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Development of a visible marker trait based on leaf sheath-specific anthocyanin pigmentation applicable to various genotypes in rice.

ABSTRACT: To overcome a limitation to the breeding of autogamous crops, recurrent selection using transgenic male sterility (RSUTMS) has been proposed. In this system, negatively or positively selectable marker traits are required along with dominant transgenic male sterility. Anthocyanin pigmentation is an excellent marker trait. Two regulatory genes for MYB and bHLH and a structural gene for DFR are required for anthocyanin pigmentation in rice. Therefore, to apply anthocyanin pigmentation as a marker trait in various rice genotypes, coordinated expression of the three genes is required. In this study, we developed a leaf sheath-specific promoter and introduced three genes-DFR and C1/Myb, driven by the 35S promoter, and OsB2/bHLH, driven by the leaf sheath-specific promoter-into the rice genome. Leaf sheath-specific pigmentation was confirmed in all seven genotypes tested, which included japonica and indica cultivars. Analysis of genome sequence data from 25 cultivars showed that the strategy of conferring leaf sheath-specific anthocyanin pigmentation by introduction of these three genes would be effective for a wide range of genotypes and will be applicable to RSUTMS.

SUBMITTER: Oshima M

PROVIDER: S-EPMC6711742 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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Development of a visible marker trait based on leaf sheath-specific anthocyanin pigmentation applicable to various genotypes in rice.

Oshima Masao M Taniguchi Yojiro Y Akasaka Maiko M Abe Kiyomi K Ichikawa Hiroaki H Tabei Yutaka Y Tanaka Junichi J

Breeding science 20190315 2

To overcome a limitation to the breeding of autogamous crops, recurrent selection using transgenic male sterility (RSUTMS) has been proposed. In this system, negatively or positively selectable marker traits are required along with dominant transgenic male sterility. Anthocyanin pigmentation is an excellent marker trait. Two regulatory genes for MYB and bHLH and a structural gene for DFR are required for anthocyanin pigmentation in rice. Therefore, to apply anthocyanin pigmentation as a marker t ...[more]

PMID: 31481833

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Project description:BACKGROUND:The cultivated potato Solanum tuberosum L. is the fourth most important crop worldwide. Anthocyanins synthesis and accumulation in potato tissues are considered as one of important traits related to stress resistance and nutritional value. It is considered that the major regulatory gene for anthocyanin biosynthesis is R2R3 MYB-encoding gene StAN1. However, the genetic control of pigmentation of different potato tissues is substantially under investigated. The development of genetic markers for breeding of potato with specific pigmentation pattern remains an actual task. RESULTS:We investigated 36 potato varieties and hybrids with different pigmentation of tubers and leaves. Sequence organization of regulatory R2R3 MYB (StAN1, StMYBA1, StMYB113), bHLH (StbHLH1, StJAF13) and WD40 (StWD40) genes potentially controlling anthocyanin biosynthesis has been evaluated. The results demonstrated a high variability in the StAN1 third exon and promoter region with the exception for 35 bp, containing elements for the transcription start and activation of gene expression in roots. The analysis of transcriptional activity of genes coding R2R3 MYBs, bHLHs and WD40 transcriptional factors in leaves of eight potato genotypes with different anthocyanin pigmentation was performed. The results showed a relation between the gene expression level and plant pigmentation only for the StAN1 and StWD40 genes, while other studied genes had either strong expression in all varieties and hybrids (StMYBA1, StbHLH1 and StJAF13) or they were not expressed at all (StMYB113). CONCLUSIONS:It was found that StAN1 is the major regulatory gene controlling potato anthocyanin synthesis. However, diagnostic markers developed for the functional StAN1 alleles (StAN1777 and StAN1816) can not be used efficiently for prediction of potato pigmentation patterns. It is likely that the sequence organization of StAN1 promoter is important for anthocyanin synthesis control and the development of additional diagnostic markers is necessary.

Project description:BackgroundPurple carrots accumulate large quantities of anthocyanins in their roots and leaves. These flavonoid pigments possess antioxidant activity and are implicated in providing health benefits. Informative, saturated linkage maps associated with well characterized populations segregating for anthocyanin pigmentation have not been developed. To investigate the genetic architecture conditioning anthocyanin pigmentation we scored root color visually, quantified root anthocyanin pigments by high performance liquid chromatography in segregating F2, F3 and F4 generations of a mapping population, mapped quantitative trait loci (QTL) onto a dense gene-derived single nucleotide polymorphism (SNP)-based linkage map, and performed comparative trait mapping with two unrelated populations.ResultsRoot pigmentation, scored visually as presence or absence of purple coloration, segregated in a pattern consistent with a two gene model in an F2, and progeny testing of F3-F4 families confirmed the proposed genetic model. Purple petiole pigmentation was conditioned by a single dominant gene that co-segregates with one of the genes conditioning root pigmentation. Root total pigment estimate (RTPE) was scored as the percentage of the root with purple color.All five anthocyanin glycosides previously reported in carrot, as well as RTPE, varied quantitatively in the F2 population. For the purpose of QTL analysis, a high resolution gene-derived SNP-based linkage map of carrot was constructed with 894 markers covering 635.1 cM with a 1.3 cM map resolution. A total of 15 significant QTL for all anthocyanin pigments and for RTPE mapped to six chromosomes. Eight QTL with the largest phenotypic effects mapped to two regions of chromosome 3 with co-localized QTL for several anthocyanin glycosides and for RTPE. A single dominant gene conditioning anthocyanin acylation was identified and mapped.Comparative mapping with two other carrot populations segregating for purple color indicated that carrot anthocyanin pigmentation is controlled by at least three genes, in contrast to monogenic control reported previously.ConclusionsThis study generated the first high resolution gene-derived SNP-based linkage map in the Apiaceae. Two regions of chromosome 3 with co-localized QTL for all anthocyanin pigments and for RTPE, largely condition anthocyanin accumulation in carrot roots and leaves. Loci controlling root and petiole anthocyanin pigmentation differ across diverse carrot genetic backgrounds.

Dataset Information

Development of a visible marker trait based on leaf sheath-specific anthocyanin pigmentation applicable to various genotypes in rice.

Publications

Development of a visible marker trait based on leaf sheath-specific anthocyanin pigmentation applicable to various genotypes in rice.

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