Transcriptome profiling of genes related to light-induced anthocyanin biosynthesis in eggplant (Solanum melongena L.) before purple color becomes evident.
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ABSTRACT: The anthocyanins are highly enriched in eggplants (Solanum melongena L.) with purple peel. However, our previous study showed that anthocyanins biosynthesis in eggplant cultivar 'Lanshan Hexian' was completely regulated by light and color becomes evident at most 2 days after exposure to light. In the present investigation, transcriptome study was made to explore the underlying molecular mechanisms of light-induced anthocyanin biosynthesis in eggplant (Solanum melongena L.) before color becomes evident.RNA-Seq was performed for four time points (0, 0.5, 4 and 8 h after bags removal) where concerted changes happened. A total of 32,630 genes or transcripts were obtained by transcriptome sequencing, from which 1956 differentially expressed genes (DEGs) were found. Gene Ontology analysis showed that the 1956 DEGs covered a wide range of cellular components, molecular functions and biological processes. All the DEGs were further divided into 26 clusters based on their distinct expression patterns. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis found out 24 structural anthocyanin biosynthesis genes which distributing in seven clusters. In addition, 102 transcription factors, which exhibited highly dynamic changes in response to light, were found in the seven clusters. Three photoreceptors, UV Resistance Locus 8 (UVR8), Cryptochrome 3 (CRY3) and UVR3, were identified as DEGs. The light signal transduction elements, COP1 and two SPAs, might be responsible for anthocyanin biosynthesis regulation.Based on the transcriptome data, the anthocyanin biosynthesis structural genes, transcription factors, photoreceptors and light signal transduction elements were quickly screened which may act as the key regulatory factors in anthocyanin biosynthesis pathway. By comparing the transcriptome data with our previous studies, 869 genes were confirmed to participate in the light-induced anthocyanin biosynthesis. These results expand our knowledge of light-induced anthocyanin biosynthesis in plants, which allowing for fruit coloration to be improved under low-light conditions in future.
Transcriptome profiling of genes related to light-induced anthocyanin biosynthesis in eggplant (Solanum melongena L.) before purple color becomes evident.
<h4>Background</h4>The anthocyanins are highly enriched in eggplants (Solanum melongena L.) with purple peel. However, our previous study showed that anthocyanins biosynthesis in eggplant cultivar 'Lanshan Hexian' was completely regulated by light and color becomes evident at most 2 days after exposure to light. In the present investigation, transcriptome study was made to explore the underlying molecular mechanisms of light-induced anthocyanin biosynthesis in eggplant (Solanum melongena L.) bef ...[more]
Project description:BACKGROUND:Anthocyanin synthesis is affected by many factors, among which temperature is an important environmental factor. Eggplant is usually exposed to high temperatures during the cultivation season in Shanghai, China. Therefore,RNA -seq analysis was used to determine the effects of high-temperature stress on gene expression in the anthocyanin biosynthetic pathway of eggplant (Solanum melongena L.). RESULTS:We tested the heat-resistant cultivar 'Tewangda'. The plants were incubated at 38?°C and 45?°C, and the suitable temperature for eggplant growth was used as a control. The treatment times were 3?h and 6?h. The skin of the eggplant was taken for transcriptome sequencing, qRT-PCR assays and bioinformatic analysis. The results showed that 770 genes were differentially expressed between different treatments. Gene Ontology (GO) database and Kyoto Encyclopedia of Genes and Genomes (KEGG) database analyses identified 16 genes related to anthocyanin biosynthesis, among which CHSB was upregulated. Other genes, including BHLH62, MYB380, CHI3, CHI, CCOAOMT, AN3, ACT-2, HST, 5MA-T1, CYP75A2, ANT17, RT, PAL2, and anthocyanin 5-aromatic acyltransferase were downregulated. In addition, the Myb family transcription factor PHL11 was upregulated in the CK 3?h vs 45?°C 3?h, CK 3?h vs 38?°C 3?h, and CK 6?h vs 38?°C 6?h comparisons, and the transcription factor bHLH35 was upregulated in the CK 3?h vs 38?°C 3?h and CK 6?h vs 38?°C 6?h comparisons. CONCLUSION:These results indicated that high temperature will downregulate most of the genes in the anthocyanin biosynthetic pathway of eggplant. Our data have a reference value for the heat resistance mechanism of eggplant and can provide directions for molecular breeding of heat-resistant germplasm with anthocyanin content in eggplant.
