HORTSCIENCE 53(12):1772–1775. 2018. https://doi.org/10.21273/HORTSCI13574-18 Naga (Purkayastha et al., 2012). ‘Bhut Jolo- kia’ was considered one of the hottest in the world; in 2007, it was registered in the Guin- Behavior of the Hottest Chili Peppers ness Book of World Records with 1,001,304 SHU. Later, the ‘Trinidad Moruga Scorpion’ in the World Cultivated in variety, originally from Trinidad and Tobago, occupied that place in the Guinness Book of Yucatan, Mexico World Records with a pungency of 2,009,231 SHU (Bosland et al., 2012). Cur- Liliana S. Mun˜oz-Ramírez, Laura P. Pen˜a-Yam, Susana A. Aviles-Vin˜as, rently, the ‘Carolina Reaper’ variety, de- Adriana Canto-Flick, Adolfo A. Guzman-Antonio, veloped by the U.S. company PuckerButt and Nancy Santana-Buzzy1 Pepper in South Carolina, is recognized as the Unidad de Bioquímica y Biología Molecular, Centro de Investigacion most pungent in the world, with 2.2 million í · SHU (PuckerButt Pepper Company, 2013), and Cient fica de Yucatan A.C., Calle 43 No. 130 32 y 34, Merida, Yucatan, is registered in the Guinness Book of World CP 97205, Mexico Records. Taking into account that the Peninsula of Yucatan is known to distinguish the Haba- Additional index words. Capsicum, HPLC, pungency, capsaicinoids, Scoville heat units nero pepper for its high level of pungency, our Abstract. The Yucatan Peninsula is recognized as the center of genetic diversity of proposal was to evaluate, under the conditions Habanero peppers (Capsicum chinense Jacq.), which can be distinguished from those of the region, the pungency behavior of three cultivated in other regions of the world by their aroma, taste, and—most of all—by their varieties internationally known for their high pungency. We evaluated three commercial varieties of chili peppers reported as being the level of pungency. hottest in the world: ‘Bhut Jolokia’, ‘Trinidad Moruga Scorpion’, and ‘Carolina Reaper’. The aim of our study was to determine the behavior of the pungency when cultivated under the edaphoclimatic conditions of Yucatan. Our results show that the Materials and Methods three varieties registered greater contents in comparison with those reported in other regions of the world. ‘Carolina Reaper’—considered to be the hottest variety in the Plant material and growth conditions. For world, with a pungency of 2,200,000 Scoville heat units (SHU)—when cultivated in this study, three varieties of commercially Yucatan, had a pungency of 3,006,330 SHU, which was greater than all the other varieties available chili peppers—‘Trinidad Moruga analyzed. Scorpion’, ‘Bhut Jolokia’, and ‘Carolina Reaper’—were obtained from different sup- pliers on the Internet [Spicegarden (n.d.) and Chili peppers form part of the identity and noids are in great demand in the pharmaceutical PuckerButt Pepper Company (2018), respec- culture of Mexico. They belong to the genus industry as a result of their application as anti- tively]. They were planted from Sept. 2016 to Capsicum, which originated in America, inflammatories (Liu and Nair 2010; Tag et al., June 2017 in greenhouses located in the seed from Mexico to South America. The word 2014), antioxidants (Reddy et al., 2010; production unit of the Scientific and Techno- chili derives from the nahuatl chilli or xilli, Rodriguez-Maturino et al., 2012), and antican- logical Park of Yucatan in Sierra Papacal, and in Mexico it is used to refer to any fruit of cer agents (Jia et al., 2013). Merida, Yucatan, Mexico, at lat. 2107’20"N the genus Capsicum (Long-Solís, 1986). Mexico is one of the centers of domesti- and long. 8943’41"W, and at an altitude of Chili pepper is an extraordinary condiment cation and diversity of chili peppers. It is also 9 m.a.s.l. During the study period, the average with a high nutritional value. It is a source of where the greatest genetic diversity of the temperature was 18 C (minimum) and 37 C vitamins A, C, and E; minerals; and natural genus Capsicum is located (Ibiza et al., (maximum), and relative humidity was 86% dyes; in addition to its multiple industrial 2012). Yucatan is well known for its culinary (Servicio Meteorologico Nacional, 2018). uses, such as flavorings, sauces, powders, and tradition, which is associated closely with the The seeds