Lapworth, A. & Royle, F. (1925). The chemical society. London: Gurney& Jackson.

The chemical society by Lapworth and Royle talk about capsaicin, the main component of chili pepper. It can help our study by showing the chemical formula and structure of capsaicin. With the knowledge of the organic composition of chili pepper, we will know its functions.

Effects of Oleoresin Capsicum Pepper Spray on Human Corneal Morphology and Sensitivity Capsaicin (8-methyl-vanilyl-6-nonenamide), the pungent component of chili peppers, has been shown to induce intense pain in humans and pseudoaffective pain reactions in animals when applied to the skin and the front of the eye, as well as neurogenic inflammation due to the release of neuropeptides contained in nerve terminals. Capsaicins effects are associated with acute stimulation of primary sensory nerve endings, which is accompanied by a depletion of their neuropeptide content. This process is followed by nerve inactivation and suppression of neurogenic inflammation in response to injury. Capsaicin also has long-lasting effects on sensory nerves and their target tissues. Neonatal injection of capsaicin into small rodents induces a selective sensory denervation followed by a slow and incomplete regeneration of the sensory fibers that is not fully compensated by increased sprouting of intact nerve fibers and persists in adult animals. Corneal and/or skin ulcers and scars are concomitantly formed and may persist for months, resembling the clinical picture of neuroparalyticneurotrophic keratitis secondary to trigeminal denervation. Capsaicin treatment in adult animals causes less prominent but still detectable alterations of sensory innervation. In vitro and in vivo studies show that nerve growth factor (NGF) reverses the decrease of transmitter content and restores the peripheral function of primary afferent neurons impaired by capsaicin treatment. In humans, the effects of capsaicin on sensory innervation are poorly known. Capsaicin injected subcutaneously produces acute pain and hyperalgesia. Topically applied capsaicin ointment (0.075%) used for pain relief results in a reduced sensitivity to cutaneous stimuli and decreased numbers of epidermal nerve fibers. Reinnervation of the skin and recovery of sensitivity have been observed after discontinuation of treatment. In the present work, we studied in human eyes the effects of topical capsaicin on corneal innervation and corneal sensitivity using in vivo confocal microscopy and noncontact esthesiometry. We also measured tear fluid NGF concentrations in subjects exposed to capsaicin. Results After OC exposure, all subjects had mild to moderate facial hyperemia. Additionally, they reported mild to moderate ocular and/or facial stinging sensations with a mean duration of 24 minutes (range, 450 minutes). Five police officers had mild nasal congestion during the first 30 minutes, but none of them had dyspnea. The mean heart rate increased significantly from a basal value of 79.7 13.3 beats/min (mean SD) to 116.0 19.1 beats/min (at 1 minute) and subsequently decreased in 10 minutes to 73.0 17.9 beats/min (one-way ANOVA, P < 0.001). Scoring of Ocular Lesions by Slit Lamp Biomicroscopy Draizes scale for scoring of ocular lesions includes signs in cornea, iris, and conjunctiva. Six corneas of four police officers showed focal corneal epithelial cell damage at 20 minutes but none of the corneas showed opacities as described in Draizes scale. The following day, the epithelial surface of all subjects was normal again. All eyes showed conjunctival hyperemia at 20 minutes. The mean duration of conjunctival injection was 9.8 hours (range, 224 hours). Mild chemosis was observed in two subjects after the exposure, but it was undetectable on the following day. Visual Acuity Best corrected visual acuity (BCVA) was not tested immediately after OC exposure, because the subjects could not keep their eyes open. Except for those in one subject, all eyes

had unchanged BCVA (20/20) throughout the study. One police officer lost one line at both 1 day and 1 week after OC. Discussion The rationale of adding capsaicin to a self-protecting spray is that this substance induces an intensive but relatively short-lasting pain leading to blepharospasm and hypersecretion of tears. Capsaicin sprayed onto the face immobilizes the individual and prevents attack or resistance against arrest by law enforcement personnel. Early-generation tear gases such as chloroacetophenone (CN) and CS frequently induced detachment of the whole corneal epithelium necessitating medical assistance for days or weeks. Pepper spray, containing OC as the active substance, is widely used in private self-protection and by the police. This study shows that the structural and functional effects of OC spray on conjunctiva and cornea are mild and temporary. This interpretation applies only to the preparation used in the present work; other solvents, such as 92% trichloroethylene, may induce severe corneal damage (Tervo et al., unpublished observations, 1999). Several studies on the ocular effects of capsaicin in animals have been published. However, only recently the effects of OC sprays on the human eye were briefly reported. These researchers described transient and moderate conjunctival and corneal changes with occasional punctate epithelial erosions. In the present study, mild and shortlasting corneal and conjunctival signs were also observed. It is possible that epithelial damage was caused by the alcohol solvent rather than by capsaicin, because similar grayish white epithelial changes can occasionally be seen after contact with isopropyl alcohol-cleaned, but wet, tonometer tips. Visual acuity was unaffected in all but one eye, in agreement with the data of Zollman et al. The acute effects of capsaicin on the sensory activity of corneal nerve afferents in animals are well documented. In the cat eye, capsaicin at high concentrations (1%, 33 mM) produced a transient excitation of polymodal nociceptive fibers followed by inactivation of most fibers to all subsequent stimuli; cold sensory fibers were weakly activated, but many remained active after capsaicin, whereas pure mechanosensory fibers were largely unaffected by this substance. The excitatory effects of capsaicin are due to activation of a vanilloid receptor (VR1) present in polymodal nociceptive neurons that acts as a nonselective cationic channel, thus depolarizing sensory nerve terminals. This strong excitatory effect on polymodal nociceptive fibers explains the intense pain experienced immediately after capsaicin application to the eye. Toxic effects of capsaicin are the consequence of a massive calcium entrance that leads to cell damage and functional inactivation, making nerve endings insensitive to further stimuli. Determinations of mechanical threshold with the CochetBonnet esthesiometer after capsaicin treatment showed an acute reduction of corneal mechanical sensitivity followed by a progressive recuperation of control values. Nevertheless, the CochetBonnet esthesiometer does not permit evaluation in detail of the degree of short- and long-term functional disturbances caused by capsaicin in the various populations of sensory fibers that sustain corneal sensitivity. Graded measurement of the responsiveness to mechanical, chemical, and thermal stimulation of the cornea with the gas esthesiometer indicate that the effect of capsaicin on the different populations of corneal nerve fibers was heterogeneous and evolved with time. With the gas esthesiometer, the reduction of mechanical sensitivity observed with CochetBonnet stimulation was confirmed. Responses to low and moderate mechanical stimulation were depressed to a varying degree among individuals 30 minutes after OC and remained below control values 1 week after OC treatment. Acute blockade of a fraction of polymodal nociceptors, preferentially those with unmyelinated axons that are highly sensitive to

capsaicin, seemed to be responsible for the immediate reduction of sensitivity to mechanical stimulation. Residual mechanical sensitivity in the first hour after OC application is attributable to activation of pure mechanosensory fibers that have a higher threshold and would be much less affected by capsaicin and to those polymodal units presumably A-delta that remained functional. The gradual return of mechanosensitivity during the ensuing hours and days may be ascribed to the recovery of those corneal polymodal fibers that were initially inactivated by capsaicin. The response to chemical stimulation with CO2, which is also mediated by polymodal fibers was enhanced immediately after OC application. Twenty-four hours later, it remained high in three subjects but was absent in two, reappearing in a depressed state in all subjects 7 days after treatment. These results confirm that a variable fraction of the polymodal fibers was acutely inactivated and that this process reached a maximum 24 hours after OC exposure. They further indicate that the fibers that remain functional became sensitized thus producing a hyperalgesic response. This phenomenon was more prominent with CO2 than with heat stimulation, although both stimuli activate polymodal nociceptive fibers, probably reflecting the fact that heat responses are mediated by VR1 (capsaicin) receptors while additional, capsaicin-insensitive ion channels participate in the responses to acid. The absence of changes in cold sensitivity during the 24 hours after OC exposure indicates that cold-sensory fibers were not immediately affected by capsaicin. Nevertheless, as occurred with the other modalities of sensation, cold sensitivity was depressed 1 week later, implying that a part of both polymodal and cold fibers were disturbed in the long term by the treatment. NGF is the prototypical member of the neurotrophin family of growth factors. It plays a critical role in the development of primary sensory neurons during embryonic life, including those that innervate the cornea. In adult animals, NGF receptors (TrkA) remain in the subpopulation of small nociceptive sensory ganglion neurons. During chemically induced inflammation with carrageenan or turpentine, increased tissue levels of NGF have been measured. Tissue NGF seems to increase the sensibility of peripheral terminals to noxious stimuli. In the present experiments two of the five police officers showed detectable levels of NGF in tears, and levels increased after OC treatment. Elevated values could still be measured 1 week later. In spite of the limited number of data, these results suggest that in the cornea, as in other tissues, NGF is released during noxious stimulation contributing to sensitization and hyperalgesia of inflamed ocular tissues. Moreover, elevated NGF levels may contribute to nerve sprouting and enhanced neuropeptide synthesis observed in the skin after injury and in the cornea after capsaicin treatment. This in turn would facilitate healing of the injured target tissues. In vivo confocal microscopy is a noninvasive method for examining tissue responses in different corneal sublayers of the human cornea. Toxicity of various substances has been evaluated in an animal model, but to our knowledge, this is the first in vivo confocal microscopy study on potential toxic effects of a substance on the human cornea. The results show that OC spray causes surface epithelial damage of short duration in some subjects. That the cell borders of the basal epithelial cells were easily visualized at 30 minutes and 1 day after OC, without signs of cell damage, suggests epithelial swelling. No changes could be ascertained in the morphology of subbasal nerves after a single pepper spray exposure. Electron microscopic observations have revealed that corneal subbasal nerves that are seen by in vivo confocal microscopy correspond to nerve bundles, because visualization of individual nerve fibers is beyond the level of resolution of confocal microscopy. In most cases the nerves were more apparent after OC exposure, probably because of swelling of the epithelial cells through which the nerves are pressed into the same focal plane. The images of the nerve fiber bundles did not vary during the study, and no signs of sprouting were apparent. It is possible that the insult to the nerves is not great enough to induce sprouting.

