Efficient transformation of somatic embryos and regeneration of cork oak plantlets with a gene (CsTL1) encoding a chestnut thaumatin-like protein

Autor: Vanesa Cano, F. J. Vieitez, Elena Corredoira, Elena Varas, José Luis Couselo, Mᵃ Teresa Martínez
Přispěvatelé: Ministerio de Asuntos Económicos y Transformación Digital (España)
Rok vydání: 2021
Předmět:
0106 biological sciences
0301 basic medicine
Somatic embryogenesis
Somatic cell
in vitro Phytophthora cinnamomi tolerance test
3103 Agronomía
Agrobacterium tumefaciens
01 natural sciences
2417.09 Fitopatología
Plantlet
lcsh:Chemistry
Quercus
2417.14 Genética Vegetal
Transgenes
lcsh:QH301-705.5
Spectroscopy
Disease Resistance
Plant Proteins
oak decline
thaumatin-like protein
Kanamycin
General Medicine
Plants
Genetically Modified
Computer Science Applications
vitrification procedure
medicine.drug
Phytophthora
pathogenesis related proteins
Biology
Article
Catalysis
Inorganic Chemistry
03 medical and health sciences
Transformation
Genetic
Quercus suber
medicine
Physical and Theoretical Chemistry
Molecular Biology
Gene
Plant Diseases
Organic Chemistry
somatic embryogenesis
Carbenicillin
Molecular biology
Transformation (genetics)
030104 developmental biology
lcsh:Biology (General)
lcsh:QD1-999
010606 plant biology & botany
Transformation efficiency
Zdroj: Investigo. Repositorio Institucional de la Universidade de Vigo
Universidade de Vigo (UVigo)
International Journal of Molecular Sciences, Vol 22, Iss 1757, p 1757 (2021)
Digital.CSIC: Repositorio Institucional del CSIC
Consejo Superior de Investigaciones Científicas (CSIC)
Digital.CSIC. Repositorio Institucional del CSIC
instname
International Journal of Molecular Sciences
Volume 22
Issue 4
Popis: We present a reproducible procedure for transforming somatic embryos of cork oak with the CsTL1 gene that codes for a thaumatin-like protein, in order to confer tolerance to Phytophthora cinnamomi. Different concentrations/combinations of the antibiotics carbenicillin and cefotaxime, as bacteriostatic agents, and kanamycin, as a selective agent, were tested. A lethal dose of 125 mg/L kanamycin was employed to select transgenic somatic embryos, and carbenicillin was used as a bacteriostatic agent at a concentration of 300 mg/L, which does not inhibit somatic embryo proliferation. The transformation efficiency was clearly genotype-dependent and was higher for the TGR3 genotype (17%) than for ALM80 (4.5%) and ALM6 (2%). Insertion of the transgenes in genomic DNA was confirmed by PCR analysis, whereas expression of the CsTL1 gene was evaluated by semi-quantitative real-time PCR (qPCR) analysis. A vitrification treatment successfully cryopreserved the transgenic lines generated. The antifungal activity of the thaumatin-like protein expressed by the gene CsTL1 was evaluated in an in vitro bioassay with the oomycete P. cinnamomi. Of the eight transgenic lines analyzed, seven survived for between one or two times longer than non-transgenic plantlets. Expression of the CsTL1 gene and plantlet survival days were correlated, and survival was generally greater in plantlets that strongly expressed the CsTL1 gene.
This report was partly funded by MINECO (Spain) through projects AGL2013-47400-C4-3-R and AGL2016-76143-C4-4-R and a predoctoral fellowship awarded to V. Cano (BES-2014-070572).
Databáze: OpenAIRE
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