were germinated in polystyrene oleoresins, among other products (Qin et al., trays with 200 cavities using commercial sub- Habanero pepper, considered to be one of the Ò 2014). It is the only plant species containing hottest chili peppers in the world. In Yucatan, strate called Peat Moss Premier (Green Forest, capsaicin, the most abundant metabolite Puebla, Mexico). The plantlets were trans- the Habanero pepper found particular condi-  among the capsaicinoids, which belongs to tions of soil and climate that allowed it to plantedtosoilafter 35 d, under greenhouse conditions, at a distance of 30 cm between the alkaloids and is responsible for the adapt and diversify in an environment very plants and 100 cm between rows. Plant man- pungency of the chili (Duelund and Mouritsen, different to that of its origin. These conditions agement and irrigation, pest and disease control, 2017). The capsaicinoids, integrated mainly by are associated with the high temperatures of the and fertilization were conducted following the capsaicin, dihydrocapsaicin, nordihydrocapsai- region, sparse rainfall, and stony soils (basically cin, and homocapsaicin, are produced in glands established standards applied to the cultivation calcareous with alkaline pH) very poor in located in the point of union of the placenta of Habanero pepper in the region (Tun, 2001). organic material (Borges-Gomez et al., 2014). with the pod wall and are accumulated in One hundred fifty plants per variety were used, In June 2010, the Designation of Origin was vacuoles of the epidermal cells of the placenta distributed in three rows of 50 plants. Each granted to ‘‘Habanero pepper of the Peninsula until they metabolize (Stewart et al., 2007; variety was evaluated based on the descriptors Suzuki et al., 1980). The total content of of Yucatan’’ (Diario Oficial de la Federacion, established for Capsicum (IPGRI,1995).The capsaicinoids is called pungency, which is 2010). Subsequently, the Official Mexican characteristics evaluated were type of epidermis, Standard (NOM-189-SCFI-2012) was gener- fruit color, weight of the fruit (measured in calculated and expressed in SHUs, derived from the organoleptic test for the pungency of ated (Diario Oficial de la Federacion, 2013). grams), pericarp thickness (measured in milli- the chili peppers (Scoville, 1912). This test has However, there are other varieties of the genus meters), fruit length (measured in centimeters), been replaced with high-performance liquid Capsicum originating from other regions of the fruit width (measured in centimeters), number chromatography (HPLC) (Collins et al., 1995; world that have been recognized for their high of fruits per plant, yield per plant (grams per Kozukue et al., 2005). Currently, the capsaici- pungency, such as ‘Bhut Jolokia’, a natural plant). hybrid of C. chinense and C. frutescens, Standards and solvents. Capsaicin (>99%) cultivated mainly in Bangladesh and India and dihydrocapsaicin (>97%) (Sigma Aldrich) (Meghvansi et al., 2010). It can be found with were used as external standards, whereas meth- Received for publication 14 Sept. 2018. Accepted other names, such as Bih Jolokia, Borbih anol, acetonitrile, and HPLC-grade water were for publication 10 Oct. 2018. Jolokia, Nagahari, Naga Jolokia, Naga Mor- used as solvents (J.T. Baker). The filtrates were 1Corresponding author. E-mail: [email protected]. ich, Naga Moresh, Raja Mirchi, and Dorset acquired using a 0.45-mm membrane filter

1772 HORTSCIENCE VOL. 53(12) DECEMBER 2018 Table 1. Morphoagronomic characteristics of the three varieties evaluated. Type of Thickness of No. of Varieties epidermis Color of fruit Length of fruit (cm) Width of fruit (cm) pericarp (mm) Wt of fruit (g) fruits/plant Yield (g/plant) Bhut Jolokia Wrinkled Red 5.28 ± 1.52 az 2.42 ± 1.03 b 1.46 ± 0.20 b 6.75 ± 0.32 b 219.7 ± 3.65 a 1482.2 ± 9.8 a Trinidad Moruga Scorpion Wrinkled Red 3.63 ± 1.64 b 2.97 ± 1.22 a 1.65 ± 0.19 a 7.77 ± 0.48 a 108.7 ± 4.09 b 846.7 ± 4.5 b Carolina Reaper Wrinkled Red 2.61 ± 2.63 c 2.37 ± 1.71 b 1.14 ± 0.10 c 3.70 ± 0.45 c 98.1 ± 2.76 c 362.9 ± 4.3 c zThe means followed by different letters in the same column are significantly different (P < 0.05) with the Tukey test, the data are shown as the mean value ± SD.

