Fine Mapping of Candidate Gene Controlling Anthocyanin Biosynthesis for Purple Peel in Solanum melongena L.

Fruit color is an intuitive quality of horticultural crops that can be used as an evaluation criterion for fruit ripening and is an important factor affecting consumers' purchase choices. In this study, a genetic population from the cross of green peel 'Qidong' and purple peel '8 guo' revealed that the purple to green color of eggplant peel is dominant and controlled by a pair of alleles. Bulked segregant analysis (BSA), SNP haplotyping, and fine genetic mapping delimited candidate genes to a 350 kb region of eggplant chromosome 10 flanked by markers KA2381 and CA8828. One ANS gene (EGP22363) was predicted to be a candidate gene based on gene annotation and sequence alignment of the 350-kb region. Sequence analysis revealed that a single base mutation of 'T' to 'C' on the exon green peel, which caused hydrophobicity to become hydrophilic serine, led to a change in the three-level spatial structure. Additionally, EGP22363 was more highly expressed in purple peels than in green peels. Collectively, EGP22363 is a strong candidate gene for anthocyanin biosynthesis in purple eggplant peels. These results provide important information for molecular marker-assisted selection in eggplants, and a basis for analyzing the regulatory pathways responsible for anthocyanin biosynthesis in eggplants.

Comparative Transcriptome Analysis Reveals Gene Expression Differences in Eggplant (Solanum melongena L.) Fruits with Different Brightness.

Fruit brightness is an important quality trait that affects the market value of eggplant. However, few studies have been conducted on eggplant brightness. In this study, we aimed to identify genes related to this trait in three varieties of eggplant with different fruit brightness between 14 and 22 days after pollination. Using RNA-Seq Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses, we found that wax- and cutin-related pathways and differentially expressed genes displayed significant differences among different development stages and varieties. Scanning electron microscopy revealed that the wax layer was thinner in '30-1' and 'QPCQ' than in '22-1'. Gas chromatography-mass spectrometry analysis revealed that wax content was significantly lower in '30-1' than in '22-1', which indicated that wax may be an important factor determining fruit brightness. We further identified and analyzed the KCS gene family, which encodes the rate-limiting enzyme of FA elongation in wax synthesis. The results provide an insight into the molecular mechanisms of fruit brightness in eggplants and further eggplant breeding programs.

Metabolomic Analysis, Combined with Enzymatic and Transcriptome Assays, to Reveal the Browning Resistance Mechanism of Fresh-Cut Eggplant.

Browning has been the primary limitation in eggplant processing. This study investigates the molecular mechanism underlying fresh-cut eggplant fruit browning by observing the physicochemical characteristics of browning-resistant ('F') and browning-sensitive ('36') eggplant cultivars. Browning-related enzyme activity and gene expression (PPO, LOX, and PLD) were significantly higher in the '36' eggplant, thereby enhancing the degree of browning, compared to the 'F' eggplant. The MDA content and O2- production rate progressively increased as browning increased, while the antioxidant capacity of the fruit decreased. The cutting injury significantly activated the expression of PAL, thereby inducing the accumulation of phenolic acids, while the PPO gene was significantly upregulated, which activated the activity of polyphenol oxidase. Our results showed that the oxidation of chlorogenic acids to chlorogenic quinones resulted in the occurrence of browning, which suggests chlorogenic acid as the main browning substrate in fresh-cut eggplant.

Transcriptome profiling reveals potential genes involved in browning of fresh-cut eggplant (Solanum melongena L.).

