In vitro organogenesis optimization and plantlet regeneration in Citrus sinensis and C. limonia

Almeida, Weliton Antônio Bastos de; Filho, Francisco de Assis Alves Mourão; Mendes, Beatriz Madalena Januzzi; Martinelli, Adriana Pinheiro

doi:10.1590/s0103-90162002000100004

Cited by 32 publications

(48 citation statements)

References 17 publications

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“…The existence of this meristem like cells around the vascular region suggests that the regeneration process in Jatropha might have a multicellular origin. Indeed we can safely assume that new tissues are regenerated from meristematic cells arising from the "de-differentiation" of fundamental parenchyma like parenchymatous bundle sheath (Almeida et al, 2002).…”

Section: Anatomical Changes During Competence Inductionmentioning

confidence: 99%

Analysis of organogenic competence of cotyledons of Jatropha curcas and their in vitro histological behavior

Nogueira¹,

Soares²,

Ibrahim³

et al. 2011

Afr. J. Biotechnol.

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Using cut pieces cotyledons from germinating zygotic embryos of Jatropha curcas, we monitored the series of anatomical events leading to the generation of shoot through organogenesis by histological analysis. 14 days old cotyledons that were pre-cultured in a half strength Murashige and Skoog (MS) media supplemented with 100 mg/L myoinositol and 10 mg/L thiamine HCl, were cultured in organogenic competence induction media (CIM) comprising of MS salts with 1.5 mg/L benzyl adenine (BA) and 0.05 mg/L indole-3-butyric acid (IBA) and incubated for the induction of organogenic competence. Following their sequential transfer to shoot induction media (containing MS + 1.5 mg/L BA, 0.05 mg/L IBA and 0.5 mg/L GA 3 ); shoot elongation media (containing MS + 0.3 mg/L BA) and rooting media (containing half strength MS + IBA at different concentrations), we selected individual explants and subjected them to histological analysis in order to study the morphological changes occurring during organogenesis. Our findings show that to induce organogenesis using cut pieces of cotyledons, these tissues must be harvested at least by the 14th day following germination. Optimal organogenic competence was attained after 21 days incubation period in the dark where most of the explants showed evidence of protruding shoots surrounded by calli with various morphological features. Up to 53.91% of the total explants cultured in this study produced well defined shoots with each explant producing an average number of 1.5 shoots bearing two to eight leaves. We recorded the highest percentage of root formation, which stood at 27.50% when the shoots were cultured in a rooting media containing 0.3 mg/L IBA. Histological analysis of the different events occurring during the process of organogenesis suggest that the protuberances arising from parenchymatous cells and perhaps bundle sheath forming meristematic centres acquiring organogenic competence have a multicellular origin, indicating that the regeneration process takes place through direct organogenesis.

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Section: Anatomical Changes During Competence Inductionmentioning

confidence: 99%

Analysis of organogenic competence of cotyledons of Jatropha curcas and their in vitro histological behavior

Nogueira¹,

Soares²,

Ibrahim³

et al. 2011

Afr. J. Biotechnol.

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“…Most citrus in vitro organogenesis protocols are based on the culture of young tissue, mainly epicotyls and intermodal segments (GARCIA-LUIS et al, 1999;MOURA et al, 2001) cultivated in culture media supplemented with cytokinin (BORDÓN et al, 2000;ALMEIDA et al, 2002). For Rangpur lime these protocols have not shown a high efficiency.…”

