Ethnic Uses and Commercial Applications of Capsicum assamicum (Bhut jolokia)

Ajitabh Bora, Khonamai S. Nakhuru, Baikuntha J. Cogoi, Pronobesh Chattopadhyay and Sanjai K. Dwivedi

1.1 INTRODUCTION

Capsicum assamicum, an extremely pungent chilli, is endemic to northeast (NE) India, particularly in the states of Assam, Nagaland, and Manipur. The NE region, lying between 20.59° latitude and 78.96° longitude, has a unique environment with a high degree of humidity, which has led to the development of extreme pungency in this chilli. It is known by different names and Bhut jolokia is the name as it is called in Assam, while it is called Naga chilli in Nagaland.

The identity of this species has been a point of controversy since its discovery, regarding its phylogenetic position. Mathur et al. (2000) reported the Bhut jolokia to be a variety of Capsicum frutescens L., while in the Guinness Book of World Records (2006), Bhut jolokia has been recorded as belonging to the species of C. chinense (Purkayastha et al. 2012b). On the other hand, Bosland and Baral (2007) reported Bhut jolokia to be a natural hybrid of C. frutescens and C. chinense based on Random Amplified Polymorphic DNA (RAPD) analysis. But the RAPD technique has several limitations, such as non-reproducibility of results, competition between different DNA fragments for amplification (Williams et al. 1993; Hallden et al. 1996), and identical mobility of fragments with different sequences (Xu 2006), which limits its uses for interpopulation or interspecies comparisons. Purkayastha et al. (2012b) undertook molecular studies of Bhut jolokia to determine the distinctness of this species from C. frutescens and C. chinenese through sequencing of the ribosomal RNA gene-internal transcribed region along with its phylogenetic analysis. The phylogenetic analysis of the internal transcribed 1,5.8S, and internal transcribed 2 sequences lead to a distinct clade for all the accessions of Bhut jolokia, while C. frutescens and C. chinenese occupied discrete lineages. Further, a unique 13 base pair deletion was observed in all the accessions of Bhut jolokia, making it distinct from all other chilli species. The morphological features of Bhut jolokia, such as profusely branched habitat, yellow green corolla, pale blue anther, orange red color, and sub-conical to conical fruits with rough, uneven, dented skin, and anatomical details of the stem make its distinct from C. frutescens and C. chinense (Purkayastha et al. 2012a). Moreover, phylogeny, based on median joining network analysis, revealed clear distinction of the Naga King Chilli from its closest relatives (C. chinense and C. frutescens) and the absence of a star-like network of hap- lotypes indicates the expansion of an ancient population of this chilli species. Besides, the differential proteomic analysis of Bhut jolokia revealed 22 differential protein spots, which appeared only in this species, but not in C. frutescens and C. chinense. Thus, Bhut jolokia has been established as a distinct species within the genus Capsicum and christened as C. assamicum Jubilee Purkayastha & L. Singh sp. nov. (Purkayastha et al. 2012a). This was further validated by Kehie et al. (2016), who also observed distinct 13 base deletions in the 5.8S gene in all the accessions of the Naga King Chilli, unlike other Capsicum species. These works clearly establish Bhut jolokia or Naga King Chilli as a distinct species of chilli endemic to northeast India. This chilli has been patented under the Geographical Indication Registry by the Government of Nagaland (Kehie et al. 2014).

1.2 BOTANY, DISTRIBUTION, AND DIVERSITY

Capsicum assamicum, belonging to the family Solanaceae, is a self-pollinated crop, though cross-pollination occurs (10%) when the insect population is high. Under optimum conditions, the plant behaves as perennials and grows to a height of 50-100 cm or sometimes even taller. The plant is the most similar to C. frutescens and C. chinense, but can be distinguished by its profusely branched habitat, yellow green corolla, pale blue anther, orange red color, and sub-conical to conical fruits with rough, uneven, dented skin, and anatomical details of the stem.

The leaves are ovate in shape and the size ranges from 5.5-7.5 cm in width and 10-14 cm in length. Flowers are pendant shaped, having creamy white corollas. The fruit is usually oblong in shape with a rough texture and is bright red or orange in color, which varies with soil and climate. A single fruit bears about 25-35 slightly wrinkled seeds and weighs around 5 gram (g) on average. A single plant produces, on average, around 15-20 full-sized fruit and 10-14 smaller ones (Borgohain and Devi 2007) (Figures 1.1 and 1.2).