Fresh-cut processing promotes enzymatic browning of fresh fruits and vegetables, which negatively affects the product appearance and impacts their nutrition. We used RNA-sequencing to analyze the transcriptomic changes occurring during the browning of fresh-cut eggplant fruit samples from both browning-sensitive and browning-resistant cultivars to investigate the molecular mechanisms involved in browning. A total of 8347 differentially expressed genes were identified, of which 62 genes were from six gene families (i.e., PPO, PAL, POD, CAT, APX, and GST) potentially associated with enzymatic browning. Furthermore, using qRT-PCR, we verified 231 differentially regulated transcription factors in fresh-cut eggplant fruits. The enzyme activities of PPO, POD, PAL, and CAT in '36' were significantly higher than those of 'F' fresh-cut for 15 min. Both PPO and POD play a major role in the browning of eggplant pulp and might therefore act synergistically in the browning process. Meanwhile, qPCR results of 18 browning related genes randomly screened in 15 eggplant materials with different browning tolerance showed variant-specific expression of genes. Lastly, gene regulatory networks were constructed to identify the browning-related genes. This work provides a basis for future molecular studies of eggplants, and lays a theoretical foundation for the development of browning-resistant fresh-cut fruits and vegetables.

Study on browning mechanism of fresh-cut eggplant (Solanum melongena L.) based on metabolomics, enzymatic assays and gene expression.

Enzymatic browning is one of the crucial problems compromising the flavor and texture of fresh-cut fruit and vegetables. In this study, an untargeted metabolomics approach based on liquid chromatography-mass spectrometry (LC-MS) was used to explore the browning mechanism in fresh-cut eggplant. Metabolomics studies showed that with the increase of fresh-cut time, the contents of 946 metabolites changed dynamically. The metabolites having the same trend share common metabolic pathways. As an important browning substrate, the content of chlorogenic acid increased significantly, suggesting that may be more important to fresh-cut eggplant browning; all 119 common differential metabolites in 5 min/CK and 3 min/CK contrastive groups were mapped onto 31 KEGG pathways including phenylpropanol metabolism, glutathione metabolism pathway, et al. In physiological experiments, results showed that the Phenylpropanoid-Metabolism-Related enzymes (PAL, C4H, 4CL) were changed after fresh-cut treatment, the activities of three enzymes increased first and then decreased, and reached the maximum value at 5 min, indicating the accumulation of phenolic substances. At the same time, ROS were accumulated when plant tissue damaged by cutting, the activities of related antioxidant enzymes (SOD, APX and CAT) changed dynamically after oxidative damage. SOD and APX content increased significantly and reached the maximum value at 10 min after cutting, and then showed a downward trend. However, CAT activity increased sharply and reached the maximum value within 3 min after cutting, then maintained the same activity, and showed a downward trend after 30 min. These data fully demonstrated that the activities of browning related enzymes and gene expression increased with the prolonging of fresh cutting time. We explained the browning mechanism of fresh-cut eggplant by combining metabolomics and physiology, which may lay the foundation for better understanding the mechanism of browning during the fruits and vegetables during processing.

Transcriptome profiling and gene expression analyses of eggplant (Solanum melongena L.) under heat stress.

Global warming induces heat stress in eggplant, seriously affecting its quality and yield. The response to heat stress is a complex regulatory process; however, the exact mechanism in eggplant is unknown. We analyzed the transcriptome of eggplant under different high-temperature treatments using RNA-Seq technology. Three libraries treated at high temperatures were generated and sequenced. There were 40,733,667, 40,833,852, and 40,301,285 clean reads with 83.98%, 79.69%, and 84.42% of sequences mapped to the eggplant reference genome in groups exposed to 28°C (CK), 38°C (T38), and 43°C (T43), respectively. There were 3,067 and 1,456 DEGs in T38 vs CK and T43 vs CK groups, respectively. In these two DEG groups, 315 and 342 genes were up- and down-regulated, respectively, in common. Differential expression patterns of DEGs in antioxidant enzyme systems, detoxication, phytohormones, and transcription factors under heat stress were investigated. We screened heat stress-related genes for further validation by qRT-PCR. Regulation mechanisms may differ under different temperature treatments, in which heat shock proteins and heat stress transcription factors play vital roles. These results provide insight into the molecular mechanisms of the heat stress response in eggplant and may be useful in crop breeding.

Chalcone synthase (CHS) family members analysis from eggplant (Solanum melongena L.) in the flavonoid biosynthetic pathway and expression patterns in response to heat stress.

