Polymorphisms in the transit peptide of phytoene synthase ZmPSY1 link to the white color of grain endosperm in maize inbred lines

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Abstract

The yellow and orange color of Zea mays L. grain is determined by the presence of carotenoids, the first enzyme of the biosynthesis pathway of which is phytoene synthase PSY. In this study, we analyzed allelic variants of the ZmPSY1 gene in accessions of yellow-grain and white-grain maize inbred lines of domestic selection. In four lines with different grain colors, full-length ZmPSY1 cDNAs were amplified and sequenced, and their variability was characterized. In the cDNA sequence of ZmPSY1 from white-grain lines, nonsynonymous SNPs were found that lead to substitutions of four amino acid residues (L47I, W52S, E53D and A54V) in the N-terminal transit peptide responsible for the plastid localization of the enzyme. A primer system has been developed for PCR identification of the ZmPSY1 allele type in maize accessions. Testing of primers on 44 maize lines showed the presence of the wild-type ZmPSY1 allele and the absence of the mutant allele in the genome of all 22 yellow-grain lines analyzed. The mutant ZmPSY1 allele was detected in the genome of 41% of the 22 tested white-grain lines. The use of the developed primer system may be promising in the selection of corn with altered carotenoid content in the grain endosperm.

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About the authors

D. K. Arkhestova

Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences; Institute of Agriculture - branch of the Kabardino-Balkarian Scientific Center of the Russian Academy of Sciences

Author for correspondence.
Email: shchennikova@yandex.ru
Russian Federation, Moscow 119071; Nalchik 360004

A. D. Khaudov

Institute of Agriculture - branch of the Kabardino-Balkarian Scientific Center of the Russian Academy of Sciences

Email: shchennikova@yandex.ru
Russian Federation, Nalchik 360004

A. V. Shchennikova

Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences

Email: shchennikova@yandex.ru
Russian Federation, Moscow 119071

E. Z. Kochieva

Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences; Lomonosov Moscow State University

Email: shchennikova@yandex.ru
Russian Federation, Moscow 119071; Moscow, 119234

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Supplementary files

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2. Fig. 1. Alignment of the ZmPSY1 cDNA region (a) and the amino acid sequence of ZmPSY1 homologues (b) in two yellow-grained (2452-2, 5580-1) and two white-grained (5254-3, 6097-1) inbred maize lines in comparison with the reference Z. mays var. B73 (LOC100136882; MN128624.1). The arrow indicates the region of the nucleotide sequence selected for the forward primer (for analysis of the allelic variant of the ZmPSY1 gene). In the amino acid sequence, the polymorphic region of the transit peptide containing the substitutions of ao L47I, W52S, E53D and A54V is highlighted in a frame, the SQS and PSY consensuses (solid lines) and the YAKTF and RAYV regions limiting the catalytically active site (double lines) are underlined, and radical substitutions of ao are indicated (arrows).

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3. Fig. 2. Electrophoresis in 2.5% agarose gel of the products (394 bp) of PCR amplification on genomic DNA of 44 inbred maize lines with primers designed for the wild-type (PSY155R/PSY155FG; top of each gel) and mutant (PSY155R/PSY155FC; bottom of each gel) alleles. a – white-grained lines (1–22). The wild-type ZmPSY1 allele was detected in nine samples, and 13 samples contained the mutant allele; b – yellow-grained lines (23–44). The presence of the wild-type ZmPSY1 allelic variant was shown in all 22 samples. The application of samples corresponds to the ordinal numbers of the lines in Table 1; M – DNA length marker GeneRuler Low Range (first four bands from top to bottom: 700, 500, 400, 300 bp).

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