Bhut jolokia is widely cultivated in northeast India, mainly in the states of Assam, Manipur, and Nagaland. This chilli is known by different names in the regions. It is popularly called Bhut jolokia in Assam. The term Bhut means ghost

FIGURE 1.1 Bhut jolokia plant.

FIGURE 1.2 Bhut jolokia fruits.

and jolokia means chilli in the Assamese language, which may be due to its ghostly bite or due to its introduction by the Bhutias from Bhutan according to another legend. It is also called Bih jolokia or Borbih Jolokia (Bih meaning poison, indicating high hotness of the chilli fruit) and by other names, such as Nagahari, Naga Morich, Naga Moresh, Naga Jolokia (named after the once ferocious “Naga” warrior tribe of northeast India), Raja Mirchi, and Dorset Naga (its derivative from Bangladesh).

In Nagaland, it is known as Naga King Chilli and in the local Angami dialect of Nagaland as “Kedi Chusi,” meaning the “King of Chillies” (Kehie et al. 2016). In Manipur, it is known as Omorok (Oo meaning tree and morok meaning chilli in the Manipuri language, probably due to the perennial growing habit of this chilli like a tree) (Talukdar et al. 2015). Besides, it is also known by several other names as Saga jolokia, Indian mystery chilli, India rough chilli (due to the rough texture of the fruits), and ghost pepper by the Western media (Meghvansi et al. 2010). Regardless of its name, it is cultivated and consumed in different states of northeast India as a spice or eaten raw with staple food.

Diversity of this chilli is seen in terms of fruit size, color, and bearing capacity, which may be termed as land races. Three colors of matured fruit can be found viz. orange, light to dark red, and dark chocolate. Two types of plants can be classified based on the immature fruit color which bears light and dark green. The color of the immature fruit is green in color, which turns to orange or red or chocolate on maturity depending on the landraces (Figures 1.3 and 1.4).

FIGURE 1.3 Different types of Bhut jolokia fruits.

FIGURE 1.4 Unripe fruits of Bhut jolokia.

1.3 ETHNIC USES

Traditionally, Bhut jolokia is used as a spice or consumed raw due to its pleasant and palatable aroma that is unique to this chilli. The chilli is an integral part of the local cuisine of most of the tribes of the NE region (Table 1.1). Since time immemorial, this chilli has been used in treating a myriad of human ailments such as asthma, gastrointestinal abnormalities, toothache, muscle pain, arthritis, and removal of puss from boils as seen in Table 1.1 (Bhagowati and Changkija 2009). Besides its use in treating various human ailments, the chilli is smeared on fences or used in smoke bombs to ward off elephants since the smell of chilli is deterrent to elephants. The elephants get repelled as this chilli is highly pungent, it is quite effective in repelling the elephants.

1.4 CHEMISTRY OF HIGH HOTNESS

The hotness and pungency of a chilli is due to the presence of capsaicinoids, a group of lipophilic alkaloids produced by the plants as a self-defense mechanism against the invasion of predators and herbivores. Among the capsaicinoids, capsaicin and dihydrocapsaicin are responsible for 90% of the pungency in most of the chilli species (Suzuki et al. 1980). These compounds are found only in the members of the genus Capsicum and are mainly localized in the placenta and flesh of the chilli fruit. The gland on the placenta of the fruit produces capsaicinoids (Bosland 1996). One fruit of Bhut jolokia can sustain intense heat sensation in the mouth for up to 30 minutes before subsiding (Purkayastha et al. 2012b).

Conventionally, an organoleptic test is being used to measure the hotness of chillies. This test was first developed by Wilber Scoville in 1912 and, hence, the unit of measure of hotness was given as Scoville Heat Unit (SHU). However, in modern times, with the advent of sophisticated instruments, the capsaicinoid content is being estimated more accurately with the help of High Performance

TABLE 1.1

Few Ethnic Uses of Capsicum assamicum in Treating Human Aliments

Liquid Chromatography (HPLC) and spectrophotometer. Pure capsaicin measures 15,000,000-16,000,000 SHUs. The higher the capsaicinoid content, the higher is the hotness (Borgohain and Devi 2007). Unlike other chilli species, cultivated in the Indian sub-continent, which contain around 1% capsaicin, Bhut jolokia has around 3%-5% capsaicin (Mathur et al. 2000; Sanatombi and Sharma 2008). When Mathur et al. (2000) reported hotness of Bhut jolokia to be 855,000 SHUs, it superseded the Red Savina habanero of Mexico whose pungency was

577,000 SHUs.

