scholarly journals Efficient in vitro plant regeneration from leaf derived callus and genetic fidelity assessment of an endemic medicinal plant of Ranunculus wallichianus Wight & Arnn using RAPD and ISSR markers

2021 ◽  
Author(s):  
Srinivasan P ◽  
David Raja H ◽  
Tamilvanan R
Keyword(s):  
Callus Formation ◽  
Leaf Explants ◽  
Genetic Fidelity ◽  
Issr Markers ◽  
Ms Medium ◽  
Genetic Purity ◽  
Fidelity Assessment ◽  
Half Strength ◽  
Rapd And Issr

Abstract Ranunculus wallichianus is a medicinally important plant and an endemic species to Western Ghats of South India. An efficient and reliable indirect regeneration protocol system for R. wallichianus was developed from leaf explants in the present investigation. Leaf explants were cultured on both full-strength and half-strength MS (Murashige & Skoog) medium supplemented with different concentrations (1.0 mg L− 1 to 3.0 mg L− 1) of 2,4-D and NAA. Among the different concentrations tested, the highest percentage of yellowish green compact nodular callus formation was observed on half-strength MS medium with 2.0 mg L− 1 of 2, 4-D. Then, the in vitro raised organogenic callus was cultured on half strength MS medium containing various concentrations (1.0 mg L− 1 to 3.0 mg L− 1) of BA, KIN and TDZ with 0.5 mg L− 1 NAA and 10% CW for in vitro shoot regeneration. The highest percentage of regeneration response (97%) and maximum number of shoots formation (11.1 ± 0.13 shoots/culture with 9.2 ± 0.35 cm mean shoot length) were obtained from MS medium containing 2.5 mg L− 1 BA with 0.5 mg L− 1 NAA and 10% CW. The well elongated in vitro raised shoots were rooted in half strength MS medium with 2.5 mg L− 1 IBA + 250 mg L− 1 activated charcoal shows high frequency of root formation. The well rooted plantlets were successfully hardened and acclimatized with the survival rate of 94%. Clonal fidelity of in vitro raised plantlets was assessed by using DNA based RAPD and ISSR molecular markers. The total of 56 and 47 monomorphic bands were obtained from RAPD and ISSR markers respectively. This present in vitro propagation protocol system could be an effective for the conservation of R. wallichianus with their genetic purity and its further investigations.

scholarly journals IN VITRO REGENERATION AND MASS PROPAGATION OF AZIMA TETRACANTHA [LAM.] FROM THE LEAF EXPLANTS THROUGH CALLUS CULTURE

2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T
Keyword(s):  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Ms Medium ◽  
Apical Leaf ◽  
Half Strength ◽  
Regenerated Plantlets ◽  
Azima Tetracantha

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.

scholarly journals Callus Induction in Baru (Dipteryx alata Vog.) Explants

2018 ◽  
Vol 47 (2) ◽  
pp. 538-543
Author(s):  
Rodrigo Kelson S. REZENDE ◽  
Ana Maria N. SCOTON ◽  
Maílson V. JESUS ◽  
Zeva V. PEREIRA ◽  
Fernanda PINTO
Keyword(s):  
Ascorbic Acid ◽  
Callus Induction ◽  
Callus Formation ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Alternative Technique ◽  
Propagation Methods ◽  
Dipteryx Alata

Baru (Dipteryx alata Vog.) is a species with great economic and environmental potential; it has popular acceptance, besides being a very productive species. Alternative propagation methods are important for species maintenance and exploration. Thus, micropropagation emerged as an alternative technique, providing genetic stability and the production of a large number of seedlings. The aim of the present investigation was to develop a callus induction protocol for in vitro baru explants. The tested explants were nodal, internodal and foliar segments. The explants were disinfected for 30 seconds in 70% alcohol (v/v) and 2 minutes in sodium hypochlorite (1.25% active chlorine). This was followed by triple washing. The inoculation was carried out in test tubes containing 15 mL MS medium (30 g L-1 sucrose, 6 g L-1 agar and 100 mg L-1 ascorbic acid) supplemented with 2.0 mg L-1 naphthalene acetic acid (NAA). The solution also contained 0.0, 2.5 or 5.0 mg L-1 of 6-benzylaminopurine (BAP) with the pH adjusted to 5.8. In the incubation phase, the explants were cultured for seven days in the dark and then subjected to a photoperiod of 16 hours (43 µmol m-2 s-1) at 25 ± 2 °C. The treatments were studied with 2.5, 5.0, 7.5 or 10.0 mg L-1 BAP additions to the MS. Callus formation, contamination and oxidation evaluations were undertaken. The results obtained when using 2.0 mg L-1 NAA concluded that such a treatment should be used to induce callogenesis from nodal explants, while for the tested baru leaf explants, the best results for callus formation were given by the combination of 2.0 mg L-1 NAA with 2.5 mg L-1 of BAP to.

