Abstract

Micronutrient deficiency is the cause of multiple diseases in developing countries. Staple crop biofortification is an efficient means to combat such deficiencies in the diets of local consumers. Biofortified lines of sweet potato (Ipomoea batata L. Lam) with enhanced beta-carotene content have been developed in Ghana to alleviate Vitamin A Deficiency. These genotypes are propagated using meristem micropropagation to ensure the generation of virus-free propagules. In vitro culture exposes micropropagated plants to conditions that can lead to the accumulation of somaclonal variation with the potential to generate unwanted aberrant phenotypes. However, the effect of micropropagation induced somaclonal variation on the production of key nutrients by field-grown plants has not been previously studied. Here we assessed the extent of in vitro culture induced somaclonal variation, at a phenotypic, compositional and genetic/epigenetic level, by comparing field-maintained and micropropagated lines of three elite Ghanaian sweet potato genotypes grown in a common garden. Although micropropagated plants presented no observable morphological abnormalities compared to field maintained lines, they presented significantly lower levels of iron, total protein, zinc, and glucose. Methylation Sensitive Amplification Polymorphism analysis showed a high level of in vitro culture induced molecular variation in micropropagated plants. Epigenetic, rather than genetic variation, accounts for most of the observed molecular variability. Taken collectively, our results highlight the importance of ensuring the clonal fidelity of the micropropagated biofortified lines in order to reduce potential losses in the nutritional value prior to their commercial release.

Highlights

  • IntroductionAs a predominantly vegetatively propagated crop, virus accumulation in vegetative propagules (i.e. vine cuttings and tubers) can cause devastating loss in yield and poor root quality in subsequent cultivation [2]

  • To determine to what extent the observed changes in Methylation Sensitive Amplification Polymorphism (MSAP) profiles could be attributed to genetic or epigenetic of the studied samples, we identified Non-Methylated Loci (NML) and Methylation Susceptible Loci (MSL) by comparing the MSAP profiles generated using HpaII/EcoRI and MspI EcoRI as implemented in msap v.3.3.1

  • We show that micropropagation reduces the nutritional value of sweet potato tubers and that micropropagated plants are both genetically and epigenetically dissimilar from field maintained plants

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Summary

Introduction

As a predominantly vegetatively propagated crop, virus accumulation in vegetative propagules (i.e. vine cuttings and tubers) can cause devastating loss in yield and poor root quality in subsequent cultivation [2] Micropropagation techniques, such as meristem or nodal tip culture, coupled with thermotherapy or cryotherapy, are currently the principal plant tissue culture (PTC) methods for producing healthy (pathogentested/disease-free) clones of planting materials [3]. Somaclonal variation refers to changes that can be induced during in vitro tissue culture and have been reported in all in vitro systems [5,6,7,8] Such changes can be genetic and/or epigenetic in nature. DNA demethylation in plants is not as well characterized but it is understood to occur through either active demethylation mediated by DEMETER (DME) [13, 14], or via passive demethylation during DNA replication (where the newly created strand lacks methylated cytosines) [15]

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