Effect of various plant growth regulators on in vitro plantlet regeneration from different explants of Citrus sinensis (L.) (original) (raw)
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— In this paper a shoot multiplication is described for Citrus latifolia Tan. (persian lime) using nodal segment explants of young one – old – year trees by two different pathways contain with and without callusing phase. The best result for multiple shoot formation and regenerated shoot formation was 3.2 and 2.6 shoots per explants with 4.44 µM BA plus 0.053 µM NAA and 4.44 µM BA plus 0.049 µM IBA respectively. Alike shoot regeneration, shoot elongation was occurred in medium with 4.44 µM BA and 0.049 µM IBA. Micropropagated and regenerated plants are under other experiments. Abbreviation: BA – 6 benzylaminopurine; IBA – Indole acetic acid; NAA – Naphtalene acetic acid; PGRs – Plant Growth Regulators.
AFRICAN JOURNAL OF BIOTECHNOLOGY, 2011
6 benzyl adenine (BA), 1-napthaleneacetic acid (NAA) and 2,4-D. 1 mg/l BA + 0.5 mg/l NAA on MS medium was the most effective in callus induction and proliferation. Maximum number of shoots (11) was recorded on the medium with 2 mg/l NAA + 3 mg/l BA. The best medium for root induction was MS together with 2.5 mg/l indole-3-acetic acid (IAA) + 2 mg/l indole-3-butyric acid (IBA), where maximum (16) plantlets were rooted. The regenerated plantlets were successfully acclimatized in jiffy pots containing sterilized soil mixture of sand, silt and clay in 1:1:1 ratio to study their response to in vivo conditions.
Scientia Horticulturae, 2001
The possible application of some less commonly used in vitro growth regulating compounds is outlined. A number of treatments were applied to determine the best way of inducing in vitro shoot proliferation and rooting on a modified Driver-Kuniyuki [HortScience 19 (1984) 507] basal medium of lemon (Citrus limon (L.) Burm, f. cv. Interdonato) seedlings. 6-Benzyladenine (BA) alone (1, 2 and 4 mg l À1) and in combination with either orange juice (10%, v/v), silver nitrate (3 mg l À1), gibberellic acid (GA 3) (0.1 mg l À1 at the establishment stage and 0.5 mg l À1 at all combinations during the proliferation stage) or abscisic acid (ABA) (0.2 mg l À1 only at the establishment stage) were used to stimulate shoot formation during the establishment and the proliferation stage. The combination of BA with ABA gave a high rate of shoot formation, while GA 3 and silver nitrate enhanced shoot elongation. When these shoots were transferred to the rooting stage, the effect of application of two different auxins (indole-3-butyric acid (IBA) and anapthaleneacetic acid) was examined, as well as different methods of application (auxin added to the basal medium and auxin application by dipping the base of the explant in auxin solution). Dipping the base of the explants in a 50% ethanol solution of IBA at 1000 mg l À1 for 5 s resulted in 80% rooting with subsequent 90% survival of these explants, during acclimatization under mist.
Pesquisa Agropecuária Brasileira, 2010
The objective of this study was to evaluate the effects of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA) combinations, basal media and beta-lactam antibiotics on in vitro organogenesis from mature stem segments of 'Pêra', 'Valência' and 'Bahia' sweet oranges and 'Cravo' rangpur lime. For induction of shoot regeneration, the segments of the four cultivars were placed on Murashige and Skoog (MS) medium containing the following BAP/NAA concentrations: 0.0/0.0; 0.25/0.0; 0.25/0.25; 0.5/0.0; 0.5/0.5; 1.0/0.0; 2.0/0.0; 2.0/0.25; 2.0/0.5; and 2.0/1.0 mg L-1. In order to test the influence of the culture media on shoot-bud induction, (MS), Murashige and Tucker (MT), and woody plant medium (WPM) formulations were evaluated, associated with the best combination of plant growth regulators obtained in the previous experiment. The influence of four beta-lactam antibiotics (timentin, cefotaxime sodium salt, meropenem trihydrate and augmentin) on shoo...
