In vitro plant regeneration from seed explants of wild groundnut species (Genus Arachis, Section Extranervosae) (original) (raw)
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This study was designed to evaluate the effects of plant growth regulators – auxin (NAA) and cytokinin (BA) – on the regeneration potential of two accessions of bambara groundnut ‘TVSu1834’ (Nigeria) and ‘TVSu255’ (Niger) through in vitro culture. Nodal vine segments from 28-days-old plantlets derived from the embryonic axis were used as explants for nodal culture. The explants were cultured on half-strength MS basal salts supplemented with different concentrations and combinations of cytokinin and auxins for primary shoot proliferation. BA at 0.45 mg/l in combination with three different concentrations of NAA, namely 0 mg/l (M4), 0.15 mg/l (M2) and 0.25 mg/l (M3) were used for nodal culture. The control medium M1, had no plant growth regulators added. The shoots, leaves, and whole plant regeneration were evaluated at 4 weeks by means of percentage of explants producing shoot, callus and rooting per explant. The best shoot proliferation of nodal cuttings of ‘TVSu1834’ was observed in M3 medium containing half-strength MS, 0.45 mg/l BA, 0.25 mg/l NAA and 100 mg/l myo-inositol which produced multiple shoots with the longest shoots and most leaves compared to ‘TVSu255’. In M1 medium, the control, taller plantlets formed than at other concentrations and was the only medium that induced rooting. There was a genotype-dependent effect (P < 0.05) on shoot proliferation, shoot length and number of leaves in the two accessions.
Plant regeneration of A. lakoocha from encapsulated nodal explants
Archives of Applied Science Research, 2015
One of the alternative methods adopted in recent ye ars is to use biotechnological approaches for impro ving the tree species. A proficient protocol for encapsulation of nodal segments of Artocarpus lakoocha Roxb has bee n developed for plant regeneration through non-embryogenic synt hetic seeds. Concentrations of sodium alginate and calcium chloride greatly affected morphology and texture en capsulating gel. 3 % sodium alginate with 100 mM Ca Cl 2 has been found to be best possible concentration for th e production of identical synthetic seeds. 25 days old in vivo plantlet of A. lakoocha was used for obtaining noda l segments, used in all experiments. Five experime nts were performed using MS medium (agar solidified and liqu id) supplemented with 1, 3 and 5 mg/l BAP. Concentr ation of 1 mg/l BAP was observed to be more effective for ac hieving highest regeneration frequency in compariso n to other concentration of BAP (3 and 5 mg/l). In third exper iment only different strength of ...
Rapid Plant Regeneration from Nodal Explants of
Excised nodal explants of Spilanthes acmella (L.) Murr., 'Toothache Plant' proliferate rapidly in vitro on MS medium containing 0.5-2.0 mg/l of BAP. Rapid and prolific shoot proliferation occurred. Regenerated shoots vary considerably in size (3-10 cm long) and relative stage of development, with some (50%) producing adventitious roots without transferal to a separate rooting medium. With maximum possibility of adventitious roots induction was induced from middle order nodes (3 rd to 5 th node from apex) obtained from 3 months old in vivo plant on full-strength MS medium supplemented with 1.0 mg/l BAP under the photoperiod of 18-h. The possibility of adventitious roots induction directly from regenerated shoot was greatly influenced by the concentration of BAP, photoperiod, age of donor plant and nodal position on stem.
Journal of Plant Biochemistry & Physiology, 2019
A reproducible regeneration protocol for ICGV 12991, CG 7 and Red Valencia groundnut genotypes using Cotyledonary Node explants has been optimized. The effect of different BAP concentrations combined with either 2,4-D or TDZ was tested to determine optimum conditions for high shoot induction. Different BAP concentrations were tested to determine an optimum concentration for shoot elongation. Different NAA concentrations were similarly evaluated to determine the best concentration for rooting. Media containing combination of 5 mg/L BAP and 1 mg/L TDZ was the best concentration for shoot induction while media containing BAP at 5 mg/L was the best for elongation of shoots. NAA concentration of 1 mg/L gave the highest number of plants with roots. This works provides a very good protocol which will be beneficial during groundnut tissue culture as well as genetic transformation of groundnuts.
