Myeong-Cheoul Cho

Transcriptome profiling and molecular marker discovery in red pepper, Capsicum annuum L. TF68.

Transcriptome from high throughput sequencing-by-synthesis is a good resource of molecular markers. In this study, we present utility of massively parallel sequencing by synthesis for profiling the transcriptome of red pepper (Capsicum annuum L. TF68) using 454 GS-FLX pyrosequencing. Through the generation of approximately 30.63 megabases (Mb) of expressed sequence tag (EST) data with the average length of 375 base pairs (bp), 9,818 contigs and 23,712 singletons were obtained by raw reads assembly. Using BLAST alignment against NCBI non-redundant and a UniProt protein database, 30% of the tentative consensus sequences were assigned to specific function annotation, while 24% returned alignments of unknown function, leaving up to 46% with no alignment. Functional classification using FunCat revealed that sequences with putative known function were distributed cross 18 categories. All unigenes have an approximately equal distribution on chromosomes by aligning with tomato (Solanum lycopersicum) pseudomolecules. Furthermore, 1,536 high quality single nucleotide discrepancies were discovered using the Bukang mature fruit cDNA collection (dbEST ID: 23667) as a reference. Moreover, 758 simple sequence repeat (SSR) motif loci were mined from 614 contigs, from which 572 primer sets were designed. The SSR motifs corresponded to di- and tri- nucleotide motifs (27.03 and 61.92%, respectively). These molecular markers may be of great value for application in linkage mapping and association mapping research.

Biosynthesis of Capsinoid is Controlled by the Pun1 Locus in Pepper

Pungency in pepper (Capsicum annuum L.) has unique characteristics due to the alkaloid compound group, capsaicinoids, which includes capsaicin. Although capsaicinoids have been proved to have pharmacological and physiological effects on human health, the application of capsaicinoids has been limited because of their pungency. Capsinoids found in non-pungent peppers share closely related structures with capsaicinoids and show similar biological effects. Previous studies demonstrated that mutations in the p-AMT gene were related to the production of capsinoids; however, the pathway of capsinoid synthesis has not yet been fully elucidated. In this study, we performed genetic analysis to determine the mechanism of capsinoid synthesis using a F6 recombinant inbred line population. In this population, the presence/absence of capsinoids co-segregated with the genotype of the Pun1 locus, without exception. In addition, we screened the patterns of capsinoid synthesis and the correlation between the Pun1 locus and capsinoid synthesis in p-AMT mutant accessions. In Capsicum germplasms, we selected amino-acid-substituted mutants in the PLP binding domain of the p-AMT gene. Capsinoids were not synthesized with the recessive pun1 gene, regardless of the p-AMT genotype, and no relationship was found between p-AMT mutant type and capsinoid content. We concluded that the Pun1 gene, which is responsible for capsaicinoid synthesis, also controls capsinoid synthesis.

SNP Marker Integration and QTL Analysis of 12 Agronomic and Morphological Traits in F8 RILs of Pepper (Capsicum annuum L.)

Red pepper, Capsicum annuum L., has been attracting geneticists’ and breeders’ attention as one of the important agronomic crops. This study was to integrate 41 SNP markers newly developed from comparative transcriptomes into a previous linkage map, and map 12 agronomic and morphological traits into the integrated map. A total of 39 markers found precise position and were assigned to 13 linkage groups (LGs) as well as the unassigned LGe, leading to total 458 molecular markers present in this genetic map. Linkage mapping was supported by the physical mapping to tomato and potato genomes using BLAST retrieving, revealing at least two-thirds of the markers mapped to the corresponding LGs. A sum of 23 quantitative trait loci from 11 traits was detected using the composite interval mapping algorithm. A consistent interval between a035_1 and a170_1 on LG5 was detected as a main-effect locus among the resistance QTLs to Phytophthora capsici at high-, intermediate- and low-level tests, and interactions between the QTLs for high-level resistance test were found. Considering the epistatic effect, those QTLs could explain up to 98.25% of the phenotype variations of resistance. Moreover, 17 QTLs for another eight traits were found to locate on LG3, 4, and 12 mostly with varying phenotypic contribution. Furthermore, the locus for corolla color was mapped to LG10 as a marker. The integrated map and the QTLs identified would be helpful for current genetics research and crop breeding, especially in the Solanaceae family.

A splicing mutation in the gene encoding phytoene synthase causes orange coloration in Habanero pepper fruits

Peppers (Capsicum spp.) display a variety of fruit colors that are reflected by the composition and amount of diverse carotenoid pigments accumulated in the pericarp. Three independent loci, c1, c2, and y, are known to determine the mature color of pepper fruits by their allelic combinations. We examined the inheritance of fruit color in recombinant inbred lines (RILs) derived from an interspecific cross between C. annuum cv. TF68 (red) and C. chinense cv. Habanero (orange). The c2 gene encodes phytoene synthase (PSY), a rate-limiting enzyme in the carotenoid biosynthesis pathway. TF68 has a dominant c2+ allele whereas Habanero is homozygous for the recessive c2 allele, which determined RIL fruit color. Here we report that the recessive c2 allele has a point mutation in the PSY gene that occurs at a splice acceptor site of the fifth intron leading to both a frame shift and premature translational termination, suggesting that impaired activity of PSY is responsible for orange fruit color. During ripening, PSY is expressed at a significantly high level in orange colored fruits compared to red ones. Interestingly, the PSY gene of red Habanero has a conserved splice acceptor dinucleotide AG. Further analysis suggests that red Habanero is a wild type revertant of the PSY mutant orange Habanero.

Genotypic variation in carotenoid, ascorbic acid, total phenolic, and flavonoid contents, and antioxidant activity in selected tomato breeding lines

Genotypic diversity of antioxidants is important for the development of tomato cultivars with high antioxidant contents. The contents and antioxidant activities of major antioxidant phytochemicals (carotenoids, ascorbic acid, total phenolics, and flavonoids) of 119 cherry and non-cherry tomato genotypes (16 commercial cultivars and 103 germplasm lines) were analyzed. Antioxidant activity was evaluated using FRAP, ABTS, and DPPH aßsays. Significant genotypic differences were observed in the content and antioxidant activity of all of the studied antioxidant phytochemicals. Relatively higher genotypic variation was found in carotenoid, ascorbic acid, and flavonoid content than in total phenolic content and antioxidant activities. Most variation in total carotenoid content of tomato fruits was caused by lycopene, followed by ß-carotene and lutein. One cherry tomato line, TG-110, and 3 non-cherry tomato germplasm lines, IT237605, IT237703, and IT237706, had much higher lycopene contents (> 1930 mg·kg -1), and thus higher total carotenoid contents, than did the other lines. The highest ascorbic acid and total flavonoid contents were found in TG-106 (388 mg·kg -1) and TC-053 (2353.0 mg·kg -1), respectively. Two cherry tomato lines, TC-019 and TC-053, and 3 non-cherry tomato lines, TG-107, TG-112, and TG-113, had significantly higher total phenolic contents and antioxidant activities than did the commercial cultivars. The genotypes selected on the basis of their phytochemical content could be used to develop new tomato cultivars with high antioxidant contents and activities.

Development of Resistance Evaluation Method for Powdery Mildew (Leveillula taurica) in Capsicum spp.

Pepper powdery mildew causes increasingly economical damage due to increased cultivation of pepper in greenhouses. To assess resistance of pepper resources against pepper powdery mildew, we developed a novel evaluation formula for pepper resistance against powdery mildew. The evaluation formula named S index is as follows; (number of the highest disease leaf/top leaf number)-(number of the lowest disease leaf/top leaf number). Positive correlation (81%, P