Progress 05/01/17 to 04/30/21
Outputs
Target Audience:Graduate and undergraduate students Biomedical Researchers Pepper Geneticists Plant Breeders and Pepper Growers Changes/Problems:Student participation is restricted during the final year of the grant, due to the pandemic of 2019-2020. What opportunities for training and professional development has the project provided?Four graduate students submitted their master's thesis on genetics /genomics of pepper phytochemicals, and effect of capsaicinoids using Drosophila melanogaster as a model system, and cell culture assays. We also published 4 peer reviewed journal articles from this research. Both Graduate and undergraduate students participated in local, regional and International conferences and presented the research results. Lav Kumar Yadav. 2017. Thesis Title: Genome-wide Identification and Characterization of Ankyrin Gene Family and Other genes involved in Capsaicin content and Fruit Weight in Pepper. Thesis submitted to the Biotechnology Graduate Program at West Virginia State University. Nirwan Tandukar. 2018. Thesis Title: Genome wide effects of body weight and triglycerides for dietary responses of various capsicum species on Drosophila melanogaster. Thesis submitted to the Biotechnology Graduate Program at West Virginia State University. Joshua Haynes. 2018. A survey of the gut microbiome diversity in response to dietary treatments of various pepper species in Drosophila melanogaster. Thesis submitted to the Biotechnology Graduate Program at West Virginia State University. Binlit Mathew. 2019. Effect of ghost pepper on epigenetic modifications in human renal cell adenocarcinoma in vitro. Thesis submitted to the Biotechnology Graduate Program at West Virginia State University. How have the results been disseminated to communities of interest?We presented the results of the project in Regional, National and International conferences. Peer reviewed publications are generated on pepper biomedicalresearch. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
As per the proposed objectives, we have estimated the content of capsaicin, dihydrocapsaicin, Vitamin C, and seven types of carotenoid compounds, namely capsanthin, zeaxanthin, lutein, β-cryptoxanthin, lycopene, α-carotene, and β-carotene in 200 accessions of Capsicum annuum, 268 accessions of C. baccatum, 225 accessions of C. chinense and 175 accessions of C. frutescens for use in GWAS. In this process, we have standardized the HPLC method for capsanthin, zeaxanthin, lutein, β-cryptoxanthin, lycopene, α-carotene, and β-carotene. We genotyped all of our pepper collections using GBS (genotyping by sequencing) technology and performed association mapping for the metabolites mentioned above. In addition, we also analyzed the transcriptome and metabolome pertaining to cutin in habanero peppers. These analyses improved our knowledge of the molecular mechanisms regulating the accumulation of cutin in habanero pepper fruits. In summary, we have archived 20,000 SNPs that are common for all the pepper species and individual species groups have about ~50,000 SNPs each with 70% call rate and 0.05 minor allele frequencies. We resolved population structures and selective sweeps important for various fruit traits and nutraceutical traits in various cultivated species. All these resources will be made available for public use. A cell culture study was conducted to examine the effect of Ghost pepper, one of the highest capsaicin containing peppers in the world, on cell proliferation, apoptosis, senescence and the global proteomic profile in human renal cell adenocarcinoma in vitro. 769-P human renal adenocarcinoma cells were cultured on RPMI-1640 media supplemented with fetal bovine serum (10%) and antibiotic-antimycotic solution (1%). Cell proliferation was tested with phenol red-free media with capsaicin (0-400 µM), dihydrocapsaicin (0-400 µM), capsaicin + dihydrocapsaicin (5:1), and dry Ghost peppers (0-3 g L-1) for 24, 48 and 72 h. Polycaspase and senescence associated-beta-galactosidase (SA-beta-gal) activities were tested with capsaicin, dihydrocapsaicin, capsaicin + dihydrocapsaicin, and Ghost pepper treatments. Global proteomic profile of cells in control and Ghost pepper treatment was analyzed after 6 h by a shotgun proteomic approach using tandem mass spectrometry. About 68,220 protein isoforms were identified by shotgun proteomic approach. Among these, about 8.2% were significantly affected by Ghost pepper. Ghost pepper exhibited antiproliferation activity by inducing apoptosis through a complex network of proteins in human renal cell adenocarcinoma in vitro. Another study was conducted by a graduate student to explore anti-proliferative and epigenetic effects of ghost pepper extracts on human renal cell adenocarcinoma in vitro. Cells treated with capsaicin, dihydrocapsaicin, capsaicin + dihydrocapsaicin and ghost pepper extract showed significant increase in methylation. Ghost pepper treatment increased DNMT (DNA methyl transferase) activity and also showed increased expression of DNMT1 and DNMT3a. The results strongly suggest that capsaicin and its related capsaicinoids can significantly reduce cell proliferation and affect the