Applied Biochemistry and Biotechnology, 1999
An efficient protocol for regeneration of groundnut plantlets from immature and mature leaflet and petiole explants excised from axenic seedlings has been developed. The highest frequency of callus induction obtained from leaflet explants was on Murashige and Skoog (MS) medium containing 2.0 mg/L of α-naphthalene acetic acid (NAA) and 0.5 mg/L of kinetin combination. MS medium containing different auxins in combination with 6-benzylaminopurine (BAP) induced shoot buds. A BAP (2.0 mg/L) and NAA (0.5 mg/L) combination resulted in the highest frequency of shoot-bud regeneration. Subsequent shoot multiplication was obtained on MS medium supplemented with either BAP or kinetin (5.0 mg/L) in combination with NAA (1.0 mg/L). Immature leaflet explants were found to be more responsive to shoot induction than mature leaflet explants. Direct shoot-bud regeneration was also observed from petiole explants on the same regeneration medium used for leaflet callus. Regenerated shoots were rooted on MS medium containing indole-3-butyric acid (2.0 mg/L) and kinetin (0.5 mg/L). Plantlets obtained were successfully established in the field, where they grew to maturity and set viable seeds.
Bambusa nutans subsp. cupulata, a perennial tall grass bamboo has hig h potential value for economic and social aspects. Due to unpredictable flowering and seeds, it is obligated to propagate vegetative ways but it is inefficient and costly as well as the destruction of the huge stock of bamboo plants. The alternative method , micropropagation enhances a large-scale production of it. For this, the nodal segment was inoculated in MS solid and liquid media with a supplement of different concentrations of growth hormones. This article is reported the optimized in vitro propagation protocol to regenerate the large scale of the plants. The maximum bud sprout was observed in liquid (100%) with 2.5±0.19 cm length of shoots and solid (98.67%) with 3±0.19 cm length of shoots in MS media supplemented with of BAP (1mg/L) and KN (0.5 mg/L) in combined. The multiplication of shoots rapidly occurred on the liquid media fortified with BAP (1mg/L) and KN (0.5 mg/L). In vitro rooting was well achieved (84.67%) in the MS liquid media than solid media with fortified combined auxins 3 mg/L NAA and 3 mg/L IBA. We obtained 92.80% in primary (15 days) and secondary (3 months) hardening which yields a 100% survival rate in field transfer. We optimized the simple protocol for in vitro propagation of B. nutans subsp. cupulata. An efficient, but simple protocol, which can be applied for mass multiplication of B. nutans subsp. cupulta plant through nodal segment using the in vitro propagation technique.
In Vitro Cellular & Developmental Biology - Plant, 2008
A rapid and efficient method for the large-scale propagation of an endemic medicinal plant, Andrographis neesiana Wight, through in vitro culture of nodal explants obtained from 30-d-old aseptic seedling has been developed. High frequency direct shoot proliferation was induced in nodal explants cultured on Murashige and Skoog's medium supplemented with thidiazuron. Amongst the various cytokinins tested (BAP, kinetin, thidiazuron and 2-isopentyl adenine), thidiazuron proved to be the most effective. The shoot forming capacity of the nodal explants was influenced by the thidiazuron concentration (1-12.5 µM) and the optimal response was observed at 10 µM thidiazuron, which induced an average of 34 shoots in 94% of the cultures within 4 wk. Significant differences were recorded in terms of average number of shoots per explant (8.6-34.1) among the different concentrations of thidiazuron investigated. Concentrations of all cytokinins tested reach a level that can be considered above the optimum level, as marked by a reduced frequency of shoot proliferation. The multiple shoots obtained on various concentrations of thidiazuron failed to elongate even after transfer to hormone-free MS medium. Elongation of the induced shoots was achieved on MS basal medium supplemented with 1.0 µM GA within 2 wk. A proliferating shoot culture was established by repeatedly 3 subculturing the original nodal explants on shoot multiplication medium after each harvest of the newly formed shoots. The explants retained their morphogenic potential even after three harvests. Therefore, in 90 d, about 60-70 shoots were obtained from a single nodal explant and the nodal explants from primary shoots further regenerated equivalent number of shoots, depicting their high frequency regeneration potential in A. neesiana. Rooting was best induced in 94% of shoots cultured on MS medium supplemented with 2.5 µM indole-acetic acid (IAA), within a wk. The plantlets were successfully transferred to soil after hardening with a 92% survival rate. The system is rapid: the initiation of shoot buds to the transplanting of regenerants to soil is completed in 8-9 wk.
