RP-HPLC and transcript profile indicate increased leaf caffeine in Coffea canephora plants by light

Avinash Kumar, P. S. Simmi, Gyanendra Kumar Naik, Parvatam Giridhar


Light is a survival quotient for all photosynthetic plants and its reception is very complex due to direct regulation by photoreceptors and their downstream transcriptional factors or indirectly by circadian rhythm. Shade-grown coffee cultivation though less productive than the sun tolerant varieties, pose high potential as benefit to the environment. Other than high nutrient soil associated with shade-cultivated coffee, light is another important difference when compared to full sun cultivation practice. It is thus important to study if light has a role in accumulation of caffeine - the most undesired compound in coffee. Light irradiation of suspension cultures of Coffea arabica enhances caffeine content. However, no such study is available on whole plants, which are anticipated to act in accord with organismal homeostasis. Moreover, the promoter of theobromine synthase-like gene involved in caffeine biosynthesis carries several light responsive motifs. In this report, it is shown that in complete darkness the caffeine content in young leaves of 1 year old seedlings is very low (0.094±0.003 mg/100 mg tissue dw.). However, it increases to 5.9 folds within 6hrs of exposure to light. In addition, caffeine content drops (0.218±0.03; mg/100 mg tissue dw.) when light exposed plants are returned to complete dark. Transcript analysis further reveals that this difference is due to regulation of the caffeine biosynthetic genes. A further discussion to the effect of dark and light on levels of caffeine is also provided. Though cup quality of shade-grown coffee is indefinite, this study clearly demonstrates the role of light in regulation of caffeine biosynthesis.


Coffea; N-methyltransferase; Light; Transcript profiling; High Performance Liquid Chromatography

Full Text:



Bou-Torrent J, Roig-Villanova I, Martínez-García JF. Light signalling: back to space. Trends Plant Sci. 2008; 13(3): 108-114. http://dx.doi.org/10.1016/j.tplants.2007.12.003

Roden LC, Carre IA. The molecular genetics of circadian rythyms in Arabidopsis. Semin Cell Dev boil. 2001; 12(4): 305-315. http://dx.doi.org/10.1006/scdb.2001.0258

Zhou D-X. Regulatory mechanism of plant gene transcription by GT-elements and GT-factors. Trends Plant Sci. 1999; 4(6): 210-214. http://dx.doi.org/10.1016/S1360-1385(99)01418-1

Charrier A, Berthaud J. 1985. Botanical classification of coffee. In: Coffee: Botany, Biochemistry and Production of beans and Beverage. Ed. Clifford MN and Willson KC. pp. 13-47. Westport, Connecticut, AVI publishing company Inc.

Gobbi JA. Is biodiversity-friendly coffee financially viable? An analysis of five different coffee production systems in western El Salvador. Ecol Econ. 2000; 33(2): 267-281. http://dx.doi.org/10.1016/S0921-8009(99)00147-0

Elmqvist T, Tuvendal M, Krishnaswamy J Hylander K. 2013. Managing trade-offs in ecosystem services. In: Values, Payments and Institutions for Ecosystem Management. Ed. Kumar P and Thiaw I. pp. 70-89. UK, Edward Elgar publishing.

Marie-Vivien D, Garcia CA, Kushalappa CG, Vaast P. Trademarks, geographical indications and environmental labelling to promote biodiversity: the case of agroforestry coffee in India. Development Policy Review. 2014; 32(4): 379-398. http://dx.doi.org/10.1111/dpr.12060

Reddy DRB, Raghuramulu Y Naidu R. 2004. Impact of diversification in Indian coffee plantations-a sustainable approach. In: 20th International Conference on Coffee Science. pp. 1129-1135. Bangalore, ASIC 2004.

Geromel C, Ferreira LP, Davrieux F, Guyot B, Ribeyre F, Scholz MB dos S, Pereira LFP, Vaast P, Pot D, Leroy T, Filho AA, Vieira LGE, Mazzaferra P, Marraccini P. Effects of shade on the development and sugar metabolism of coffee (Coffea Arabica L.) fruits. Plant Physiol. Biochem. 2008; 46(5-6): 569-579. http://dx.doi.org/10.1007/s13580-012-0078-3

Joёt T, Salmona J, Laffargue A, Descroix F, Dussert S. Use of the growing environment as a source of variation to identify the quantitative trait transcripts and modules of co-expressed genes that determine chrologenic acid accumulation. Plant Cell Env. 2010; 33(7): 1220-1233. http://dx.doi.org/10.1111/j.1365-3040.2010.02141.x

Schmitt-Harsh M, Evans TP, Castellanos E, Randolph JC. Carbon stocks in coffee agroforests and mixed dry tropical forests in the western highlands of Guatemala. Agroforest Syst. 2012; 86(2): 141-157. http://dx.doi.org/10.1007/s10457-012-9549-x

Ricci M dos SF, Rouws JRC, de Oliveira NG, Rodrigues MB. Vegetative and productive aspects of organically grown coffee cultivars under shaded and unshaded systems. Sci. Agric. 2011; 68(4): 424-430. http://dx.doi.org/10.1590/S0103-90162011000400006

James JE, Crosbie J. Somatic and psychological health implications of heavy caffeine use. Brit J Addict. 1987; 82(5): 503-509. http://dx.doi.org/10.1111/j.1360-0443.1987.tb01507.x

Pichersky E, Lewinsohn E. Convergent evolution in plant specialized metabolism. Annu Rev Plant Biol. 2011; 62: 549-566. http://dx.doi.org/10.1146/annurev-arplant-042110-103814

Schimpl FC, Kiyota E, Mayer JLS, Gonҫalves JF de C, da Silva JF, Mazzafera P. Molecular and biochemical characterization of caffeine synthase and purine alkaloid concentration in guarana fruit. Phytochemistry. 2014; 105: 25-36. http://dx.doi.org/10.1016/j.bbrc.2014.09.043

Suzuki T, Ashihara H, Waller GR. Purine and purine alkaloid metabolism in Camellia and Coffea plants. Phytochemistry. 1992; 31(8): 2575-2584. http://dx.doi.org/10.1016/0031-9422(92)83590-U

Kato M, Mizuno K. Caffeine synthase and related methyltransferases in plants. Front biosci. 2004; 9: 1833-1842.

