Variegation in Vitis hybrids was investigated to confirm the inheritance as a single, recessive gene as previously proposed and commonly observed in breeding programs. Variegated leaves have ornamental appeal, but the phenotype is sublethal in some environments. Twenty-nine grape families were characterized for variegation including F1, S1, and S2 populations. The majority segregated 3 wild type (WT):1 variegated and were supported by chi-square tests. Four populations had segregation ratios supporting 15:1 or 1:1 models, and a unique flecking phenotype was identified in a Landot 4511 S1 population that suggested the interaction of two recessive loci. A variegated parent was selfed to produce progeny with no WT offspring and was segregated 0:1. Marker trait associations including bulk segregant analysis (BSA), genome-wide association mapping, and quantitative trait loci (QTL) mapping was used on three populations. On chromosome 14, Lvar1 was identified and mapped to 24.5 to 29.5 Mb and associated closely with rhAmpSeq marker 14_27607541. Lvar2 was associated with rhAmpSeq marker 11_18433819 on chromosome 11 at 12.2 to 18.4 Mb. The identification of two loci and the segregation data in some populations suggest that grape breeding germplasm segregates for two recessive loci. The pedigree records suggest that 'Frontenac' inherited one of these loci, and that Landot 4511, an ancestor of many populations tested in this experiment, may carry two loci. A total of 252 candidate genes were identified at these loci, including a key target-adenosine triphosphate (ATP)-dependent zinc metalloprotease FtsH6, involved in photosystem II and similar to the var2 mutant in Arabidopsis. This knowledge can help breeders select for ornamental grapevines or eliminate variegation from their breeding programs.
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