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Marker-free transgenic rice lines with a defensin gene are potentially active against phytopathogenic fungus Sarocladium oryzae

Pérez-Bernal, M. and Delgado, M. and Cruz, A. and Abreu, D. and Valdivia, O. and Armas, R. (2017) Marker-free transgenic rice lines with a defensin gene are potentially active against phytopathogenic fungus Sarocladium oryzae. Acta Phytopathologica et Entomologica Hungarica, 52 (2). pp. 135-144. ISSN 0238-1249

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Abstract

In this work it was developed marker-free transgenic indica rice plants (cv J-104) by biolistic co-transformation and segregation approach. We attempted to express the NmDef02 antifungal defensin. Primary transformants were regenerated from embryogenic callus on culture medium with 50 mg/L hygromycin. Screening of hpt-marker-free transgenic lines was made by PCR in T<sub>1</sub> progeny lines, germinated on semisolid medium without hygromycin. Relative expression of NmDef02 mRNA was examined by quantitative RT-PCR in marker-free T<sub>1</sub> plants. In vitro antifungal test was performed by disk diffusion assay against Sarocladium oryzae. PCR assay verified that 15.12% of T<sub>1</sub> plants were marker-free (NmDef02+/hpt−). RT-PCR analysis indicated that NmDef02 gene was successfully transcribed and the transgenic lines displayed different expression levels of the NmDef02 cDNA. Protein extracts of marker-free lines with high relative expression of NmDef02 inhibited fungus mycelial growth around disks. In contrast, it was confirmed fungus proliferation on disks impregnated with protein extracts of non-transgenic plants. The results of the present work demonstrated that the expression of the NmDef02 defensin in transgenic rice plants is effective against the phytopathogenic fungus Sarocladium oryzae under in vitro conditions. Thus, NmDef02 defensin could be a useful tool for J-104 rice improvement.

Item Type: Article
Subjects: Q Science / természettudomány > QH Natural history / természetrajz > QH540 Ecology / ökológia
Q Science / természettudomány > QK Botany / növénytan > QK10 Plant physiology / növényélettan
Q Science / természettudomány > QL Zoology / állattan > QL01 Systematic zoology / állatrendszertan
Depositing User: Ágnes Sallai
Date Deposited: 12 Dec 2017 08:20
Last Modified: 31 Dec 2018 00:18
URI: http://real.mtak.hu/id/eprint/70970

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