epigenome of renal cell adenocarcinoma, in vitro. One of the greatest impacts on the gastrointestinal microbiome is diet because the host and microbiome share the same food source. In addition, the effect of diet can diverge depending on the host genotype. Diets supplemented with phytochemicals found in peppers might cause shifts in the microbiome. Thus, understanding how these interactions occur can reveal potential health implications associated with such changes. We analyzed the gut microbiomes of threeDrosophila melanogastergenetic backgrounds (Canton-S, Oregon-RC, and Berlin-K) reared on control and pepper-containing diets (bell, serrano, and habanero peppers).Sequencing libraries of the 16S rRNA gene were constructed and sequenced on an Illumina NextSeq 500 platform. Pairwise comparisons revealed that microbial communities of flies reared on serrano-containing diet are significantly different from microbial groups of flies under the control diet. Likewise, our data revealed that specific microbial taxa were abundantly different depending on the diet. Firmicutes showed the highest abundance across all Drosophila populations reared on serrano-containing diets being represented principally by the Lactobacillaceae family. It is imperative to highlight that serrano peppers reported the highest level of capsaicin (4054 µg/g) among the peppers utilized in this study. These results may imply that the capsaicin content can stimulate Lactobacillaceae abundance, shifting the microbial communities in the gut of Drosophila. To understand the effect of Capsaicinoids on body weight and to control obesity, we selected Drosophila melanogaster as a model organism. We analyzed the effects of pepper containing diet in the transcriptome response of Drosophila and identified genes that may relate to pathogenesis accompanying an obesity-like state. We used the wild type D. melanogaster Berlin-K strain reared on control and 7.5% (w/w) pepper-containing diets. Experiments were initiated by placing approximately 10 males and 10 females onto vials containing the different diets. Adults were allowed to lay eggs for 72 h before being removed. The larvae were fed and once the adult stage was achieved, these flies were selected for bodyweight, triglyceride, and glucose level determination and RNAseq analysis. We observed a significant weight reduction in female flies on a pepper diet compared with those reared on control diet. Similarly, triglycerides level showed a high reduction at both sexes under pepper treatment. Although glucose levels did not show a significant difference, we noticed a slightly reduction in male and female flies. RNA-seq revealed 539 differentially expressed genes between control and pepper diets. This transcriptome study provided a comprehensive understanding of various molecular mechanisms underlying pepper diet effects in Drosophila. Outcomes/impacts For the first time, the current research provided the complete metabolite profile of 1200 Capsicum accessions belonging to C. annuum, C. baccatum, C. frutescens and C. chinense. The major carotenoids found in large amounts in peppers were the carotenes α-carotene, β-carotene, and lycopene; the xanthophylls, β-cryptoxanthin, capsanthin, lutein and zeaxanthin. The major flavonoids in peppers are apigenin, luteolin, myricetin, and quercetin and kaempferol. This information will be very useful for developing pre breeding lines with enhanced nutrition and for functional food research in future. We also noticed weight reduction in flies with capsaicin diet. We analyzed the effects of pepper containing diet in the transcriptome response of Drosophila and identified genes that can be used for anti-obesity research in the future. Drug development targeting the protein of these genes might also be useful for treating obesity. We found thatDrosophilagut microbiome can be shifted by pepper-containing diets, but the response varied according to the genotype and pepper type. Restoration or promotion of standard members of the gut microbiome appeared to be mediated by the pepper diet. This effect might be attributed to the various metabolites of the peppers. Four graduate students submitted their master's thesis on genetics /genomics of pepper phytochemicals, and effect of capsaicinoids using Drosophila melanogaster as a model system, and cell culture assays. We also published 4 peer reviewed journal articles from this research. Both Graduate and undergraduate students participated in local, regional and International conferences and presented the research results.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Perla, V., Nadimi, M., Reddy, R., Hankins, G.R., Nimmakayala, P., Harris, R.T., Valluri, J., Sirbu, C. and Reddy, U.K., 2018. Effect of ghost pepper on cell proliferation, apoptosis, senescence and global proteomic profile in human renal adenocarcinoma cells. PloS one, 13(10), p.e0206183.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Lopez-Ortiz, C., Dutta, S.K., Natarajan, P., Pe�a-Garcia, Y., Abburi, V., Saminathan, T., Nimmakayala, P. and Reddy, U.K., 2019. Genome-wide identification and gene expression pattern of ABC transporter gene family in Capsicum spp. PloS one, 14(4), p.e0215901.
