Direct and Indirect Shoot Organogenic Pathways in Epicotyl Cuttings of Troyer Citrange Differ in Hormone Requirements and in their Response to Light

Moreira-Dias, J.M.; Molina, Rosa Victoria; Bordón, Y.; Guardiola, J. L.; García-Luis, Amparo

doi:10.1006/anbo.2000.1001

Cited by 64 publications

(63 citation statements)

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“…The concentration of the cytokinin used had a significant effect only on the organogenesis percentage achieved (p < 0.01), as every cytokinin tested proved to be more efficient at its lowest concentration. The inhibitory effect of high cytokinin concentration (especially that of BA) has been reported with other citrus species too (Moreira-Dias et al, 2000;Molina et al, 2007;Cervera et al, 2008). The control treatment resulted in nearly 5% successful organogenesis.…”

Section: -Cppu Enhances Sour Orange Epicotyl Explant Direct Shoot Ormentioning

confidence: 71%

“…When the shoots were excised and cultured in medium supplemented with either IBA or α-NAA, the rooting percentages reached 90% and 75%, respectively, while the rooted explants were successfully acclimatized under mist (85% successful acclimatization). The responsiveness of citrus epicotyl explants to direct organogenesis has been reported to range from very high percentages, such as 100% for grapefruit (Costa et al, 2004), 90-95% for Troyer and Carrizo citranges Moreira-Dias et al, 2000) to values around 70% for sweet orange (Duran-Vila et al, 1992). However some genotypes seem to be very recalcitrant, exhibiting very low organogenesis eff iciency, such as 45 to 60% for C. limonia (Costa et al, 2004) to the extreme low responsiveness of 4% for sour orange .…”

Section: -Cppu Enhances Sour Orange Epicotyl Explant Direct Shoot Ormentioning

confidence: 99%

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Short communication. N-(2-chloro-4-pyridyl)-N-phenylurea(4-CPPU) enhances in vitro direct shoot organogenesis of Citrus aurantium L. epicotyl segments compared to other commonly used cytokinins

Roussos¹,

Dimitriou²,

Voloudakis³

2011

Span. j. agric. res.

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The effect of three concentrations of five different cytokinins, i.e. 6-benzylamino purine, 2-isopentyl adenine, kinetin (Kin), thidiazuron and N-(2-chloro-4-pyridyl)-N-phenylurea (4-CPPU), was evaluated on the in vitro direct shoot organogenesis of epicotyl explants of sour orange (Citrus aurantium L.). The basal medium used was that of Murashige and Tucker and epicotyl explants were incubated in medium supplemented with the prementioned cytokinins for 45 days. The addition of Kin and 4-CPPU in the medium enhanced the direct shoot organogenesis of sour orange epicotyl segments. The concentration of each of these two cytokinins which gave the best results, was combined with indole-3-acetic acid (IAA) or α-naphthalene acetic acid (α-NAA) at concentrations ranging from 0. presented equally high organogenesis percentages (91.7%). The incubation of the produced shoots, in medium supplemented with either indole-3-butyric acid or α-NAA resulted in high rooting percentages (up to 90%) and the rooted explants were successfully acclimatized under mist (85%). Although 4-CPPU has been used in in vitro culture of various species, this is the first report on its use in the direct shoot organogenesis of citrus species and could be of great value in citrus genetic transformation protocols using epicotyl segments, since this cytokinin resulted in the absolute organogenesis percentage. Additional key words: 2-isopentyl adenine; 6-benzylamino purine; explants; kinetin; sour orange; thidiazuron. Resumen Comunicación corta. El N-(2-cloro-4-piridil)-N-fenilurea (4 CPPU) mejora la regeneración in vitro vía organogénesis directa de segmentos de epicótilo de Citrus aurantium L. en comparación con otras citoquininas de uso comúnSe evaluó el efecto de tres concentraciones de cinco diferentes citoquininas [6-bencilaminopurina; 2-isopentil adenina; kinetina (Kin), tidiazurón y N-(2-cloro-4-piridil)-N-fenilurea (4-CPPU)], en la organogénesis directa in vitro de explantes de epicótilo de naranjo amargo (Citrus aurantium L.). Se incubaron durante 45 días explantes de epicótilo en medio Murashige y Tucker suplementado con las cinco citoquininas. La adición de Kin y 4-CPPU en el medio aumentó la organogénesis directa de segmentos de epicótilo de naranjo amargo. Se combinó la concentración que dio mejores resultados de cada uno de estas dos citoquininas con ácido indol-3-acético (IAA) o con ácido α-naftaleno acético (α-NAA) en concentraciones desde 0,01 hasta 0,2 mg L -1 . La inclusión de IAA a 0,2 mg L -1 en el medio con 4-CPPU a 0,05 mg L -1 resultó en un 100% de éxito en la organogénesis directa, mientras que la combinación de Kin a 0,25 mg L -1 con IAA o α-NAA, ambas a 0,01 mg L -1 , resultó en un 91,7% de organogénesis. La incubación de los

