scholarly journals Genetic transformation of European chestnut somatic embryos with a native thaumatin-like protein (CsTL1) gene isolated from Castanea sativa seeds

2012 ◽  
Vol 32 (11) ◽  
pp. 1389-1402 ◽  
Author(s):  
E. Corredoira ◽  
S. Valladares ◽  
I. Allona ◽  
C. Aragoncillo ◽  
A. M. Vieitez ◽  
...  
2021 ◽  
Vol 12 ◽  
Author(s):  
Vera Pavese ◽  
Andrea Moglia ◽  
Elena Corredoira ◽  
Mª Teresa Martínez ◽  
Daniela Torello Marinoni ◽  
...  

CRISPR/Cas9 has emerged as the most important tool for genome engineering due to its simplicity, design flexibility, and high efficiency. This technology makes it possible to induce point mutations in one or some target sequences simultaneously, as well as to introduce new genetic variants by homology-directed recombination. However, this approach remains largely unexplored in forest species. In this study, we reported the first example of CRISPR/Cas9-mediated gene editing in Castanea genus. As a proof of concept, we targeted the gene encoding phytoene desaturase (pds), whose mutation disrupts chlorophyll biosynthesis allowing for the visual assessment of knockout efficiency. Globular and early torpedo-stage somatic embryos of Castanea sativa (European chestnut) were cocultured for 5 days with a CRISPR/Cas9 construct targeting two conserved gene regions of pds and subsequently cultured on a selection medium with kanamycin. After 8 weeks of subculture on selection medium, four kanamycin-resistant embryogenetic lines were isolated. Genotyping of these lines through target Sanger sequencing of amplicons revealed successful gene editing. Cotyledonary somatic embryos were maturated on maltose 3% and cold-stored at 4°C for 2 months. Subsequently, embryos were subjected to the germination process to produce albino plants. This study opens the way to the use of the CRISPR/Cas9 system in European chestnut for biotechnological applications


New Forests ◽  
2016 ◽  
Vol 47 (5) ◽  
pp. 669-684 ◽  
Author(s):  
E. Corredoira ◽  
M. C. San José ◽  
A. M. Vieitez ◽  
I. Allona ◽  
C. Aragoncillo ◽  
...  

Author(s):  
Enrico Marcolin ◽  
Mario Pividori ◽  
Fernanda Colombari ◽  
Maria Chiara Manetti ◽  
Francesco Pelleri ◽  
...  

2006 ◽  
Vol 226 (1-3) ◽  
pp. 199-207 ◽  
Author(s):  
Cécile Robin ◽  
Olivier Morel ◽  
Anna-Maria Vettraino ◽  
Charikleia Perlerou ◽  
Stephanos Diamandis ◽  
...  

2009 ◽  
Vol 45 (3) ◽  
pp. 275-282 ◽  
Author(s):  
Mónica Sebastiana ◽  
Andreia Figueiredo ◽  
Bartolomeu Acioli ◽  
Lisete Sousa ◽  
Fernando Pessoa ◽  
...  

2017 ◽  
pp. 79-82
Author(s):  
Gabriella Kovács ◽  
László Radócz

The most destructive pathogen for the European chestnut is the blight fungus Cryphonectria parasitica (Murr.) Barr. The spread of the fungus was very fast in Europe within a few decades in the second half of the past century. During the tree-health checking in the chestnut andwalnut plantation in Romania, Hargita county, next to Homoródkarácsonyfalva village, we especially concentraded on the signs of blight disease occurrence. The grove is laying on a western slope, under a pine forest. This favourable geographical space protects it not only from pathogen attacts, but it has a special, mild microclimate for nut and chestnut trees. The European chestnut could be a valuable member of local forests, opening a new perspective under conditions of climate changes.


2017 ◽  
Vol 2 (6) ◽  
pp. 599 ◽  
Author(s):  
Tifa R. Kusumastuti ◽  
Rizkita R. Esyantia ◽  
Fenny M. Dwivany

Banana is one of the major fruit crops, though its conventional breeding has limitations, such as sterility and high polyploidy  levels.  Biotechnological  approach  using genetic  transformation  crop for improvement  offers  an alternative  solution.  In  this  study  a  protocol  was developed  for  establishing genetic  transformation  from embryogenic callus and somatic embryos of the banana cv Ambon Lumut . Embryogenic callus was obtained in ID4 medium (MS-based medium) supplemented with 1 mg L-1 IAA, 4 mg L-1 2,4D, and 0.03 g L-1 active charcoal. Embryogenic callus was transferred into liquid mediu m to establish somatic embryos. Embryogenic callus and somatic embryos were used for Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain A GL1, containing pART-TEST7 p lasmid with gfp gene as a reporter and CaM V35S as a promoter, was used for transformations. The embryogenic callus and somatic embryos were transformed using heat-shock method followed by centrifugation  (2000 rpm) and co-cult ivation in liquid medium containing acetosyringone (100 M) for 3 days. Results of the GFP analysis showed transient expression from gfp gene reporter in transformed embryogenic callus and somatic embryos. Transformation efficiency in somatic embryos (85,9%) was higher than  that in embryogenic callus (32.09%). PCR analysis using CaMV primer showed bands that compatible with CaMV35S promoter at 507 bp. This is a report showing establisment of embryogenic callus and somatic embryo culture transformation by using A. tumefaciens-mediated transformation protocol of the local banana cv Ambon Lumut. This study proved  the huge potential for genetic transformation of banana cv Ambon Lumut for crop improvement, such as pest or disease  resistance and abiotic factor stress tolerance. Keywords: banana; embryogenic callus; somatic embryos.


2014 ◽  
pp. 87-90
Author(s):  
I. Prgomet ◽  
D. Torello Marinoni ◽  
D. Donno ◽  
Z. Prgomet ◽  
I. Mujić ◽  
...  

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