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Garcia-Lozano, M.; Haynes, J.; Lopez-Ortiz, C.; Natarajan, P.; Pe�a-Garcia, Y.; Nimmakayala, P.; Stommel, J.; Alaparthi, S.B.; Sirbu, C.; Balagurusamy, N.; Reddy, U.K. Effect of Pepper-Containing Diets on the Diversity and Composition of Gut Microbiome of Drosophila melanogaster. International journal of molecular sciences 2020, 21, 945.
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Natarajan, P.; Akinmoju, T.A.; Nimmakayala, P.; Lopez-Ortiz, C.; Garcia-Lozano, M.; Thompson, B.J.; Stommel, J.; Reddy, U.K. Integrated metabolomic and transcriptomic analysis to characterize cutin biosynthesis between low-and high-cutin genotypes of Capsicum chinense Jacq. International journal of molecular sciences 2020, 21, 1397
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Tandukar N, Saminathan T; Alaparthi S, Nimmakayala P, Hankins G and Reddy U.2018. Genome Wide Association studies on nutraceutical effects of various chili peppers on Drosophila melanogaster. 59th Annual Drosophila Research
Conference , April 11-15, 2018, Philadelphia, PA.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Ortiz C, Abburi V., Saminathan T., Shinde S., Nimmakayala P. and Reddy U. Regulation of novel and conserved miRNAs associated with flowering and fruit development across different Capsicum species. The Donald Danforth Plant Science
Center 19th Annual Fall Symposium in Saint Louis, Missouri September 26- 28, 2018
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Lopez-Ortiz, C., Crummett, G., Edwards, M., Natarajan, P., Nimmakayala, P., Adjeroh, D., Sirbu, C., and Reddy, U.K. Transcriptome Response to a Habanero Pepper-Containing Diet in Drosophila melanogaster. 62nd Annual Drosophila Research Conference (March 23-April 1, 2021).
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Lopez-Ortiz, C., Edwards, M., Natarajan, P., Nimmakayala, P., Adjeroh, D., Sirbu, C., and Reddy, U.K. Transcriptome and Metabolome Response to diet containing different pepper species with high-low heat levels in Drosophila melanogaster. 10th Annual and 1st Virtual ACMAP Conference.
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Progress 05/01/19 to 04/30/20
Outputs
Target Audience:Graduate and undergraduate students, Biomedical Researchers, Pepper Geneticists, Plant breeders and Pepper growers. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Threegraduate students submitted their thesis research. Two undergraduate students did research with drosophila feedling trials. We have trained one technician to work at various levels of the project. How have the results been disseminated to communities of interest?Conference presentations, research publications, classroom teacing and emails. What do you plan to do during the next reporting period to accomplish the goals?We will estimate various important phytochemicals,volatiles and flavor specific compounds for 200 Capsicum frutescens germplasm lines . GWAS and gene annotation will be performed for carotenoid and flavonoid levels in C.frutescens. We plan to generate biparental mating to fine map genes for cutin biosynthesis and cutin variation in germplasm accessions of hot peppers.
Impacts
What was accomplished under these goals?