Enzymes of the chalcone synthase (CHS) family participate in the synthesis of multiple secondary metabolites in plants, fungi and bacteria. CHS showed a significant correlation with the accumulation patterns of anthocyanin. The peel color, which is primarily determined by the content of anthocyanin, is an economically important trait for eggplants that is affected by heat stress. A total of 7 CHS (SmCHS1-7) putative genes were identified in a genome-wide analysis of eggplants (S. melongena L.). The SmCHS genes were distributed on 7 scaffolds and were classified into 3 clusters. Phylogenetic relationship analysis showed that 73 CHS genes from 7 Solanaceae species were classified into 10 groups. SmCHS5, SmCHS6 and SmCHS7 were continuously down-regulated under 38°C and 45°C treatment, while SmCHS4 was up-regulated under 38°C but showed little change at 45°C in peel. Expression profiles of key anthocyanin biosynthesis gene families showed that the PAL, 4CL and AN11 genes were primarily expressed in all five tissues. The CHI, F3H, F3'5'H, DFR, 3GT and bHLH1 genes were expressed in flower and peel. Under heat stress, the expression level of 52 key genes were reduced. In contrast, the expression patterns of eight key genes similar to SmCHS4 were up-regulated at a treatment of 38°C for 3 hour. Comparative analysis of putative CHS protein evolutionary relationships, cis-regulatory elements, and regulatory networks indicated that SmCHS gene family has a conserved gene structure and functional diversification. SmCHS showed two or more expression patterns, these results of this study may facilitate further research to understand the regulatory mechanism governing peel color in eggplants.

Transcriptome analysis revealed expression of genes related to anthocyanin biosynthesis in eggplant (Solanum melongena L.) under high-temperature stress.

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.

Alleviation of exogenous 6-benzyladenine on two genotypes of eggplant (Solanum melongena Mill.) growth under salt stress.

Cytokinins were recently shown to control plant adaptation to environmental stresses. To characterize the roles of cytokinins in the tolerance of eggplant (Solanum melongena Mill.) to salt stress, the protective effects of 6-benzyladenine (6-BA) on the growth, photosynthesis, and antioxidant capacity in the leaves of two eggplant cultivars Huqie12 (salt-sensitive) and Huqie4 (salt-tolerant) were investigated. Under 90 mM NaCl stress, Huqie4 showed higher biomass accumulation and less oxidative damage compared to the Huqie12. Application of exogenous 10 µM 6-BA significantly alleviated the growth suppression caused by salt stress in two eggplant genotypes. In parallel with the growth, 6-BA application in salt-stressed plants resulted in enhanced chlorophyll contents, as well as photosynthetic parameters such as net CO2 assimilation rate (P n), stomatal conductance (g s), transpiration rate (E), and intercellular CO2 concentration (C i). Furthermore, exogenous 6-BA also significantly reduced the O2 (-) production rate and malondialdehyde content and markedly increased the antioxidant enzymes superoxide dismutase and peroxidase, the antioxidant metabolites ascorbate and reduced glutathione (GSH), and proline in both genotypes under salt stress. The results indicate that exogenous 6-BA is useful to improve the salt resistance of eggplant, which is most likely related to the increase in photosynthesis and antioxidant capacity.

Yeast heat-shock protein gene HSP26 enhances freezing tolerance in Arabidopsis.

In the yeast Saccharomyces cerevisiae, the molecular chaperone Hsp26 is one component of the heat-shock response. Hsp26 has the remarkable ability to directly sense increases in temperature and switch from an inactive state to a chaperone-active state. In this study, we report a functional analysis of Hsp26 in Arabidopsis thaliana and its response to freezing stress. After freezing stress, the HSP26 transgenic plants exhibited stronger growth than the wild-type plants. We found that over-expression of HSP26 in Arabidopsis increased the amounts of free proline and soluble sugars, elevated the expression of stress defense genes, and enhanced Arabidopsis tolerance to freezing stress. Taken together, our results indicate that Hsp26 may play an important role in the response of transgenic Arabidopsis plants to freezing stresses.