Abstract
Potato (Solanum tuberosum) is among the best producers of edible biomass in terms of yield per hectare and a variety of different regional cultivars are used as a staple commodity in many countries. However, this crop is attacked by several diseases, with the worst being the late blight disease caused by Phytophthora infestans. Stacking of resistance (R) genes from wild Solanum relatives are interesting prospects for the sustainable control of late blight. Therefore, we optimized methods for the efficient generation and screening of R-gene-containing transformants in tetraploid and diploid hybrid potato genotypes. Using these methods, a high transformation efficiency was achieved for the transformation of tetraploid and diploid potato lines with a triple resistance (3R) gene construct. Transformation efficiencies were improved by optimizing several factors affecting regeneration, including the quality of the starting plant material, and the composition of the plant growth regulators used during selective regeneration. A refreshment protocol was designed to alleviate in vitro related stress in stock plants, which significantly improved the growth vigor and resulted in a 4- to 10-fold increase in transformation efficiency. Furthermore, long-term exposure to exogenous Indole-3-butyric acid that is usually used for the initiation of roots in vitro, was found to cause aberrant morphological phenotypes in potato.
Highlights
Potato is a good staple source of carbohydrate in many countries, even though worldwide and regional differences in cultivar preference exist [1]
All resistance genes were covered by the genomic PCR analysis (Figure S1A)
The improvement in growth vigor contributed to an increase in the number of shoots recovered from leaf explants of King Edward and B101, during regeneration on shoot induction media (Figure 5A)
Summary
Potato is a good staple source of carbohydrate in many countries, even though worldwide and regional differences in cultivar preference exist [1]. Biotechnology-based stacking of R-genes provides a good solution for sustained late blight control and has the benefit of producing much quicker results, compared to conventional breeding methods [7,8,9]. An efficient and versatile pipeline for the transformation, regeneration, selection, and screening of R-gene transformants is required in order to evaluate their potential usefulness in potato. Such a pipeline would allow genes for many other traits to be integrated directly into different elite potato lines, and to be field-tested in less than a year. Techniques for the ‘refreshment’ of in vitro-maintained potato lines were discussed, together with their merits for improving transformation efficiency in potatoes
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