Alternatively, sprouting may be beyond the level of resolution or is limited to the peripheral cornea, as described for experimental animals, which was out of the range of observation in the present experiment in which explorations were limited to the central cornea. A surprising finding was the spirallike nerve fiber bundle arrangement of the subbasal plexus in the eyes of a police officer repeatedly exposed to OC or CS. A similar organization has been observed in the nerves of the cornea of an alkaline phosphatase transgenic mice (Belmonte and Raviola, unpublished observations, 1999), but its significance is obscure. Because of the mild and transient signs of tissue injury, it can be concluded that single exposure of human eyes to OC is relatively harmless to the cornea and conjunctiva. However, one should be cautious in repeated OC exposures, because long-lasting changes in corneal sensitivity could occur. These changes are possibly associated with damage of nerve terminals of mainly unmyelinated polymodal nociceptive fibers. Vesaluoma, M., Muller, L., Gallar, J., Lambiase, A., Moilanen, J, Hack, T., Tervo, T. (2000). Effects of Oleoresin Capsicum Pepper Spray on Human Corneal Morphology and Sensitivity. Investigative Opthalomology & Visual Science. 41(8), 2138-2147. Vesalluoma, Muller, Gallar, etc. examine the potential harmful effects on corneal structure, innervation, and sensitivity of a spray containing the neurotoxin capsaicin or oleoresin capsicum. They conducted an experiment to ten police officers who volunteered for the study. Clinical signs were assessed on the human volunteers. It discusses capsaicins long-lasting effects on sensory nerves and tissues. It can help our study by showing the effects of chili pepper spray on humans. Its effects include pseudoaffective pain reactions, neurogenic inflammation, acute stimulation of primary sensory nerve endings, depletion of neuropeptide content, nerve inactivation and suppression of neurogenic inflammation in response to injury.

Capsaicin, The Active Principle of Capsicum Fruits Isolating the pungent principle of cayenne The first step is to obtain the oily fluid named by Buohheim (capsicol, by treating the powdered fruit with ether, distilling off the ether, dissolving the residual extract in boiling caustic alcoholic lye, diluting with water and precipitating with barium chloride ; this precipitate is washed, dried, and treated with ether, and upon evaporation the oily capsicol is obtained, which may be purified by a repetition of the process. From capsicol; capsaicin may be obtained in two ways: (a) Capsicol is dissolved in twice its volume of almond oil, and agitated with three successive portions of proof spirit, the alcoholic solution is separated, and upon evaporation leaves a redbrown fatty residue, which when dissolred in dilute solution of potash, and treated with dilute ammonia, deposits, on standing pearly white crystals of Capsaicin. (6) The capsicol is dissolved in dilute potash, precipitated by ammonium chloride, the coloured precipitate re-dissolved in potash, and re-precipitated at 120 F by ammonium chloride in excess-in a few day8 an abundant crop of capsaicin crystals will be the result. Capsaicin may also be obtained by dialysing the tincture of capsicum-the dialysed solution has an acid reaction. Capsaicin is powerfully pungent, the most minute portion, if volatilised, causing severe fits of coughing. It dissolves slightly in cold water, more readily in boiling water, a portion at that temperature becoming volatilised, and causing long continued fits of sneezing, the excess of what is taken up by the water melts and floats on the surface of the fluid 8s a colourless oil. The hot solution precipitated by the addition of a strong acid deposits crystals. Capsaicin dissolves readily in proof spirit, giving, when not too dilute, white precipitates with barium and calcium chlorides, both soluble in ether. Silver nitrate gives a precipitate which dissolves in dilute ammonia, and the solution when boiled darkens in colour and deposits a curdy brown-black precipitate. Capsaicin is volatilised slowly at 212oF. and may be obtained as a sublimate of fatty globules, if mixed with water and distilled. The distillate has a distinctly pungent taste. Jacquemin, E. (1876). Capsaicin, the active principle of capsicum fruits. Pharmaceutical Journal, 315. 21. Jacquemin discusses the different techniques of extracting capsaicin in capsicum fruits. It thoroughly discusses the procedure of extraction icluding the materials and the set up. It can help our study by guiding us on the experimental technique that is viable. With the knowledge of the different methods, we can compare and conduct our own experiment.

The Pun1 Gene for Pungency in Pepper Encodes a Putative Acyltransferase Pungency in Capsicum fruits is due to the accumulation of the alkaloid capsaicin and its analogs. The biosynthesis of capsaicin is restricted to the genus Capsicum and results from the acylation of an aromatic moiety, vanillylamine, by a branched-chain fatty acid. Many of the enzymes involved in capsaicin biosynthesis are not well characterized and the regulation of the pathway is not fully understood. Based on the current pathway model, candidate genes were identified in public databases and the literature, and genetically mapped. A published EST colocalized with the Pun1 locus which is required for the presence of capsaicinoids. This gene, AT3, has been isolated and its nucleotide sequence has been determined in an array of genotypes within the genus. AT3 showed significant similarity to acyltransferases in the BAHD superfamily. The recessive allele at this locus contains a deletion spanning the promoter and first exon of the predicted coding region in every non-pungent accession tested. Transcript and protein expression of AT3 was tissue-specific and developmentally regulated. Virus-induced gene silencing of AT3 resulted in a decrease in the accumulation of capsaicinoids, a phenotype consistent with pun1. In conclusion, gene mapping, allele sequence data, expression profile and silencing analysis collectively indicate that the Pun1 locus in pepper encodes a putative acyltransferase, and the pun1 allele, used in pepper breeding for nearly 50 000 years, results from a large deletion at this locus. Pepper (Capsicum) was among the earliest domesticated plant genera, based upon archeological evidence from Central America dating back at least 7000 years (Basu and Krishna De, 2003). In 1876, the alkaloid capsaicin was identified as the compound responsible for the characteristic pungency in pepper (Suzuki et al., 1980; Thresh, 1876). Within the pepper fruit, capsaicin and its analogs, known collectively as capsaicinoids (Bennett and Kirby, 1968), are synthesized in the epidermal cells of the placental dissepi-ment beginning approximately 20 days post-anthesis (dpa), and accumulate in pockets or blisters along the epidermis (Iwai et al., 1979; Ohta, 1962; Suzuki et al., 1980; Zamski et al., 1987). Biosynthesis of this group of compounds is unique to the Capsicum genus, and has driven the domes-tication of several species including C. annuum, C. frutes-cens and C. chinense, which are now valued for use as vegetables, spices, and for medicinal and industrial purpo-ses (Andrews, 1984; Walsh and Hoot, 2001). Although pungency is highly desirable or essential for many uses, non-pungent peppers have also been selected for use as a vegetable and as the spice known as paprika. Bell peppers were first described nearly 500 years ago; the earliest reference to a bell variety specifically selected for cultivation occurred in 1774 (Boswell, 1937). Recent years have seen an explosion of information about the directed applications of capsaicin in food and medicine and the evolutionary role these compounds play in Capsi-cum. The nociceptor responsible for perception of these compounds, the vanilloid receptor 1 (VR1), is localized to peripheral pain-sensing nerve fibers (Caterina et al., 1997, 2000). Birds, the primary dispersal agent of Capsicum seeds, lack a functional VR1 receptor, hence are oblivious to the presence of capsaicin (Jordt and Julius, 2002; Tewksbury and Nabhan, 2001). In humans, VR1 is involved in pain sensing pathways. Capsaicin has long been used in topical analgesic prepara-tions and clinical trials have supported capsaicin as an effective treatment for various types of nerve pain and arthritis, although the mechanism is not fully understood (Deal et al., 1991; Watson et al., 1993). Beyond this use as an analgesic, capsaicin has also been described in the treatment of diverse bladder and digestive syndromes, and as a tumorigenesis chemopreventive agent in cancer treatment through its effects on intracellular signaling pathways (Chancellor and De Groat, 1999; Cruz, 2004; Han et al., 2001; Srinivasan, 2005; Surh, 2002, 2003). Evidence also supports an antimicrobial activity. Capsaicin used in spice has been reported to inhibit or kill microorganisms that contribute to food spoilage (Billing