To understand the genetic architecture of natural variation in body weight and Triglyceride (TG) level due to chili pepper diet we have used Drosophila melanogaster as a model system. To have a better understanding of the genetic mechanism, we exploited naturally occurring polymorphisms in the Drosophila Genetic Reference Panel (DGRP), wild-derived, fully inbred, fully sequenced model suitable for genome wide association studies. DGRP fly lines were used to perform genome-wide association studies (GWAS) for body weight and Triglyceride (TG) level in various chili pepper (Habanero, Serrano, and Bell pepper) supplemented diet. The effect of peppers on body weight and TG level for the DGRP lines were found to be genotype-dependent and sex-dependent. Several candidate genes were revealed from genome association studies which had functions relating to body weight and TG level. Some of these genes include CG6847, htl, klar, vn, CG31317, CG12512, CG31729, Egfr, and FAD2. We functionally validated several of these genes using RNAi based mutants. Some of the candidate genes that showed promising results are bib, vn, RIC-3, Liprin-gamma, rg, CG9674, CG6847, and SP1. Gene ontology (GO) analysis revealed important biological processes including axon guidance, cell-cell adhesion and nucleotide biosynthesis that may explain the underlining mechanism of how peppers affect body weight and TG. Chili peppers are an important constituent of many foods and contain medicinally valuable compounds, such as capsaicin and dihydrocapsaicin. As various dietary botanicals have anticancer properties, this study was aimed to examine the effect of Ghost pepper (Bhut Jolokia), one of the hottest chili peppers in the world, on cell proliferation, apoptosis, senescence and the global proteomic profile in human renal cell adenocarcinoma in vitro. 769-P human renal adenocarcinoma cells were cultured on RPMI-1640 media supplemented with fetal bovine serum (10%) and antibiotic-antimycotic solution (1%). Treatment stock solutions were prepared in ethanol. Cell proliferation was tested with phenol red-free media with capsaicin (0-400 µM), dihydrocapsaicin (0-400 µM), capsaicin + dihydrocapsaicin (5:1), and dry Ghost peppers (0-3 g L-1) for 24, 48 and 72 h. Polycaspase and senescence associated-beta-galactosidase (SA-beta-gal) activities were tested with capsaicin (400 µM), dihydrocapsaicin (400 µM), capsaicin (400 µM) + dihydrocapsaicin (80 µM), and Ghost pepper (3 g L-1) treatments. Global proteomic profile of cells in control and Ghost pepper treatment (3 g L-1) was analyzed after 6 h by a shotgun proteomic approach using tandem mass spectrometry. At 24 h after treatment (24 HAT), relative to control, cell proportion with capsaicin (400 µM), dihydrocapsaicin (400 µM), capsaicin (400 µM) + dihydrocapsaicin (80 µM), and Ghost pepper (3 g L-1) treatments was reduced to 36%, 18%, 33% and 20%, respectively, and further reduced at 48 and 72 HAT. All treatments triggered an early polycaspase response. SA-beta-gal activity was normal or suppressed with all treatments. About 68,220 protein isoforms were identified by shotgun proteomic approach. Among these, about 8.2% were significantly affected by Ghost pepper. Ghost pepper regulated various proteins involved in intrinsic and extrinsic apoptotic pathways, Ras, Rb/E2F, p53, TGF-beta, WNT-beta catenin, and calcium induced cell death pathways. Ghost pepper also induced changes in proteins related to methylation, acetylation, genome stability, cell cycle check points, carbohydrate, protein and other metabolism and cellular mechanisms. Ghost pepper exhibited antiproliferation activity by inducing apoptosis through a complex network of proteins in human renal cell adenocarcinoma in vitro.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Garcia-Lozano, Marleny, Joshua Haynes, Carlos Lopez-Ortiz, Purushothaman Natarajan, Yadira Pe�a-Garcia, Padma Nimmakayala, John Stommel et al. "Effect of Pepper-Containing Diets on the Diversity and Composition of Gut Microbiome of Drosophila melanogaster." International Journal of Molecular Sciences 21, no. 3 (2020): 945.
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Ortiz C, Dutta SK, Natarajan P, Pena Garcia Y, Abburi V, Nimmakayala P and Reddy UK (2020). The ankyrin repeat gene family in Capsicum spp.: Genome-wide survey, characterization and gene expression profile. Nature Scientific Reports: 10.1038/s41598-020-61057=-098*4
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Progress 05/01/18 to 04/30/19
Outputs
Target Audience:Graduate and undergraduate students, Biomedical Researchers, Pepper Geneticists, Plant breeders and Pepper growers. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Two graduate students submitted their thesis research. Two undergraduate students did research with drosophila feedling trials. We have trained one technician to work at various levels of the project. How have the results been disseminated to communities of interest?Conference presentations, research publications, classroom teacingand emails. What do you plan to do during the next reporting period to accomplish the goals?We plan to generate biparental mating to fine map genes for cutin biosynthesis and cutin variation in germplasm accessions of hot peppers. We willestimate various important phytochemicals,volatiles and flavor specific compounds for 180 Capsicum chinenese germplasm lines . GWAS and gene annotation will be performed for carotenoid and flavonoid levels in pepper.
