiated only from leaf base explants after 60 days of culture while other parts of leaves failed to response in all the treatments. Medium supplemented with 1.0 mg/L TDZ produced protocorm like bodies (PLBs) at the leaf base. Shoot tip explants of A. odorata showed different morphogenetic responses in different phytohormone treatments. Calli were initiated only in the medium containing α-naphthalene acetic acid (NAA). Highest calli frequency was observed in the medium containing 2 mg/L (NAA) (85.71±0.21) which indicates the importance of exogenous auxin in embryonic callus proliferation. Direct shoot regeneration on the other hand was observed in all the treatments. Highest number of shoot was obtained in higher concentration of NAA (2 mg/L) and BAP (4 mg/L) (4.80±0.18), showing combined effect of BAP and NAA, which may be due to the synergistic effect of cytokinin and auxin. Among the different rooting phytohormones, addition of NAA (0.5 mg/L) in ½ MS medium shows highest frequency of root induction. More than 95% in vitro plants survived during acclimatization under ex vitro conditions. This phytohormones and explants based micropropagation system can open up the route for in vitro clonal multiplication of this commercially important Aerides species.

HortScience ◽  
2010 ◽  
Vol 45 (5) ◽  
pp. 797-800 ◽  
Author(s):  
Xiuli Shen ◽  
William S. Castle ◽  
Frederick G. Gmitter

This study examined the effects of plant growth regulators, explant types, and their orientations on in vitro shoot proliferation of Casuarina cunninghamiana Miq. and also the subsequent rooting ability of shoots. Results showed that shoot proliferation occurred only in shoot tip explants cultured vertically on Murashige and Skoog (MS) medium supplemented with 2 or 4 μM thidiazuron (TDZ). Neither 6-benzylaminopurine alone nor in a combination with 1-naphthalene acetic acid (NAA) or gibberellic acid had any effect on shoot proliferation. TDZ at 4 μM resulted in the greatest percentage of axillary bud sprouting (70%) and mean number of sprouts per explant (2.3). Additionally, no shoot proliferation was observed from detipped or single-node explants or from horizontally placed shoot tip explants when cultured on the same TDZ-containing medium. The induced shoots produced adventitious roots on MS medium supplemented with 2.5, 5, or 10 μM indole-3-butyric acid (IBA), not with indole-3-acetic acid and NAA. Although the mean number of roots per explant was not significantly different between 2.5 and 5 μM IBA, the highest rooting percentage (68%) and mean length of roots per explant (0.7 cm) was achieved at 5 μM IBA. The current study provided preliminary information toward commercial in vitro propagation of Casuarina cunninghamiana male plants.


2016 ◽  
Vol 12 (1) ◽  
pp. 103
Author(s):  
Lazarus Agus Sukamto

Nepenthes albomarginata Lobb ex Lindl. is a carnivorous plant, distributes in several regions in Indonesia. The plant population decreases drastically because of over exploitation and ruining nature habitat. Plant propagation by nature and cutting are not enough to rehabilitation its population. In vitro culture of N. albomarginata was carried out using plantlets grown from the seeds in vitro. Plantlets were cut to became two part explants, consisted of shoot tip and under-shoot tip cuttings. These cutting explants were grown on Murashige & Skoog (MS) media with addition of plant growth regulators of 6-benzylaminopurine (BA), combined with or without-naphthalene acetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D) at 1 mg/l. Shoot tip cuttings of N. albomarginata formed double multiple shoot 25,00% on control; formed triple multiple shoots 25,00% onBA 1 mg/l treatment; formed callus 37,50%, triple or quartet shoots 25,00% and rooted plantlets 25,00% on BA 1 mg/l + NAA 1 mg/l treatment. The under-shoot tip cuttings ofN. albomarginata formed double – triple shoots 25,00% and rooted plantlets 37,50% on control; formed double – triple shoots 25,00% and rooted plantlets 12,50% on BA 1 mg/ltreatment; formed callus 12,50%, double - pentacle shoots 37,50% and rooted plantlets 25,00% on BA 1 mg/l + NAA 1 mg/l treatment. 2,4-D 1 mg/l or its combined with BA 1mg/l treatment caused deadly shoot tip or under-shoot tip explants. The combination of BA 1 mg/l + NAA 1 mg/l was the best treatment for producing callus, multiple shootsand rooted plantlets of N. albomarginata.


2009 ◽  
Vol 18 (1) ◽  
pp. 1-6 ◽  
Author(s):  
H. David Raja ◽  
D.I. Arockiasamy

A simple micropropagation method from nodal and shoot tip explants is reported here for Mentha viridis, an economically important medicinal plant. High frequency of microshoots was obtained from these two explants on MS supplemented with various concentrations of BAP (1.0 - 4.0 mg/l) and Kn (1.0 -4.0 mg/l). Maximum number of shoots was obtained from nodal explants in the medium containing 3.0 mg/l BAP. The root induction was carried out by using IBA and IAA (0.5 - 2.0 mg/l). Among these, high frequency of root proliferation was achieved in the medium containing 1.5 mg/l of IBA. The rooted plantlets were hardened and transferred to the field. The survival rate was 90 - 95% after 25 days. Key words: Micropropagation, Mentha viridis, Node, Shoot tip, Multiple shoots D.O.I. 10.3329/ptcb.v18i1.3243 Plant Tissue Cult. & Biotech. 18(1): 1-6, 2008 (June)


1970 ◽  
Vol 7 (2) ◽  
pp. 253-258 ◽  
Author(s):  
A Ferdausi ◽  
UK Nath ◽  
BL Das ◽  
MS Alam

