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Differences in acidity of apples are probably mainly caused by a malic acid transporter gene on LG16

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Abstract

Acidity has profound effects on the taste of apples (Malus × domestica). Malic acid is the predominant organic acid in apples. Differences in malic acid content are caused by differences in accumulation of malic acid in the vacuole. This accumulation may be caused by a gene that is responsible for transport of malic acid from the cytosol into the vacuole. Here, we provide evidence that a malic acid transporter gene at the top of chromosome 16 caused significant differences in malic acid concentration and pH of apples. The pH of apples in a segregating F1 population was mapped and at the pH locus (named henceforth Ma locus for malic acid), two putative malic acid transporter genes were detected. These genes show high homology to AtALMT genes that code for malate channel proteins located in vacuolar membrane in Arabidopsis. The expression of one of the candidate genes (Ma1) cosegregated clearly with malic acid content. The inheritance of at least one dominant allele of this gene sufficed for an increased expression level that likely caused the observed threefold increase of the malic acid concentration and the reduction of the pH from 4 to 3 in mature apples, compared to the presence of the recessive, lowly expressed allele only. Our results show that differences in fruit acidity were probably caused by differences in expression levels of alleles of a malic acid transporter gene.

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Acknowledgments

This project was financially supported by PiDON, Inova Fruit B.V., and the Higher Education Commission (HEC) of Pakistan. We are thankful to Chris Maliepaard from Plant Breeding Department (Wageningen UR) for providing pH data and Aranka van der Burgh for helping in cDNA synthesis. We also thank Harry Jonker for technical assistance during the malic acid analysis.

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Authors and Affiliations

  1. Wageningen University and Research Centre, Plant Breeding, P.O. Box 386, 6700 AJ, Wageningen, The Netherlands

    Sabaz Ali Khan, Jan G. Schaart, Jose Miguel Soriano, Evert Jacobsen & Henk J. Schouten

  2. Wageningen University and Research Centre, Plant Research International, Business Unit Bioscience, P.O. Box 16, 6700 AA, Wageningen, The Netherlands

    Jules Beekwilder & Roland Mumm

  3. Inova Fruit BV, Postbus 222, 4190 CE, Geldermalsen, The Netherlands

    Jose Miguel Soriano

Authors
  1. Sabaz Ali Khan
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  2. Jules Beekwilder
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  7. Henk J. Schouten
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Correspondence to Henk J. Schouten.

Additional information

Communicated by A. Dandekar

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Supplementary Table 1

Genotypes used for measuring malic acid content and expression of three candidate genes. Average sizes of eight fruits per tree of a genotype are given at each stage. The malic acid relative content at these three stages is also given (DOC 53 kb)

Supplementary Table 2

Primer pairs developed along Ma1 used for checking allele specific information between Ma and ma haplotypes. Start and Stop positions are referred to the genomic sequence of Ma1 (DOCX 15.7 KB)

Supplementary Figure 1

Examples of single nucleotide polymorphisms in the Ma gene for the two parental cultivars, and of two progeny genotypes. In the progeny the parental alleles can be recognized (DOCX 201 KB)

Supplementary Table 3

Genome annotation of the scaffold 16 of apple (Velasco et al. 2010) within the genetic window for the Ma locus between markers CH02a03 and CH05e04z (download from http://www.rosaceae.org). The two malic acid transporters found within the genetic window are highlighted in grey. The start and stop positions and the lengths (bp) of the putative transcripts are given (DOCX 82.7)

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Khan, S.A., Beekwilder, J., Schaart, J.G. et al. Differences in acidity of apples are probably mainly caused by a malic acid transporter gene on LG16. Tree Genetics & Genomes 9, 475–487 (2013). https://doi.org/10.1007/s11295-012-0571-y

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