Impacts
What was accomplished under these goals?
We have analyzed the metabolome and transcriptome profile of three habanero peppers to understand cutin accumulation and metabolism. The cuticle plays crucial roles in plant survival and physiology. For this study, three habanero genotypes (Naga morich, PI 222448 and PI 257145) were selected based on the waxy or glossy phenotype of their fruit surfaces. Our results showed a huge variation in cutin composition across the three genotypes with about 6-fold range between the extreme genotypes. A total of 146,569 unigenes were obtained from the transcriptome analysis and 1,689 unigenes were differentially expressed between the high and low cutin pepper genotypes. Our study revealed 42 unigenes involved in the cutin biosynthesis and these genes showed significant differential gene expression. Genes such as cutin synthase, glycerol-3 phosphate acyltransferase 6, long chain acyltransferase 2, long chain acyltransferase 1 and CYP86A4, cytochrome P450, family 86, subfamily A, polypeptide 4 were expressed at higher levels in PI 257145 which was consistent with the metabolome data. The results obtained by RNA-seq were further validated by RT-qPCR carried out on the pepper genotypes accumulating different amount of cutin in their fruits. Network analysis revealed the relationship between the cutin genes and transcription factors while those of the DEGs showed enrichment in the acetyl CoA metabolic process while the MapMan pathway analysis showed upregulation of the genes in the Lipid pathway. These findings can be incorporated into breeding programs for targeting pepper fruit quality improvement. Microbial communities in the gut of animals create a complex environment composed mainly of bacteria, archaea, fungi and viruses. These microorganisms interact with each other and play a central role in the host by mediating different processes including metabolism, physiology, and immune responses. These functions can have significant impacts on the overall health of the host, and significant changes in the microbiome diversity can alter these functions for positive or negative effects as well. One of the greatest impacts on the gastrointestinal microbiome is diet, as the food source for the host is also the food source for the microbiome. Dietary compounds such as capsaicin found in peppers can cause shifts in the microbiome, hence it is important to understand how these interactions occur to reveal potential health implications associated with such changes. This study aimed to explore the effects of dietary pepper treatments on the gastrointestinal bacterial flora for several genotypes of Drosophila melanogaster. To achieve this goal, gut microbiome of three Drosophila genotypes (Canton-S, Oregon-RC and Berlin-K) reared on control and pepper-containing diets (bell, serrano and habanero peppers) were analyzed. Sequencing libraries of the 16S rRNA gene were constructed and sequenced on an Illumina NextSeq 500 platform. Results revealed that Drosophila microbiome is strongly influenced by genotype based on PERMANOVA of unweighted UniFrac distance which analyzes presence/absence data. Moreover, when abundance was considered in weighted UniFrac distance, pepper-containing diets also showed a significant effect on gut microbiome of the different Drosophila genotypes. Pairwise comparisons revealed that microbial communities of flies reared on serrano-containing diet are significantly different from microbial groups of flies under the control diet. Likewise, our data revealed that specific microbial taxa were abundantly different depending on the diet. Firmicutes showed the highest abundance across all Drosophila populations reared on serrano-containing diets being represented principally by the Lactobacillaceae family. It is imperative to highlight that serrano peppers reported the highest level of capsaicin (4054.08 µg/g) among the peppers utilized in this study. These results may imply that the capsaicin content can stimulate Lactobacillaceae abundance, shifting the microbial communities in the gut of Drosophila. This family was mainly composed by Lactobacillus brevis which has been shown to reduce colitis symptoms in aged mice. Additionally, this microbial group has been demonstrated to produce probiotic effects in humans and has been employed as a supplement principally to counter gastro intestinal problems including irritable bowel syndrome. In our study, pepper