scholarly journals Micropropagation of Two Selected Male Kiwifruit and Analysis of Genetic Variation with AFLP Markers

HortScience ◽  
2005 ◽  
Vol 40 (3) ◽  
pp. 740-746 ◽  
Author(s):  
M.J. Prado ◽  
M.T. Herrera ◽  
R.A. Vázquez ◽  
S. Romo ◽  
M.V. González

A simple and reliable protocol for micropropagation during 12 subcultures of two field growth male plants of kiwifruit [Actinidia deliciosa (A.Chev.) Liang and Ferguson] is described. The best results of shoot multiplication and elongation were obtained in Cheng's K(h) medium in the presence of 0.5 μm NAA, 22 μm BA and 1.4 μm GA3 for `Tomuri' explants, and of 0.1 μm NAA, 4.4 μm BA, and 0.3 μm GA3 for clone A explants. In addition, the cytokinin compounds TDZ and mT were also tested allowing improving the multiplication rate in `Tomuri' explants. For rooting, `Tomuri' and clone A developed shoots were treated by basal immersion in a 5 mm IBA solution for 15 seconds. Treated shoots were then cultured in half-strength K(h) medium without growth regulators showing 100% rooting after 30 days. Regenerated plantlets were successfully transplanted to soil (90% survival) and they are actively growing in the field. Somaclonal variation analysis by AFLP was carried out using 15 primer combinations, yielding reproducible and well-resolved bands with a 57% of polymorphism. AFLP markers showed to be effective to discriminate genetic variation in this species, being greater in clone A than `Tomuri'. Chemical names used: N6-benzyladenine (BA); gibberellic acid (GA3); indole-3-butyric acid (IBA); meta-topolin (mT); naphthaleneacetic acid (NAA); thidiazuron (TDZ).

1990 ◽  
Vol 8 (4) ◽  
pp. 177-179
Author(s):  
S. Yusnita ◽  
R. L. Geneve ◽  
S. T. Kester

Abstract A white flowering Eastern redbud (Cercis canadensis var. alba L.) has been successfully micropropagated. Two node explants collected from the initial flush of spring growth were cultured on woody plant medium (WPM). Increased shoot multiplication occurred at 10,15 and 20 μM (2.3, 3.4 and 4.5 ppm) benzyladenine (BA). Microshoots were rooted in vitro on half strength WPM with a 15-day treatment of 100 and 300 μM (18.6 and 55.9 ppm) α-naphthaleneacetic acid (NAA) or 100 and 300 μM (20.3 and 60.9 ppm) indolebutyric acid (IBA) prior to being moved to full strength WPM without growth regulators. Percentage rooting and the mean number of roots per cutting were comparable between NAA and IBA treated microcuttings, however, the subsequent root morphology differed between the two treatments. NAA treated plants developed a coarse, unbranched root system, while IBA treated cuttings developed a more desireable fine, branched root system. Rooted microshoots were successfully acclimated to greenhouse conditions.


1970 ◽  
Vol 43 (2) ◽  
pp. 215-222 ◽  
Author(s):  
AKM Sayeed Hassan ◽  
Farhana Afroz ◽  
Laila Shamroze Bari ◽  
John Liton Munshi ◽  
Miskat Ara Akhter Jahan ◽  
...  

A protocol was established for mass propagation of a valuable medicinal herb, Eclipta alba (Linn.) Hassk (Asteraceae) through in vitro culture. Apical and axillary buds of young sprouts from selected plants were used as explants. Best shoot induction was observed on MS basal medium supplemented with 0.5 mgl-1 BAP + 0.1 mgl-1 NAA, in which 94% of the explants produced 18 shoots per culture. Repeated subcultures in the same medium, resulted rapid shoot multiplication with 26 shoots per culture. In vitro raised shoots rooted on half strength MS medium with 1.0 mgl-1 IBA +1.0 mgl-1 NAA. For acclimatization and transplantation, the plantlets in the rooting culture tubes were kept in normal room temperature for 7 days before transplanting in pots where plantlets were reared for three weeks. The survival rate of regenerated plantlets was 80%. Key words: Eclipta alba, Medicinal plant, Shoot proliferation, Micropropagation, Acclimatization   DOI: 10.3329/bjsir.v43i2.965 Bangladesh J. Sci. Ind. Res. 43(2), 215-222, 2008 