and Sherman, 1998). Recently, a physiological basis for the paradoxical attraction between humans and hot peppers has been proposed. Natural capsaicin analogs identified in mammalian brains presumably function as neurotransmitters related to positive sensations (Appendino et al., 2002; Chu et al., 2003; Huang et al., 2002). The ecology and physiology of capsaicin therefore both explains the selective advantage of these compounds, presumably evolved to deter mammalian herbi-vory, as well as the human desire to consume spicy foods. Capsaicinoids are produced by the condensation of vanillylamine, derived from phenylalanine, with a branched-chain fatty acid, derived from either valine or leucine (Bennett and Kirby, 1968; Leete and Louden, 1968; Sukrasno and Yeoman, 1993; Suzuki et al., 1981). A vast array of Capsicum species and cultivars are now available with varying degrees of pungency (Zewdie-Tarekegen, 1999). Further variation in capsaicinoid content results from the influences of the plant growth environment (Blum et al., 2003; Harvell and Bosland, 1997; Zewdie and Bosland, 2000). Considering the economic and agricultural importance of this pathway, it is surprising that relatively little is known, particularly at the molecular level, concerning the genetics, biosynthesis, subcellular localization and cellular structures required for pungency accumulation in peppers (Blum et al., 2003). Nearly 100 years ago, Webber reported that the absence of pungency was controlled by a single recessive gene, pun1 (formerly known as c), epistatic to all other pungency-related genes (Boswell, 1937; Webber, 1911). At present, the pun1 allele is the only confirmed mutation that has a qualitative affect on the presence/absence of capsaici-noids (Blum et al., 2002 and references therein). Acleaved-amplified polymorphic sequence marker linked at a small genetic distance was developed to assist in breeding programs. However, Pun1, often presumed to be a master regulator of the pathway, itself has remained elusive (Blum et al., 2002). Here we report the identity of Pun1 and show that in contrast to earlier predictions, this gene encodes a putative acyltransferase important in capsaicin biosynthesis. We describe the sequence of a very famous allele at this locus that is responsible for the absence of pungency in pepper fruit. Finally, we describe the expression profile of the wild-type transcript and its gene product through fruit development and demonstrate that reducing this transcript and gene product in planta reduces accumulation of capsai-cinoids in fruit. Capsicum originated in South America, most likely in the area now known as Bolivia (Andrews, 1984). The explorer Columbus played a key role in dispersing peppers throughout the world, returning to Spain with several pun-gent forms of Capsicum, most of which were members of the C. annuum species (Walsh and Hoot, 2001). From here, peppers spread throughout the world and eventually were reintroduced to the Americas. Asingle genetic source for non-pungency within C. annuum is suggested by the early identification of a non-pungent pepper in the 1500s that was widely distributed (Boswell, 1937), and documented and inferred pedigrees of non-pungent pepper varieties released by breeding programs in the United States, South America and Europe (Cook, 1984a,b; Homma, 1986; Millet and Jones, 1982; Prashar and Enevoldsen, 1984; Smith et al., 1987; Vil-lalon, 1986; Villalon et al., 1986). This dispersal pattern and subsequent selective propagation of lines containing favo-rable mutations such as non-pungency likely resulted in a progressive narrowing of the genetic base of subsequent populations, in effect creating genetic bottlenecks. Similar effects on genetic diversity have been documented in soy-bean and wheat (Duvick, 1977; Harlan, 1987). An early gen-etic event in C. annuum domestication such as the deletion responsible for non-pungency at the Pun1 locus, identified in this study, would be expected to be present broadly within the species. While it is probable that there is more than one mutation that results in non-pungency, the presence of the deletion in Pun1 across many C. annuum genotypes tested indicates that it likely arose early in the domestication of C. annuum.

HPLC analysis of capsaicinoids One gram of dried, ground pepper samples were extracted into 10 ml of acetonitrile for 4 hours at 80oC according to the method of Collins et al. (1995). The extract was syringe filtered and 20 llofthe filtered extract was injected for HPLC quantification using Beckman 126 pumps and a Beckman 166 detector set at 280 nm or 227 nm (Beckman Coulter Inc., Fullerton, CA, USA). An isocratic mobile phase consisting of 50% acetonitrile: 45% water: 5% tetrahydrofuran )1 at 1.5 ml min (Phenomenex method ALF-519) was used on 150 mm 4.6 mm Phenomenex Synergi Hydro RP columns (Phe-nomenex, Torrance, CA, USA). External standards were prepared by dissolving commercial capsaicinoids (Sigma M3403; Sigma-Aldrich, St. Louis, MO, USA) in acetonitrile and making a dilution series. Stewart, Jr., C., Kang, B., Liu, K., Mazourek, M., Moore, S., Yoo, E., Jahn, M. (2005). The Pun1 gene for pungency in pepper encodes a putative acyltransferase. The Plant Journal, 42, 675-685. The article, Pun1 gene for pungency in pepper encodes a putative acyltransferase studies about acyltransferase. It is a compound found in chili pepper. The history of chili pepper is narrated including its origin, effects and applications. It can help our study by showing the facts regarding the pungency of chili pepper that can affect animals.

Evolutionary Ecology of Pungency in Wild Chilies The evolution of fruit, a reward for animal dispersal of seeds, is a commonly cited example of a key innovation in the radiation of angiosperms. However, the nutritional qualities of fruit pulp that are responsible for attracting beneficial dispersers also attract consumers that are detrimental to plant fitness. These consumers range from vertebrate and invertebrate seed predators to microbial consumers of fruits and seeds that reduce the likelihood of dispersal and the viability of seeds. Fruit chemistry is commonly thought to mediate these interactions, either by deterring seed predators or reducing microbial attack of fruits and seeds. These mechanisms are not mutually exclusive, but chemicals that deter fruit consumption often affect a wide range of species, and defensive chemistry in ripe fruit must be sufficiently targeted toward detrimental organisms to allow consumption by vertebrate seed dispersers. Fruit secondary compounds that deter microbial consumers without reducing seed dispersal by vertebrates are thought to be far more plausible than secondary compounds that selectively deter vertebrate predators, because microbial fruit consumers are uniformly negative in their impacts on plant fitness and are farther removed in their morphology, physiology, and mode of consumption from vertebrate seed dispersers than are other unwanted consumers. Microbial deterrence is thus a primary hypothesis explaining the presence of noxious, bitter, and sometimes toxic chemicals in many ripe fruits; the negative effects these chemicals often have on vertebrate dispersers are assumed to be balanced by the benefits of deterring microbial consumers. Unfortunately, this hypothesis remains largely untested, because no work to date has shown that variance in microbial pathogen pressure is related to variance in the chemistry of ripe fruits in wild populations. A strong test would require a species in which fruit chemistry is well known, likely to protect against microbial pathogens, unique to the fruit, and highly variable. The most famous plants with these qualities are chilies (genus Capsicum). Chilies were one of the first plants domesticated in the New World, and they are now consumed by one in four humans daily, largely because of the pungency produced by capsaicinoids. Capsaicinoids are well characterized and broadly antimicrobial. In fact,early humans likely selected chilies for use and domestication expressly because of their antimicrobial properties. Finally, because capsaicinoids are found only within the fruit of Capsicum species and their concentrations increase during fruit ripening, the function of these chemicals is likely restricted in the fruit itself, not attributable to alternative functions in other parts of the plant. Chilies thus provide an exceptionally clear window into the function of fruit chemistry, and our recent rediscovery of a polymorphism for capsaicinoid production in wild populations of multiple chili species provides the variability we need to explicitly examine the function of these chemicals in wild populations. We have studied this polymorphism most intensively in Capsicum chacoense Hunz., which is native to the Chaco region of Bolivia, Argentina, and Paraguay. In polymorphic populations, C. chacoense plants producing fruits that contain capsaicinoids grow alongside plants with fruits that are nutritionally similar but completely lack capsaicinoids. In addition, the proportion of plants producing capsaicinoids varies widely among populations. At the southwestern end of our 300-km-long study area in southeastern Bolivia, the polymorphism is virtually absent; most populations contain only pungent plants. To the north and east of this area, nonpungent plants gradually increase in frequency, until 70% of individuals lack capsaicinoids, and the few plants that do produce pungent fruit have capsaicinoid concentrations barely one-third the level found in completely pungent populations. We use this geographic gradient as a tool to study the impact of microbial pathogens on fruit chemistry, and we made the following predictions: (i)Microbial fruit pathogens will have a large negative impact on nonpungent chilies, (ii) capsaicinoids will reduce microbial damage to chili fruits and seeds, and (iii) among populations, the proportion of plants producing capsaicinoids will increase as the intensity of microbial attack increases.

Impact of Microbial Pathogens Across all populations in this system, the only significant cause of predispersal fruit and seed damage is microbial infection. This damage appears to be caused primarily by a single fungal species, Fusarium semitectum Berkeley and Ravenel (hereafter Fusarium). Fusarium infection of seeds causes discoloration that is easy to score, and we found Fusarium infection in 90% of all ripe fruits sampled across our populations (305 fruits). The vast majority (95%) of these infections were provisionally attributable to Fusarium, which rots chili fruits and kills seeds. Even at low levels of infection, Fusarium causes substantial reductions in seed survival. Its entry into fruits is facilitated by hemipteran bugs that pierce the pericarp of fruits with their proboscises. This piercing introduces Fusarium into the fruit and seeds, leaving visible scars on the fruit surface, which turns black as the fungus invades. We randomly selected single ripe fruits from pungent and nonpungent plants in our primary study site (called San Julian), counted foraging scars on the fruit, and scored all seeds in each fruit for degree of Fusarium infection. Fungal infection of seeds increased with the number of foraging scars on the fruit, and seeds from fruits without signs of insect damage showed no signs of fungal infection. Capsaicinoids and Microbial Damage The same data gathered to assess the impact of hemipteran foraging on fungal infection also suggest a strong antifungal role for capsaicinoids. Although fungal infection of seeds increased with the number of hemipteran-foraging scars in both nonpungent and pungent fruits, the slope of this relationship was significantly steeper in nonpungent fruits. Thus, for a given level of hemipteran foraging pressure, seed infection rates in nonpungent fruits are almost twice as high as in pungent fruit. We experimentally verified this susceptibility of nonpungent fruits to Fusarium by placing cages over randomly selected pungent and nonpungent plants in the same polymorphic population such that birds were prevented from removing fruits, but Fusarium transmitting hemipterans had free access. We let these fruits mature naturally, then removed and scored their seeds for Fusarium infection. Degree of infection was more than twice as high on seeds from nonpungent plants than on seeds from pungent plants. Nonpungent and pungent fruits are visually indistinguishable in the field, and their nutritional profiles are virtually identical. Nonetheless, the large difference in seed infection we observed between pungent and nonpungent plants could be because of a factor other than the presence or absence of capsaicinoids. Fungal loads in pungent fruit were 4555% lower than in nonpungent fruit. If this reduction were caused by the presence of capsaicinoids in the pungent fruit, we should be able to generate a similar effect size in a more controlled experimental setting where capsaicinoid content is the sole independent variable. To test this we created artificial fruit media that mimicked the nutritional composition of C. chacoense fruit, differing only in the presence and concentration of the two primary capsaicinoids, capsaicin and dihydrocapsaicin. Inoculating these media with Fusarium isolates cultured from C. chacoense seeds from the same population showed that both capsaicin and dihydrocapsaicin cause strong dosedependent inhibition of Fusarium growth, and quadratic fits. More importantly, at the capsaicinoid levels found in our focal population, capsaicin reduced Fusarium growth by 33%, and dihydrocapsaicin reduced Fusarium growth by 16%. Together, these chemicals fully account for theobserved reduction in Fusarium seed infection in pungent fruit (predicted reduction based on capsaicinoid concentrations. Fruit Chemistry and Fungal Selection Capsaicinoids thus protect chili fruits and seeds from a fungal pathogen that severely reduces seed viability. If this process shapes the chemistry of chili fruits, changes in fungal selection pressure among chili populations should lead to parallel changes in the chemical defense of chili fruits, explaining among-population variation in capsaicinoid production. This prediction was supported. We surveyed seven chili populations, distributed across a 1,600 km2

