Callogenesis and plant regeneration of sweet orange cv. Washington Navel from floral organ cultures (original) (raw)
Related papers
In vitro Propagation of Citrus Species through Callus Induction and Regeneration: A Review
International Journal of Current Microbiology and Applied Sciences
Citrus, one of the most important group of fruit crops around the world, are propagated at large scale with many difficulties. Propagation through seeds is challenging because of Phytophthora foot rot together with recalcitrance of citrus seeds. Vegetative propagation of Citrus species is mainly performed now-a-days by budding on seedling rootstocks. As heavy losses are experienced among the susceptible seedlings due to Phytophthora and Citrus tristeza virus (CTV), the interest in resistant rootstocks has greatly increased. The potential of conventional methods of citrus plant breeding of rootstocks are limited by physiological factors such as heterozygosity, inbreeding depression, nucellar polyembryony and juvenility. Under such conditions advanced tissue culture techniques provide best possible alternative for producing large number of resistant progenies from elite citrus genotypes. Plant tissue culture provides reliable and economical method of maintaining pathogen free plants that allows rapid multiplication and international exchange of germplasm. Generally, when in vitro propagation protocols are developed for any specific plant species, specialized conditions for individual genotypes, elite species and even various developmental stages of the explants plants are selected via error-andtrial experiments. Because large diversity is observed in Citrus plant family, it takes many months to develop protocols for most suitable culture medium, best concentrations and combinations of plant growth regulators and other supplements for better development of explant cultures. Therefore, in this review, we tried to put together results from difficultto-find literatures and listed all the identified findings, in which callus induction or somatic organogenesis was used to develop citrus plants. Successful protocols of surface sterilization method, culture establishment, shoot regeneration, in vitro rooting and acclimatization are presented systematically.
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.
Regeneration of Citrus via Shoot Apecies
HortScience
Viral damage is a major problem in citrus. As most citrus are asexually propagated, it is necessary to have an alternative way of regenerating virus-free plants from infected plants. Shoot apicies are the most suitable explant material for this purpose because that part of the plant is virus-free. Fifty sour orange shoot tips and 22 Swingle shoot tips, 1 mm - 1.5 mm long, were excised from in vitro germinated seedlings and cultured on semisolid Murashige and Skoog medium, without growth regulators, containing 0.2 % Gelrite. After 8-10 weeks, shoots and leaves developed in 68'% of the sour orange explants, and in 77% of the Swingle explants. Some explants produced roots, after 11-12 weeks, and could be removed from culture and established in soil medium.
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.
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.
Agricultural Sciences, 2019
Citrus reticulata (Mandarin Orange), commonly known as "Sweet Orange", is one of the most difficult plants to improve through traditional breeding approaches as it poses various biological limitations that greatly hinder the cultivar improvement. In the present study, using the fresh seed of native orange as explant, an efficient, reproducible, regeneration method was developed through in vitro organogenesis. Mature, healthy and dehusked seeds were treated with Murashige and Skoog, (MS) media containing 3% sucrose, 0.7% agar supplemented with different concentrations and combinations of phytohormones. The highest calli initiation (93.3% ± 0.5%) responses were observed on MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) at 3.0 mg/L followed by 2,4-D at 3.5 mg/L (86.7% ± 1.75%) in this experiment. Maximum shoot regeneration (86.7% ± 3.35%) responses were reported using MS medium supplemented with the combination of 6-benzylaminopurine (BAP) at 3.0 mg/L and 1-naphthaleneacetic acid (NAA) at 2.0 mg/L. MS medium supplemented with NAA at 1.0 mg/L showed the best rooting (80% ± 2.89%) response in comparison to (70% ± 5.20%) indole-3-butyric acid (IBA) at 1.0 mg/L. The regenerated plantlets were acclimatized in pots containing sterile garden soil mixture to examine their response in natural conditions.
ESTABLISHMENT OF TISSUE CULTURE TECHNIQUES IN CITRUS SPECIES
THESIS , 2020
An efficient system for in vitro plant regeneration by multiple shoot formation from shoot tip and epicotyl explants of Citrus limon, Citrus clementine, Citrus reticulata, Citrus aurantifolia and Poncirus trifoliata was developed. The optimum concentration of sodium hypochlorite solution for surface sterilization of nodal segments of Citrus limon was 4.5% for 15 minutes. The highest number of axillary shoots per explant were obtained when the shoot tip explants cultured on MS (1962) medium supplemented with 4.4µM BAP and 2.7µM NAA for Citrus limon and MS (1962) medium supplemented with 4.4µM BAP, 2.3µM Kinetin and 2.7µM NAA for Citrus reticulata, Citrus aurantifolia and Poncirus trifoliata. The best treatment, in terms of "response frequency" and "mean number of de novo shoots per epicotyl explant" was on MS (1962) medium with B5 vitamins supplemented with 33.3µM BAP and 5.4µM NAA for Citrus reticulata, Citrus aurantifolia, Citrus limon, Citrus clementine and Poncirus trifoliata.
International Journal of Sciences: Basic and Applied Research, 2016
A method for in-vitro propagation of wild type Indian orange ( Citrus chrysocarpa L.) was developed by shoot organogenesis from seed. Mature seed embryos were used as explants and treated with different hormones and plant growth regulators on MS medium for callus, shoots and roots induction. For callus induction seed embryos were treated with different concentration of 2, 4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) and a-naphthalene acetic acid (NAA) and maximum 90.90% callus was observed on MS medium supplemented with 2, 4 -D at 16.0 µM concentration. Approximately 85% of the callus was granular, while 15% was smooth and compact. Maximum 83.33% shoot regeneration were observed on MS medium supplemented with BA at 13µM concentration. Regenerated shoots were rooted on MS medium supplemented with different hormones and maximum 80% roots were observed in MS medium supplemented with 10 µM Indole-3-butyric acid (IBA). The regenerated plantlets were successfully acclim...