Fig. 1. Varieties evaluated include (A) ‘Bhut Jolokia’, (B) ‘Trinidad Moruga Scorpion’, and (C) ‘Carolina Reaper’.

gies, USA) column (4.6 mm i.d. · 250 mm) at a temperature of 25 C and an injection volume of 20 mL. The wavelengths used for the de- tection were 280 nm (excitation) and 338 nm (emission). The mobile phase was isocratic with 70% of solvent B (100% methanol) and 30% of solvent A (10% methanol solution v/v). The operating conditions of the HPLC for determining total capsaicinoids were a temper- ature of 25 C, a flow rate of 1 mL/min, and a runtime of 15 min. HPLC calibration. The standards of cap- saicin and dihydrocapsaicin were used to obtain a calibration curve based on the relation- ship of the maximum area for the known concentrations of external standards. The stock solution was prepared in 100% (v/v) methanol, and a total of five different concentrations—20, 60, 100, 200, and 300 ppm—were used to generate the calibration curve. Quantification of capsaicinoids. The con- Fig. 2. High-performance liquid chromatography chromatograms of capsaicin (1) and dihydrocapsaicin (2) (A) standard. (B) Sample of ‘Trinidad Moruga Scorpion’. centration of the two major capsaicinoids (capsaicin and dihydrocapsaicin) was esti- mated using the calibration curve obtained. The quantified concentration of capsaicinoids is reported in milligrams per gram dry weight (HVHP0470; Millipore) and degasified before at 80 C for 4 hours with periodic agitation. (DW). The capsaicin contents obtained in the use. The extracts were allowed to cool at room three varieties were converted to SHU to classify Extraction of capsaicinoids. The extrac- temperature, after which they were centrifuged them according to their different levels of tion of capsaicinoids was performed accord- (Sigma 2-16kl) at 17,968 gn for 10 min at 4 C. pungency. This conversion to SHU was per- ing to the methodology reported by Collins The supernatant was collected and filtered formed by multiplying the capsaicin content by et al. (1995), with slight modifications (100 through syringe filters with a polytetrafluoro- the coefficient corresponding to the pungency mg/40 mL, w/v). Mature fruit was picked ethylene (PTFE) membrane of 0.45 mm(no. value for pure capsaicin (1.6 · 107)(Sanatombi randomly per row of each variety, cut verti- 721–1345; Thermo Scientific) in 2-mL vials of and Sharma, 2008). cally, and transferred to an aluminum tray to amberglassandfinallystoredat4C until the Statistical analysis. The experiment was a be dried in an oven with a circulation of air chromatographic analysis was performed. random block design, with three repetitions. for 48 h, after which the dried fruit were Analysis of capsaicinoids. Separation and The data obtained are shown as the mean ± ground until a fine homogeneous powder was quantification of the capsaicinoids was carried SD. To detect significant differences among obtained. The powder was stored at –80 C. out using HPLC (Agilent series 1200) equipped capsaicinoid profiles of the chili varieties To extract the capsaicinoids, three replicates with an automatic injector and a fluorescence evaluated, an analysis of variance and a of 100 mg fruit powder were mixed with 40 detector. The capsaicinoids were separated by a Tukey test (P < 0.05) were carried out using mL acetonitrile and maintained in water bath Zorbax Eclipse Plus C18 (Agilent Technolo- the software SPSS (version 22) for Windows.

