Novel and efficient transformation of wild passion fruit (Passiflora cincinnata Mast.) using sonication-assisted Agrobacterium-mediated transformation

Silva, Maurecilne Lemes da; Pinto, Daniela Lopes Paim; Passos, Andreia Barcelos; Marcelino-Guimarães, Francismar Corrêa; Rossi, Ana Aparecida Bandini; Krause, W; Carvalho, Ilio Fealho de; Batista, Diego Silva; Rocha, Diego Ismael; Otoni, Wagner Campos

doi:10.1007/s11627-020-10134-4

Cited by 11 publications

(9 citation statements)

References 26 publications

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“…Tissue culture mediated regeneration and transformation have been proven to be disadvantageous in several plant species due to low transformation efficiencies, genotype dependence and are time consuming. 55–57 However, the in planta transformation approach does not demand any sterile growth conditions, phyto-hormones, is less time-consuming, needs low-cost inputs and also genotype independent.…”

Section: Resultsmentioning

confidence: 99%

Amenability of an Agrobacterium tumefaciens-mediated shoot apical meristem-targeted in planta transformation strategy in Mango (Mangifera indica L.)

et al. 2022

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Mango ( Mangifera indica L.) is one of the most popular tropical fruits in the world owing to its rich taste, flavor, color, production volume and diverse end usage. Conventional mango breeding practices are unable to withstand the demand for improved varieties as it is time consuming and requires heavy investment. However, problems associated with traditional plant breeding can be curtailed through genetic transformation. Nevertheless, major limitation of transgenic development has been its recalcitrant nature toward tissue culture practices involving latent microbial infection, phenol exudation, etc. This opens wide scope for tissue culture-independent in planta transformation approaches These strategies have proved to be easy to execute and cost effective in producing large number of transformants. One such apical meristem targeted in planta approach was successfully exploited to demonstrate its utility in transforming a tree species. Mango variety Amrapali was transformed with two visual marker gene vectors GFP::hptII in pCAMBIA1302 and GUS::nptII in pCAMBIA2301 individually, to demonstrate its amenability. Preliminary confirmations identified 65.0% of GFP and 57.14% of GUS plants to be transformed. Further, molecular characterization of these primary transformants demonstrated transgene integration at genomic and transcript level in some of the plants. This established protocol holds good for functional gene validation and knock in/out studies and aid in mango improvement programs.

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Section: Resultsmentioning

confidence: 99%

Amenability of an Agrobacterium tumefaciens-mediated shoot apical meristem-targeted in planta transformation strategy in Mango (Mangifera indica L.)

et al. 2022

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“…To date, some regeneration and genetic transformation systems of chrysanthemum have been reported [ 9 , 10 , 11 , 12 , 13 ]. However, the most important factors affecting the success of Agrobacterium -mediated transformation are the characteristics of the varieties, and the factors affecting the transformation efficiency vary among varieties and species [ 14 , 15 , 16 , 17 ]. The explant type and age of chrysanthemum are the key factors affecting in vitro plant regeneration, and the use of young explants, such as newly expanded leaves, is a good material for transgenic chrysanthemum [ 18 , 19 ].…”

Section: Resultsmentioning

confidence: 99%

Development of a Transformation System and Locus Identification Pipeline for T-DNA in Chrysanthemum seticuspe, A Model Species for Hexaploid Cultivated Chrysanthemum

Zhang

et al. 2022

IJMS

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Chrysanthemum is one of the most popular flowers worldwide and has high aesthetic and commercial value. However, the cultivated varieties of chrysanthemum are hexaploid and highly heterozygous, which makes gene editing and gene function research difficult. Gojo-0 is a diploid homozygous line bred from a self-compatible mutant of Chrysanthemum seticuspe and is expected to become a model plant of the genus Chrysanthemum. After assessment of different growth regulator combinations, the optimal concentrations of α-naphthaleneacetic acid (NAA) and 6-benzyladenine (6-BA) in the regeneration system were 1.0 mg·L−1 and 0.2 mg·L−1, respectively. In the genetic transformation system, the selected concentrations of kanamycin, hygromycin and glufosinate-ammonium were 10 mg·L−1, 2.5 mg·L−1 and 0.6 mg·L−1 for bud generation and 12 mg L−1, 1.5 mg·L−1 and 0.5 mg·L−1 for rooting. The transgenic plants were verified by not only PCR detection and GUS staining, but also identification of the T-DNA insertion locus using high-throughput sequencing. Our results lay the foundation for gene functional research on chrysanthemum and will help with the identification of transgenic plants.