HortScience ◽  
2019 ◽  
Vol 54 (9) ◽  
pp. 1558-1564 ◽  
Author(s):  
Georgia Vlachou ◽  
Maria Papafotiou ◽  
Konstantinos F. Bertsouklis

Seed ecophysiology and micropropagation of Clinopodium nepeta, an aromatic Mediterranean plant with pharmaceutical and horticultural uses was investigated. The optimum germination temperature of seeds stored at room temperature for 0, 6, or 12 months was 15 to 20 °C (100% germination completed in 10 to14 days) and cardinal temperatures were defined at 10 and 30 °C (80% to 82% and 62% to 76% germination, respectively). Six or 12 months of storage did not seem to affect germination, although 12-month-old seeds germinated at higher percentage and completed germination earlier at 15 °C than at 20 °C. Concerning micropropagation, shoot multiplication at subcultures of both adult plant- and seedling-origin nodal explants was tested on Murashige and Skoog (MS) medium supplemented with various cytokinin types, i.e., zeatin (ZEA), 6-benzyladenine (BA), kinetin (KIN), and 6-γ-γ-(dimethylallylamino)-purine (2IP), at various concentrations from 0.0 to 8.0 mg·L−1. Both explant types presented a rather similar response during in vitro culture. Increasing concentration of all cytokinin types resulted in an increase in shoot number per responding explant (1.1–5.3) and in most cases a decrease in shoot length (0.6–3.4 cm). Increasing cytokinin concentration induced hyperhydricity to a number of shoots (0.1–6.5) per explant, mostly when ZEA and BA were used. Supplementing the MS medium with 8.0 mg·L−1 BA combined with 0.1 mg·L−1 1-naphthaleneacetic acid (NAA) led to almost elimination of hyperhydricity and very satisfactory shoot production (80%/88% explant response and 6.5/7.5 shoot number per responding explant for seedling- / adult-origin explants, respectively). Alternatively, increasing the agar concentration to 12.0 g·L−1 and supplementing the medium with 8.0 mg·L−1 BA only, resulted in the same effect on eliminating hyperhydricity, such as the addition of NAA, and in the best shoot multiplication response achieved in this study (100% explant response, 9.4/9.9 shoots per explant for seedling-/adult-origin explants, respectively). Microshoots rooted abundantly (92% to 100%) on half-strength MS medium, either Hf or supplemented with 0.5 mg·L−1 to 4.0 mg·L−1 indole-3-butyric acid (IBA). The addition of IBA to the rooting medium, regardless of its concentration, affected only the root length by increasing it 2- to 3-fold. Microshoot clusters produced on multiplication media rooted at 96% when cultured on Hf half-strength MS medium. Rooted microshoots and shoot clusters survived at 80% to 100%, respectively, after ex vitro acclimatization in peat:perlite 1:1 (v/v).


1970 ◽  
Vol 44 (3) ◽  
pp. 341-346
Author(s):  
AKM Sayeed Hassan ◽  
Laila Shamroze Bari ◽  
Rebeka Sultana ◽  
Nadira Begum ◽  
Rahima Khatun

An efficient protocol was established for in vitro clonal propagation of the perennial medicinal herb Scoparia dulcis L. (Family. Scrophulariaceae) through in vitro culture. Apical and axillary buds of young sprouts from selected plants were used as explants. Best shoot induction was observed on MS basal medium supplemented with 0.1 mg/l BAP, in which 94% of the explants produced 12 shoots per culture. Repeated subcultures in the same medium, resulted rapid shoot multiplication with 16 shoots per culture. The half strength MS medium with 0.5 mg/l IBA +0.5 mg/l NAA the highest percentage (85.20) and maximum number (13.40) of roots were initiated within four weeks of culture. For acclimatization and transplantation, the plantlets in the rooting culture tubes were kept in normal room temperature for 7 days before transplanting in pots where plantlets were reared for three weeks. The survival rate of regenerated plantlets was 85%. Key words: Scoparia dulcis, Medicinal plant, Shoot proliferation, Micropropagation, Acclimatization, IAA (indoleacetic acid), IBA(indolebutanoic acid), NAA(α-naphthaleneacetic acid), BAP(benzylamino purine) DOI: 10.3329/bjsir.v44i3.4408 Bangladesh J. Sci. Ind. Res. 44(3), 341-346, 2009