Bhut jolokia was officially recognized as the world’s hottest chilli in the year 2006, measuring 1,001,304 SHUs after testing its hotness by the New Mexico State University, thereby replacing the Red Savina, which had a SHU of 248,566 (Guinness Book of World Records 2006; Bosland and Baral 2007). However, after holding its glory for 5 years, Bhut jolokia has been superseded by other chilli species in subsequent years, as in 2011 by the Infinity chilli, in 2012 by the Naga Viper, in 2012 by the Trinidad Moruga Scorpion, and in 2013 by the Carolina Reaper (Kehie et al. 2016) until now. Presently, Bhut jolokia holds seventh position in the list of world’s hottest chilli—2018.

The pungency in a Bhut jolokia is affected by the climatic conditions and the time of cultivation. High rainfall and humidity w'ith the moderate climate of northeast India favors the development of its extreme pungency. When this chilli was cultivated elsewhere, lower levels of pungency were recorded. For example, when it was grown in Gwalior, there was a reduction of capsaicin and dihyrocapsaicin by 50% (Tiwari et al. 2005), and when cultivated in Pithoragarh, in Uttarakhand, the pungency level came down to 254,896 SHUs (Pandey et al. 2009).

1.5 NUTRITIVE VALUE

The anti-oxidant and anti-radical potentials of Bhut jolokia were investigated in vitro to validate the claims of its uses by the local folks. It was found to have immense potential as a natural source of anti-oxidant and anti-radical agents, as given in Figure 1.5 with half maximum inhibitory concentration (IC₅₀) values of 74.42±2.31, 25.00±0.31, and 2.15±0.04 against 2, 2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2'-Azino-bis-3- ethylbenzothiazoline-6-sulfonic acid (ABTS), and hydroxyl, respectively (Nakhuru et al. 2018). Therefore, this chilli is a potential candidate for exploitation as a source of natural antioxidant for dietary supplements and for pharmaceutical/cos- metic uses.

1.6 COMMERCIAL APPLICATIONS

Bhut jolokia has tremendous potential for its use as a value- added product in the form of dry chilli, as whole, pow'dered, or flakes, paste, distilled oil for flavor, pickles, sauce, etc., which needs to be explored. This chilli is gaining popularity in the Western world and is a popular ingredient of several recipes of restaurants worldwide. In recent times, Bhut jolokia sauce,

FIGURE 1.5 Anti-oxidant and radical scavenging activities of aqueous methanol extract of Blmt jolokia fruit in different concentrations. Each value represents mean±SD. (a) DPPH, (b) ABTS, and (c) hydroxyl.

prepared from the chilli paste, is gaining much popularity for its off-the-shelf uses in various cuisines apart from the traditional recipes. The sauce is available in online marketing platforms, which has given it the much-needed hype and popularity. The sauce is being produced by Naga Spices & Herbs in Senapati, Manipur under the brand name Poumai and marketed by Lunar Agro Chemicals. Another start-up firm, Zizira, based in Meghalaya, is marketing a couple of products based on В hut jolokia, while another firm, East by Northeast (ENE), has launched three variants of the Bhut jolokia sauce—sweet, hot, and extra hot to cater to different degrees of endurance of the taste buds. Frontech Agritech Pvt. Ltd. in Jorhat, an agri-business company, is involved in the exporting of Bhut jolokia to markets in Europe and the United States (Figure 1.6).

FIGURE 1.6 Dried fruits of Bliut jolokia.

