NPK rates, packaging materials and storage duration on anthocyanin pigmentation and postharvest quality characteristics of lemongrass (Cymbopogon

citratus)
1

Tajidin, N.E.1 Mislan, A.1, Ahmad, S. H.1, Rosenani A. B 2and Munirah, M1 Crop Science Department and 2Land Management Department, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia.

Introduction Lemongrass (Cymbopon citratus) is a perennial turf grass with a stalk formed by layers of leaf sheaths (pseudo stem). Three popular accessions of lemongrass, Galah, Paha Ayam and Hijau are planted commercially in Malaysia. Hijau lemongrass is highly demanded as additive ingredients in soft drinks and cosmetics. The lemongrass suffers from red pigmentation on the pseudo-stem during postharvest, thus, reducing its market value. According to Shaikh et al. (2008) phosphorus level played a greater role than nitrogen level in controlling leaf purple pigmentation and forage yield. Deficiencies of P and N induce the accumulation of anthocyanin in many plant species (P-Teresa and SBallesta, 2008). Excess N can adversely affect red color development in Fuji apple (Rudell et al., 2002). An increase application of N resulted in a decrease content of red pigment in cranberry. Bean plants cultured on P-deficient media had higher concentrations of anthocyanin the leaves to protect the plant from oxidative stress (P-Teresa and S-Ballesta, 2008). K fertilization appears to enhance anthocyanin accumulation and red coloration of apples (Ritenour and Khemira, 1997). K application also can reduce the effects of excess N (Rudell et al., 2002). According to Lancaster et. al. (1997) the formation of anthocyanin is mainly influenced by cultivar, genetic, light, temperature and cultural practices. Anthocyanin accumulation is also influenced by different packaging methods (Ritenour and Khemira, 1997). The formation of anthocyanin can occur rapidly in presence of light. Pietrini et. al. (2002) reported that the formation of anthocyanin in the fruit and vegetables will be reduced under storage at low temperature. Temperature of 35 oC speed destruction of anthocyanin resulting in production of chalkons which are responsible for brown color development on Berberis spp. (Laleh et. al., 2006). According to Siomos et. al. (2000) combination of low temperature and low light intensity reduced or slowed down anthocyanin synthesis in stored white asparagus. However, specific recommendation for different types of fertilizer application rate, type of packaging for lemongrass and storage duration is lacking. Thus, the objective of this study was to determine the lemongrass quality characteristics and anthocyanin pigmentation as affected by different types of fertilizer application rates, type of packaging materials and storage duration.

Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.

Materials and Methods Plant materials Hijau lemongrass was planted on an inland soil in Kuala Selangor, Selangor, Malaysia. Three rates of N:P:K fertilization, F1 = 400 kg N/ha, 50 kg P 2O5/ha and 50 kg K2O/ha (control treatment based on farmers practice); F2 = 300 kg N/ha,100 kg P 2O5/ha and 100 kg K2O/ha; F3 = 200 kg N/ha, 100 kg P2O5/ha and 100 kg/ha were used. The fertilizers were applied around the basal of each lemongrass cluster at 1.5, 3 and 4.5 months after planting and weeding was carried out every 2-3 weeks as required. The plants were harvested at seven months after planting. Each plant was trimmed to 30 cm to remove the leaf blades. After trimming, well formed and damaged and diseased free lemongrass stalks were selected and transported to the Postharvest Laboratory, Faculty of Agriculture, Universiti Putra Malaysia and stored in a cold room (10-12 oC) for 12 hr. The next day, the lemongrass stalks were cut to 25 cm length and two to three outer layers of leaf sheaths (pseudo stem) were discarded until a white pseudo stem appeared. Then, the lemongrass stalks were packed into two types of packaging materials (black and clear plastic bags) with five lemongrass stalks per bag per replication. The stalks were stored under full florescence light at 10 oC for 0, 5, 10 and 15 days. Measurements Total anthocynin was measured by adjusting lemongrass extracts to buffer solutions of pH 1.0 and 4.5. Absorbances were read at 520 and 700 nm using a visible spectrophotometer (Prim light 230V, Secomam, France). Firmness of the equatorial diameter of five stalks was measured with a penetrometer (Bishop FT 327, Italy) results were expressed as Newton (N). Soluble solids concentrations were determined with a digital refractometer (Baush Lomb Abbe 3 L, Rochester, NY) and the results were expressed in % soluble solids concentration. Juice pH was determined by using a pH meter (Crison Micro pH 2000, Crison Instruments, Spain) calibrated using buffer solutions of pH 4 and 7. Titratable acidity was analyzed using the titration with 0.1 % NaOH. The results were expressed as percentage of citric acid per 100 g fresh weight. Ascorbic acid (vitamin C) was determined using the 2,6-dichlorophenol-indophenol dye method and expressed as mg-1 .100g of fresh stalk. Experimental design The three nutrient combination of N, P205 and K20 2 packaging material (clear and black plastic bags) 4 storage days (0, 5, 10 and 15 days) factorial experiment was arranged in a randomized complete block design with four replications. Data were analyzed by using analysis of variance of the main and interaction effects. The mean differences were determined by LSD values with significant level of 5%. The significant interaction effects were further evaluated by relationship between dependent and independent variables using regression analysis.

Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.

Results and Discussion Effect on anthocyanin content. NPK fertilization had no significant effect on anthocyanin content (Table 1). There was a significant (P0.05) quadratic relationship between anthocyanin content and storage duration at 10 oC of lemongrass packed in clear packaging material (Fig. 1). During the first 5 days of storage, there was a linear increase of anthocyanin, followed by a gradual increase from 5 to 10 days of storage. During 10 to 15 days of storage, anthocyanin in lemongrass decreased. The maximum anthocyanin content was formed around 10 days. However, there were no significant quadratic relationships between anthocyanin content and storage duration of lemongrass packed in black packaging material. Thus the black packaging material and storage temperature of 10 oC reduced the anthocyanin content of lemongrass. Anthocyanin content of lemongrass packed in black packaging material was not significantly different throughout the storage duration (Table 1). By using black packaging material, lemongrass was not exposed to white light. White light was reported to induce anthocyanin synthesis in sorghum seedlings (Oelmuller and Mohr, 1985). According to Siomos et. al. (2000), combination of low temperature and low light intensity reduced or slowed down the anthocyanin synthesis in stored white asparagus. Effects on titratable acidity, pH, soluble solids concentration and ascorbic acid. There were no significant interaction effects of the treatment combinations on titratable acidity and pH of lemongrass (Table 1). However, there were significant interaction effects between packaging x storage duration on soluble solids concentration and ascorbic acid content of the lemongrass. There were significant interaction effects between fertilizer rates x storage duration and packaging x storage duration on soluble solids concentration of lemongrass (Table 1). There was a significant quadratic relationship between soluble solids concentration and storage duration of lemongrass treated with fertilizer rate 1 (F1) that contained the highest rate of N i.e. 400 kg N/ha. (Fig. 2A). During 5 days of storage, soluble solids concentration of lemongrass increased gradually, followed by a rapid increase of soluble solids concentration from 5 to 15 days of storage. According to Wang et al. (2007) by increasing N and K rates increased the concentrations of soluble solids concentration. But, within a moderate range of N supply, increasing N rates thus increased soluble solids concentration. These might be related to enhanced photosynthesis, due to an increased leaf area under high N supply, and, to some degree, postponing the process of leaf senescence. More photosynthates are produced under high N application, some of which can be stored as reducing sugars, leading to the accumulation of soluble solids concentration in tomato fruits. Delgado et al., (2004) reported that N stimulates plant growth to the detriment of sugar accumulation in the grapes during ripening. The delayed sugar accumulation in fruits from vines with higher N levels does not seem to be due to reduce photosynthesis as a result of changes in canopy structure and/or the light microclimate. This result is relevant in this study, whereby high a N rate increased the concentration of soluble solids concentration. From the experiment, F1 was the higher rates of N, thus high soluble solids concentration was obtained from this treatment.
3 Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.