content appeared to promote L. brevis presence in Oregon-RC since its abundance was 4-fold higher in flies reared on pepper containing diets than in guts of flies raised on the control diet. This effect may be attributed to the chemical properties of peppers including the capsaicin content of serrano and habanero pepper and the polyphenol content in all peppers. Furthermore, Acetobacter presence in Oregon-RC and Canton-S appeared to be stimulated by these pepper properties. It has been reported that colonization of Drosophila gut with Acetobacter species decreased significantly the triglyceride content compared to axenic flies. Thus, metabolic advantages of dietary pepper consumption can be inferred.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Ortiz C, Abburi V., Saminathan T., Shinde S., Nimmakayala P. and Reddy U. Regulation of novel and conserved miRNAs associated with flowering and fruit development across different Capsicum species. The Donald Danforth Plant Science Center 19th Annual Fall Symposium in Saint Louis, Missouri September 26- 28, 2018
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Belcher S., Jones S., Tandukar N., Reddy U., Nimmakayala P., Katam R. Study on genome wide capsaicin effects for body weight and triglycerides using Drosophila melanogaster diversity panel. ARD Research Symposium 2019. Jacksonville, FL.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Akinsipe T., Abburi L.., Nimmakayala P., Reddy U. GWAS for seedling root growth variation in acidic conditions across the cultivated pepper species complexes. ARD Research Symposium 2019. Jacksonville, FL.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Perla, V., Nadimi, M., Reddy, R., Hankins, G.R., Nimmakayala, P., Harris, R.T., Valluri, J., Sirbu, C. and Reddy, U.K., 2018. Effect of ghost pepper on cell proliferation, apoptosis, senescence and global proteomic profile in human renal adenocarcinoma cells. PloS one, 13(10), p.e0206183.
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Progress 05/01/17 to 04/30/18
Outputs
Target Audience:Graduate and undergraduate students, Pepper growers, Biomedical Researchers, Pepper Geneticists and Plant breeders. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Two graduate students submitted their thesis research. Two undergraduate students worked in the project. We have trained one technician to work at various levels of the project. How have the results been disseminated to communities of interest?Two papers are in review for publication in peer review journals. We made 5 presentations in Plant and Animal Genome conference, American Society of Horticulture Science, Drosophila Research conference and ARD conference. Two seed companies were interested in our nutraceutical research in peppers and associated SNPs to fruit colors. What do you plan to do during the next reporting period to accomplish the goals?We plan to estimate various important volatiles and flavor specific compounds. GWAS and gene annotation will be performed for carotenoid and flavonoid levels in pepper. Crosses involving various species are currently under evaluation. Segregating populations will be further advanced and selections will be made based on nutraceutical values.
Impacts
What was accomplished under these goals?
We have estimated carotenoids, flavonoids, vitamin C, and capsaicinoids levels at the immature and mature stages for 800 different collections belonging to C. annuum, C. frutescence, C. baccatum and C. chinense. We employed genotyping by sequencing to generate ~30,000 SNPs for each species and assayed all the 800 collections for 120 genome-wide SSR markers. In our collections, α-carotene content ranged from 1.08 to 198.65 µg/g, β-carotene content from 10.48 to 538.92 µg/g and capsathin content from 7.01 to 184.12 µg/g in mature fruits. β-cryptoxanthin content ranged from 4.26 to 203.28 µg/g, zeaxanthin content from 1.33 to 221.38 µg/g, and lutein content from 1.87 to 480.92 µg/g. Lycopene content in orange and yellow peppers ranged from 1.11 to 308.77 µg/g DW. Means for α Carotene, β Carotene, Capsanthin, Lutein, Zeaxanthin, β Cryptoxanthin, and Lycopene for red pepper fruit accessions were 29.71, 23.44, 53.80, 54.15, 41.37, 25.33, and 63.91 µg/g respectively, in the mature (red) peppers. The α-carotene content ranged from 0.15 to 66.03 µg/g DW in ripe fruits and β-carotene content from 0.57 to 86.01 µg/g DW. Zeaxanthin content was in the range of 11.24 to 