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Tytuł pozycji:

A simple and fast Agrobacterium-mediated transformation system for passion fruit KPF4 (Passiflora edulis f. edulis × Passiflora edulis f. flavicarpa)

Tytuł :
A simple and fast Agrobacterium-mediated transformation system for passion fruit KPF4 (Passiflora edulis f. edulis × Passiflora edulis f. flavicarpa)
Autorzy :
Lydia K. Asande
Richard O. Omwoyo
Richard O. Oduor
Evans N. Nyaboga
Pokaż więcej
Temat :
Passiflora edulis Sims
Agrobacterium-mediated transformation
Transient gusA expression
Transformation efficiency
Plant culture
SB1-1110
Biology (General)
QH301-705.5
Źródło :
Plant Methods, Vol 16, Iss 1, Pp 1-12 (2020)
Wydawca :
BMC, 2020.
Rok publikacji :
2020
Kolekcja :
LCC:Plant culture
LCC:Biology (General)
Typ dokumentu :
article
Opis pliku :
electronic resource
Język :
English
ISSN :
1746-4811
Relacje :
http://link.springer.com/article/10.1186/s13007-020-00684-4; https://doaj.org/toc/1746-4811
DOI :
10.1186/s13007-020-00684-4
Dostęp URL :
https://doaj.org/article/d1b090efd3124a9d84915a673bb4294f
Numer akcesji :
edsdoj.1b090efd3124a9d84915a673bb4294f
Czasopismo naukowe
Abstract Background Passion fruit (Passiflora edulis Sims) is an important horticultural crop in the tropics and subtropics, where it has great commercial potential due to high demand for fresh edible fruits and processed juice as well as source of raw materials in cosmetic industries. Genetic engineering shows great potential in passion fruit improvement and can compensate for the limitations of conventional breeding. Despite the success achieved in genetic modification of few passion fruit varieties, transgenic passion fruit production is still difficult for farmer-preferred cultivars. Therefore, it is important to establish a simple and fast Agrobacterium-mediated cell transformation of commercial hybrid passion fruit KPF4 (Passiflora edulis f. edulis × Passiflora edulis f. flavicarpa). Results In the present study, we have developed a simple and fast Agrobacterium-mediated transformation system for hybrid passion fruit KPF4 using leaf disc explants. Factors affecting the rate of transient beta (β)-glucuronidase (gusA) expression and consequently transformation efficiency were optimized as follows: Agrobacterium cell density with an OD600 of 0.5, 30 min infection time, 3 days of co-cultivation duration and the incorporation of 200 µM acetosyringone into Agrobacterium infection suspension medium. Using the optimized conditions, transgenic plants of KPF4 were produced within 2 months with an average transformation efficiency of 0.67%. The β-glucuronidase (GUS) histochemical staining confirmed the expression and integration of an intron-containing gusA gene into transformed leaf discs and transgenic plant lines of KPF4. The presence of gusA gene in the transgenic plants was confirmed by polymerase chain reaction (PCR). The results confirmed that the gusA gene was efficiently integrated into the passion fruit genome. Conclusions The developed transformation protocol is simple and rapid and could be useful for functional genomic studies and transferring agronomically important traits into passion fruit hybrid KPF4. This study developed a method that can be used to transfer traits such as resistance to viral diseases, low fruit quality and short storage life. To the best of our knowledge, this is the first report on genetic transformation system for commercial passion fruit hybrid KPF4.
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