Due to the high capsaicin content of Bhut jolokia, it has immense potential for commercial extraction of the oleoresin, which has wide applications in pharmaceutical as well as in defense sectors. Oleoresin contains capsaicin, which is in great demand both in domestic as well in international markets. The capsaicin compound has several pharmacological activities, such as hepatoprotective, anti-inflammatory, analgesic, anti-obesity, anti-cancer, anti-oxidants, anti-microbial, anti-pyretic, anti-diabetic, etc. (Meghvansi et al. 2010; Roy 2016). Defence Research Laboratory, DRDO, Tezpur (Assam) has found unusual applications of this chilli for mob dispersal and self-defense. The oleoresin extracted from Bhut jolokia is being used in chilli grenades and chilli spray in different versions and forms. The chilli grenade is mainly meant for use by law enforcement agencies as a non-lethal mob dispersal agent in lieu of tear gas, which is harmful to human eyes. The chilli grenade has been tested by state police of different states and has been inducted in their armory. The Indian Army has also used it for expelling terrorists from their hideouts in Jammu & Kashmir, which proved to be very useful in such circumstances. The chilli spray is meant for use by ladies and senior citizens for their self-defense, which they can carry along with them and can spray in the eyes of assaulters (Figure 1.7).

1.7 AGRO-TECHNOLOGY

The climatic condition of northeast India favors the natural growth of Bhut jolokia and the subsequent development of its unique extreme hotness without much care. Traditionally, it was grown as a sporadic intercrop with paddy under Jhum cultivation in the hills, as well as in homestead gardens. In the NE region, there are two planting seasons for this chilli, Kharif and Rabi. In the hilly states, Kharif cultivation is followed where planting is done in the month of February/ March and harvesting starts from May/June onward, while Rabi is followed in the plains where planting is done

FIGURE 1.7 Capsigrenade (a), capsispray for personal protection (b), and mob dispersal (c).

in August/September and harvesting from November/ December. In the homestead garden, it is cultivated under partial shade and the plants are kept for 2-3 years, beyond w'hich, there is a reduction in fruit size and yield. The plants grow to a height of 12—13 feet at the age of 3 years. The plants are reported to be infested by aphid and mealy bugs in traditional fields. The farmers usually sprinkle sieved ash on the leaves and the base of the plant as an indigenous crop management practice (Borgohain and Devi 2007; Bhagowati and Changkija 2009). Though no package of practices for Bhut jolokia could be found, Singh (2009) reported highest cost: benefit ratio (1:8.7) on sowing the crop in September 15 and planting at a spacing of 75 cm X 75 cm followed by October 15 sowing (1:7.3).

The average yield of fresh fruits of Bhut jolokia under rainfed conditions is estimated to be around 80-100 q/ha, w'hich after dehydration comes to around 10-12 q/ha (Borgohain and Devi 2007). The price per kilogram of the fruit of Bhut jolokia in local markets is Rs. 100-150, as reported by Borgohain and Devi (2007), while the current price in the Tezpur region ranges from Rs. 400- 600 depending upon the season. Since the cultivation of Bhut jolokia is an unorganized sector spread out in different pockets of northeast India, there is no documented data available on area, production, and yield loss due to diseases. As per a rough estimate, 1000 tons of Bhut jolokia is produced per year in different regions of northeast India, which may increase up to five times, if environmental factors are conducive (Meghvansi et al. 2010).

1.8 CONCLUSION

Bhut jolokia or Naga King Chilli is a chilli species of northeast India with a high potential for commercial applications. This chilli crop brought fame and glory to this region by being the world’s hottest chilli and, if explored judiciously with proper post-harvest handling and good market network, can bring about an economic upsurge for the farmers of this region. With the increase in popularity and demand for this native chilli of NE India, the first King Chilli Festival was organized in the Tamenglong District of Manipur in 2018 to attract tourists as well as entrepreneurs for setting up post-harvest processing units for export of this chilli in the form of value- added products.

Due to the rising demand of this chilli, both in the domestic and international markets, this chilli is being cultivated on a wide scale in different regions of northeast India. The chilli is being sold in the open market at the price of Rs. 300-400 per kg in Assam depending upon the season and size of the fruit. They are also sold loose at 3-4 fruits for ten rupees. This reflects the high demand of this chilli for domestic consumption, since the supply is not adequate as the cultivation is restricted to only a few areas owing to the suitability of the agro-climate. Since the availability of the chilli is restricted to only a few months of the year, the demand is high in the market. Moreover, extraction of capsaicin is limited to the time of availability of the fruit, for which, R&D efforts are being made by the Defence Research Laboratory, Tezpur for in vitro production of capsaicin through cell suspension culture. Breakthroughs have been made at laboratory scale, which need to be scaled-up, for which further studies are being carried out.

REFERENCES

Bhagowati, R.R., and Changkija, S. 2009. Genetic variability and traditional practices in Naga King Chilli landraces of Nagaland. Asian Agri-History 13: 171-180.