Effect of Phytohormones on In Vitro Morphogenesis of Citrus Cultivars Using Shoot Tip Explant
Annals of Plant Sciences, 2021
An attempt was made to examine the effect of varied concentrations and combinations of phytohormones on morphogenic response of four citrus cultivars viz., Rough lemon (C. Jambhiri), Kinnow (C. reticulata), Feutrell's early (C. reticulate × C. sinensis) and Musambi (C. sinensis) from shoot tip explants. To achieve in vitro shoot regeneration, in vitro maintained healthy shoot tips of four cultivars were excised and positioned on modified MS (Murashige and Skoog, 1969) medium fortifying the following benzylaminopurine (BAP)/Indole acetic acid (IAA) concentrations: 0.5/0.4, 1.0/0.4, 1.5/0.4, 2.0/0.4 and 2.5 mg L-1/0.4 mg L-1. Synergetic effect of BAP with IAA was best found on MS medium containing 1.0/0.4 mg L-1 concentration of BAP to IAA for shoot number/plant (2.25), shoot length (2.52cm) and leaf number/plant (10.58). The shoot multiplication rate was decreased in all four cultivars when BAP concentration was increased from 1.0-2.5 mg L-1. In order to assess the impact of different concentrations of auxins, well proliferated shoots were shifted to the rooting medium ((MS micro & macro elements, 100 mg L-1 myo-inositol, MS vitamins, 2 mg L-1 glycine, 30 g L-1 sucrose and 7.0 g L-1 agar) augmented with varied naphthalene-1-acetic acid (NAA)/ indole-3-butyric acid (IBA) concentrations i.e. 0.5/0.3, 1.0/0.3, 1.5/0.3, 2.0/0.3 and 2.5/0.3 mg L-1. Regenerated shoots started to rooting within 22.25 days, had more number of roots/plant (4.25) and root length (3.35cm) on medium containing 1.0 mg L-1 NAA and 0.3 mg L-1 IBA. Sub or supra optimal concentrations of phytohormones resulted in low plant regeneration in all the four cultivars assessed. It was also found that the morphogenic response was genotype dependent in citrus cultivars.
Effect of various factors on shoot regeneration from citrus epicotyl explants
Journal of Applied Horticulture
The effect of various treatments on shoot organogenesis from seedling epicotyl explants from various scion and rootstock polyembryonic citrus types was determined. Treatments included water source, gelling agent, explant insertion, seed size, light intensity, malachite green, nonionic surfactants, and sodium sulphate. Tap water, with the highest levels of SO 4 2-, Ca 2+ , K + , Mg 2+ , and Na + , resulted in the most shoots compared to the other 5 sources, suggesting a mineral nutrient effect. Carrageenan produced fewer shoots than agar and gellan gum. Explants inserted into the medium produced more shoots than those cultured on the surface, presumably because of better exposure to water and nutrients. Seed size, light intensity, malachite green, and sodium sulphate had no effect on the number of shoots
American Journal of Plant Sciences, 2019
In-vitro callus induction and regeneration method was developed using different plant growth regulators (PGRs), and basal media (Murashige and Skoog (MS), CHU (N6) and Gamborg (B5) media) of Citrus sinensis (L.) Osbeck. Observations of the effect of PGRs were carried out using different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D),1-naphthalene acetic acid (NAA) and combinations of 2,4-D and NAA using different basal media. This study found Citrus sinensis (L.) Osbeck exhibited a high frequency of callus induction on MS medium supplemented with 3 mg/L 2,4-D and callus induction frequency was 86.7% ± 3.4% whereas N6 and B5 showed lower callus induction frequency of 83.3% ± 8.8% and 82.2% ± 1.9% respectively compared to that of MS media with supplementation of the same hormone. Among the induced calli, the morphological analysis showed only 40%-50% was embryogenic calli. Regeneration of plantlets from calli was done using different concentrations and combinations of auxin and cytokinin. The study showed that 3 mg/L 6-benzylaminopurine (BAP) supplemented medium has the maximum potential to promote regeneration of Citrus sinensis (L.) Osbeck from embryogenic calli with the frequency of 89.3% ± 8.8% but no regeneration occurred from the non-embryogenic calli. The regenerated plantlets were rooted on MS medium with supplementation of 5 mg/l NAA. These observations in Citrus sinensis (L.) Osbeck regeneration will be helpful for genetic improvement with desired traits.
