Highly Efficient and Rapid Plant Regeneration in Citrus sinensis

Singh, Sandeepa; Rajam, Manchikatla Venkat

doi:10.1007/bf03263340

Cited by 10 publications

(4 citation statements)

References 35 publications

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“…Citrus epicotyls show a good in vitro morphogenic response, and therefore have been mostly used for the standardization of regeneration protocols [30]; considering mature shoots, internodes of 1 cm have been used for the regeneration and the transformation of adult tissues in citrus [20,31,32].…”

Section: Source Of Explant Typementioning

confidence: 99%

“…The majority of Citrus species are recalcitrant to Agrobacterium -mediated transformation; in fact, this genus is not a natural host for A. tumefaciens, and so their mutual interaction has not evolved at the optimum level, as for other species [ 30 ]. To increase the rate of success, the disarmed hypervirulent A. tumefaciens strain EHA105, a derivative of the most virulent strain, A281 [ 26 , 62 , 63 ], or AGL-1 [ 56 ] were used, and the insertion of additional copies of vir genes from A. tumefaciens enhanced the transformation efficiency [ 20 , 63 ].…”

Section: Citrus Transformation Protocolsmentioning

confidence: 99%

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Recent Advances of In Vitro Culture for the Application of New Breeding Techniques in Citrus

Poles

Licciardello

Distefano

et al. 2020

Plants

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Citrus is one of the most important fruit crops in the world. This review will discuss the recent findings related to citrus transformation and regeneration protocols of juvenile and adult explants. Despite the many advances that have been made in the last years (including the use of inducible promoters and site-specific recombination systems), transformation efficiency, and regeneration potential still represent a bottleneck in the application of the new breeding techniques in commercial citrus varieties. The influence of genotype, explant type, and other factors affecting the regeneration and transformation of the most used citrus varieties will be described, as well as some examples of how these processes can be applied to improve fruit quality and resistance to various pathogens and pests, including the potential of using genome editing in citrus. The availability of efficient regeneration and transformation protocols, together with the availability of the source of resistance, is made even more important in light of the fast diffusion of emerging diseases, such as Huanglongbing (HLB), which is seriously challenging citriculture worldwide.

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Section: Source Of Explant Typementioning

confidence: 99%

Section: Citrus Transformation Protocolsmentioning

confidence: 99%

Recent Advances of In Vitro Culture for the Application of New Breeding Techniques in Citrus

Poles

Licciardello

Distefano

et al. 2020

Plants

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“…Because the majority of citrus species are not natural hosts for A. tumefaciens and are resistant to Agrobacterium-mediated transformation techniques, their mutual interaction has not evolved to its full potential, as it has in other species. (Singh and Rajam, 2010). The first effort at citrus transformation via direct DNA transfer was reported by Kobayashi and Uchimiya (1989).…”

Section: Introductionmentioning

confidence: 99%

Estimation of Heterosis and Combining Ability for Some White Maize Inbred Lines Using Half-Diallel Crossing

El-Zaher¹

2021

Journal of Plant Production

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Traditional breeding techniques of citrus are complicated and time intensive. In this study, an effective and simple method for the production of sour orange transgenic plants incorporating salt tolerance gene (Pr10a) has been established. Transgenic plants were produced by Agrobacterium tumefaciens -mediated gene transfer using epicotyl explants under the selection pressure of phosphinothericin and NaCl as selective agents. Epicotyl segments were excised from in vitro seedlings and incubated with Agrobacterium tumefaciens EHA105 strain harboring the transgene (Pr10a) under the control of mannopine synthase promoter (p-MAS). In addition, the used dicistronic binary vector contained luciferase (Luc) gene linked to internal ribosome entry site elements in the T-DNA region. MS medium containing 1.5 mg/l 6-benzylaminopurine (BA) and 0.2 mg/l indole butyric acid (IBA) proved optimal for shoot regeneration (97%) from sour orange epicotyl. The number of marker gene (Luc) expressing plants increased as the inoculation time and bacterial density (OD600) increased up to20 min and 0.4, respectively. Under this condition, the transformation efficiency of epicotyl segments was 24.2%. PCR analysis using Pr10a and Luc primers confirmed the existence of the transgenes in transgenic plants where the expected band size of 480 and 837 bp were obtained for both transgenes, respectively. The integration of T-DNA containing Pr10a and Luc genes in the transgenic sour orange was revealed by Southern blot analysis and verified the results of luciferase enzyme activity and PCR analysis of Pr10a and Luc genes. A simple integration pattern for all the transgenes including the single copy integration was predominantly observed.

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“…However, for single gene transfer into elite backgrounds, the development and standardization of a high frequency, efficient plant regeneration and genetic transformation protocol is the utmost pre-requisite. A number of studies had been carried out in the past to develop suitable regeneration and genetic transformation protocol in many horticultural species including apple (Rustaee et al 2007), pomegranate (Parmar et al 2012(Parmar et al , 2013(Parmar et al , 2015, chilli (Sharma et al 2006;Khan et al 2011a), cucumber (Vasudevan et al 2007), lily (Kathryn and Han 2008), sweet orange (Singh and Rajam 2010), broccoli (Kumar and Srivastava 2015), datepalm (Aslam et al 2015), chrysanthemum (Naing et al 2016), etc.…”

Section: Introductionmentioning

confidence: 99%

Genetic engineering strategies for biotic and abiotic stress tolerance and quality enhancement in horticultural crops: a comprehensive review

et al. 2017

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Genetic engineering technique offers myriads of applications in improvement of horticultural crops for biotic and abiotic stress tolerance, and produce quality enhancement. During last two decades, a large number of transgenic horticultural crops has been developed and more are underway. A number of genes including natural and synthetic Cry genes, protease inhibitors, trypsin inhibitors and cystatin genes have been used to incorporate insect and nematode resistance. For providing protection against fungal and bacterial diseases, various genes like chitinase, glucanase, osmotin, defensin and pathogenesis-related genes are being transferred to many horticultural crops world over. RNAi technique has been found quite successful in inducing virus resistance in horticultural crops in addition to coat protein genes. Abiotic stresses such as drought, heat and salinity adversely affect production and productivity of horticultural crops and a number of genes encoding for biosynthesis of stress protecting compounds including mannitol, glycine betaine and heat shock proteins have been employed for abiotic stress tolerance besides various transcription factors like DREB1, MAPK, WRKY, etc. Antisense gene and RNAi technologies have revolutionized the pace of improvement of horticultural crops, particularly ornamentals for color modification, increasing shelf-life and reducing post-harvest losses. Precise genome editing tools, particularly CRISPR/Cas9, have been efficiently applied in tomato, petunia, citrus, grape, potato and apple for gene mutation, repression, activation and epigenome editing. This review provides comprehensive overview to draw the attention of researchers for better understanding of genetic engineering advancements in imparting biotic and abiotic stress tolerance as well as on improving various traits related to quality, texture, plant architecture modification, increasing shelf-life, etc. in different horticultural crops.

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Highly Efficient and Rapid Plant Regeneration in Citrus sinensis

Cited by 10 publications

References 35 publications

Recent Advances of In Vitro Culture for the Application of New Breeding Techniques in Citrus

Recent Advances of In Vitro Culture for the Application of New Breeding Techniques in Citrus

Estimation of Heterosis and Combining Ability for Some White Maize Inbred Lines Using Half-Diallel Crossing

Genetic engineering strategies for biotic and abiotic stress tolerance and quality enhancement in horticultural crops: a comprehensive review

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