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“…Similar results were obtained by Tuhaise, Nakavuma [15] and Asande, Omwoyo [9] between 0.456% and 0.67% with the KPF4 variety of P. edulis f. flavicarpa using leaf disk as explant for in vitro regeneration and transformation technique. Using an in vitro somatic embryogenesis sonication-assisted transformation system, da Silva, Pinto [16] achieved a transformation efficiency of 28.3% in Passiflora cincinnata anthers. In vitro transformation and regeneration systems are applicable for various plant species, but they are time-consuming and labor-intense, require aseptic cultivation conditions, and achieve low regeneration rates [16,[20][21][22][23][24].…”

Section: Discussionmentioning

confidence: 99%

“…Using an in vitro somatic embryogenesis sonication-assisted transformation system, da Silva, Pinto [16] achieved a transformation efficiency of 28.3% in Passiflora cincinnata anthers. In vitro transformation and regeneration systems are applicable for various plant species, but they are time-consuming and labor-intense, require aseptic cultivation conditions, and achieve low regeneration rates [16,[20][21][22][23][24]. Furthermore, it is a high cost technology due to the requirement of sterile culture media, controlled sterile regeneration conditions, and long regeneration periods [44].…”

Section: Discussionmentioning

confidence: 99%

“…Tuhaise, Nakavuma [15] established an in vitro transformation protocol for Uganda's yellow passion fruit and Asande, Omwoyo [9] generated transgenic hybrid passion fruit KPF4 with efficiencies below 1%. Only recently, da Silva, Pinto [16] established a sonication-assisted A.-mediated transformation protocol for anthers of wild passion fruit (P. cincinnata Mast.) with a transformation efficiency of 28%.…”

Section: Introductionmentioning

confidence: 99%

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Establishment of a Novel and Efficient Agrobacterium-Mediated in Planta Transformation System for Passion Fruit (Passiflora edulis)

Rizwan

Ahmed

Zhang

et al. 2021

Plants

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Passion fruit (Passiflora edulis) is an important fruit crop with high economic value. Genetic engineering plays an important role in crop improvement with desired traits and gene functional studies. The lack of a simple, efficient, and stable transformation system for passion fruit has greatly limited gene functional studies. In this study, a simple and efficient Agrobacterium-mediated in planta transformation system for passion fruit was established, using Agrobacterium virulent strain EHA105 harboring the binary vectors pCAMBIA1301 and pCAMBIA1302 with GUS and GFP reporter genes. The system requires less time and labor costs than conventional transformation systems, and no additional phytohormones and sterile conditions are required. Regeneration efficiency of 86% and transformation efficiency of 29% were achieved, when the wounds were wrapped with Parafilm and the plants were kept in darkness for 15 days. Approximately 75% of the regenerated plants had a single shoot and 26% multiple shoots. The transformation was confirmed at the DNA and RNA levels as well as by GUS staining and GFP fluorescent measurements. The developed protocol will contribute to the genetic improvement of passion fruit breeding.

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Novel and efficient transformation of wild passion fruit (Passiflora cincinnata Mast.) using sonication-assisted Agrobacterium-mediated transformation

Cited by 11 publications

References 26 publications

Amenability of an Agrobacterium tumefaciens-mediated shoot apical meristem-targeted in planta transformation strategy in Mango (Mangifera indica L.)

Amenability of an Agrobacterium tumefaciens-mediated shoot apical meristem-targeted in planta transformation strategy in Mango (Mangifera indica L.)

Development of a Transformation System and Locus Identification Pipeline for T-DNA in Chrysanthemum seticuspe, A Model Species for Hexaploid Cultivated Chrysanthemum

Establishment of a Novel and Efficient Agrobacterium-Mediated in Planta Transformation System for Passion Fruit (Passiflora edulis)

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