Mapping and validation of the epistatic D and P genes controlling anthocyanin biosynthesis in the peel of eggplant (Solanum melongena L.) fruit

Abstract Fruit color is an important trait influencing the commercial value of eggplant fruits. Three dominant genes (D, P and Y) cooperatively control the anthocyanin coloration in eggplant fruits, but none has been mapped. In this study, two white-fruit accessions (19 141 and 19 147) and their F2 progeny, with 9:7 segregation ratio of anthocyanin pigmented versus non-pigmented fruits, were used for mapping the D and P genes. A high-density genetic map was constructed with 5270 SNPs spanning 1997.98 cM. Three QTLs were identified, including two genes on chromosome 8 and one on chromosome 10. Gene expression analyses suggested that the SmANS on chromosome 8 and SmMYB1 on chromosome 10 were the putative candidate genes for P and D, respectively. We further identified (1) a SNP leading to a premature stop codon within the conserved PLN03176 domain of SmANS in 19 141, (2) a G base InDel in the promoter region leading to an additional cis-regulatory element and (3) a 6-bp InDel within the R2-MYB DNA binding domain of SmMYB1, in 19 147. Subsequently, these three variations were validated by PARMS technology as related to phenotypes in the F2 population. Moreover, silencing of SmANS or SmMYB1 in the purple red fruits of F1 (E3316) led to inhibition of anthocyanin biosynthesis in the peels. Conversely, overexpression of SmANS or SmMYB1 restored anthocyanin biosynthesis in the calli of 19 141 and 19 147 respectively. Our findings demonstrated the epistatic interactions underlying the white color of eggplant fruits, which can be potentially applied to breeding of eggplant fruit peel color.

[1]  G. Giuliano,et al.  Improved genome assembly and pan‐genome provide key insights into eggplant domestication and breeding , 2021, The Plant journal : for cell and molecular biology.

[2]  Dandan Li,et al.  A high‐quality genome assembly of the eggplant provides insights into the molecular basis of disease resistance and chlorogenic acid synthesis , 2021, Molecular ecology resources.

[3]  Huoying Chen,et al.  Functional characterization of SmMYB86, a negative regulator of anthocyanin biosynthesis in eggplant (Solanum melongena L.). , 2021, Plant science : an international journal of experimental plant biology.

[4]  Huoying Chen,et al.  Functional Characterization of SmbHLH13 in Anthocyanin Biosynthesis and Flowering in Eggplant , 2020 .

[5]  A. Kilchevsky,et al.  Identification of DNA Markers of Anthocyanin Biosynthesis Disorders Based on the Polymorphism of Anthocyanin 1 Tomato Ortholog Genes in Pepper and Eggplant , 2020 .

[6]  T. Hu,et al.  Construction of a SNP-Based Genetic Map Using SLAF-Seq and QTL Analysis of Morphological Traits in Eggplant , 2020, Frontiers in Genetics.

[7]  Margaret H. Frank,et al.  TBtools - an integrative toolkit developed for interactive analyses of big biological data. , 2020, Molecular plant.

[8]  Jinfang Chu,et al.  Natural variation in the promoter of Awn Length Inhibitor 1 (ALI-1) is associated with awn elongation and grain length in common wheat. , 2020, The Plant journal : for cell and molecular biology.

[9]  Y. Liu,et al.  Anthocyanin Biosynthesis and Degradation Mechanisms in Solanaceous Vegetables: A Review , 2018, Front. Chem..

[10]  Guoping Wang,et al.  Fine Mapping Identifies SmFAS Encoding an Anthocyanidin Synthase as a Putative Candidate Gene for Flower Purple Color in Solanum melongena L. , 2018, International journal of molecular sciences.

[11]  D. Fu,et al.  Virus-Induced Gene Silencing of the Eggplant Chalcone Synthase Gene during Fruit Ripening Modifies Epidermal Cells and Gravitropism. , 2018, Journal of agricultural and food chemistry.

[12]  E. Portis,et al.  QTL analysis reveals new eggplant loci involved in resistance to fungal wilts , 2018, Euphytica.

[13]  Huoying Chen,et al.  Molecular cloning and characterization of anthocyanin biosynthesis genes in eggplant (Solanum melongena L.) , 2016, Acta Physiologiae Plantarum.

[14]  M. Fibiani,et al.  Mapping Quantitative Trait Loci Affecting Biochemical and Morphological Fruit Properties in Eggplant (Solanum melongena L.) , 2016, Front. Plant Sci..

[15]  Giorgia Batelli,et al.  Phenylpropanoids Accumulation in Eggplant Fruit: Characterization of Biosynthetic Genes and Regulation by a MYB Transcription Factor , 2016, Front. Plant Sci..

[16]  H. Fukuoka,et al.  Detailed mapping of a resistance locus against Fusarium wilt in cultivated eggplant (Solanum melongena) , 2016, Theoretical and Applied Genetics.

[17]  Luyan Zhang,et al.  QTL IciMapping: Integrated software for genetic linkage map construction and quantitative trait locus mapping in biparental populations , 2015 .

[18]  A. von Haeseler,et al.  IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies , 2014, Molecular biology and evolution.

[19]  Jinfeng Chen,et al.  An SNP-based saturated genetic map and QTL analysis of fruit-related traits in cucumber using specific-length amplified fragment (SLAF) sequencing , 2014, BMC Genomics.

[20]  E. Portis,et al.  Linkage disequilibrium and genome-wide association analysis for anthocyanin pigmentation and fruit color in eggplant , 2014, BMC Genomics.