This study was aimed to establish an in vitro propagation method of Artocarpus chaplasha Roxb. trees. Shoot proliferation was induced from shoot apices and lateral bud explants of Artocarpus chaplasha Roxb. cultured on MS medium containing different concentrations of BAP, NAA and Kinetin either alone or in various combinations. Coconut water (CW) and casein hydrolysate (CH) were used as additives to the medium to determine their effects on shoot growth and development. Polyvinyl pyrrolidone (PVP) were used in medium as an antioxidant and antibrowning agent. Results showed that 2.5 mg/L BAP + 0.5 mg/L NAA showed highest 75% shoot induction with multiple shoots of 6.4 shoots per explants and 5.0 cm longest shoot, while 3.0 mg/L KIN + 0.5 mg/L NAA showed (64%) shoot induction with 5.8 shoots per explants and 4.80 cm longest shoot. Addition of 20% (v/v) CW and 300 mg/l CH to the medium enhanced the number of shoots up to 10.5 per explants. The length of shoots also was found to be enhanced with these additives. 0.7% PVP was used in the medium as antibrowning agent. In vitro grown shoots were cultured to root on half strength MS medium containing either of 3 auxins, namely IAA, IBA or NAA at concentrations of 1.5, 1.0 or 0.5 mg/L. Concerning rooting potentiality, addition of 1.0 mg/L IBA + 0.5 mg/L NAA to half strength MS medium gave 80% rooting percentage. Successful transplanting was obtained when rooted plantlets were transferred to a mixture of soil, sand and compost (1:1:1). Plantlets were transferred to a tray containing soil, sand and compost and covered by polythene sheets. After two weeks they were transplanted individually in the small poly bags and kept in open place with indirect sunlight where 75% of the plants survived.
Journal of Animal & Plant Sciences, 2020
This study was carried out to optimize in vitro regeneration system of groundnut through tissue culture technology that can provide good environment in terms of food security. The two varieties of groundnut (Golden and BARI 2001) were used and an effective in vitro regeneration system was established for selecting the best responsive variety to tissue culture using embryo as an explant sources. MS media having 5.5 mg/l BAP and 1.5 mg/l NAA was found to be best by producing the highest callus induction frequencies (86 & 78%) in Golden and BARI 2001, respectively. Similarly, the maximum multiple shoots/ plant (9 & 6) with optimum length of shoots (4.8 & 4.1 cm) was obtained with the application of 4 mg/l BAP along with 1 mg/l NAA and 1.1 mg/l TDZ in Golden and BARI 2001, respectively. In case of roots initiation, the highest root frequency (90 & 76%) having optimum roots/shoots (8.3 & 6.2) and substantial root length (8.1 & 7.0 cm) was obtained in Golden followed by BARI 2001 variety on MS media having 1 mg/l phytohormone IBA. Results showed that callus and multiple shoot induction and in vitro rooting, varied with different hormonal combinations in the media and the genotype that might be due to genetic variations between the varieties. This study delivers a base line for efficient in vitro propagation and genetic transformation in elite groundnut genotypes for desirable characteristics.
Murraya koenigii (L.)Spreng, commonly known locally as “kurry patta” or “mitha neem” in India, is a valuable medicinal plant known for its biochemical and aromatic properties. Adventitious regeneration, which is a pre-requisite in most genetic transformation studies using Agrobacterium and ballistics, needs to be developed as a protocol for micropropagation of M. koenigii.This paper presents a procedure for the rapid, high frequency regeneration of M. koenigii plantlets from internode explants via adventitious shoot formation. The concentration of plant growth regulators (PGRs) in liquid MS medium exhibited a discrete role in the efficacy of adventitious shoot induction. N6-benzyle adenine (BA), kinetin, adenine sulphate and indole-3-acetic acid (IAA) in combination were the most effective PGRs for adventitious shoot induction. Murashige and Skoog (MS) liquid medium with 9.29 µM kinetin, 13.317 µM BA, 2.854 µM IAA and 70 mg/l adenine sulphate yielded the maximum number (18) of shoot buds from internode explants. The number of shoots was further increased (27.30) after sub-culturing them into semi-solid (containing 8 g/l agar-agar) MS medium fortified with similar concentrations and combinations of PGRs. Most in vitro shoots (2.5-3.0 cm long), rooted (90%) on semi-solid MS medium containing 19.68 µM indole-3-butyric acid within 28-30 days. The rooted plantlets were transplanted into pots containing a mixture of soilrite (mixture of peat moss + vermiculite + perlite in a 1: 1: 1 ratio that was mixed with natural soil in the ratio of 1: 1) at 70-80% relative humidity and 28 ± 2°C for hardening. 85% of in vitro-raised plantlets survived under field conditions.