Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat

Khadija Ahansal; Rabha  Abdelwahd; Sripada  Udupa; Hanane  Aadel; Fatima Gaboun; Mohammed Ibriz; Driss Iraqi

doi:10.14393/BJ-v38n0a2022-54513

Autores

Khadija Ahansal Ibn Tofail University Kenitra https://orcid.org/0000-0001-9045-7415
Rabha Abdelwahd National Institute of Agronomic Research
Sripada Udupa International Center for Agriculture Research in the Dry Areas
Hanane Aadel Ibn Tofail University Kenitra
Fatima Gaboun National Institute of Agronomic Research https://orcid.org/0000-0003-2018-8031
Mohammed Ibriz Ibn Tofail University Kenitra
Driss Iraqi National Institute of Agronomic Research

DOI:

https://doi.org/10.14393/BJ-v38n0a2022-54513

Palavras-chave:

Agrobacterium tumefaciens, Drought Tolerance, Genetic Transformation, HVA1 Gene, Triticum turgidum L.

Resumo

A seca é uma das principais restrições na produção de trigo da bacia do Mediterrâneo. A fim de superar este problema, a transformação genética é uma das opções para a melhoria da tolerância à seca. O objetivo deste estudo é investigar o efeito do tipo de explante e concentração de acetosinganona para o desenvolvimento eficiente do gene mediado por Agrobacterium em três variedades de trigo duro (Amria, Chaoui, Marouane). Embriões maduros foram inoculados com Agrobacterium tumefaciens, cepa EHA101, contendo o vetor binário pTF101.1 contendo o gene de tolerância à seca HVA1, e um gene de resistência (bar) marcado com a fosfinotricina (PPT). Os explantes foram inoculados com densidade de células de A. tumefaciens (OD650 a 0,7) em quatro diferentes concentrações de acetoseringona (0, 100, 200 e 400 µM). De acordo com os resultados de regeneração e seleção de plântulas, os calos embriogênicos apresentaram maior eficiência de regeneração e seleção do que as metades e partes de embriões maduros. A integração do transgene foi confirmada por amplificação por PCR utilizando primers específicos para os genes bar, 2x35S e HVA1. A eficiência de transformação variando de 0,33% a 2,33% foi obtida utilizando calos embriogênicos. A concentração de acetoseringona de 100 e 400 µM mostrou uma eficiência de transformação de 0,26%, que é maior do que o valor observado para 200 µM (0,04%). A integração bem como a herança do transgene foi confirmada por amplificação por PCR na geração T0 e T1. Este é o primeiro relatório descrevendo uma transformação genética do trigo duro marroquino via Agrobacterium tumefaciens.

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