There was also a significant interaction effect between packaging materials x storage duration on soluble solids concentration (Table 1) followed by a significant quadratic relationship between soluble solids concentration and storage duration of lemongrass packed in clear packaging material (Fig. 2B). There was a gradual decrease of soluble solids concentration during 5 days of storage, followed by a rapid increase of soluble solids concentration from 5 to 15 days of storage. This indicated that there was an increase of 18.6% soluble solids concentration from 0 to 15 days of storage. There was no significant quadratic relationship between soluble solids concentration and storage duration of lemongrass packed in black packaging material. . According Bartz and Brecht (2003) vegetables contain storage reserves such as fructosan or starch that can be degraded into soluble carbohydrates, thus maintaining nearly constant concentrations of soluble sugars for a long period. According to Cia et al. (2006) soluble solids concentration increased with storage time, similar to the result obtained in this study. The higher soluble solids concentration indicated higher degradation rate of storage carbohydrates of lemongrass. According to Deng et al. (2005) modified atmosphere occurs when fruits are sealed in polyethylene bags with a relatively low permeability to gases. Consequently, as the fruit respires, the O2 level decreases, and the CO2 level increases in the bags. Under these atmospheric conditions, the respiration rate of the fruit is decreased, and as a direct effect, the consumption of respiration substrates of sugars is retarded. According to Cia et al. (2006) soluble solids concentration increased with storage days, similarly the same result was obtained in this study. There was a significant quadratic relationship between ascorbic acid content and storage duration of lemongrass packed in clear packaging material (Fig. 3). During 5 days of storage, ascorbic acid content of lemongrass decreased gradually, followed by a rapid increase from 5 to 15 days of storage. The clear packaging material exposed the lemongrass stalks to light causing photo-reduction of the ascorbic acid. There was no significant quadratic relationship between ascorbic acid and storage duration of lemongrass packed in black packaging material indicating that the ascorbic acid remained constant during storage. According to Siomos et al. (2000) packaging was effective in ascorbic acid retention of white asparagus spears during storage. On the contrary, the spears stored in air lost up to 83% of their initial ascorbic acid after 6 days, depending on the storage temperature. In many vegetables, modified atmosphere storage resulted in greater ascorbic acid retention than storage in air. This beneficial effect is the result of the modified atmosphere inside the package. The lower O2 concentration of the storage atmosphere was associated with smaller losses of ascorbic acid, probably due to the inhibition of ascorbic acid breakdown. Effect on color values (L*, C* and hO) and firmness. There were no significant interaction effects among the treatment combinations (fertilizer rates, packaging material and storage duration) on L*, and hO colour values and firmness of lemongrass (Table 2). The lemongrass stalks were 24% softer after 15 days of storage. The significant increase in L* after 10 days of storage followed by a significant decrease after 15 days of storage indicate that the stalks had become lighter in colour during storage. The increase in L* corresponded with the increase in C* after 10 days of storage, while the values of hO remained constant through out storage. However, during
4 Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.

these periods there were no differences in anthocyanin content of the stalks (Table 1). Thus, expression of colours using the colour values (L*, C* and hO) is not a good indicator of the amount of anthocyanin in the stalks. In fact the values of hO when combined with C* indicated that the stalks were lighter green. A significant benefit of modified atmosphere packaging (MAP) is its ability to prevent loss or degradation of chlorophyll. Loss of chlorophyll and biosynthesis of carotenoids are diminished in fruit kept in MAP (Cia et al., 2006). Jia et al. (2005) However, this study showed that lemongrass that were put in clear packaging that allowed light through it only affected the C* values while the L* and h0 values were not affected. Conclusion The NPK fertilizers did not influence anthocyanin pigmentation, color, firmness, ascorbic acid, titratable acidity and pH of lemongrass (cymbopogon citratus). The soluble solids concentration was significantly higher when the lemongrass was treated with 400 kg/ha N. Packaging with clear plastic bag had significant effects on anthocyanin, soluble solids concentration, ascorbic acid, firmness, and C* value. These quality characteristics were also affected by the storage duration. There was an interaction effect of fertilizer rates x storage days on soluble solids concentration. Anthocyanin content was higher when the lemongrass was packed with clear packaging compared to black packaging. Ascorbic acid was higher when the lemongrass was packed in black packaging. By packaging in black plastic bag, light was prevented from penetrating the lemongrass. Thus, reduce the formation of anthocyanin.

Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.

