Regeneration of Carica papaya L. plants from callus transformed by crispr/cas9 and gateway for resistance to the papaya sticky disease


Publication date: 25/10/2023

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Summary: This work aimed to achieve the complete regeneration of Carica papaya L. plants
for resistance to the papaya meleira virus complex from calli transformed with the
empty plasmid for control (TGV), for the silencing of the ­1,3­glucanase gene
(TBG) through the CRISPR/Cas9 and for the overexpression of the chitinase
protein (TQT) through GATEWAY. Callus from TQT transformation showed the
best results regarding callus growth and shoot regeneration. White colored and
friable type callus showed the highest growth rates and highest number of shoots.
Seedlings from TQT transformation showed the best results for stem growth and
number of leaves. The survival rate during the developmental stage was 100%
for the TQT, 96% for TGV and 92% for TBG. TQT showed the highest number
and average length of roots. For TBG transformed plants 80% did not amplify the
941 base pair (Bp) PCR fragment of the ­1,3­glucanase gene, possibly silenced
or deleted. None of the TQT transformed plants amplified the GFP reporter gene
region. During the stage prior to acclimatization, plants from the TQT
transformation had a better development in height and number of leaves. Among
the 20 transformed plant TQT ones were the only that did not present infection
by PMeV and PMeV2. Only 10% of the TGV transformed plants were infected
with PMeV2, while of the TBG transformation plants, 10% were infected with
PMeV and 70% with PMeV2.

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