area in eastern Bolivia. In each population, we randomly selected fruit and counted hemipteranforaging scars. Because foraging pressure is positively correlated with fungal attack on seeds, the number of scars provides an index of variation in Fusarium pressure on fruits across populations. As predicted, the mean number of hemipteran scars on fruits in a population was a strong predictor of the proportion of plants producing capsaicinoids. Insect-mediated fungal attack on seeds thus appears at least partially responsible for driving phenotypic evolution in chili fruits; as fungal pressure increases, selection for protection against fungal attack should also increase, favoring pungent phenotypes. Yet pungency does not appear to come free of costs: Our previous work shows that tradeoffs between capsaicinoid production and seed-coat thickness can favor nonpungent plants because seeds from these plants have thicker seed coats and are better protected as they pass through the digestive tract of seed dispersers. Teasing apart the relative importance of these potential selection pressures will require direct experimental evidence for specific adaptive functions. We suggest that the ratio of pungent to nonpungent plants in a given population reflects a long-term averaging of multiple benefits and costs of capsaicinoid production, creating a mosaic of evolutionary outcomes. These findings provide strong support for the role of microbes in shaping fruit chemistry in wild species as increases in microbial pressure are met by concomitant increases in the frequency of chemically protected fruit. Though the focus of our work has been the chemical response of chilies to microbial attack, the antimicrobial properties of capsaicinoids extend well beyond Fusarium and have captured the interests of food scientists, ethnopharmacologists, and evolutionary biologists interested in historical and geographic patterns of how chilies are used by humans. For example, it has been postulated that the capsaicinoids in chilies may have had a profound influence on the domestication and use of chilies as a spice because of humans harnessing capsaicinoids antimicrobial benefits for food preservation. Before the advent of refrigeration, microbial contamination of food was a common cause of illness and death in many cultures, and the consumption of chilies with food may have reduced the risk of microbial infection, providing an adaptive reason to eat pungent food. If the antimicrobial properties of chilies are truly responsible for their early domestication and spread, our research provides an evolutionary foundation for this relationshiphuman use of chilies may mirror the evolutionary function of these compounds in the fruits that produce them. Tewksbury, J., Reagan, K., Machniki, N., Carlo, T., Haak, D., Penaloza, A., & Levey, D. (2008). Evolutionary ecology of pungency in wild chilies. Proceedings of the National Academy of Sciences of the United States of America, 105, 11808-11811. Evolutionary ecology of pungency in wild chilies uses the geographic gradient as a tool to study the impact of microbial pathogens on fruit chemistry. It studies the relationship of chili pepper and microbial pathogens. Microbial pathogens are found in chili peppers on a specific situation. It can help our study by showing that chili pepper can deter bacteria, fungi and other microbial dirt. It can prove the effectivity of the chili pepper pesticide.

Bhut Jolokia, The Worlds Hottest Known Chile Pepper is a Putative Naturally Ocurring Interspecic Hybrid Chile peppers (CapsicumL. spp.) are known for causing the sensation of heat or burning when consumed. The heat sensation is incited by the type and the amount of a group of capsaicinoids, the alkaloids found only in chile pepper pods (Zewdie and Bosland, 2001). The amount of capsaicinoids in a chile pepper pod is dependent on the genetic makeup of the plant and the environment where it is grown (Harvell and Bosland, 1997; Zewdie and Bosland, 2000). The capsaicinoids have evolved in chile peppers as a defense mechanism against mammalian predators (Tewksbury and Nabhan, 2001); nevertheless, this trait is an important fruit quality attribute and one of the most important reasons chile peppers are consumed. Chile peppers were rst introduced to Europe by Christopher Columbus. Shortly after the voyage of Columbus, Portuguese traders introduced chile peppers along their trade routes in Africa and Asia, including India (Andrews, 1999). By 1542, three varieties of chile peppers were recognized to be growing in India (Purseglove, 1968). Today, numerous landraces of chile pepper differing in shape, size, color, and heat level can be found in India as farmers selected chile peppers to t their needs. The northeastern region of India claims that the chile peppers grown in this region are the hottest in the world. Genetic resources of chile pepper landraces in northeastern India have not been well documented, but a few names mentioned include Naga Jolokia, Bhut Jolokia, and Bih Jolokia. The Assamese word jolokia means the Capsicum. Mathur et al. (2000) reported the Naga Jolokia to be a variety of C.frutescens L. and to have a very high heat level, i.e., 855,000 Scoville heat units (SHUs). The hottest chile pepper on record is the C.chinense Jacq. cultivar Red Savina with heat level of 577,000 SHUs (Guinness Book of World Records, 2006). The growing season in 2005 was favorable for the production of fruits on all three chile pepper cultivars. The environment is known to affect the heat level of chile pepper cultivars (Harvell and Bosland, 1997). Having a replicated eld trial with standard control cultivars allows for a better comparison of heat levels among cultivars. The HPLC analysis revealed that orange habanero had a mean heat level of 357,729 SHUs, which is in the range normally seen for this cultivar in Las Cruces, N.M.. The results of the analysis for Bhut Jolokia indicated that it possessed an extremely high heat level, 1,001,304 SHUs, whereas Red Savina recorded a heat level of 248,556 SHUs. Independent tests conrmed this high level of heat for Bhut Jolokia with 927,199 SHUs and 879,953 SHUs from Southwest Bio-Laboratories and AgBiotech, respectively. The leaf surface of Bhut Jolokia has the characteristic crinkle look like in others. A widely used heat measurement of chile peppers is the SHU (Scoville, 1912). This measurement is the highest dilution of a chile pepper extract at which heat can be detected by a taste panel. Alternative instrumental methods have been developed since Scovilles test. HPLC is the most accurate and efcient method (Wall and Bosland, 1998). The extreme high heat level of Bhut Jolokia was consistent over the past 3 years in non-replicated tests. In addition,

the heat measurements from the commercial testing laboratories conrm its high heat level. The results demonstrate that Bhut Jolokia is indeed the worlds hottest known chile pepper. Bosland, P. & Baral, J. (2007). Hortscience. New Mexico: New Mexico State University. Bosland and Baral discuss in Hortscience, Bhut Jolokia. Bhut Jolokia is part of the capsicum family. It researches about the history and location of the chili pepper. Bhut Jolokia is known to be the hottest chili pepper last 2006 with a heat level of 577,000 heat Scoville units (HSU). It can help our study regarding the chili peppers heat level. The heat level describes how pungent the chili pepper is and how it affects pests.

Nutritional Information Fresh Green Chile Peppers The following information is for one serving of hot green chile peppers. That would be about one half cup of hot green chile peppers, or 75 grams. This general information is for any variety of raw green chile peppers. Macronutrients: Water: 65.80g, Calories: 30Protein: 1.50 g Carbohydrates: 7.10 g, Fiber: 1.1 g, Sugars: 3.82 g, Total Fat: 0.15 g, Saturated Fat: 0.016 g, Monounsaturated Fat: 0.008 g, Polyunsaturated Fat: 0.082 g, Cholesterol: 0 mg Micronutrients: Calcium: 14 mg Iron: 0.90 mg Magnesium: 19 mg Phosphorus: 35 mg Potassium: 255 mg Sodium: 5 mg Zinc: 0.22 mg Vitamin C: 181.9 mg Thiamin: 0.068 mg Riboflavin: 0.068 mg Niacin: 0.712 mg Pantothenic Acid: 0.046 mg Vitamin B6: 0.209 mg Vitamin B12: 0 mcg Folate: 17 mcg Vitamin A, IU: 884 IU Vitamin E: 0.52 mg Vitamin K: 10.7 mcg Phytonutrients: beta Carotene: 503 mcg beta Cryptoxanthin: 38 mcg Lycopene: 0 mcg Lutein and Zeaxanthin: 544 mcg Fresh Red Chile Peppers The following information is for one serving of red sweet peppers. That would be about one cup of sliced red sweet peppers, or 92 grams. This general information is for any variety of sweet red peppers. Macronutrients: Water: 84.83 g Calories: 24 Protein: 0.91 g Carbohydrates: 5.55 g Fiber: 1.8 g Sugars: 3.86 g Total Fat: 0.28 g Saturated Fat: 0.054 g Monounsaturated Fat: 0.006 g Polyunsaturated Fat: 0.144 g Cholesterol: 0 mg Micronutrients: Calcium: 6 mg Iron: 0.40 mg Magnesium: 11 mg Phosphorus: 24 mg Potassium: 194 mg Sodium: 2 mg Zinc: 0.23 mg Vitamin C: 174.8 mg Thiamin: 0.050 mg Riboflavin: 0.078 mg Niacin: 0.901 mg Pantothenic Acid: 0.292 mg Vitamin B6: 0.268 mg Vitamin B12: 0 mcg Folate: 17 mcg Vitamin A, IU: 2881 IU Vitamin E: 1.45 mg Vitamin K: 4.5 mcg Phytonutrients: beta Carotene: 1494 mcg beta Cryptoxanthin: 451 mcg Lycopene: 283 mcg Lutein and Zeaxanthin: 47 mcg Important Vitamins and Minerals found in Chile Peppers Calcium Importance: Builds and maintains bones and teeth; regulates heart rhythm; eases insomnia; helps regulate the passage of nutrients in & out of the cell walls; assists in normal blood clotting; helps maintain proper nerve and muscle function; lowers blood pressure; important to normal kidney function and in current medical research reduces the incidence of colon cancer, and reduces blood cholesterol levels. Deficiency Symptoms: May result in arm and leg muscles spasms, softening of bones, back and leg cramps, brittle bones, rickets, poor growth, osteoporosis ( a deterioration of the bones), tooth decay, depression.