Brinjal is the second most important vegetable crop after potato in Bangladesh in respect of total areas and third in production. It also plays a vital role in the national economy as a cash crop. An experiment was conducted with two cultivars of brinjal namely Jhumky and Islampuri to observe the callus induction ability of different explants-shoot tip, hypocotyl and midrib in MS basal media supplemented with different concentrations and combinations of growth regulators. The rate of callus induction from shoot tip, hypocotyl and midrib were 82.78%, 74.88% and 78.71%, respectively. Highest rate of callus induction was found in shoot tip. Variety Islampuri showed higher rate of callus induction (80.62%). Among the treatments 2mg/l NAA showed the best performance in callus proliferation. Cytokinin (0.5 mg/l BAP) showed highest percentage of shoot regeneration (57.13%). For root induction, MS basal medium was proved to be better treatment for average number (12-15) of roots. The survival rate of transferred regenerated plantlets after hardening was higher in Jhumky (80%). Regenerated plantlets from callus of both the varieties exhibited 4-9 times higher proline, 2-3 times lower vitamin C and 2-3 times higher iron (Fe) content compared to their seed derived seedlings. This experiment showed that it is possible to develop shoot and fruit borer tolerance brinjal genotypes through somatic embryogenesis that was selected based on biochemical markers within the very short period of time. These findings will be helpful for further selection of the selected variants in field condition in the next phase of the study. Keywords: Somaclonal variants; Biochemical markers; Brinjal DOI: 10.3329/jbau.v7i2.4731 J. Bangladesh Agril. Univ. 7(2): 253-258, 2009


Author(s):  
R. Anandan ◽  
T. Deenathayalan ◽  
R. Bhuvaneshwari ◽  
M. Merlin Monisha ◽  
M. Prakash

Here an efficient protocol for micropropagation of black gram [Vigna mungo (L.) Hepper] cv. VBN 3 is reported. The deembryonated cotyledonary explants were cultured on MS medium containing different concentrations of plant growth regulators. The maximum frequency (72%) of direct shoot regeneration (devoid of callus phase), multiple shoot induction and shoot elongation was achieved from culturing the explants on MS medium containing 3.0 mg/l of 6-benzylaminopurine (BAP). Up to 65% of the regenerated shoots were rooted on MS medium containing 0.25 mg/l of á-naphthalene acetic acid (NAA) within 3 weeks after subculturing. The in vitro-raised plantlets were successfully hardened first under culture room conditions with 62% survival rate and then in greenhouse. The identified regeneration system could be efficiently used in various in vitro manipulation studies in black gram as well.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Pooja Taak ◽  
Siddharth Tiwari ◽  
Bhupendra Koul

Abstract Stevia rebaudiana Bertoni is a commercially important zero calorie natural-sweetener herb which produce sweet compounds known as steviol glycosides. Rising demands of steviol glycosides by food and beverage industries has led to an increase in its cultivation in various countries. Unfortunately, stevia cultivation faces 2–25% yield penalty due to weeds which further adds to its cultivation cost. To resolve this major challenge, Agrobacterium-mediated genetic transformation of in vitro derived stevia-nodal explants using herbicide resistance gene (bar) has been optimized, for the production of stable transgenic stevia plants. Several parameters including explant type, pre-incubation duration, acetosyringone (As) concentration, Agrobacterium cell density, Agro-inoculation duration, co-cultivation duration, selection regime and plant growth regulators (PGRs) combination and concentration, have been successfully optimized. Among the two types of explants used, nodal explants showed a higher regeneration response of 82.85%, with an average of 25 shoots/explant. The best PGRs combination and concentration for shoot-induction, shoot-elongation and root-induction was found to be 6-benzyladenine (1.0 mg l−1) + naphthalene acetic acid (0.5 mg l−1), gibberellic acid (1.0 mg l−1), and half-strength MS medium, respectively. The two-step selection (phosphinothricin) regime resulted in an average transformation efficiency of 40.48% with nodal explants. Molecular characterization of putative transformants through PCR, RT-PCR, qRT-PCR and Southern-blot hybridization confirmed the presence, stability, expression as well as copy number of bar gene respectively. Compared to the non-transgenic plants, the T0 transgenic plants successfully tolerated 8 mg l−1 glufosinate ammonium sprays. Thus, the optimized protocol can be useful for the introduction of other genes (inter-kingdom transfer) into stevia genome.


2015 ◽  
Vol 10 (1) ◽  
Author(s):  
Xia Yu ◽  
Jiajing Sheng ◽  
Lingling Zhao ◽  
Ying Diao ◽  
Xingwen Zheng ◽  
...  

AbstractIn this study, an efficient and reproducible plant regeneration system for lotus (Nelumbo nucifera Gaertn.) was established using shoot apical meristems from the buds and one-week-old aseptically germinated embryos as explants. Multiple shoot clumps were induced on Murashige and Skoog (MS) basal medium supplemented with various combinations of N6-Benzylaminopurine (6-BA) and α-Naphthalene acetic acid (NAA). The maximum response was obtained with 2.22 μM 6-BA, and produced 21.33 shoots per explant after four weeks. After five subcultures, multiple shoot clumps were transferred to MS basal medium supplemented with various combinations of 3-Indolebutyric acid (IBA), NAA and sucrose for root induction. After four weeks, plantlets with well-developed roots were achieved on MS basal medium supplemented with 0.54 μM NAA and 30 g/L sucrose with 100% rooting rate. The successfully acclimated plantlets were transferred to pots with the addition of 2 g/L KMnO4 into the soils, and finally fertile plants with much bigger leaves were obtained in the greenhouse. The survival rate was 97.33%.


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