Denoeud F, Carretero-Paulet, Dereeper A, et al. The coffee genome provides insight into convergent evolution of caffeine biosynthesis. Science. 2014; 345(6201): 1181-1184. http://dx.doi.org/10.1126/science.1255274

Satyanarayana KV, Kumar V, Chandrashekar A, Ravishankar GA. Isolation of promoter for N-methyltransferase gene associated with caffeine biosynthesis in Coffea canephora. Electron J Biotechn. 2005; 119(1): 20-25. http://dx.doi.org/10.1016/j.jbiotec.2005.06.008

Kurata H, Matsumura S, Furusaki S. Light irradiation causes physiological and metabolic changes for purine alkaloid production by a Coffea arabica cell suspension culture. Plant Sci. 1997; 123(1-2): 197-203. http://dx.doi.org/10.1016/S0168-9452(96)04588-8

Mishra MK, Sandhyarani N, Suresh S, Kumar S, Soumya PR, Yashoda MH, Bhat A, Jayarama. 2012; Journal of Crop Improvement. 2012; 26 (6): 727-750. http://dx.doi.org/10.1080/15427528.2012.696085

Ashihara H, Monteiro AM, Moritz T, Gillies FM, Crozier A. Catabolism of caffeine and related purine alkaloids in leaves of Coffea arabica L. Planta. 1996; 198(3): 334-339. http://dx.doi.org/10.1007/BF00620048

Thippeswamy R, Gouda KGM, Rao DH, Martin A, Gowda LR. Determination of theanine in commercial tea by liquid chromatography with fluorescence and diode array ultraviolet detection. J Agric Food Chem. 2006; 54(19): 7014-7019. http://dx.doi.org/10.1021/jf061715

Ogita S, Uefuji H, Morimoto M, Sano H. Application of RNAi to confirm theobomine as the major intermediate for caffeine biosynthesis in coffee plants with potential for construction of decaffeinated varieties. 2004; Plant Mol Biol 54(6):931-941. http://dx.doi.org/10.1007/s11103-004-0393-x

Pompelli MF, Pompelli GM, de Oliveira AFM, Antunes WC. The effect of light and nitrogen availability on the caffeine, theophylline and allantoin contents in leaves of Coffea arabica L. Aims Environmental Science. 2014; 1(1): 1-11.


Coelho GC, Rachwal MFG, Dedecek RA, Curcio GR, Nietsche K, Schenkel EP. Effect of light intensity on methylxanthine contents of Ilex paraguariensis A. St. Hil. Biochem Syst Ecol. 2007; 35(2): 75-80. http://dx.doi.org/10.1016/j.bse.2006.09.001

Song R, Kelman D, Johns KL, Wright AD. Correlation between leaf age, shade levels, and characteristic beneficial natural constituents of tea (Camelia sinensis) grown in Hawaii. Food Chem. 2012; 133(3): 707-714. http://dx.doi.org/10.1016/j.foodchem.2012.01.078

Koshiishi C, Ito E, Kato A, Yoshida Y, Crozier A, Ashihara H. Purine alkaloid biosynthesis in young leaves of Camellia sinensis in light and darkness. J Plant Res. 2000; 113(2): 217-221. http://dx.doi.org/10.1007/PL00013902

Vaast P, Bertrand B, Perriot J-J, Guyot B, Génard M. Fruit thinning and shade improve bean characteristics and beverage quality of coffee (Coffea arabica L.) under optimal conditions. J Sci Food Agric. 2006; 86(2): 197-204. http://dx.doi.org/10.1002/jsfa.2338

Sridevi V, Giridhar P. Changes in caffeine content during fruit development in Coffea canephora P. ex. Fr. grown at different elevations. Journal of Biology and Earth Sciences. 2014; 4(2): B168-B175.

Sridevi V, Giridhar P. Influence of Altitude Variation on Trigonelline Content during Ontogeny of Coffea Canephora Fruit. J of Food Studies. 2013; 2(1): 62-74. http://doi:10.5296/jfs.v2i1.3747

Mazzafera P, Crozier A, Sandberg G. Studie s on the metabolic control of caffeine turnover in developing endosperms and leaves of Coffea arabica and Coffea dewevrei. Journal of Agricultural and Food Chemistry. 1994; 42(7): 1423-1427. http://dx.doi.org/10.1021/jf00043a007

Perrois C, Strickler SR, Mathieu G, Lepelley M, Bedon L, Michaux S, Husson J, Mueller L, Privat I. Differential regulation of caffeine metabolism in Coffea arabica (Arabica) and Coffea canephora (Robusta). 2014; Planta. (In press). http://dx.doi.org/10.1007/s00425-014-2170-7


  • There are currently no refbacks.