HORTSCIENCE VOL. 53(12) DECEMBER 2018 1773 Table 2. Concentration of capsaicinoids analyzed in milligram per gram of dry weight and pungency of the chili varieties investigated. Varieties Capsaicin (mg·g–1 DW) Dihydrocapsaicin (mg·g–1 DW) Total capsaicinoids (mg·g–1 DW) Pungency (SHU) Bhut Jolokia 92.83 ± 5.53z by 27.55 ± 1.79 b 120.38 ± 7.33 c 1,938,089 ± 118,030 c Trinidad Moruga Scorpion 116.40 ± 5.15 a 30.71 ± 1.26 b 147.11 ± 6.42 b 2,368,534 ± 103,395 b Carolina Reaper 123.43 ± 1.77 a 63.30 ± 0.97 a 186.73 ± 2.75 a 3,006,330 ± 44,384 a z Each value is the average of three samples ± the SD of the mean. yAverages followed by different letters in the same column are significantly different (P < 0.05) with the Tukey test. DW = dry weight; SHU = Scoville heat units.

iability, with a range of 11.95 to 72.05 mg·g–1 DW and a pungency of 191,135 to 1,152,832 SHU. In our study, the content of capsaici- noids registered for ‘Bhut Jolokia’ was 120.38 mg·g–1 DW, which corresponds to a pungency of 1,938,089 SHU, surpassing by 48% that reported by Bosland and Baral (2007) (Fig. 3). In 2012, Bosland et al., working with ‘Trinidad Moruga Scorpion’ cultivated in New Mexico, reported a pun- gency of 2,009,231 SHU. ‘Trinidad Moruga Scorpion’, cultivated under the conditions of Yucatan, registered a pungency of 2,368,534 SHU (147.11 mg·g–1 DW), which represents a 15% increase in the content of capsaicinoids (Fig. 3). However, the most significant pun- gency was observed in ‘Carolina Reaper’ cultivated in the state of Yucatan. This cultivar registered 3,006,330 SHU, with a content of 186.73 mg·g–1 DW total capsaicinoids, which Fig. 3. Pungency [Scoville heat unit (SHU)] of the varieties analyzed in this study compared with those represents a 27% increase in the content of reported by PuckerButt Pepper Company (2013) (•), Bosland et al. (2012) (••), and Bosland and Baral capsaicinoids in comparison with those re- (2007) (•••). ported in the Guinness Book of World Records (PuckerButt Pepper Company, 2013). Results and Discussion standards, for each compound. The chro- It has been documented by different matograms shown in Fig. 2 correspond to authors that the accumulation of capsaici- Table 1 shows the results of the analysis the standard and the extract of ‘Trinidad noids and the pungency of the pepper is of the characteristics evaluated for ‘Bhut Moruga Scorpion’, for which it was possible influenced by different factors, including Jolokia’, ‘Trinidad Moruga Scorpion’, and to identify capsaicin and dihydrocapsaicin, genotype, environment conditions, nutri- ‘Carolina Reaper’. As can be seen, they with retention times of 7.46 min and 10.62 tional regime, and ripeness of the fruit differ significantly in all characteristics min, respectively. The calibration curves (Barbero et al., 2014; Zewdie and Bosland, evaluated. ‘Bhut Jolokia’ presented the showed a linear behavior over the concentra- 2000). Tiwari et al. (2005) compared the greatest fruit length (5.28 cm), which dif- tion, in a range of 20 to 300 ppm of capsaicin capsaicin and dihydrocapsaicin content in fered significantly from ‘Trinidad Moruga and dihydrocapsaicin; the coefficient of cor- Bhut Jolokia cultivated in two regions of Scorpion’ and ‘Carolina Reaper’ (3.63 cm India—Tezpur (Assam) and Gwalior (Madhya relation (r) was 0.99912 and 0.99931, re- and 2.61 cm, respectively); ‘Trinidad Mor- Pradesh)—with different climatic conditions. spectively. uga Scorpion’ outperformed ‘Bhut Jolokia’ Their results showed that the pungency of the The data obtained for pungency (Table 2) and ‘Carolina Reaper’ significantly with pepper was reduced by more