[21]  Hideki Hirakawa,et al.  Draft Genome Sequence of Eggplant (Solanum melongena L.): the Representative Solanum Species Indigenous to the Old World , 2014, DNA research : an international journal for rapid publication of reports on genes and genomes.

[22]  Zongli Hu,et al.  Anthocyanin accumulation and molecular analysis of anthocyanin biosynthesis-associated genes in eggplant (Solanum melongena L.). , 2014, Journal of agricultural and food chemistry.

[23]  D. Lewis,et al.  A Conserved Network of Transcriptional Activators and Repressors Regulates Anthocyanin Pigmentation in Eudicots[C][W][OPEN] , 2014, Plant Cell.

[24]  G. Valè,et al.  QTL Mapping in Eggplant Reveals Clusters of Yield-Related Loci and Orthology with the Tomato Genome , 2014, PloS one.

[25]  Dongyuan Liu,et al.  SLAF-seq: An Efficient Method of Large-Scale De Novo SNP Discovery and Genotyping Using High-Throughput Sequencing , 2013, PloS one.

[26]  G. Valè,et al.  A RAD Tag Derived Marker Based Eggplant Linkage Map and the Location of QTLs Determining Anthocyanin Pigmentation , 2012, PloS one.

[27]  S. Isobe,et al.  Development of gene-based markers and construction of an integrated linkage map in eggplant by using Solanum orthologous (SOL) gene sets , 2012, Theoretical and Applied Genetics.

[28]  H. Fukuoka,et al.  Development of selective markers linked to a major QTL for parthenocarpy in eggplant (Solanum melongena L.) , 2012, Theoretical and Applied Genetics.

[29]  Qian Qian,et al.  Genome-wide association study of flowering time and grain yield traits in a worldwide collection of rice germplasm , 2011, Nature Genetics.

[30]  D. Lewis,et al.  Members of an R2R3-MYB transcription factor family in Petunia are developmentally and environmentally regulated to control complex floral and vegetative pigmentation patterning. , 2011, The Plant journal : for cell and molecular biology.

[31]  R. Hellens,et al.  Identification of Mendel's White Flower Character , 2010, PloS one.

[32]  M. DePristo,et al.  The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. , 2010, Genome research.

[33]  Zhen-xing Li,et al.  AFLP and SCAR Markers Associated with Peel Color in Eggplant (Solanum melongena) , 2009 .

[34]  M. Bodis,et al.  The potato developer (D) locus encodes an R2R3 MYB transcription factor that regulates expression of multiple anthocyanin structural genes in tuber skin , 2009, Theoretical and Applied Genetics.

[35]  R. Halitschke,et al.  The potato R locus codes for dihydroflavonol 4-reductase , 2009, Theoretical and Applied Genetics.

[36]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[37]  Richard Durbin,et al.  Sequence analysis Fast and accurate short read alignment with Burrows – Wheeler transform , 2009 .

[38]  R. Salaman The inheritance of colour and other characters in the potato , 1910, Journal of Genetics.

[39]  Bernard R. Baum,et al.  Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components , 1997, Plant Molecular Biology Reporter.

[40]  S. Tian,et al.  Effect of oxalic acid on control of postharvest browning of litchi fruit , 2006 .

[41]  J. Mol,et al.  PH4 of Petunia Is an R2R3 MYB Protein That Activates Vacuolar Acidification through Interactions with Basic-Helix-Loop-Helix Transcription Factors of the Anthocyanin Pathway[W] , 2006, The Plant Cell Online.

[42]  Richard G. F. Visser,et al.  SMOOTH: a statistical method for successful removal of genotyping errors from high-density genetic linkage data , 2005, Theoretical and Applied Genetics.

[43]  R. Koes,et al.  Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. , 2005, Trends in plant science.

[44]  N. Ramsay,et al.  MYB-bHLH-WD40 protein complex and the evolution of cellular diversity. , 2005, Trends in plant science.

[45]  A. Frary,et al.  Eggplant (Solanum melongena) fruit colour: pigments, measurements and genetics. , 2004 .

[46]  S. Dinesh-Kumar,et al.  Virus-induced gene silencing in tomato. , 2002, The Plant journal : for cell and molecular biology.

[47]  S. Tanksley,et al.  Conservation of gene function in the solanaceae as revealed by comparative mapping of domestication traits in eggplant. , 2002, Genetics.

[48]  I. Day,et al.  An efficient procedure for genotyping single nucleotide polymorphisms. , 2001, Nucleic acids research.

[49]  Masashi Hirai,et al.  Mapping of fruit shape and color development traits in eggplant (Solanum melongena L.) based on RAPD and AFLP markers , 2001 .

[50]  J. Mol,et al.  Molecular Analysis of the anthocyanin2 Gene of Petunia and Its Role in the Evolution of Flower Color , 1999, Plant Cell.

[51]  E. Grotewold,et al.  How genes paint flowers and seeds , 1998 .

[52]  T. Holton,et al.  Genetics and Biochemistry of Anthocyanin Biosynthesis. , 1995, The Plant cell.

[53]  E. C. Tigchelaar,et al.  The Genetics of Anthocyanin Coloration in Eggplant (SOLANUM MELONGENA L.). , 1968, Genetics.

[54]  D. D. Kosambi The estimation of map distances from recombination values. , 1943 .

[55]  G. Mendel,et al.  Mendel's Principles of Heredity , 1910, Nature.