scholarly journals Comparative analyses of stevioside between fresh leaves and in-vitro derived callus tissue from Stevia rebaudiana Bert. using HPLC

2015 ◽  
Vol 49 (4) ◽  
pp. 199-204 ◽  
Author(s):  
S Mahmud ◽  
S Akter ◽  
IA Jahan ◽  
S Khan ◽  
A Khaleque ◽  
...  
Keyword(s):  
Retention Time ◽  
Callus Tissue ◽  
Callus Formation ◽  
Leaf Explants ◽  
Stevia Rebaudiana ◽  
Poor Performance ◽  
The Other ◽  
Ms Medium ◽  
Green Callus

A protocol was developed to produce large amount of callus in short a period of time from leaf explants of Stevia rebaudiana Bert. The highest amount of white callus was obtained on MS medium supplemented with 2.5 mg/l 2, 4-D and 0.5 mg/l BAP after 3 weeks of inoculating leaf segments. On the other hand, 0.5 mg/l BAP and 1.0 mg/l Kn exhibits poor performance towards callus formation while after using 1.0 mg/l Kn alone did not develop any callus. In this experiment, highest amount of green callus was obtained when MS medium supplemented with 2.5 mg/l NAA and 10% coconut water was used. An improved analytical method HPLC was applied to analyze stevioside extracted from the leaf and callus of Stevia rebaudiana. The stevioside in each sample were analyzed by comparing their retention times with those of the standards. The retention time (RT) of stevioside for leaves were found 14.96 and for callus 13.81 mins. The percentage of stevioside content from leaves and callus was 12.19% and 12.62% respectively DOI: http://dx.doi.org/10.3329/bjsir.v49i4.22621 Bangladesh J. Sci. Ind. Res. 49(4), 199-204, 2014

scholarly journals In vitro adventitious shoot regeneration from leaf explants of some apricot cultivars

2019 ◽  
Vol 43 ◽  
Author(s):  
Olga Vladimirovna Mitrofanova ◽  
Irina Vjacheslavovna Mitrofanova ◽  
Tatyana Nikolaevna Kuzmina ◽  
Nina Pavlovna Lesnikova-Sedoshenko ◽  
Sergey Vladimirovich Dolgov
Keyword(s):  
Callus Formation ◽  
System Development ◽  
Low Frequency ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Limiting Factor ◽  
Ms Medium ◽  
Regeneration System ◽  
Propagation Methods

ABSTRACT Apricot is one of the most valuable commercial fruits. In vitro propagation of apricot is very important for rapid multiplication of cultivars with desirable traits and production of cleaning up and virus-free plants. Low frequency of multiplication is the main limiting factor for traditional propagation methods. In this regard, the objective of our investigation was to study the morphogenetic capacity of apricot leaf explants of the promising cultivars ‘Iskorka Tavridy’, ‘Magister’ and ‘Bergeron’ for regeneration system development and solving some breeding questions. The source of explants was in vitro plants regenerated and cultured on QL medium. Leaves were maintained in the dark at 24±1 °C in thermostat for three-four weeks. Morphogenic callus and structures were mainly formed at the central and proximal parts of leaves on MS, QL and WPM media with 1.5 or 2.0 mg L-1 BAP and 1.5 or 2.0 mg L-1 IAA in different combinations, or TDZ (0.6 and 1.3 mg L-1). Callus with adventive buds was transferred to regeneration medium and placed into a growth chamber at 24±1 °C and 16-hour photoperiod with a light intensity of 37.5 μmol m-2 s-1. The best results were obtained when adaxial leaf surface was in contact with the culture medium. Frequency of leaf callus formation on MS medium with 1.5 mg L-1 BAP and 1.5 mg L-1 IAA was higher in the explants of ‘Iskorka Tavridy’ (80.0%) than in - ‘Bergeron’ (50.0%) and ‘Magister’ (36.7%). The best results of callogenesis for ‘Magister’ was obtained on MS medium with 1.3 mg L-1 TDZ (53.3%). Active microshoot regeneration in ‘Iskorka Tavridy’ cultivar was shown on MS medium with BAP and IAA and in ‘Magister’ cultivar - on MS medium with TDZ. Rhizogenesis was obtained on half strength MS medium with 2.0 mg L-1 IBA.

scholarly journals Transformation of Nicotiana alata Link and Otto. with an Autoregulatory Senescence-inhibitor Gene Construct

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 617c-617
Author(s):  
Kenneth R. Schroeder ◽  
Dennis P. Stimart
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Nicotiana Alata ◽  
Ms Medium ◽  
Gene Encoding ◽  
Half Strength ◽  
Cytokinin Synthesis ◽  
Senesced Leaves