than 50% in show the concentration of capsaicin and regard to fruit weight, fruit width, pericarp Gwalior, a dry region of India, which demon- dihydrocapsaicin, as well as the pungency thickness, number of fruits per plant, and strates the effect of environmental conditions expressed in SHU of the three cultivars yield per plant. ‘Carolina Reaper’ had the on pepper pungency. Duelund and Mouritsen lowest values for all characteristics evalu- analyzed. The total content of capsaicinoids (2017) quantified the content of capsaicinoids ated. Figure 1 shows the distinctive charac- obtained in this study was within a range of in ‘Carolina Reaper’ cultivated in Denmark. · –1 teristics of the fruit analyzed. There are 120.38 to 186.73 mg g DW, differing sig- They obtained a pungency of 1,046,000 SHU, differences among the three cultivars in nificantly from each other. which was low compared to our result terms of the shape of the fruit; however, ‘Bhut Jolokia’ has been considered one of (3,006,330 SHU). In a study conducted by they have in common the rugged nature of the most pungent chili varieties in the world, Canto-Flick et al. (2008), the content of the pericarp. Values similar to those of this although this characteristic has shown a high capsaicinoids from 18 accessions of Habanero study have been reported for fruit weight, variation depending on the region where it is pepper collected in the Yucatan, Mexico, was fruit length, and fruit width in ‘Bhut Jolokia’ cultivated. Mathur et al. (2000), in a study determined. They reported pungency values (Bhagowati and Changkija, 2009; Kundu conducted with this variety in India, reported between 405,228 SHU and 892,719 SHU, et al., 2015). a pungency of 855,000 SHU, whereas Bos- which are significantly greater than the Identification and quantification of land and Baral (2007) reported 1,001,304 reports made for Habanero pepper grown capsaicinoids. To separate the two major SHU when it was grown in New Mexico, in other regions of Mexico (Orellana- capsaicinoids (capsaicin and dihydrocapsai- which allowed them to record it in the Guin- Escobedo et al., 2013). Similarly, our results cin) responsible for 90% of the pungency ness Book of World Records as the most spicy allow inferring that the capsaicinoids are (Manirakiza, 2003) using HPLC, the capsai- variety in the world, displacing ‘Red Savina’ highly influenced by the environment and cinoid content in the chili fruit was deter- (577,000 SHU). Subsequently, Islam et al. that the edaphoclimatic conditions of the mined from the retention time and the peak (2015) evaluated the content of capsaicinoids Yucatan Peninsula exert a favorable effect size for each capsaicinoid present, in relation in 92 accessions of ‘Bhut Jolokia’ collections for the increase of these compounds in these to the time of retention of the commercial in northwestern India. They found high var- species.

1774 HORTSCIENCE VOL. 53(12) DECEMBER 2018 Conclusions Origen Chile Habanero de la Península de World Record: Smokin’ Ed’s ‘Carolina Yucatan. 1 Mar. 2018. hottest chili pepper Given that the Habanero pepper culti- detalle.php?codigo=5145315&fecha=04/06/2010>. in the world! 15 May 2018. . flavor—attributes conferred by the edapho- prueba. 30 Nov. 2012. . puckerbuttpeppercompany.com/collections/ of this study corroborate that pungency is Duelund, L. and O.G. Mouritsen. 