Iron Importance: Its major function is to combine with protein and copper in making hemoglobin. Hemoglobin transports oxygen in the blood from the lungs to the tissues which need oxygen to maintain basic life functions. Iron builds up the quality of the blood and increases resistance to stress and disease. It is also necessary for the formation of myoglobin which is found only in muscle tissue. Myoglobin supplies oxygen to muscle cells for use in the chemical reaction that results in muscle contraction. Iron also prevent fatigue and promotes good skin tone. Deficiency Symptoms: May result in weakness, paleness of skin, constipation, anemia. Magnesium Importance: Plays an important role in regulating the neuromuscular activity of the heart; maintains normal heart rhythm; necessary for proper calcium & Vitamin C metabolism; converts blood sugar into energy. Deficiency Symptoms: May result in calcium depletion, heart spasms, nervousness, muscular excitability, confusion; kidney stones. Phosphorous Importance: Works with calcium to build strong bones and teeth. Helps in metabolism. Potassium Importance: Works with sodium to regulate the body's waste balance and normalize heart rhythms; aids in clear thinking by sending oxygen to the brain; preserves proper alkalinity of body fluids; stimulates the kidneys to eliminate poisonous body wastes; assists in reducing high blood pressure; promotes healthy skin. Deficiency Symptoms: May result in poor reflexes, nervous disorders, respiratory failure, cardiac arrest, muscle damage. Vitamin C : is essential in wound healing and in the formation of collagen, a protein important in the formation of healthy skin, tendons, bones, and supportive tissues. Deficiency results in defective collagen formation and is marked by joint pains, irritability, growth retardation, anemia, shortness of breath, and increased susceptibility to infection. Vitamin A : has a direct role in vision and is a component of a pigment present in the retina of the eye. It is essential for the proper functioning of most body organs and also affects the functioning of the immune system. Riboflavin or Vitamin B2: is required to complete several reactions in the energy cycle. Reddening of the lips with cracks at the corners of the mouth, inflammation of the tongue, and a greasy, scaly inflammation of the skin are common symptoms of deficiency.

Niacin or nicotinic acid : helps the metabolism of carbohydrates. Prolonged deprivation leads to pellagra, a disease characterized by skin lesions, gastrointestinal disturbance, and nervous symptoms. Vitamin B6 : is a coenzyme for several enzyme systems involved in the metabolism of proteins, carbohydrates, and fats. No human disease has been found to be caused by a deficiency of this vitamin. Chronic use of large doses of vitamin B6 can create dependency and cause complications in the peripheral nervous system. Folate or folic acid : is necessary for the synthesis of nucleic acids and the formation of red blood cells. Folic-acid deficiency most commonly causes folic-acid-deficiency anemia. Symptoms include gastrointestinal problems, such as sore tongue, cracks at the corners of the mouth, diarrhea, and ulceration of the stomach and intestines. Large doses of folic acid can cause convulsions and other nervous-system problems. Tryptophan (Essential Amino Acid) A natural relaxant, helps alleviate insomnia by inducing normal sleep; reduces anxiety & depression; helps in the treatment of migraine headaches; helps the immune system; helps reduce the risk of artery & heart spasms; works with Lysine in reducing cholesterol levels. Lysine (Essential Amino Acid) Insures the adequate absorption of calcium; helps form collagen ( which makes up bone cartilage & connective tissues); aids in the production of antibodies, hormones & enzymes. Recent studies have shown that Lysine may be effective against herpes by improving the balance of nutrients that reduce viral growth. A deficiency may result in tiredness, inability to concentrate, irritability, bloodshot eyes, retarded growth, hair loss, anemia & reproductive problems Phenylalaine (Essential Amino Acid) Used by the brain to produce Norepinephrine, a chemical that transmits signals between nerve cells and the brain; keeps you awake & alert; reduces hunger pains; functions as an antidepressant and helps improve memory.

CHILE NUTRITION Hot Peppers : Red, Canned Nutrient Proximates Protein Units 1 pepper 45g g 0.657

Carbohydrates Fiber Minerals Calcium Iron Magnesium Phosphorous Potassium Sodium Vitamins Vitamin C Riboflavin Niacin Vitamin B-6 Folate Vitamin A, IU Vitamin A, RE Vitamin E Amino Acids Tryptophan Lysine Phenyalanine

g g mg mg mg mg mg mg mg mg mg mg mg mg mg mg g g g

0.949 0.949 5.110 0.365 10.22 12.41 136.51 856.29 49.640 0.037 0.584 0.112 7.300 8681.160 867.970 0.504 0.009 0.029 0.020

Red Chile and Paprika Production in New Mexico New Mexico is the leading state in chile acreage (non-bell pepper, Capsicum annuum), and red chile and paprika represent approximately 40% of the states overall production. Red chile is derived from New Mexican-type chile varieties that are harvested when fruit have turned red or when they have reached physiological maturity. Paprika is a type of red chile and is the designation used for low (or no) heat, red Capsicum annuum varieties (Wall, 1994). Paprika varieties are also distinguished by their high levels of red pigments (capsanthin and capsorubin) in the pericarp (walls) of the fruit. The majority of the red chile and paprika crop is dehydrated and crushed into flakes or powder for use in a wide variety of products. Approximately 15% of the paprika crop is further processed into oleoresin paprika, a natural red food colorant (Walker, 2007). Cultivar Selection The larger red chile processing operations in the Southwest maintain in-house breeding programs, and all preferentially process their own proprietary cultivars (Bosland and Walker, 2004). Cultivars specifically bred to display attributes such as superior drying are preferred. For smaller operations and home gardeners, many of the commercially available New Mexican-type green chile cultivars also provide an excellent red, ripe, dry product. New Mexico 6-4 is a mild cultivar that works well as either a green or red chile. Sandia is a hot variety that can be used for either green or red chile, and is commonly used in the manufacture of red chile ristras

because of its excellent drying characteristics. Land Preparation A well-balanced crop rotation plan is essential for optimum red chile production. Chile, as well as other crops in the Solanaceae family (e.g., tomatoes, potatoes, eggplant), should not be planted in the same field more than once every three years. Rotation with a monocot (e.g., corn, wheat, oats, sudan grass) aids in reducing many of the disease and pest pressures associated with chile production (Pennock, 2003). Water accumulation or pooling in any part of the field encourages rapid reproduction and spread of chile wilt caused by the soilborne pathogen Phytophthora capsici. To reduce potential disease incidence, fields should be prepared for thorough drainage. Laser leveling fields at a grade of 0.01 to 0.03%, in one or both directions, is optimum (Bosland and Walker, 2004). In addition, digging drainage holes at the low end of the field is a best management practice to quickly drain large amounts of water from a field at times of high rainfall. Virtually all paprika grown in the Southwest for commercial processing is direct seeded and irrigated by furrow or overhead pivot. Because of the long growing period for red chile and paprika (approximately 165 days) and the desire to begin processing as early as possible, the crop is usually direct seeded early in the season (Bosland and Walker, 2004). Red chile and paprika fields are planted beginning in early March, and the majority of the crop has been seeded by mid-April. Seed should be obtained from a trusted source to ensure that it is clean and free from pathogens, such as bacterial leaf spot (Xanthomonas campestris), that can be spread via seed. Many early season diseases and pests impact chile seedlings, and many producers will therefore plant a high rate of seed (4 lb/ac or greater) to ensure strong stand establishment. Red chile and paprika fields are thinned or blocked to clumps of 1 to 3 plants, with the clumps spaced approximately 4 to 6 inches apart. A final stand of 40,000 plants per acre is optimum, but up to 80,000 plants per acre will not adversely affect quality or yield (Paroissien and Flynn, 2004). Fertilization Nitrogen and phosphorus are the two nutrients routinely added to the soil of red chile and paprika in New Mexico. A soil test prior to planting is recommended to determine nutritional needs. Phosphorus is relatively immobile in the soil, and so is typically applied pre-plant or banded into the soil at planting. Approximately 50 to 100 lb/ac of P2O5 are incorporated, depending on existing levels. Approximately 200 lb/ac of nitrogen are used by a chile crop over the course of the season. The optimum application window for nitrogen begins when plants reach the reproductive period (first bloom) and continues through early fruit development. Fertilization is usually stopped in mid-August to limit new growth and encourage ripening of fruit in anticipation of a red chile harvest. Harvest Harvest begins in late September and continues until the harvest quota is completed or all red chile is harvested. Mechanical harvest of the red chile and paprika crops has been widely adopted, with more than 80% of the commercial crop currently harvested by machine (Bosland and Walker, 2004). There are several different types of mechanical harvesters currently being used for red chile and paprika in the Southwest. Their performance is greatly affected by field and crop