Borgohain, R., and Devi, J. 2007. The hottest chilli: A new horizon in agri-entrepreneurship. Entrepreneur August 2007. www. assamagribusiness.nic.in

Bosland, RW. 1996. Capsicums: Innovative uses of an ancient crop. In Janick, J., (Ed.), Progress in New Crops. ASHS Press, Arlington, VA. 479-487.

Bosland. P.W.. and Baral, J.B. 2007. ‘Bhut jolokia’—The world’s hottest known chille pepper is a putative naturally occurring interspecific hybrid. HortScience 42: 222-224.

Guinness Book of World Records. 2006. http://en.wikipeia.org/wiki/ Bhut_jolokia.

Hallden, C„ Hansen. M.. Nilsson. N.O.. Hjerdin. A., and Sail. T. 1996. Competition as a source of errors in RAPD analysis. Theoretical and Applied Genetics 93: 1185-1192.

Kehie, M., Kumaria, S., Devi, K.S., and Tandon, P. 2016. Genetic diversity and molecular evolution of Naga King Chilli inferred from internal transcribed spacer sequence of nuclear ribosomal DNA. Meta Gene 7: 56-63.

Kehie, M., Kumaria, S. and Tandon, P. 2014. Manipulation of culture strategies to enhance capsicin biosynthesis in cell cultures of Capsisum Chinense Jacq. cv Naga King Chilli. Bioprocess Biosyst. Bioprocess Eng. 37: 1055-1063.

Mathur, R.. Dangi, R.S., Das. S.C., and Malhotra, R.C. 2000. The hottest chilli variety in India. Current Science 79: 287-288.

Meghvansi, M.K.. Sidiqui, S.. Khan, H., Gupta. V.K., Vairale, M.G., Gogoi, H.K., and Singh, L. 2010. Naga chilli: A potential source of capsaicinoids with broad-spectrum ethnopharmacological applications. Journal of Ethnopharmacology 132: 1-14.

Pandey, V., Pandey, H.K.. Dayal, D.. Joshi. U.C., Pant, T., and Ahmed, Z. 2009. Studies on antioxidant constituents of some domesticated capsicums in the middle hill conditions of western Himalayas. Horticultural Science 36: 26-30.

Purkayastha, J., Alam, S.I., Gogoi, H.K., and Singh. L. 2012a. Capsicum assamicum sp. Nov. (Solanaceae) from Assam, Northeastern India. Ozean Journal of Applied Science 5:55-66.

Purkayastha, J., Alam, S.I., Gogoi, H.K., Singh, L., and Veer, V. 2012b. Molecular characterization of ‘Bhut jolokia’ the hottest chilli. Journal of Biosciences 37: 757-768.

Roy, A. 2016. Bhut jolokia (Capsicum chinense Jacq.): A review. International Journal of Pharmaceutical Sciences and Research 7: 882-889.

Sanatombi, K., and Sharma, G.J. 2008. Capsaicin content and pungency of different Capsicum spp. Cultivars. Notulae Botanicae Horti. Agrobotanici Cluj-Napoca 36: 89-99.

Singh, M.S. 2009. Standardization of sowing time and plant population of Bhut Jolokia (Capsicum chinense). MSc Thesis. Assam Agricultural University.

Suzuki, T., Fujiyake, H., and Iwai, K. 1980. Intracellular localization of capsaicin and its analogues, capsaicinoid in Capsicum fruit 1. Microscopic investigation of the structure of the placenta of Capsicum annuum cv. Karayatsubusa. Plant and Cell Physiology 21: 839-853.

Talukdar, J., Saikia, A., and Borah, P. 2015. Survey and detection of the diseases of Bhut jolokia (Capsicum chinense Jacq.) in Assam. Journal of Crop and Weed 11: 186-192.

Tiwari, A., Kaushik, M.P.. Pandey, K.S., and Dangi. R.S. 2005. Adaptability and production of hottest chilli variety under Gwalior agro-climatic conditions. Current Science 88: 1545-1546.

Williams, J.G.K., Hanafey. M.K., Rafalski, J.A.. and Tingey. S.V. 1993. Genetic analysis using random amplified polymorphic DNA markers. Methods in Enzymology 218: 705-740.

Xu, J. 2006. Fundamentals of fungal molecular population genetic analyses. Current Issues in Molecular Biology 8: 75-90.