References:
Bartz, J.A. and Brecht, J.K. 2003. Postharvest physiology and pathology of vegetables. USA. Marcel Dekker Inc. New York, USA. 1. Cia, P., Benato, E.A., Sigrist, J.M.M., Sarantopoulos, C., Oliveira, L.M. and Padula, M. 2006. Modified atmosphere packaging for extending the storage life of Fuyu persimmon. Postharvest. Biol. Technol. 42: 228234. 2. Delgado, R., Martin, P., Alamo, M.D. and Gonzalez, M.R. 2004. Changes in the phenolic composition of grape berries during ripening in relation to vineyard nitrogen and potassium fertilization rates. J. Sci. Food Agric. 84:623-630. 3. Delgado, R., Gonzlez, M. and Martin, P. 2006. Interactive effects of nitrogen and potassium fertilization on anthocyanin composition and chromatic features of Temperanillo grape. Intl. J. Vine Wine Sci. 40(3):141-150. 4. Deng, Y., Wu, Y. and Li, Y. 2005. Changes in firmness, cell wall composition and cell wall hydrolases of grapes stored in high oxygen atmospheres. Food Research Intl. 38:769776. 5. Jia, H.J., Arakib, A. and Okamoto, G. 2005. Influence of fruit bagging on aroma volatiles and skin coloration of Hakuho peach (Prunus persica Batsch). Postharvest Biol. and Technol. 35:6168. 6. Lancaster, J.E., Lister, C.E., Reay, P.F. and Trigs, C.M. 1997. Influence of pigment composition on skin colour in a wide range of fruits and vegetables, J. Amer. Soc. of Hort. Sci. 122:594598. 7. Laleh, G.H., Frydoonfar, H., Heidary, R., Jameei, R. and Zare, S. 2006. The effect of light, temperature, pH and species on stability of Anthocyanin pigments in Four Berberis species. Pakistan J. of Nutr. 5(1):90-92. 8. Oelmuller, R. and Mohr, H. 1985. Mode of coactions between blue/UV light and light absorbed by phytochrome in light mediated anthocyanin formation in the milo (Sorghum vulgare Pers.) seedling. Proc. Natl. Acad. Sci. 82:6124-6128. 9. Pietrini, F., Iannelli, M.A. and Massacci, A. 2002. Anthocyanin accumulation in the illuminated surface of maize leaves enhances protection from photo-inhibitory risks at low temperature, without further limitation to photosynthesis. Plant Cell Envir. 25:1251-1259. 10. P-Teresa, S.D. and S-Ballesta, M.T. 2008. Anthocyanins: from plant to health. Phytochem. Rev. 7: 281-299. 11. Ritenour, M. and Khemira, H. 1997. Red color development of apple: A literature review. http://postharvest.tfree.wsu.edu/REP2007A.pdf . 12. Rudell, D.R., Mattheis, J.P., Fan, X. and Fellman, J.K. 2002. Methyl jasmonate enhances anthocyanin accumulation and modifies production of phenolics and pigments in Fuji apples. J. Amer. Soc. Hort. Sci. 127(3):435-441. 13. Siomos, A.S., Sfakiotakis, E.M. and Dogras, C.C. 2000. Modified atmosphere Packaging of white asparagus spears: composition, color and textural quality responses to temperature and light. Sci. Hort. 84:1-13. 14. Shaikh, N.P., Adjei, M.B and Scholberg, J.M. 2008. Interactive effect of phosphorus and nitrogen on leaf anthocyanins, tissue nutrient concentrations and dry-matter yield of Floralta Limpograss during short day length. Comm. in Soil Sci. and Plant Analysis. 39:1006-1015.

Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.

15. Wang, Y.T., Huang, S.W., Liu, R.L. and Jin, J.Y. 2007. Effects of nitrogen application on
flavor compounds of cherry tomato fruits. J. Plant Nutr. Soil Sci. 170:461-468.

Storage duration

Fig. 1. Relationship between anthocyanin content (mg. L-1) and storage duration of Hijau lemongrass packed in clear plastic bag (), y = -0.086x2 + 1.620x - 0.904, R2 = 0.3936 and black plastic bag (). Solid line indicates a significant relationship. Each point corresponds to the average of 5 stalks. A B

Storage duration

Storage duration

Fig. 2. Relationship between soluble solids concentration (%) and storage duration of Hijau lemongrass. (A) Treated with different N:P:K fertilization, F1 = 400 kg N/ha, 50 kg P 2O5/ha and 50 kg K2O/ha (control treatment based on farmers practice) () y = 0.009x2 - 0.016x + 5.258, R2 = 0.6747; F2 = 300 kg N/ha,100 kg P2O5/ha and 100 kg K2O/ha (); F3 = 200 kg N/ha, 100 kg P2O5/ha and 100 kg/ha (). (B) Packed in clear plastic bag (), y = 0.013x2 - 0.095x + 5.407, R2 = 0.6322 and black plastic bag (). Solid line indicates a significant relationship. on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 7 Paper presented at International seminar Each point corresponds to the average of 5 stalks.
Febuary 2010. Yogyakarta, Indonesia.