Leaf explants of Nicotiana alata Link and Otto. were surface disinfested and cultured on Murashige and Skoog (MS) medium containing 2.66 μm N6-benzyladenine (BA) to promote shoot proliferation. After 5 weeks, proliferated shoots were removed and remaining callus saved. Callus was inoculated with Agrobacterium tumefaciens encoding a senescence-specific promoter SAG12 cloned from Arabidopsis thaliana fused to a Agrobacterium tumefaciens gene encoding isopentenyl transferase which catalyzes cytokinin synthesis. Following inoculation, the callus was cocultivated for 6 days on BA medium. Selection for transgenics was done on BA medium plus 100 mg Kanamycin and 400 mg Ticarcillin (antibiotics) per liter. Proliferating shoots were rooted on MS medium containing antibiotics. Rooted cuttings were transplanted to soil, acclimated and flowered in the greenhouse. Transgenics were outcrossed to a commercial N. alata hybrid. Seed was germinated in vitro on half-strength MS medium plus antibiotics. Segregation of transgenics to nontransgenics was 1:1. Evaluation of leaf senescence on 5-month-old plants showed 2 to 14 times fewer senesced leaves on the transgenic than the nontransgenic plants.

scholarly journals In vitro Propagation of Solanecio biafrae and Determination of Genetic Stability of Plantlets Using RAPD and ISSR Markers

2016 ◽  
Vol 24 (1) ◽  
pp. 29-36 ◽  
Author(s):  
Jelili Opabode ◽  
Oluyemisi Akinyemiju
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Issr Markers ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Germplasm Exchange ◽  
Indole Butyric Acid ◽  
Half Strength ◽  
Micropropagated Plants

Abstract An efficient and reproducible micropropagation protocol of Solanecio biafrae (Oliv. & Hiern) C. Jeffrey has been developed from nodal stem segments. Shoot development was obtained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP) alone and in combination with zeatin and 1-naphthaleneacetic acid (NAA). Elongated shoots were rooted in the presence of zeatin or 3-indole-butyric acid (IBA) alone or in combinations. The highest number of explants forming shoots (100%) as well as the highest number of shoots per explant (3.4) and the longest shoots (22 mm) were recorded on medium containing 4.0 mg·dm−3 BAP, 2.0 mg·dm−3 NAA, and 1.0 mg·dm−3 zeatin. About 76% of shoots formed roots on half-strength MS medium free of plant growth regulators. The best root formation (approximately 88%) was recorded on the medium containing 1.0-1.5 mg·dm−3 IBA. The micropropagated shoots with well-developed roots were efficiently acclimatized under greenhouse conditions. The random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) amplification products were monomorphic in micropropagated plants and similar to those of mother plant showing their genetic uniformity. This is the first report of micropropagation of S. biafrae, which will facilitate in vitro mass propagation, conservation, and germplasm exchange of this endangered African vegetable.

scholarly journals Germplasm Conservation of Economically Important Medicinal Plant Nyctanthes Arbor-Tristis L. Through Encapsulation Technique and Maintenance Under Slow Growth Condition

2021 ◽  
Author(s):  
Awadhesh Kumar Mishra ◽  
Kavindra Nath Tiwari ◽  
Pallavi Mishra ◽  
Sunil Kumar Mishra ◽  
Shailesh Kumar Tiwari
Keyword(s):  
Growth Condition ◽  
Slow Growth ◽  
Germplasm Conservation ◽  
Start Codon ◽  
Ms Medium ◽  
Mobility Patterns ◽  
Fidelity Assessment ◽  
Half Strength ◽  
Banding Profile

Abstract An efficient encapsulation and germplasm conservation protocol were developed for Nyctanthes arbor-tristis L. In this study the gel matrix containing three percent sodium alginate (SA) and 100 mM calcium chloride (CaCl2. 2H2O) was found best for the formation of encapsulated seeds from node explant of this economically valuable species. The viability of encapsulated seeds and shoot sprouting potential was optimized. Encapsulated seeds stored at 4ºC and 24 ºC maintained its viability up to 90 days and showed sprouting potential 42.89±6.04 and 33.53±7.15 percent respectively. Node explant maintain under slow growth condition up to 180 days on one-eighth (1/8th) strength MS medium supplemented with 0.5 percent sucrose found suitable to maintain high span viability percent (40.28±2.04) with average number of shoots/ node (1.61±0.28) and shoots length (1.12±0.32 cm) respectively. One-eighth (1/8th) strength MS medium supplemented with 0.5 percent sucrose and enriched with 0.5 mg/l abscisic acid (ABA) prolonged the viability up to 40.36±1.01 percent of explant. The best rooting response was achieved on half (½) strength MS medium enriched with 4 mg/l indole-3-acetic acid (IAA). The rooted plant shows 65 percent survivability in open field condition. The true-to-type clonal fidelity assessment of tissue culture recovered acclimated plants with start codon targeted (SCoT) primer profile shows same banding mobility patterns as with source parent mother plant. The maximum banding profile is monomorphic and consistent. Hence on this basis it confirmed the true-to-type clonal stability among them. The protocols display the novel method for conservation of this species under in-vitro condition and facilitate easy exchange of plant germplasm.

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