conditions. The three most common picking heads are the finger-type, the belt-type, and the double helix. The finger-type head is based on a design incorporating a series of counterrotating bars with fingers. The fingers comb the plants to strip the pods off the plants and onto conveyor belts. The mechanism is very aggressive with the plants and can result in an excessive amount of harvested trash, especially after a freeze. The belt-type harvester has two sets of counter-rotating vertical belts imbedded with fingers that comb either side, from the bottom to the top, of the plant. If the belts are set low enough, fruit can be harvested off the ground with this machine. This machine also tends to be very aggressive with the plants and can pick up an excessive amount of dirt and debris if set to sweep the ground. The most widely used picking head, and most consistent for picking over the entire red chile harvest season, is the double helix model (Marshall, 1997). The helices may be vertical or oriented at an angle. The helices rotate in opposite directions to each other, snapping pods off of the plants and flipping them onto conveyor belts on either side of the helices. Typically, whether by hand or machine, the crop will undergo a once-over, destructive harvest. Maintaining fruit on the plants in the field until they are ready to be processed keeps post-harvest deterioration to a minimum; this has been the most cost-effective storage method for red chile (Cotter and Dickerson, 1984), and allows for some natural drying to occur. Following harvest from the plants, fruit must soon be dried to between 8 and 12% moisture to prevent decomposition while maximizing quality (Wall, 1994). Maximum yield and quality for a paprika crop are obtained from material harvested from o October through November, before the seasons first freeze (<28 F) (Kahn, 1992b; Palevitch, et al., 1975; Wall, 1994). While paprika continues to be harvested after the first freeze, quality and yields decline significantly (Walker, 2007). Red chile experiences a 0.5% yield loss for every day that harvest is delayed past the optimum harvest window. Yield losses are attributed to preharvest fruit abscission as well as fruit deterioration (Cotter and Dickerson, 1984; Kahn, 1992a). Processing Historically, red chile and paprika were dehydrated through sun drying. Ripe, red chile fruit were laid on rooftops, canal banks, or the ground and left in place until adequate drying was achieved. Although this technique is still commonly practiced in many chile-producing countries, only the smallest operations in the United States still utilize this method. Larger, commercial operations employ either tunnel or belt dryers. Both systems are more reliable and sanitary than sun drying (Wall, 1994). These methods also use a lot of energy, especially early in the harvest season when very succulent pods are dried. Another drawback to tunnel and belt dryers is their fixed capacity. Because of the finite capacity of the dehydration equipment, only a limited amount of chile fruit can be processed through a dehydration facility over a set period of time. Early in the season, when chile pods are more succulent, less material can be processed. Later in the season, throughput increases because drier raw material is entering the processing stream. While it would therefore seem that it is in the best interest of processors to harvest late in the season and take advantage of the rapid throughput, advantages are outweighed by disadvantages. As the plants begin to senesce, yield per plant dramatically declines and brittleness increases. Harvested trash tends to increase with increasing brittleness (Cotter and Dickerson, 1984; Walker, 2007). Harvest timing and efficiency can affect the profitability of both red chile growers and processors. Red chile grower reimbursements are based on the amount of dried, processed product. Processors measure weight lost during dehydration by dividing the incoming fresh weight by the dry, processed weight to obtain what is termed shrink. Because of the critical relationship between pod succulence and throughput efficiency for red chile dehydrators, shrink is carefully monitored during the course of the harvest season. Typically, shrink is much greater early in the harvest season and decreases steadily as the chile plants continue to mature and senesce. Early in the season, the very succulent fruit have a shrink ratio of 7:1 or 8:1 (Wall,

1994). Late in the season, when fruit and plants are very dry, shrink ratio drops to around 1:1. In addition to date of harvest, chile cultivars have a large impact on shrink values. The development of cultivars with increased dry yield and lower shrink is an ongoing priority of paprika breeding programs (Walker et al., 2004). Harvest Aids Virtually all red chile and paprika grown for processing plants and harvested in the succulent stage undergoes application of salt in the form of sodium chlorate before harvest (Wall, 1994). The main purpose of salting the plants is defoliation to provide a cleaner harvest. The removal of leaves from the plants also hastens senescence and therefore encourages ripening of the fruit. A maximum rate of 10 lb/ac of salt is applied, with reduced rates for less succulent material. The chile is harvested between 10 to 14 days after the application of the salt. The treatment is relatively inexpensive and effective at removing much of the foliage that would otherwise interfere with a clean harvest of the material. Fields harvested in the last half of the season are not sprayed because the fruit dry too fast, and by late in the season, adequate natural defoliation and ripening have occurred (Walker, 2007). Ethylene is sometimes applied along with the salt (Wall, 1994). Ethylene is a naturally occurring hormone that induces ripening in fruit and can also assist in defoliation of the plants. The use of ethylene products for paprika production has been limited because fruit drop is often an unintended consequence of application. The treatments effectiveness is greatly affected by environmental conditions (Conrad and Sundstrom, 1987; Kahn, 1992; Wall et al., 2003), so it is difficult to gauge the proper timing or concentration of ethylene needed to obtain the desired effect. Red chile and paprika are used extensively in a wide variety of food products, and New Mexicos crop is known for its consistent quality. Through careful cultivar selection and use of best production practices, growers can continue to profitably produce this crop that is a critical component of New Mexicos chile industry. Walker, S. (2004). Red Chile and Paprika Production in New Mexico. New Mexico: New Mexico State University. Walker describes in Red Chile and Paprika Production in New Mexico the management of chili pepper in Mexico. It gives an overview of chili pepper regarding its cultivar selection, land preparation, fertilization, harvest, harvest aids and nutritional content. This information can help us find the proper location of chili pepper. It also gives us an idea of the environment of chili pepper. Its environment can affect its chemical and biological composition. The composition proves the effectivity of the chili pepper pesticide.

Capsicums: Innovative Uses of an Ancient Crop The Capsicum genus, chile, represents a diverse plant group, from the well known sweet green bell pepper to the fiery hot, recently exploited habanero. When Columbus tasted the small red berries he found on his voyage, he believed he had reached India and called them red pepper. Red pepper, Capsicum, has been domesticated for 7,000 years. How then, can the most consumed spice in the world be considered a new crop? One reason is that Capsicum, chile, is an essential ingredient in the fastest growing food sector in the United States, "Mexican or Southwestern food." In addition, many of the new uses of chile are hidden within manufactured products. Chile is being used as a food flavoring, a coloring agent, a pharmaceutical ingredient, and in other innovative ways. The use, and uses, of the numerous cultivars within the five domesticated species has grown exponentially. History Capsicum has been known since the beginning of civilization in the Western Hemisphere. It has been a part of the human diet since about 7500 BC (MacNeish 1964). It was the ancient ancestors of the native peoples who took the wild chile piquin and selected for the many various types known today. Heiser (1976) states that apparently between 5200 and 3400 BC, the Native Americans were growing chile plants. This places chiles among the oldest cultivated crops of the Americas. As opposed to most domesticated crops, the wild ancestral chiles are not looked upon as worthless or inferior by farming people who cultivate their domestic decedents. The wild Capsicum annuum var. aviculare is harvested and sold in the marketplace along side the larger-fruited domesticated chiles. Capsicum was domesticated at least five times by prehistoric peoples in different parts of South and Middle America. The five domesticated species are C. annuum L., C. baccatum L., C. chinense Jacq., C. frutescens L., and C. pubescens R. & P.. Chile is historically associated with the voyage of Columbus (Heiser 1976). Columbus is given credit for introducing chile to Europe, and subsequently to Africa and to Asia. On his first voyage, he encountered a plant whose fruit mimicked the pungency of the black pepper, Piper nigrum L. Columbus called it red pepper because the pods were red. The plant was not the black pepper, but a heretofore unknown plant that was later classified as Capsicum. Capsicum is not related to the Piper genus. In 1493, Peter Martyr (Anghiera 1493) wrote that Columbus brought home "pepper more pungent than that from the Caucasus." Chile spread rapidly across Europe into India, China, and Japan. The new spice, unlike most of the solanums from the Western Hemisphere, was incorporated into the cuisines instantaneously. Probably for the first time, pepper was no longer a luxury spice only the rich could afford. Since its discovery by Columbus, chile has been incorporated into most of the world's cuisines. It has been commercially grown in the United States since at least 1600, when Spanish colonists planted seeds and grew chile using irrigation from the Rio Chama in northern New Mexico (DeWitt and Gerlach 1990). Capsicum terminology is confusing. Pepper, chili, chile, chilli, aji, paprika, and Capsicum are used interchangeably for plants in the genus Capsicum. Capsicum investigators use chile, pepper, or aji, as vernacular terms. Capsicum is reserved for taxonomic discussion. The word "chile" is a variation of "chil"

derived from the Nahuatl (Aztec) dialect which referred to plants now known as Capsicum, whereas "aji" is a variation of "axi" from the extinct Arawak dialect of the Caribbean. This brings up the point of the correct way to spell "chile" (Domenici 1983). The "e" ending in chile is the authentic Hispanic spelling of the word, whereas English linguists have changed the e to an i. Chile pepper has come to mean pungent chile cultivars. However, chile means pepper (Capsicum) whether the fruits are pungent or not. Generally, chili is used to identify the state dish of Texas, which is a combination of pungent chile cultivars and meat (Domenici 1983). Bell pepper generally refers to non-pungent blocky chile types. Additional confusion is present within species designation, because C. annuum was sometimes called C. frutescens in the scientific literature (Heiser and Smith 1953). Botany The genus Capsicum is a member of the Solanaceae family that includes tomato, potato, tobacco, and petunia. The genus Capsicum consists of approximately 22 wild species and five domesticated species (Bosland 1994): C. annuum, C. baccatum, C. chinense, C. frutescens, and C. pubescens. Capsicum is endemic to the western hemisphere and the preColumbian distribution extended from the southernmost border of the United States to the temperate zone of southern South America (Heiser 1976). It is a perennial small shrub in suitable climatic conditions, living for a decade or more in tropical South and Central America. Capsicum probably evolved from an ancestral form in the Bolivia/Peru area (Heiser 1976). Chile fruits are considered vegetables, but are berries botanically. Chile types usually are classified by fruit characteristics, i.e. pungency, color, shape, flavor, size, and their use (Smith et al. 1987; Bosland 1992). Despite their vast trait differences most chile cultivars commercially cultivated in the world belong to the species, C. annuum. The tabasco (C. frutescens) and habanero (C. chinense), are the best-known exceptions. Pod-types and Cultivars