235.82 µg/g DW, and lutein content from 4.82 to 553.44 µg/g DW. C. chinense genotypes were contained more carotenoid levels than C. baccatum genotypes. The flavonoid concentrations were higher in immature stage than ripened fruits. The apigenin content ranged from 2.36 to 63.31 µg/g, myricetin content from 3.07 to 42.88 µg/g and quercetin content was in range of 19.16 to 559.77 µg/g DW in mature (red) fruits. Luteoline content ranged from 27.45 to 414.78 µg/g DW, kaempferol content ranged from 0.40 to 90.75 µg/g DW in the ripened peppers. In the green fruits, apigenin content ranged from 4.50 to 116.67 µg/g DW, myricetin content from 6.90 to 63.68 µg/g DW and quercetin content from 28.76 to 1464.52 µg/g DW. Luteoline content ranged from 29.00 to 355.79 µg/g DW, kaempferol content ranged from 1.17 to 174.32 µg/g DW in green peppers. Ripened fruits accumulated significantly higher amount of vitamin C than the green fruits. Vitamin C concentrations ranged from 6.091 to 13118.42 µg/g DW in mature (red) stage and 24.71 to 7769.16 µg/g DW in immatured (green) stage. Capsaicin range was from 9.51 to 32910.23 µg/g DW and dihydrocapsaicn was in range of 10.01 to 10088.97 µg/g DW. A cell culture study was conducted to examine the effect of Ghost pepper, one of the highest capsaicin containing peppers in the world, on cell proliferation, apoptosis, senescence and the global proteomic profile in human renal cell adenocarcinoma in vitro. 769-P human renal adenocarcinoma cells were cultured on RPMI-1640 media supplemented with fetal bovine serum (10%) and antibiotic-antimycotic solution (1%). Treatment stock solutions were prepared in ethanol. Cell proliferation was tested with phenol red-free media with capsaicin (0-400 µM), dihydrocapsaicin (0-400 µM), capsaicin + dihydrocapsaicin (5:1), and dry Ghost peppers (0-3 g L-1) for 24, 48 and 72 h. Polycaspase and senescence associated-beta-galactosidase (SA-beta-gal) activities were tested with capsaicin (400 µM), dihydrocapsaicin (400 µM), capsaicin (400 µM) + dihydrocapsaicin (80 µM), and Ghost pepper (3 g L-1) treatments. Global proteomic profile of cells in control and Ghost pepper treatment (3 g L-1) was analyzed after 6 h by a shotgun proteomic approach using tandem mass spectrometry. At 24 h after treatment (24 HAT), relative to control, cell proportion with capsaicin (400 µM), dihydrocapsaicin (400 µM), capsaicin (400 µM) + dihydrocapsaicin (80 µM), and Ghost pepper (3 g L-1) treatments was reduced to 36%, 18%, 33% and 20%, respectively, and further reduced at 48 and 72 HAT. All treatments triggered an early Polycaspase response. SA-beta-gal activity was normal or suppressed with all treatments. About 68,220 protein isoforms were identified by shotgun proteomic approach. Among these, about 8.2% were significantly affected by Ghost pepper. Ghost pepper regulated various proteins involved in intrinsic and extrinsic apoptotic pathways, Ras, Rb/E2F, p53, TGF-beta, WNT-beta catenin, and calcium induced cell death pathways. Ghost pepper also induced changes in proteins related to methylation, acetylation, genome stability, cell cycle check points, carbohydrate, protein and other metabolism and cellular mechanisms. Association mapping of various metabolites is in progress. Crosses made involving high metabolite peppers with cultivars are currently under evaluation.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Nimmakayala P. 2017. Genome-wide Divergence and Linkage Disequilibrium Analyses for Capsicum baccatum Revealed by Single Nucleotide Polymorphisms. Annual conference of the American Society for Horticultural Science, 1922 September 2017, Waikoloa, Hawaii.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Tandukar N, Saminathan T; Alaparthi S, Nimmakayala P, Hankins G and Reddy U.2018. Genome Wide Association studies on nutraceutical effects of various chili peppers on Drosophila melanogaster. 59th Annual Drosophila Research Conference , April 11-15, 2018, Philadelphia, PA.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Reddy UK, Nimmakayala P, Abburi VL, Ortiz C, Akinmoju T, Akinspe T, Suresh B. Alaparthi, Nadimi M, Vajja G, Kim S, Choi D, Stommel J. 2018. Genomewide chromosomal rearrangements and impact of positional effect of associated SNPs for capsaicinoids and fruit weight in various Capsicum spp. International Plant & Animal Genome XXVI Conference. January 8-13, 2018, San Diego, CA.
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