Isolation and Cloning of 1-Aminocyclopropane-1-Carboxylate Oxidases From Carica Papaya Var. Eksotika I and Eksotika Ii

Sew, Yun Shin; Lam, Peggy; Bakar, Umi Kalsom Abu

doi:10.17660/actahortic.2007.740.17

Cited by 3 publications

(2 citation statements)

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“…This might be due to different insertional chromosal locations of the transgene, which affect its expression. Such variability in transgene expression level due to different integration positions is a common phenomenon in plant transformation [23,24]. Other possible factors that might contribute to this different expression include the copy number and transgene construct fidelity [24,25].…”

Section: Resultsmentioning

confidence: 99%

“…Such variability in transgene expression level due to different integration positions is a common phenomenon in plant transformation [23,24]. Other possible factors that might contribute to this different expression include the copy number and transgene construct fidelity [24,25]. A comparison of the total soluble solid between RNAi transgenic and wild type seed-derived control fruit showed similar profile of 11–14° Brix.…”

Section: Resultsmentioning

confidence: 99%

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RNA Interference of 1-Aminocyclopropane-1-carboxylic Acid Oxidase (ACO1 and ACO2) Genes Expression Prolongs the Shelf Life of Eksotika (Carica papaya L.) Papaya Fruit

et al. 2014

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Abstract:The purpose of this study was to evaluate the effectiveness of using RNA interference in down regulating the expression of 1-aminocyclopropane-1-carboxylic acid oxidase gene in Eksotika papaya. One-month old embryogenic calli were separately transformed with Agrobacterium strain LBA 4404 harbouring the three different RNAi pOpOff2 constructs bearing the 1-aminocyclopropane-1-carboxylic acid oxidase gene. A total of 176 putative transformed lines were produced from 15,000 calli transformed, selected, then regenerated on medium supplemented with kanamycin. Integration and expression of the targeted gene in putatively transformed lines were verified by PCR and real-time RT-PCR. Confined field evaluation of a total of 31 putative transgenic lines planted showed a knockdown expression of the targeted ACO1 and ACO2 genes in 13 lines, which required more than 8 days to achieve the full yellow colour (Index 6). Fruits harvested from lines pRNAiACO2 L2-9 and pRNAiACO1 L2 exhibited about 20 and OPEN ACCESSMolecules 2014, 19 8351 14 days extended post-harvest shelf life to reach Index 6, respectively. The total soluble solids contents of the fruits ranged from 11 to 14° Brix, a range similar to fruits from non-transformed, wild type seed-derived plants.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

RNA Interference of 1-Aminocyclopropane-1-carboxylic Acid Oxidase (ACO1 and ACO2) Genes Expression Prolongs the Shelf Life of Eksotika (Carica papaya L.) Papaya Fruit

et al. 2014

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1-Aminocyclopropane-1-Carboxylate Oxidase 2 Reduction Effects on Physical and Physiological Responses of Eksotika Papaya

Sekeli

Abdullah

Muda

et al. 2013

Journal of Crop Improvement

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The Malaysian Eksotika papaya (Carica papaya L.) has poor keeping quality due to its fast ripening attribute, which leads to post-harvest losses. This study is aimed at extending the shelf life of this perishable Eksotika fruit using antisense technology. A total of 6,000 Eksotika somatic embryogenic calli was transformed with the antisense 1aminocyclopropane-1-carboxylic acid oxidase 2 gene (ACO2) construct, and 46 PCRpositive putative transformants were obtained. Gene expression analysis using real-time PCR on the 46 regenerated putative transgenic lines revealed that 42 showed down-regulation of the ACO2 gene with two-to five-fold differences among the lines. Out of 22 independently selected transformed lines grown under net house conditions, 16 harbored a single copy of the transgene. Physical stature of the transgenic plants was not significantly different from that of the non-transformed seed-derived papaya plants. Physiological evaluations of the transgenic fruits showed a 15-day delay in ripening compared with 4 days of the nontransformed seed-derived papaya fruits. The total soluble solid (TSS) of the transgenic fruits was comparable to that of the non-transformed seed-derived fruits with similar 11-15°Brix, implying the transgenes did not affect the TSS content.

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Molecular and Phenotypic Characterization of Colletotrichum Species Associated with Anthracnose Disease of Papaya in Trinidad

Rampersad

2011

Plant Disease

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Anthracnose disease is a major limiting factor to papaya production worldwide. Accurate identification of the pathogens responsible for this disease is important to developing disease management strategies. One hundred and three (103) isolates of Colletotrichum were collected from infected papaya fruits cvs. Red lady and Tainung No. 2 - F1 hybrid in Trinidad. Of all isolates, 79% were C. gloeosporioides and 21% were C. truncatum. Spore morphology, cultural characteristics, differential reaction to benomyl in addition to ITS1 and β-tubulin gene sequence comparisons unequivocally identified and separated the two species of Colletotrichum. Certain characteristics enabled discrimination between the two species and may be used for provisional identification of these species isolated from papaya. Isolates of C. gloeosporioides grew at a significantly faster rate than those of C. truncatum. C. gloeosporioides isolates were sensitive to benomyl at 1.0 μg/ml, but C. truncatum isolates were resistant. Pathogenicity tests revealed that Colletotrichum species and papaya cultivar had no significant effect on lesion diameter. In cross-infection studies, isolates of C. gloeosporioides from mango and C. truncatum from sweet pepper were able to infect and cause symptoms in wounded mature papaya fruit under controlled conditions. There was no evidence of anthracnose infection in seeds of infected fruits based on the results of growing-on tests. Phylogenetic analyses were based on comparisons of ITS1 and β-tubulin gene sequences. Both neighbor-joining and maximum parsimony methods resolved all Colletotrichum isolates from papaya into species-specific clusters with high bootstrap support. Additionally, a pair of species-specific primers were developed (Ct-TUB-F/R) which allowed reliable detection of C. truncatum isolates from papaya.

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Isolation and Cloning of 1-Aminocyclopropane-1-Carboxylate Oxidases From Carica Papaya Var. Eksotika I and Eksotika Ii

Cited by 3 publications

References 0 publications

RNA Interference of 1-Aminocyclopropane-1-carboxylic Acid Oxidase (ACO1 and ACO2) Genes Expression Prolongs the Shelf Life of Eksotika (Carica papaya L.) Papaya Fruit

RNA Interference of 1-Aminocyclopropane-1-carboxylic Acid Oxidase (ACO1 and ACO2) Genes Expression Prolongs the Shelf Life of Eksotika (Carica papaya L.) Papaya Fruit

1-Aminocyclopropane-1-Carboxylate Oxidase 2 Reduction Effects on Physical and Physiological Responses of Eksotika Papaya

Molecular and Phenotypic Characterization of Colletotrichum Species Associated with Anthracnose Disease of Papaya in Trinidad

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