2013 ◽  
Vol 65 (1) ◽  
pp. 17-22 ◽  
Author(s):  
Marija Markovic ◽  
Marija Popovic ◽  
Dragica Vilotic

Micropropagation (shoot tip and nodal cuttings culture) was used for the rapid propagation of the non-invasive, decorative, native plants of maiden pink (Dianthus deltoides L.) in order to preserve their genetic diversity. In vitro culture was successfully established on Murashige and Skoog medium (MS) using seeds as the initial material. In the shoot multiplication phase, the explants were cultured on MS medium supplemented with different concentrations of 6-benzylaminopurine (BAP) and naphthaleneacetic acid (NAA). The highest multiplication rate was achieved on a medium containing 0.1 mgL-1 of BAP and 0.1 mgL-1 of NAA. The rooting was successful on a hormone-free medium (100%), and the highest percentage of microplant acclimatization (97%) was recorded in a 4: 1 mixture of peat and sand.


Author(s):  
Vikas Sharma ◽  
Barkha Kamal ◽  
Nidhi Srivastava ◽  
Anoop Kumar Dobriyal ◽  
Vikash Singh Jadon

Enhanced in vitro caulogenesis has been tested in Swertia chirayita on MS supplemented with BAP, IAA, IBA, NAA and additives like adenine sulfate and d?glutamine with 2.5% sucrose. The best in vitro caulogenesis was observed in MS fortified with 4.43 ?M BAP in combination with IAA (0.8 ?M) that resulted increase in shoot multiplication rate. The multiplication and elongation of shoots were further enhanced by the addition of adenine sulfate (0.007%) that resulted in the further increase in multiplication fold. The addition of adenine sulfate reduced the use of other cytokinins with different auxins reported in the previous studies on Swertia chirayita. The study suggests adenine sulfate as a primary assimilable reduced nitrogen source for enhancing the shoot multiplication and elongation in Swertia chirayita. RAPD markers were employed to check the genetic variation among the clonal stock that resulted in 97% similarity.Plant Tissue Cult. & Biotech. 23(1): 11?19, 2013 (June) DOI: http://dx.doi.org/10.3329/ptcb.v23i1.15555


2013 ◽  
Vol 31 (4) ◽  
pp. 246-252 ◽  
Author(s):  
Jason D. Lattier ◽  
Darren H. Touchell ◽  
Thomas G. Ranney ◽  
Jeremy C. Smith

Protocols were developed for micropropagation and induction of autopolyploids in a fastigiate cultivar of Norway maple (A. platanoides L. ‘Crimson Sentry’). Murashige and Skoog (MS) medium, woody plant medium (WPM), and Quoirin and Lepoivre medium were supplemented with 2 μM 6-benzylaminopurine (BA), meta-Topolin, 6-(γ,γ-dimethylallylamino) purine, kinetin, or thidiazuron to evaluate microshoot proliferation. Murashige and Skoog medium with 2 μM BA yielded the most microshoots (3.2) and longest microshoots (30.6 mm) per subsample after 5 weeks. The influence of BA concentration on proliferation was evaluated at 0, 2, 4, 8, or 16 μM. Optimal multiplication rate was achieved at 2 or 4 μM BA producing approximately 2.8 microcuttings per subsample after 5 weeks. To induce in vitro rooting, half-strength WPM was supplemented with 0, 5, 10, 20, 40, or 80 μM indole-3-butyric acid (IBA). Optimal in vitro rooting (70%), number of roots (2.5), and root length (15 mm) per subsample were achieved with 10 μM IBA after 8 weeks. To induce polyploidy, microcuttings were pretreated for 7 days on MS medium with 4 μM BA alone or combined with 1 μM IBA, indole-3-acetic acid (IAA), or 1-naphthaleneacetic acid prior to treatment in liquid MS medium containing 15 μM oryzalin for 3 days. Homogenous tetraploids were only obtained from shoots pretreated with IAA. This research provides optimized protocols for micropropagation and autopolyploid induction of A. platanoides ‘Crimson Sentry’ and demonstrates the development of tetraploid lines for use in future improvement programs.