Project description:The basic helix-loop-helix (bHLH) superfamily is considered the second largest transcription factor (TF) family. It plays regulatory roles in the developmental processes of plants and in their defense responses. In recent years, many bHLH superfamily genes have been identified and characterized in herbaceous and woody plants. However, the comprehensive genomic and functional analyses of these genes in eggplant (Solanum melongena L.) have not been reported. In this study, 121 bHLH TFs were identified in the recently released eggplant genome. The phylogeny, gene structure and conserved motifs of the SmbHLH gene were comprehensively studied. Subsequently, the phylogenetic relationship between the bHLH of eggplant and the bHLH of other species was analyzed, and the proteins were classified into 17 subfamilies. Among these protein sequences, 16 subgroups were clustered into the functional clades of Arabidopsis. Two candidate genes (SmbHLH1, SmbHLH117) that may be involved in anthocyanin biosynthesis were screened. The tissue specificity or differential expression of the bHLH genes in different tissues and under various light and temperature conditions suggested the differential regulation of tissue development and metabolism. This study not only provides a solid foundation for the functional dissection of the eggplant bHLH gene family but may also be useful for the future synthesis of anthocyanins in eggplant.
Project description:Eggplant varieties rich in bioactive chlorogenic acid along with less browning are preferred by consumers. Therefore, genetics of fruit phenolics, fruit flesh colour, and browning related traits were studied in the genotypes of eggplant, comprising of nine cultivated varieties and one accession of eggplant's primary genepool wild relative Solanum insanum (INS2). These accessions were genotyped based on the 7335 polymorphic single-nucleotide polymorphisms (SNP) markers. After that, genotypes were crossed in half diallel fashion to produce 45 hybrids. The INS2 displayed the highest values for the total phenolics and chlorogenic acid content (CGA). For all of the biochemical traits studied, significant values of general and specific combining ability (GCA and SCA) effects were determined. The baker ratio estimates were high (>0.75) for all of the traits. Highly significant and positive heterosis (%) was determined for the dry matter, total phenolics, CGA, and area (%) of CGA content. The phenolics content of the fruit (total phenolics and CGA) was not significantly correlated with flesh colour and browning related traits. However, when the path coefficient analysis was performed considering the CGA as a dependent variable, it was determined that the flesh colour related traits most considerably affected the CGA. The genetic distance showed a diminutive correlation with the hybrid means, heterosis, and SCA values. Overall, this study provides important information regarding the underlying genetics of important biochemical traits of eggplant fruit.
Project description:Rind color is an economically important agronomic trait in eggplant that impacts consumer preferences. In this study, bulked segregant analysis and competitive allele-specific PCR were employed to identify the candidate gene for eggplant rind color through constructing a 2794 F2 population generated from a cross between "BL01" (green pericarp) and "B1" (white pericarp). Genetic analysis of rind color revealed that a single dominant gene controls green color of eggplant peel. Pigment content measurement and cytological observations demonstrated that chlorophyll content and chloroplast number in BL01 were higher than in B1. A candidate gene (EGP19168.1) was fine-mapped to a 20.36 Kb interval on chromosome 8, which was predicted to encode the two-component response regulator-like protein Arabidopsis pseudo-response regulator2 (APRR2). Subsequently, allelic sequence analysis revealed that a SNP deletion (ACT→AT) in white-skinned eggplant led to a premature termination codon. Genotypic validation of 113 breeding lines using the Indel marker closely linked to SmAPRR2 could predict the skin color (green/white) trait with an accuracy of 92.9%. This study will be valuable for molecular marker-assisted selection in eggplant breeding and provides theoretical foundation for analyzing the formation mechanism of eggplant peel color.
Project description:Heterosis (or hybrid vigor) results in a hybrid's phenotypic superiority over its founder parents for quantitative and qualitative traits. Hybrid vigor is defined by mechanisms such as dominant complementation, over-dominance, and epistasis. Eggplant (Solanum melongena L.) is an essential vegetable crop and a good source of dietary minerals, vitamins, and anthocyanins, with a high oxygen radical absorbance capacity and low caloric value. Given the economic and nutritional significance of eggplants, breeding efforts focus on developing high-yielding varieties-mostly F1 hybrids-with important traits. Studies indicate the successful exploitation of heterosis in the eggplant for a considerable improvement with respect to quantitative traits. In this direction, estimating heterosis for yield-related traits could well be useful for examining the most beneficial hybrid mix with the exploitation of top-quality hybrid. This review examines the current perception of the breeding and molecular aspects of heterosis in eggplants and cites several studies describing the mechanisms. Rendering and combining recent genomics, epigenetic, proteomic, and metabolomics studies present new prospects towards the understanding of the regulatory events of heterosis involved in the evolution and the domestication of the eggplant ideotype.