2017. Contents of carolina-reaper-worlds-hottest-pepper/products/ greatly affected by the environment, includ- capsaicinoids in chillies grown in Denmark. smokin-eds-carolina-reaper>. ing climate and soil conditions. We analyzed Food Chem. 221:913–918. Purkayastha, J., S.I. Alam, H.K. Gogoi, L. Singh, the three pepper cultivars—Bhut Jolokia, Ibiza, V.P., J. Blanca, J. Canizares,~ and F. Nuez. and V. Veer. 2012. Molecular characterization Trinidad Moruga Scorpion, and Carolina 2012. Taxonomy and genetic diversity of do- of ‘Bhut Jolokia’, the hottest chilli. J. Biosci. Reaper—that have been published in the mesticated Capsicum species in the Andean 37:757–768. Guinness Book of World Records, at different region. Genet. Resources Crop Evol. 59(6):1077– Qin, C., C. Yu, Y. Shen, X. Fang, L. Chen, J. Min, times, for being the hottest peppers in the 1088. and Y. Yang. 2014. Whole-genome sequencing world. When these cultivars were grown in IPGRI (International Plant Genetic Resources In- of cultivated and wild peppers provides insights the Yucatan, their capsaicinoid content in- stitute). 1995. Descriptores para Capsicum spp. into Capsicum domestication and specializa- Instituto Internacional de Recursos Fitogeneticos, tion. Proc. Natl. Acad. Sci. USA 111:5135– creased by 48%, 15%, and 27%, respectively. Roma, Italia; Centro Asiatico para el Desarrollo 5140. ‘Carolina Reaper’, currently considered the y la Investigacion Relativos a los Vegetales, Reddy, K.K., T. Ravinder, R.B. Prasad, and S. hottest in the world (2,200,000 SHU), when Taipei, Taiwan y Centro Agronomico Tropical Kanjilal. 2010. Evaluation of the antioxidant cultivated in Yucatan reached a pungency of de Investigacion y Ensenanza,~ Turrialba, Costa activity of capsiate analogues in polar, non- 3,006,330 SHU. We consider it important to Rica. 52 p. polar, and micellar media. J. Agr. Food Chem. emphasize that Yucatan is a particularly Islam, M.A., S.S. Sharma, P. Sinha, M.S. Negi, B. 59(2):564–569. privileged region in the world for the culti- Neog, and S.B. Tripathi. 2015. Variability in Rodriguez-Maturino, A., A. Valenzuela-Solorio, vation of Habanero pepper, which is distin- capsaicinoid content in different landraces of R. Troncoso-Rojas, D. Gonzalez-Mendoza, O. guished by its high level of pungency. Our Capsicum cultivated in north-eastern India. Grimaldo-Juarez, M. Aviles-Marin, and L. results provide solid evidence that, not only is Scientia Hort. 183:66–71. Cervantes-Diaz. 2012. Antioxidant activity Jia, S.S., G.P. Xi, M. Zhang, Y.B. Chen, B. Lei, and bioactive compounds of Chiltepin (Capsi- capsaicin production stimulated, but also it is X.S. Dong, and Y.M. Yang. 2013. Induc- cum annuum var. glabriusculum) and Habanero potentiated at significantly greater levels, tion of apoptosis by D-limonene is mediated (Capsicum chinense): A comparative study. J. even in cultivars that have been recognized by inactivation of Akt in LS174T human Med. Plants Res. 6(9):1758–1763. for their high level of pungency. colon cancer cells. Oncol. Rpt. 29(1):349– Sanatombi, K. and G.J. Sharma. 2008. Capsaicin 354. content and pungency of different Capsicum Literature Cited Kozukue, N., J.S. Han, E. Kozukue, S.J. Lee, J.A. spp. cultivars. Not. Bot. Hort. Agrobot. Cluj- Kim, K.R. Lee, and M. Friedman. 2005. Anal- Napoca 36:89–90. Barbero, G.F., A.G. Ruiz, A. Liazid, M. Palma, J.C. ysis of eight capsaicinoids in peppers and Scoville, W.L. 1912. Note on capsicums. J. Pharm. Vera, and C.G. Barroso. 2014. Evolution of pepper-containing foods by high-performance Sci. 1:453–454. total and individual capsaicinoids in peppers liquid chromatography and liquid chromatography– Servicio Meteorologico Nacional. 2018. Climato- during ripening of the cayenne pepper plant logía: Datos Estadísticos. 8 Mar. 2018. . pounds in pepper due to agro-climatic Spicegarden. n.d. Spicegarden, la tienda online de King Chili landraces of Nagaland. Asian Agrihist. 