International Journal of Agronomy
Orange is one of the most horticulturally important and widely cultivated Citrus species. Citrus cultivar improvement via conventional breeding strategies is normally impeded by factors related to its reproductive biology. The present study was undertaken to investigate the optimization of growth regulators’ composition of media in in vitro propagation of orange cultivar from nodal segment explants. The nodal segment explants were collected from sweet orange cultivars. The MS medium supplemented with sucrose and different concentrations of growth regulators were used for shoot proliferation and root induction. The optimum compositions of growth regulators in MS medium were assessed. The result indicated that the highest shoot response was recorded for Washington naval orange with maximum shoot proliferation rate (99.75%), shoot number per explant (3.10), shoot length (10.70 cm), leaf number per explants (12.50) after three weeks of culture. In all experiments, no growth was observed...
2010
In order to evaluate the formation of adventitious buds and in vitro regeneration of sour orange plants (Citrus aurantium L.) two organogenesis-inducing experiments were conducted. In the first experiment, the induction and in vitro regeneration of adventitious buds were tested on epicotyl and internodal segments under the influence of BAP or KIN associated with NAA. The second experiment evaluated the in vitro regeneration of sour orange plants related to different explant types (epicotyl segments, internodal segments of in vitro germinated plantlets and internodal segments of greenhouse cultivated plants). Data collected on both experiments included the percentage of responsive explants (explants that formed buds), and the number of buds per explant. The addition of BAP showed the best organogenic response. In vitro germinated epicotyl segments and internodal segments are recommended as explants for sour orange in vitro organogenesis. Rooting of regenerated shoots was achieved without the need for auxin in the medium. Index terms: Citrus aurantium, organogenesis, epicotyl segment, internodal segment Regeneração de gemas de laranja-azeda e desenvolvimento in vitro de plantas em função da composição do meio de cultura e tipo de explante Resumo -Com o objetivo de avaliar a formação de gemas adventícias e regeneração in vitro de plantas de laranja-azeda (Citrus aurantium L.), foram realizados dois experimentos de indução à organogênese. No primeiro experimento, a indução e a regeneração in vitro de gemas adventícias foram investigadas a partir de segmentos internodais e segmentos de epicótilo sob o efeito de BAP ou CIN associados com ANA. O segundo experimento avaliou a regeneração in vitro de plantas de laranja-azeda em função do tipo de explante (segmentos de epicótilo, segmentos internodais de plantas germinadas in vitro e segmentos internodais de plantas 1 (Trabalho cultivadas em casa de vegetação). Os dados coletados em ambos os experimentos incluíram a porcentagem de explantes responsivos (explantes que formaram gemas) e número de gemas por explante. A adição de BAP revelou a melhor resposta organogenética. Segmentos de epicótilo e segmentos internodais são explantes recomendados para a indução de organogênese in vitro de laranja-azeda. Enraizamento das brotações foi alcançado sem a adição de auxinas ao meio de cultura.
In vitro regeneration of sour orange (Citrus aurantium L.) via direct organogenesis
2013
Low cell competency for regeneration and transformation is the main cause of so-called recalcitrance to transform a species or a genotype. A research was conducted to determine the optimum conditions for in vitro plant regeneration involving organogenesis in Citrus aurantium, which is an important rootstock worldwide. Seeds with peeled teguments were germinated in vitro, either kept in dark for 6 weeks or maintained in the absolute dark for 4 weeks followed by 10 days in 16-h photoperiod (56 µmol m-2 s-1) at 27 ± 2°C. Epicotyl-originated explants were cultured in MS medium supplemented with 6-benzylaminopurine (BAP) (0, 1, 2 and 3 mg L-1) and Naphthaleneacetic acid (NAA) (0, 0.05, 0.1 and 0.2 mg L-1) to induce organogenesis. Effects of Pre-culture in liquid MS medium (0, 1 and 2 days) on the number of responsive explants (RE) have been also evaluated. In the next step, explants having buds were transferred to MS medium containing Gibberellic Acid (GA 3) (0, 0.5 and 1 mg L-1) and the size and number of shoots, which have been produced by RE are then measured. The highest percentage of responsive explants (90%) obtained by using 2.5 mg L-1 BAP in combination with the 0.05 mg L-1 NAA which had 2 days pre-culture period of epicotyls for allowing to grow in the absolute darkness for 4 weeks, followed by 10 days in 16-h photoperiod (56 µmol m-2 s-1). The highest number of well-developed shoots was 4.2 shoots per explant and obtained with medium containing 0.5 mg L-1 GA 3. These protocols are suitable in association with Citrus aurantium Agrobacterium-mediated genetic transformation.