Storage duration

Fig. 3. Relationship between ascorbic acids (mg. 100-1) and storage duration of Hijau lemongrass treated packed in clear plastic bag (), y = 0.007x2 - 0.061x + 2.766, R2 = 0.3375 and black plastic bag ().Solid line indicates a significant relationship. Each point corresponds to the average of 5 stalks.

Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.

Table 1: Main and interaction effects of different fertilization NPK fertilizer rates, packaging materials and storage duration on anthocyanin (Antho), soluble solids concentration (SSC), titratable acidity (TA), ascorbic acid (AA) and pH on Hijau lemongrass (Cymbopogon citratus). Treatment Antho (mg. L-1) SSC (%) AA (mg. 100 g-1) TA (% citric acid) pH

Fertilizer rate (F) kg/ha 1z 4.412 ay 5.952 a 3.039 a 0.165 a 5.407 a 2 4.521 a 5.530 b 2.988 a 0.173 a 5.423 a 3 4.426 a 5.608 b 2.834 a 0.164 a 5.417 a Packaging (P) Clear 3.699 b 5.895 a 2.981 a 0.179 a 5.393 a Black 5.207 a 5.499 b 2.926 a 0.156 a 5.439 b Storage duration (SD) Days 0 2.110 b 5.280 c 2.727 b 0.163 a 5.355 b 5 5.094 a 5.447 bc 2.355 c 0.152 a 5.421 a 10 5.749 a 5.572 b 3.275 a 0.185 a 5.446 a 15 4.860 a 6.488 a 3.456 a 0.170 a 5.441 a Interaction FxP NS NS NS NS NS F x SD NS * NS NS NS P x SD * * * NS NS F x P x SD NS NS NS NS NS z 1= 400 kg N/ha, 50 kg P2O5/ha and 50 kg K2O/ha (control treatment based on farmers practice); 2 = 300 kg N/ha, 100 kg P2O5/ha and 100 kg K2O/ha; 3 = 200 kg N/ha, 100 kg P2O5/ha and 100 kg K2O/ha. y Means followed by the same letter within columns and treatment are not significantly different using LSD test, P0.05. NS, * Not significant or significant at P0.05.

Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.

Table 2: Main and interaction effects of different fertilization NPK rates, packaging materials and storage duration on firmness and stem color values of Hijau lemongrass (Cymbopogon citratus). Treatment Fertilizer rate (F) kg/ha 1z 2 3 Packaging (P) Clear Black Storage duration (SD) Days 0 5 10 15 Interaction FxP F x SD P x SD F x P x SD
z

Firmness (N) L*

Color valuesx C* h0

149.838 a 149.918 a 153.761 a 152.850 a 149.495 b

66.468 a 68.543 a 67.444 a 67.266 a 67.704 a

23.660 a 23.000 a 22.524 a 23.367 a 22.755 a

108.138 a 107.366 a 106.934 a 106.827 a 108.131 a

170.004 b 177.688 a 127.182 c 129.815 c NS NS NS NS

67.739 b 66.325 bc 70.915 a 64.962 c NS NS NS NS

22.934 b 22.728 b 24.513 a 22.070 b NS NS NS *

108.538 a 106.579 a 108.604 a 106.196 a NS NS NS NS

1= 400 kg N/ha, 50 kg P2O5/ha and 50 kg K2O/ha (control treatment based on farmers practice); 2 = 300 kg N/ha, 100 kg P2O5/ha and 100 kg K2O/ha; 3 = 200 kg N/ha, 100 kg P2O5/ha and 100 kg K2O/ha. y Means followed by the same letter within columns and treatment are not significantly different using LSD test, P0.05. x Color values: L* = Lightness, C* = Chroma and h0 = hue NS, * Not significant or significant at P0.05.

10

Paper presented at International seminar on Development of Coastal Sandy Area Towards Sustainable Agriculture. 13-14 Febuary 2010. Yogyakarta, Indonesia.