Several hundred chile pod-types are grown worldwide. Chile pod-types are a subspecific category that allows for distinguishing among the specific horticultural varieties. Types such as ancho, bell, jalapeo, pasilla, New Mexican, yellow wax, are distinct pod-types that have specific traits for processing and fresh use, flavor and pungency. The most economically important species in the world is C. annuum(Greenleaf 1986; Bosland et al. 1988). Many podtypes are central ingredients in ethnic dishes such as ancho and New Mexican for chile rellenos, pasilla for mole sauce, and serrano for pico de gallo. Like wine-tasting, one can discern between the subtle flavors of chiles after a few years of experience; ancho is sweetish, mulatto is chocolaty, mirasol is fruity, and chilpotle is smokey. Grinding the pods produces one flavor, toasting before grinding produces another, and soaking the chiles in water produces yet another. For a more detailed description and use of the various pod-types the reader is referred to Bosland et al. (1988); Bosland (1992); and DeWitt and Bosland (1993). By the time the Spanish arrived in Mexico, Aztec plant breeders had already developed dozens of pod-types. According to historian Bernardino de Sahagn, who lived in Mexico in 1529, "hot green chiles, smoked chiles, water chiles, tree chiles, beetle chiles, and sharppointed red chiles" existed (DeWitt and Gerlach 1990). Undoubtedly, these chiles were the

precursors to the large number of pod-types found in Mexico today. Even though many podtypes exist, improvement continues today because of specific needs in the industry. Improvement in quality and yields are occurring, along with refinement of the pod-types to fit special niches of the industry. Fresh market types such as bell peppers, cuban, and squash, are usually non-pungent, and eaten raw, stir-fried, or prepared in some fashion to season a culinary dish. Bell apparently refers to the fruit's blocky shape, with four lobes preferred in the United States. Bell types were first mentioned in 1699 by the English pirate, Wafer, who found them growing in Panama (Wafer 1699). The conventional green bell pepper has given way to innovative colored pods that can ripen to shades of red, orange, yellow, and brown. A survey in the 1992 issue of Fresh Trends stated that 95% of the people who tried a colored bell liked them and 5 out of 6 preferred them over the green bell. More people are trying colored bell peppers than any other produce category. There has been an increase of 75% in the consumption of colored bells in the United States. In 1992, colored bells were tried by as many consumers as leaf lettuce, fresh asparagus, peas, squash, and red potatoes. Consumers bought more colored bells than yellow beans, artichokes, or romaine lettuce. The innovative fruit colors have brought increased consumption and sales. Chile processing pod-types can be grouped into two main categories. The first consists of a fresh product that is frozen, canned, or pickled Some of the pod-types included in this group are the pimento, jalapeo, serrano, pepperoncini, yellow wax, and the New Mexican. The second category consists of those used as a dehydrated product. Dehydrated pod-types are New Mexican, cayenne, ancho, pasilla, mirasol, piquin, and de arbol (Bosland 1992). A relatively recent pod-type is the New Mexican, also called long green or 'Anaheim' type. Actually, the pod-type is New Mexican, and 'NuMex Big Jim' and 'Anaheim' are cultivars within this type. The New Mexican pod-type was developed in 1894 when Fabian Garcia at New Mexico State University began improving the local chiles grown by the Hispanic gardeners around Las Cruces, New Mexico. This type, characterized by long green pods that turn red, is the chile of choice for Mexican-style cooked sauces in the United States. Green and red chile represent two developmental states of the same fruit. 'Anaheim' seed originated in New Mexico and was brought to Anaheim, California, where it was widely cultivated. Thus, until Dr. Garcia released the New Mexican pod-type, it did not exist. If pods are left on the plant to be harvested at the red stage, they are usually dried and ground into chile powder (paprika if non-pungent). A green, New Mexican chile pod contains three times the vitamin C of a 'Valencia' orange and provides the minimum daily requirement. As green pods turn red, pro-vitamin A content increases until they contain twice the pro-vitamin A of a carrot (Lantz 1943). A one half tablespoon of red chile powder furnishes the minimum daily requirement of vitamin A. A peculiar chile category is paprika. It is not a pod-type in the United States, but it is a product. In Europe, there are chile pod-types that are paprikas. This is because in the Hungarian language "paprika" meansCapsicum (Somos 1984). Paprika is defined in the United States as a sweet, dried, red powder. This mild powder can be made from any type of C. annuum that is non-pungent and has brilliant red color. Paprika may be pungent in Hungary, but paprika is always non-pungent in international trade. A small group of chiles can be classified as ornamental. Although edible, ornamentals are grown primarily for their unusual pod shapes or for their dense foliage and colorful fruits. Ornamental chiles can have all the colors of the rainbow, often displaying pods in four or five

colors on the same plant at the same time (Bosland et al. 1994). In the past, they have been called Christmas peppers because of the bright red fruits during the holiday season. Decorations, such as wreaths, made with chile that can be dehydrated are popular in the southwestern United States, and are a major tourist product. A tradition in New Mexico is to harvest mature red chiles and string them into colorful strings (ristras). The ristra is hung near the entrance of the house as a symbol of hospitality. Ornamental chiles have become an innovative way for small farmers to produce a high-value alternative crop. Exotic Germplasm

Exotic germplasm provides a bountiful source of extraordinary genetic diversity with which to improve the commercial chiles. For the past decade, it has played a significant role in the New Mexico State University chile breeding and genetics program. Because exotic germplasm inhabits a vast array of ecological zones, it can serve as a reservoir of useful and needed genes. However, the potential value of exotic germplasm for improvement of commercial chile is under exploited. Such genetic resources clearly deserve more intensive investigation. The five domesticated species have close wild relatives with which they cross readily, producing viable and fertile hybrids. These relatives have not been evaluated extensively, and may contain useful sources of resistance to viruses, bacteria, and fungi. The C. chinense species, like all Capsicum species, originated in the New World. However, the French taxonomist who named this species in 1776 got his seed from China (Smith and Heiser 1957). Habanero, or Scotch Bonnet, is the best known chile of this species. One accession of Habanero has the distinction of being the world's hottest chile (in excess of 200,000 scoville heat units). Fruit shape can vary from long and slender to short and obtuse. Fruit can be extremely pungent and aromatic, with persistent pungency when eaten. C. chinense is popular in all tropical regions. The C. frutescens species is represented by two cultivars, tabasco and malagueta. Tabasco is the most common cultivar of C. frutescens. The red fruit is the ingredient in Tabasco sauce. The malagueta is a popular cultivar in Brazil. It is not related to Aframomum melegueta, the melegueta or Guinea pepper, from Africa which is related to ginger. In South America, C. baccatum is the most commonly grown species, where it is called aji, not chile. Three botanical varieties of C. baccatum are recognized: C. baccatum var. baccatum, C. baccatum var.pendulum, and C. baccatum var. microcarpum. C. baccatum flowers have yellow, brown, or dark green spots on the corolla. As many different pod-types of chiles (in relation to shape, color, and size) exist within C. baccatum as in C. annuum. Fruits vary in pungency from non-pungent to very hot. They embody unique aromatics and flavors that can be overpowering to some people. C. baccatum is the chile of choice when making ceviche (marinated fish). Another of the five species, C. pubescens, is a relatively unknown chile. It is found from Mexico to Peru, growing in the Andean South America and the Central American highlands. Common names includerocoto or locoto in South America. Other common names are manzano and peron because the fruits can be apple- or pear-shaped. Instead of white flowers, it has purple flowers with large nectaries. The presence of conspicuous leaf

pubescence and black seeds readily distinguish this chile from any of the other species. This chile is adapted to cooler temperatures, 4.4 to 21.1C (40 to 70F), but does not tolerate frost. The other approximately 20 Capsicum species lack extensive study on their biology. It is interesting to note that all wild chiles have small fruits which are eaten with ease by birds, the natural dispersal agent forCapsicum. Many of the known wild species have restricted distribution. These species may contain genes for adaptation to unusual environmental conditions as well as disease resistance. Exotic germplasm will continue to be an important asset for breeding improved commercial chiles. Not only will it be useful in breeding for disease resistance, but it will be used to increase the nutritional quality, yield, and efficiency for machine harvesting of the crop. The enhancement of commercial cultivars by exotic germplasm is dependent on the availability of living material. This may be impossible because the natural habitat for these species is in dire straits. Collection may never be accomplished before their extinction. Tropical deforestation is among the most massive and urgent environmental problems facing Capsicumgermplasm resources. The United States National Plant Germplasm System houses an extensive Capsicum germplasm collection at the Southern Plant Introduction Station located in Experiment, Georgia. This collection contains approximately 3000 Capsicum accessions that includes lines from all over the world. Passport data are recorded upon arrival of the seed at the facility and a USDA Plant Introduction (PI#) Number is assigned. Evaluation data is subsequently entered in the Germplasm Resources Information Network (GRIN) after it has been received. The material held in the USDA collection is the source of germplasm for breeding and research programs throughout the world. Many have been evaluated for descriptors. Preserving genetic diversity in germplasm is of paramount importance. The genetic diversity of Capsicum can be saved only through the use of several strategies. The genetic resources of Capsicum need to be enlarged and conserved in base and active genebanks. The improvement of appropriate storage facilities for germplasm and the financial support of those operations is very urgent. It is especially imperative to aid the active collections of Latin America, because Capsicum is native to this region. When possible it is desirable to set up Capsicum genetic resources reserves in conjunction with relevant biosphere resources and other protected areas. Breeding In breeding chiles, the choice of breeding method depends on the breeding objective and the plant material being used as parents (Greenleaf 1986). The strategy of the chile breeder is to assemble into a cultivar the superior genetic potential for yield, protection against production hazards, and improved quality. Chile cultivars have been developed using selection within introductions and hybridization followed by selection. Hybridization is usually always within a species, but interspecific crosses, especially C. annuum by C. chinense have been accomplished successfully. Selection methods have included mass, single plant, backcross, and pedigree. Single seed descent and haploid breeding have also been applied to chiles. The only breeding technique so far not applicable to chiles is genetic transformation. This is because the technique to regenerate whole chile plants from single cells has not been ascertained. With