Project description:Eggplant berries are rich in anthocyanins like delphinidin-3-rutinoside (D3R) and nasunin (NAS), which are accumulated at high amounts in the peel. NAS is derived by D3R through acylation and glycosylation steps. The presence of D3R or NAS is usually associated with black-purple or lilac fruit coloration of the most cultivated varieties, respectively. Building on QTL mapping position, a candidate gene approach was used to investigate the involvement of a BAHD anthocyanin acyltransferase (SmelAAT) in determining anthocyanin type. The cDNA sequence comparison revealed the presence of a single-base deletion in D3R-type line '305E40' (305E40_aat) with respect to the NAS-type reference line '67/3'. This is predicted to cause a frame shift mutation, leading to a loss of SmelAAT function and, thus, D3R retention. RT-qPCR analyses confirmed SmelAAT and 305E40_aat expression during berry maturation. In D3R-type lines, '305E40' and 'DR2', overexpressing the functional SmelAAT allele from '67/3', the transcript levels of the transgene correlated with the accumulation of NAS in fruit peel. Furthermore, it was also found a higher expression of the transcript for glucosyltransferase Smel5GT1, putatively involved with SmelAAT in the last steps of anthocyanin decoration. Finally, an indel marker matching with anthocyanin type in the '305E40' × '67/3' segregating population was developed and validated in a wide number of accessions, proving its usefulness for breeding purposes.
Project description:Heterostyly is a common floral polymorphism, but the proteomic basis of this trait is still largely unexplored. In this study, self- and cross-pollination of L-morph and S-morph flowers and comparison of embryo sac development in eggplant (Solanum melongena L.) suggested that lower fruit set from S-morph flowers results from stigma-pollen incompatibility. To explore the molecular mechanism underlying heterostyly development, we conducted isobaric tags for relative and absolute quantification (iTRAQ) proteomic analysis of eggplant pistils for L- and S-morph flowers. A total of 5,259 distinct proteins were identified during heterostyly development. Compared S-morph flowers with L-morph, we discovered 57 and 184 differentially expressed proteins (DEPs) during flower development and maturity, respectively. Quantitative real time polymerase chain reactions were used for nine genes to verify DEPs from the iTRAQ approach. During flower development, DEPs were mainly involved in morphogenesis, biosynthetic processes, and metabolic pathways. At flower maturity, DEPs primarily participated in biosynthetic processes, metabolic pathways, and the formation of ribosomes and proteasomes. Additionally, some proteins associated with senescence and programmed cell death were found to be upregulated in S-morph pistils, which may lead to the lower fruit set in S-morph flowers. Although the exact roles of these related proteins are not yet known, this was the first attempt to use an iTRAQ approach to analyze proteomes of heterostylous eggplant flowers, and these results will provide insights into biochemical events taking place during the development of heterostyly.
Project description:Eggplant Solanum melongena L. is one of the most economically important vegetable crops. Here, we report the complete chloroplast (cp) genome of eggplant. The cp genome size was 154,289?bp that contained a pair of IR regions of 25,566?bp, one large single-copy (LSC) of 84,749?bp and a small single-copy (SSC) of 18,408bp, respectively. It encoded 125 predicted unique functional genes, including 84 tRNA genes, 85 protein-coding genes and 8 rRNA genes. The GC content was 37.86%. Phylogenetic analysis clearly showed a close evolutionary relationship between S. melongena and other species in the genus Solanum. The complete chloroplast genome of S. melongena provides valuable data for genetic improvement and allele mining of eggplant germplasm.
Project description: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.
Project description:Polyphenol oxidases (PPOs) in different Solanum species including eggplant have been studied. PPOs have been implicated in undesirable enzymatic browning of eggplant fruit and also in plant defense. The main objective of this study was to identify and accelerate the further functional characterization of additional eggplant PPOs that are involved in food biochemistry and defense-related functions. Eggplant PPOs identified earlier were used in "Basic local alignment search tool (BLAST)" search against expressed sequence tag and nucleotide databases. We have identified seven additional sequences which were almost complete in length. The sequences of the PPOs were aligned and their phylogenetic and evolutionary relationships established. The sequences are quite diverse, broadly falling into two major clusters; three PPOs form a separate branch/minor cluster. The thirteen sequences had conserved copper A binding sites but copper B binding sites differed considerably in two new PPO sequences (AFJ79642 and ACR61398). A third conserved 'Histidine-rich' region has been identified at the 'C' terminus of the eggplant PPOs. In addition, all the seven new PPOs exhibited at least one glycosylated sequon in the mature PPO sequence. Identification of additional PPO genes will further help in functional and biological characterization of these PPOs.