1970 ◽  
Vol 45 (1) ◽  
pp. 69-74 ◽  
Author(s):  
AKM Sayeed Hassan ◽  
Nadira Begum ◽  
Miskat Ara Akhter Jahan ◽  
Rahima Khatun

A consequency was obtained for mass propagation of a valuable ayurvedic medicinal herb, Heliotropium indicum Linn. (Boraginaceae) through in vitro culture. Apical and axillary buds of young sprouts from selected plants were used as explants. Best shoot induction was observed on MS basal medium supplemented with 0.5 mg/l BAP + 0.1 mg/l GA3, in which 92% of the axillary buds explants produced 12 shoots per culture. Repeated subcultures in the same medium, resulted rapid shoot multiplication with 18 shoots per culture. In vitro raised shoots rooted on half strength MS medium with 0.5 mg/l IBA. For acclimatization and transplantation, the plantlets in the rooting culture tubes were kept in normal room temperature for 7 days before transplanting in pots where plantlets were reared for three weeks. The survival rate of regenerated plantlets was 85%. Key words: Heliotropium indicum; Medicinal plant; Shoot proliferation; Micropropagation; Acclimatization. DOI: 10.3329/bjsir.v45i1.5185 Bangladesh J. Sci. Ind. Res. 45(1), 69-74, 2010


Author(s):  
Soumya Sucharita Singh ◽  
Cinmaya Pradhan ◽  
Dhaneswar Swain ◽  
Gyana Ranjan Rout

An efficient in vitro protocol was developed for the mass multiplication of Vigna mungo vars. PU30 and PU31, an important legume crop through apical meristems and cotyledonary nodal explants. Both apical meristems and cotyledonary nodal explants were cultured on Murashige and Skoog (MS, 1962) medium fortified with 0.5 – 1.50 mg/L 6- benzyl aminopurine (BA) or Kinetin and 3 % (w/v) sucrose. The rate of multiplication was higher when the cultures were incubated under continuous light (24h) than the 16h photoperiod. The average number of multiple shoots per culture was enhanced from 2.43 to 5.46 in the case of var. PU30 and 3.12 to 5.82 in the case of var.PU31 within 4 weeks of culture under 24h photoperiod. The multiplication rate was enhanced till 5th subculture declined thereafter. Rooting was readily achieved upon transferring the shoots to half-strength MS basal semisolid medium supplemented with 0.1– 1.0 mg/L indole-3-butyric acid (IBA) and 2% (w/v) sucrose after 2 weeks of culture.  The average number of roots per explants ranged from 3.12 to 5.76.  About 80% of regenerated plantlets were hardened in the greenhouse and successfully established in the soil. There is no morphological variation among the regenerated plantlets. This protocol can be used for genetic transformation study for crop improvement of black gram.


2013 ◽  
Vol 41 (2) ◽  
pp. 370 ◽  
Author(s):  
Marija MARKOVIĆ ◽  
Mihailo GRBIĆ ◽  
Matilda DJUKIĆ

During past decades, great attention has been paid to propagation of endangered plant taxa in order to preserve biodiversity. The aim of this study was to optimize a protocol for in vitro propagation of the critically endangered and decorative species Dianthus serotinus Waldst. et Kit. The effects of different concentration of MS salt (Murashige and Skoog) of the culture, medium pH and different carbohydrates (sucrose, glucose, and fructose) on shoot multiplication were examined. The best results were obtained on half-strength MS (Murashige and Skoog) medium, whose pH was 5.8, with sucrose supplied at a concentration of 3%, when shoots with 1-2 nodes or shoot tips (with terminal buds only) were used as explants. The shoots were rooted (76.7%) on half-strength MS medium containing 0.5 mg∙L-1 NAA (1-naphthaleneacetic acid). The obtained plantlets were successfully acclimatized (89%) in a 4:1 mixture of peat and sand and they flowered the following year. Presented protocol enables successful in vitro propagation of D. serotinus.


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