13:171–180. discrepancy: A case study with Capsicum semillas, especias y plantas aromaticas. 18 June Borges-Gomez, L., C. Moo-Kauil, J. Ruiz-Novelo, chinense ‘Bhut Jolokia’ fruit. Intl. J. Pharm. 2016. . Yam-Chimal, and F. Can-Puc. 2014. Soils used Liu, Y. and M.G. Nair. 2010. Capsaicinoids in the Stewart, C., M. Mazourek, G.M. Stellari, M. for habanero chili production in Yucatan: Pre- hottest pepper ‘Bhut Jolokia’ and its antioxi- O’Connell, and M. Jahn. 2007. Genetic control dominant physical and chemical characteris- dant and antiinflammatory activities. Nat. Prod. of pungency in C. chinense via the Pun1 locus. tics. Agrociencia (Montecillo) 48:347–359. Commun. 5(1):91–94. J. Expt. Bot. 58:979–991. í Bosland, P.W. and J.B. Baral. 2007. ‘Bhut Jolokia’: Long-Sol s, J. 1986. Capsicum y cultura: La Suzuki, T., H. Fujiwake, and K. Iwai. 1980. In- The world’s hottest known chile pepper is a historia del chilli. Fondo de Cultura Econo- tracellular localization of capsaicin and its putative naturally occurring interspecific hy- mica, Mexico. analogs, capsaicinoid, in Capsicum fruit: 1. brid. HortScience 42:222–224. Manirakiza, P.A. 2003. Pungency principles in Cap- Microscopic investigation of the structure of Bosland, P.W., D. Coon, and G. Reeves. 2012. sicum: Analytical determinations and toxicology, the placenta of Capsicum annuum var. annuum ‘Trinidad Moruga Scorpion’ pepper is the p. 71–86. Taylor and Francis, London, UK. cv. Karayatsubusa. Plant Cell Physiol. 21:839– world’s hottest measured chile pepper at more Mathur, R., R.S. Dangi, S.C. Das, and R.C. 853. than two million Scoville heat units. HortTech- Malhotra. 2000. The hottest chilli variety in Tag, H.M., O.E. Kelany, H.M. Tantawy, and A.A. nology 22:534–538. India. Curr. Sci. 79:287–288. Fahmy. 2014. Potential anti-inflammatory ef- Canto-Flick, A., E. Balam-Uc, J.J. Bello-Bello, C. Meghvansi, M.K., S. Siddiqui, M.H. Khan, V.K. fect of lemon and hot pepper extracts on Lecona-Guzman, D. Solis-Marroquin, S. Aviles- Gupta, M.G. Vairale, H.K. Gogoi, and L. adjuvant-induced arthritis in mice. J. Basic Vinas, E. Gomez-Uc, G. Lopez-Puc, N. Santana- Singh. 2010. Naga chilli: A potential source Appl. Zool. 67(5):149–157. Buzzy, and L.G. Iglesias-Andreu. 2008. Capsaicinoid of capsaicinoids with broad-spectrum ethno- Tiwari, A., M.P. Kaushik, K.S. Pandey, and R.S. content in Habanero pepper (Capsicum chinense pharmacological applications. J. Ethnopharma- Dangi. 2005. Adaptability and production of Jacq.): Hottest known cultivars. HortScience 43: col. 132:1–14. hottest chilli variety under Gwalior agro- 1344–1349. Orellana-Escobedo, L., L.E. Garcia-Amezquita, climatic conditions. Curr. Sci. 88:1545–1546. Collins, M.D., L.M. Wasmund, and P.W. Bosland. G.I. Olivas, J.J. Ornelas-Paz, and D.R. Sepulveda. Tun, D.J. 2001. Chile habanero características y 1995. Improved method for quantifying cap- 2013. Capsaicinoids content and proximate com- tecnología de produccion, p. 13–24. INIFAP, saicinoids in capsicum using high-performance position of Mexican chili peppers (Capsicum spp.) Merida, Yucatan, Mexico. liquid chromatography. HortScience 30:137– cultivated in the State of Chihuahua. CYTA J. Zewdie, Y. and P.W. Bosland. 2000. Evaluation of 139. Food 11:179–184. genotype, environment, and genotype-by- Diario Oficial de la Federacion. 2010. Declaratoria PuckerButt Pepper Company. 2013. Smokin’ environment interaction for capsaicinoids in General de Proteccion de la Denominacion de Ed’s ‘Carolina Reaper’Ò wins Guinness Capsicum annuum L. Euphytica 111:185–190.

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