Section: Introductionmentioning

confidence: 99%

In vitro organogenesis of rangpur lime

et al. 2012

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-Rangpur lime (Citrus limonia Osbeck) in vitro organogenesis was studied based on explant type and cytokinin culture media supplementation. Four explants types collected from epicotyl or hypocotyl regions of in vitro germinated seedlings were evaluated. The epicotyl-derived explants consisted of epicotyl segments and the hypocotyl-derived explants consisted of the entire hypocotyl segment, the hypocotyl segment attached to a cotyledon fragment, and the hypocotyl segment divided longitudinally. The explants were cultured on EME culture medium supplemented with benzylaminopurine (0, 0.5, 1.0, or 1.5 mg L -1 ). The evaluation was performed after 6 weeks. Best results considering both the explant responsiveness and number of shoots developed per explants were obtained when epicotyl segments-derived explants were evaluated. Considering the explant responsiveness of hypocotyl segments-derived explants no difference was detected between the entire hypocotyl segment and the hypocotyl segment attached to a cotyledon fragment. Moreover, the percentage of responsive explants decreased when hypocotyl segments divided longitudinally were tested. No difference was detected for the number of shoots developed per explant considering the three types of hypocotyl-derived explants. Culture media supplementation with BAP was not essential for Rangpur lime in vitro organogenesis. However, adventitious shoot development was stimulated in concentrations between 0.5 -1.0 mg L -1 . Index terms: adventitious shoot; Citrus limonia; tissue culture. ORGANOGÊNESE IN VITRO DE LIMÃO CRAVORESUMO -A organogênese in vitro de limão Cravo (Citrus limonia Osbeck) foi estudada em função do tipo de explante e da suplementação do meio de cultura com citocinina. Foram avaliados quatro tipos de explantes coletados da região do epicótilo e do hipocótilo de plântulas germinadas in vitro. Os explantes consistiram em segmentos de epicótilo, segmentos de hipocótilo, segmentos de hipocótilo ligado a um fragmento do cotilédone e segmentos de hipocótilo cortado longitudinalmente. Os explantes foram cultivados em meio de cultura EME suplementado com benzilaminopurina (0; 0,5; 1,0 e 1,5 mg L -1 ). As avaliações foram realizadas após seis semanas de incubação, e os melhores resultados considerando-se tanto a porcentagem de explantes responsivos como o número de gemas desenvolvidas por explante foram obtidos com o uso de segmentos de epicótilo como explante. Considerando os explantes derivados da região do hipocótilo, não foi detectada diferença na porcentagem de explantes responsivos com o uso de segmentos de hipocótilo ou de segmentos de hipocótilo ligado a um fragmento do cotilédone. A porcentagem de explantes responsivos diminuiu com o uso de segmentos de hipocótilo cortado longitudinalmente como explante. Não foi detectada nenhuma diferença no número de gemas adventícias desenvolvidas por explante considerando-se os três tipos de explantes derivados da região do hipocótilo. A suplementação do meio de cultura com BAP não é essencial para a organogênese ...

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“…In agreement with our results, Zhi-Ying et al (2011) reported that 3/4 strength MS medium with the addition of 0.35 mg L -1 IBA was effective for root formation from regenerated adventitious buds in Vitis vinifera. IBA is commonly used in tissue culture procedures for various research on different plant species such as Citrus sinensis (Almeida et al, 2002), Centella asiatica (Moghaddam et al, 2011), Camellia sinensis (Sirous et al, 2012) and Prunus empyrean (Sadeghi et al, 2015) to stimulate cell enlargement, elongation and initiate roots from explants or callus. In multiple studies,rooted plant acclimatized readily to the greenhouse with survival rates of 85, 90 and 93% (Debnath, 2005;Fang et al, 2007;Zhi-Ying et al, 2011) and these values agree with our data which showed 96% survival and normal growth and development.…”

Section: Discussionmentioning

confidence: 99%

Producing Transgenic Thompson Seedless Grape (Vitis vinifera) Plants using Agrobacterium tumefaciens

Metwali¹,

Soliman²,

Almaghrabi³

et al. 2016

IJAB

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Under osmotic stress plant avoid it by different modification in their metabolisms such as increasing the synthesis of mRNA of delta 1-pyrroline-5-carboxylate synthase (P5CS), where P5CS is a target gene for increasing proline production and expect to improve the resistance to abiotic stress. To test this hypothesis, grape (Vitis vinifera L.) cv. Thompson Seedless was conducted to develop a protocol for high frequency regeneration and Agrobacterium mediated P5CS gene transfer. Callus induction was achieved by culture leaf explants on Nitsch and Nitsch (NN) basal medium including 2.0 mg L -1 2,4-di-chloro-phenoxy-aceticacid + 0.5 mg L -1 Thiadiazuron solidified with 2.5 g L -1 phytagel. While for in vitro proliferation of plant, the calli were cultured on NN medium supplemented with 3.0 mg L -1 Zeatin riboside + 0.5 mg L -1 Thiadiazuron. Agrobacterium-mediated transformation using the strain LB4404 harbouring the binary vector pBI121 with the P5CS gene under CaMV 35S promoter and the bar gene as a plant selectable marker was used for transforming grape explants. A P5CS specific band (2100 bp) was amplified from DNA extracted only from the transgenic grape plants and 24% for P5CS gene was positive for the bar gene. Over expression of the abiotic stress P5CS gene was enhanced synthesis of proline to over 6 times higher in transgenic plants compared to controls. The successful transformation of genetically diverse grape cv. Thompson Seedless indicated that the transformation system may have general application to an even wider range of grapes cultivars.

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In vitro organogenesis optimization and plantlet regeneration in Citrus sinensis and C. limonia

Cited by 32 publications

References 17 publications

Analysis of organogenic competence of cotyledons of Jatropha curcas and their in vitro histological behavior

Analysis of organogenic competence of cotyledons of Jatropha curcas and their in vitro histological behavior

In vitro organogenesis of rangpur lime

Producing Transgenic Thompson Seedless Grape (Vitis vinifera) Plants using Agrobacterium tumefaciens

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