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Section: -Cppu Enhances Sour Orange Epicotyl Explant Direct Shoot Ormentioning

confidence: 71%

Section: -Cppu Enhances Sour Orange Epicotyl Explant Direct Shoot Ormentioning

confidence: 99%

Short communication. N-(2-chloro-4-pyridyl)-N-phenylurea(4-CPPU) enhances in vitro direct shoot organogenesis of Citrus aurantium L. epicotyl segments compared to other commonly used cytokinins

Roussos¹,

Dimitriou²,

Voloudakis³

2011

Span. j. agric. res.

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“…It has been shown that regeneration of shoots from the apical end of epicotyl explants inserted longitudinally in semi solid culture medium follows direct regeneration pathway, whereas shoot development at the basal end follows indirect organogenesis after callus formation (GarciaLuis et al, 1999) and regeneration of adventitious buds relies heavily on BAP and IAA in Troyer citrange. Moreira-Dias et al (2000) observed that differentiation of buds took place in direct organogenic manner from the exposed surface of vertically placed explants and did not require hormone supplement, although number of adventitious buds formed was significantly increased when explants were cultured in presence of BAP. On the other hand, addition of BAP and NAA was absolutely required for indirect regeneration.…”

Section: Plant Regeneration In Citrusmentioning

confidence: 96%

Citrus biotechnology: Achievements, limitations and future directions

Singh

Rajam

2009

Physiol Mol Biol Plants

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Citrus is one of the most important commercial and nutritional fruit crops in the world, hence it needs to be improved to cater to the diverse needs of consumers and crop breeders. Genetic manipulation through conventional techniques in this genus is invariably a difficult task for plant breeders as it poses various biological limitations comprising long juvenile period, high heterozygosity, sexual incompatibility, nucellar polyembryony and large plant size that greatly hinder cultivar improvement. Hence, several attempts were made to improve Citrus sps. by using various in vitro techniques. Citrus sps are widely known for their recalcitrance to transformation and subsequent rooting, but constant research has led to the establishment of improved protocols to ensure the production of uniformly transformed plants, albeit with relatively low efficiency, depending upon the genotype. Genetic modification through Agrobacterium-mediated transformation has emerged as an important tool for introducing agronomically important genes into Citrus sps. Somatic hybridization has been applied to overcome self and cross-incompatibility barriers and generated inter-specific and inter-generic hybrids. Encouraging results have been achieved through transgenics for resistance against viruses and bacteria, thereby augmenting the yield and quality of the fruit. Now, when major transformation and regeneration protocols have sufficiently been standardized for important cultivars, ongoing citrus research focuses mainly on incorporating such genes in citrus genotypes that can combat different biotic and abiotic stresses. This review summarizes the advances made so far in Citrus biotechnology, and suggests some future directions of research in this fruit crop.

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“…Some researchers have gotten better results by first transferring the shoots to hormone-free media to eliminate the cytokinin benzyl aminopurine (BA) from the regeneration media before placing on NAA containing media. Different combinations of BA, NAA and another auxin, indole 3-butyric acid (IBA), NAA and IBA only or just IBA and BA in the rooting medium have been tested so as to improve rooting efficiency in citrus cultivars such as mandarin, lemon, 'Troyer' citrange and lime (Al-Bahrany, 2002;Jajoo, 2010;Moreira-Dias et al, 2000;Singh et al, 1994). Again, the concentrations of these phytohormones vary depending on the citrus genotype.…”

Section: Recovery Of Transgenic Shootsmentioning

confidence: 99%