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Enhanced Fusarium oxysporum f. sp. tuberosi Resistance in Transgenic Potato Expressing a Rice GLP Superoxide Dismutase Gene

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Abstract

Germin like proteins (GLPs) are a large group of related and ubiquitous plant proteins which are considered to be involved in different processes important for plant development and defense. Multiple functional copies of this gene family have been reported in a number of species (wheat, barley, rice, soybean mosses and liverwort), and their role is being evaluated by gene regulation studies and transgenic approaches. To analyze the role of a rice (Oryza sativa) root expressed germin like protein1 OsRGLP1, for its antifungal activity, transgenic potato plants were developed. These transgenic potato plants were molecularly characterized and biologically assessed after inoculation with Fusarium oxysporum f. sp. tuberosi. Functional analysis showed high accumulation of H2O2, increased Superoxide Dismutase (SOD) activity and no oxalate oxidase activity (OxO) in transgenics in comparison to nontransformed control. This increased SOD activity, resistance to heat and sensitivity to H2O2 suggest it is a Fe-like SOD. OsRGLP1 expression in potato plants exhibited enhanced resistance in comparison to nontransformed wild type plants suggesting its role in providing protection against Fusarium oxysporum f. sp. tuberosi through elevated SOD level. Overall, results suggest that OsRGLP1 is a candidate for the engineering of potato plants with increased fungal tolerance however, the greater height and tuber number was observed. This phenotype associated with the resistance needs to be evaluated to determine if this is a positive or negative feature.

Resumen

Las proteínas tipo germins (GLP) son un grupo grande de proteínas relacionadas de plantas y ubicuas, que se les considera que estan involucradas en diferentes procesos importantes para el desarrollo y defensa de la planta. Se han reportado múltiples copias funcionales de esta familia de genes en muchas especies (trigo, cebada, arroz, soya, musgos, plantas hepáticas), y su papel esta siendo evaluado mediante estudios de regulación génica y enfoques transgénicos. Para analizar la función de una proteína 1 tipo germins, OsRGLP1, expresada en la raíz de arroz (Oryza sativa), por su actividad antifúngica, se desarrollaron plantas de papa transgénicas. Éstas fueron caracterizadas molecularmente y evaluadas biológicamente después de la inoculación con Fusarium oxysporum f. sp. tuberosi. El análisis funcional mostró alta acumulación de H2O2, aumento de la actividad de la superóxido-dismutasa (SOD) e inactividad de la oxalato-oxidasa (OxO) en las transgénicas en comparación con las testigos no transformadas. Este aumento en la actividad de la SOD, resistencia al calor y sensibilidad a H2O2 sugiere que es una Fe-SOD. La expresión de OsRGLP1 en plantas de papa exhibió incremento en la resistencia en comparación con las plantas tipo silvestre no transformadas, lo que sugiere que su función es proporcionar protección contra Fusarium oxysporum f. sp. tuberosi mediante un nivel elevado de SOD. En general, los resultados sugieren que OsRGLP1 es una candidata para la ingeniería de plantas de papa con aumento de tolerancia a los hongos, no obstante, se observó la mayor altura y número de tubérculos. Este fenotipo asociado con la resistencia necesita evaluarse para determinar si esta es una característica positiva o negativa.

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Abbreviations

GLPs:

Germins and germin-like proteins

SOD:

Superoxide dismutase

H2O2 :

Hydrogen peroxide

PDA:

Potato dextrose agar

NBT:

Nitroblue Tetrazolium

OXO:

Oxalate Oxidase

DAB:

3, 3′-diaminobenzidine

ANOVA:

Analysis of variance

DMRT:

Duncan’s multiple range test

ROS:

Reactive oxygen species.

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Authors and Affiliations

  1. Department of Biochemistry, PMAS Arid Agriculture University, Rawalpindi, Pakistan

    Nadia Majeed, Bushra Javaid & Syed Muhammad Saqlan Naqvi

  2. Department of Biochemistry and Biotechnology, The Women University, Multan, Pakistan

    Farah Deeba

  3. Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI, USA

    David S. Douches

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  1. Nadia Majeed
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  2. Bushra Javaid
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  3. Farah Deeba
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  4. Syed Muhammad Saqlan Naqvi
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  5. David S. Douches
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Correspondence to Nadia Majeed.

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Majeed, N., Javaid, B., Deeba, F. et al. Enhanced Fusarium oxysporum f. sp. tuberosi Resistance in Transgenic Potato Expressing a Rice GLP Superoxide Dismutase Gene. Am. J. Potato Res. 95, 383–394 (2018). https://doi.org/10.1007/s12230-018-9639-z

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