the success of other solanaceous crops, e.g. tomato and petunia, the parameters necessary for successful regeneration of chile will be forth coming. Chile plants are considered a self-pollinating crop (Allard 1960). However, the rates of out-crossing (7% to 91%) recorded by several investigators argue that Capsicum should be considered to be facultative cross-pollinating species in field research (Odland and Porter 1941; Franceschetti 1971; Tanksley 1984). The out-crossing is associated with natural insect pollinators. The amount of cross-pollination has an effect not only on the precautions needed for seed production, but also on the breeding methodologies used by the plant breeder (Bosland 1993). Natural pollinators such as insects must be excluded to insure self-pollination. The commercial production of hybrid chiles has been successful accomplished using hand-emasculation, genic male-sterility, and cytoplasmic male-sterility. Male-sterile chile plants are found as spontaneous mutants in commercial fields. Cytoplasmic male sterility (CMS) in chiles was first discovered by Peterson (1958) in C. annuum PI 164835 from India. Unfortunately, Peterson's CMS line is unstable in fluctuating environments, producing pollen at cool temperatures. Some commercial seed companies use the CMS system to produce F1 hybrids. Other genetic systems that may assist the chile breeder are the use of trisomic and chromosome mapping. Most Capsicum species have 2n = 24 chromosomes. Meiosis is surprisingly regular within interspecific crosses. The similarity in karyotype of the species was illustrated by Ohta (1962). However, cryptic structural hybridity as defined by Stebbins (1971) appears to be functioning in Capsicum. Pochard (1970) identified primary trisomics that could be distinguished from secondary and tertiary trisomics by their phenotype, frequency, and fertility and by their chromosome configurations in meiosis. Eleven of the 12 possible trisomics could be identified by foliage color, anther/stamen color, and fruit characteristics. Cytologically, only three of the 12 chromosomes in C. annuum differ significantly in length. The other nine are metacentric and have lengths too close to permit identification. The recent development of molecular marker-assisted selection techniques may provide new tools in the breeding of chiles. Isozyme and molecular markers have been applied to chiles. Usefulness of isozyme markers is restricted to the insufficient polymorphic bands and a limited number of detectable loci. Molecular based DNA markers such as Restriction Fragment Length Polymorphism (RFLP) and Random Amplified Polymorphic DNA (RAPD) are unlimited in numbers. A saturated isozyme and RFLP map of chile has been reported (Prince et al. 1993). The map contains 192 chile and tomato genomic cDNA clones with 19 linkage groups with a total coverage of 720 cM. However, specific map positions of 26 RFLP markers in seven linkage groups were not determined and vast regions of the chile genome remain unmapped. This area of research is rapidly adding to our knowledge of chiles and will undoubtedly be important in the future. A peculiar aspect of chiles is the inability to be regenerated from protoplasts. This limits the technique of genetic transformation. Within other solonaceous crops such as tomato, tobacco, and petunia, excellent progress has been made regenerating plants using genetic transformation to introduce novel genes into the genome. For unexplained reasons, chile has been recalcitrant to being regenerated but many laboratories are addressing this issue. Once

this perplexing problem is solved, genetic transformation will be available to introduce novel genes into chiles. Pungency The one attribute most typical of chiles is pungency and must be considered one of its most important traits. Some have argued that pungency should be one of the five main taste sensories, along with bitter, sweet, sour, and salty. Chile pungency is a desirable attribute in many foods. In most parts of the world, pungency increases the acceptance of the insipid basic nutrient foods. Many innovative uses of pungency are being studied. Besides new medicinal applications, it has been tried as a barnacle repellent, to repel mice from gnawing on underground electrical cables, and to keep squirrels from eating bird seed. Anti-mugger aerosols with chile pungency as the active ingredient have replaced mace and tear gas in more than a thousand police departments in the United States. The spray will cause attackers to gasp and twitch helplessly for 20 minutes. Pungency is produced by the capsaicinoids, alkaloid compounds, that are found only in the plant genus, Capsicum. The nature of the pungency has been established as a mixture of seven homologous branded-chain alkyl vanillylamides (Hoffman et al. 1983). They often are called capsaicin after the most prevalent one. Dihydrocapsaicin is usually the second most prevalent capsaicinoid, while the other five compounds, norcapsaicin, nordihydrocapsaicin, nornordihydrocapsaicin, homocapsaicin, homodihydrocapsaicin, are considered minor capsaicinoids because of their relative low abundance in most natural products. Capsaicin is a powerful and stable alkaloid that can be detected by human taste buds in solutions of ten parts per million. Capsaicin's composition (C18H27NO3) is similar to peperin (C17H19NO3),that gives black pepper its bite. The capsaicinoids are produced in glands on the placenta of the fruit. While seeds are not the source of pungency, they occasionally absorb capsaicin because of their proximity to the placenta. No other plant part produces capsaicinoids. Chile pungency is expressed in Scoville Heat Units (Scoville 1912). The Scoville Organoleptic Test was the first reliable measurement of the pungency of chiles. This test used a panel of five human representatives, who tasted a chile sample and then recorded the heat level. A sample was diluted until pungency could no longer be detected. The organoleptic method or taste test has been the standard method for pungency analysis. Although this method is widely used, it has limitations. Tasters must be trained and their ability to test many samples is restricted by the heat of the test solution. Taster fatigue is a real phenomenon and tasters are also not able to distinguish between the different capsaicinoids. Therefore, the Scoville Organoleptic Test has been replaced with instrumental methods. The most common instrumental method is high-performance liquid chromatography (HPLC). It provides accurate and efficient analysis of content and type of capsaicinoids present in a chile sample (Woodbury 1980; Collins et al. 1995). HPLC analysis has become the standard method for routine analysis by the processing industry. The method is rapid and can handle a larger number of samples. A common practice today is to multiply capsaicinoid ppm by 15 to convert to SHU.

The chile pungency level has genetic and environmental components. The capsaicinoid content is affected by the genetic make-up of the cultivar, weather conditions, growing conditions, and fruit age. Plant breeders can selectively develop cultivars with varying degrees of pungency. Also, growers can somewhat control pungency by the amount of stress to which they subject their plants. Pungency is increased with increased environmental stress (Lindsay and Bosland 1995). More specifically, any stress to the chile plant will increase the amount of capsaicinoid level in the pods. A few hot days can increase the capsaicinoid content significantly. In New Mexico, it has been observed that even after a furrow irrigation, the heat level will increase in the pods. Anthropopathically, the plant has sensed the flooding of its root zone as a stress, and has increased the capsaicinoid level in its pods. If the same cultivar was grown in both a hot semi-arid region and a cool coastal region, the fruit harvested from the hot semi-arid region would be higher in capsaicinoids than that the fruits harvested in the cool coastal climate. The medicinal applications of capsaicinoids have brought innovative ideas for their use. Medicinal use of Capsicums has a long history, dating back to the Mayas who used them to treat asthma, coughs, and sore throats. The Aztecs used chile pungency to relieve toothaches. The pharmaceutical industry uses capsaicin as a counter-irritant balm for external application (Carmichael 1991). It is the active ingredient in Heet and Sloan's Liniment, two rubdown liniments used for sore muscles. The capsaicin is being used to alleviate pain. Its mode of action is thought to be from nerve endings releasing a neurotransmitter called substance P. Substance P informs the brain that something painful is occurring. Capsaicin causes an increase in the amount of substance P released. Eventually, the substance P is depleted and further releases from the nerve ending are reduced. Creams containing capsaicin have reduced pain associated with post-operative pain for mastectomy patients and for amputees suffering from phantom limb pain. Prolonged use of the cream has also been found to help reduce the itching of dialysis patients, the pain from shingles (Herpes zoster), and cluster headaches. Further research has indicated that capsaicin cream reduces pain associated with arthritis. The repeated use of the cream apparently counters the production of substance P in the joint, hence less pain. Reducing substance P also helps by reducing long-term inflammation, which can cause cartilage break down. Pigments Color is very important in paprika and chile powder. Paprika and paprika oleoresin are currently used in a wide assortment of foods, drugs, and cosmetics, as well as for improving the feather color of flamingoes in zoos or koi in aquariums. Carotenoids control pod color with approximately 20 carotenoids contributing to the color of the powder. Carotenoid compounds are yellow to red pigments of aliphatic or alicyclic structures composed of isoprene units, which are normally fat-soluble colors (Bunnell and Bauernfeind 1962). The keto-carotenoids, capsanthin, capsorubin, and cryptocapsin are unique Capsicum carotenoids. The major red color in chile comes from the carotenoids capsanthin and capsorubin, while the yellow-orange color is from beta-carotene and violaxanthin. Capsanthin, the major carotenoid in ripe fruits, contributes up to 60% of the total carotenoids. Capsanthin and capsorubin increase proportionally with advanced stages of ripeness; with capsanthin being the more stable of the two (Harkay-Vinkler 1974; Kanner et al. 1977). The amount of carotenoids in fruit tissue depends on factors such as cultivar, maturity stage, and growing conditions (Reeves 1987). The reader is referred to Wall and Bosland (1993) for further information on chile pigments.

Conclusion Innovative uses are still being found for chile. For example, the Black Mountain Brewing Co. in Arizona developed a chile beer. The idea was to produce a spicy beer for a local Mexican restaurant. The idea worked, and now, the beer company is one of the top 20 selling microbreweries in the United States. The chile industry has grown from a regional food for tourists to an industry competing on the international market. At present, there is an expanding chile market for dietary consumption. At the same time, chile is also becoming more important in manufacturing other commodities. Chile will become more important as a food coloring agent and this may be the leading use of chile in the future. The most innovative use of chile by humankind may yet be discovered. The College of Agriculture and Home Economics at New Mexico State University has founded the Chile Institute. The Chile Institute is a non-profit, international organization devoted to the study of Capsicum. Some of the goals of The Chile Institute are to assist in preserving Capsicum germplasm, provide an international information clearinghouse and be an archive of chile publications. It has already published a bibliography containing more than 6,000 references (Bosland 1995). Bosland, P. (1996). Capsicums: Innovative uses of an ancient crop. Arlington: ASHS Press. In Capsicums: Innovative Uses of an Ancient Crop, it illustrates the history, botany, podtypes, cultivars, exotic germplasms, breeding, pungency and pigments of capsicum. Capsicum is found in chili pepper. It can help our study by giving us an idea about the application of chili pepper. It presents capsicum in a microscopic and macroscopic view. Analytical procedures regarding capsicum are tackled like high-performance liquid chromatography (HPLC). This can be utilized to gain more feasible data on chili pepper pesticide effectivity.

ANNOTATED BIBLIOGRAPHY

4ChEA Buenafe, Mia Ariela I. Ferrer, Lara Melissa V. Dy, Paulo L.