Total Protein

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ISBN: 978-81-909047-9-7, p-ISSN: 2249-2569, e-ISSN: 2320-2955

INTERNATIONAL RESEARCH JOURNAL OF HUMANITIES, ENGINEERING & PHARMACEUTICAL SCIENCES (An International Registered Research Journal)

Paper Code- Int./JUNE14/H873

Volume-02

Issue-08

Year-04,

June,2014

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“THINK BEYOND TO CREATE MIRACLES” Promoted By: Association for Innovation (Under the Society Registration Act, 1860, a government registered organization with registration no. 2241)

Year-4/Vol-2/Issue-8

ISBN:978-81-909047-9-7, p-ISSN:2249-2569, e-ISSN:2320-2955

International Research Journal of Humanities, Engineering & Pharmaceutical Sciences Promoted By: Association for Innovation

ROLE OF TOTAL PROTEIN IN PETAL SENESCENCE OF Chrysanthemum coronarium L. UNCUT FLOWER PETALS

Ruby G Patel*, Yogeshkumar B Patel & Archana U Mankad Department of Botany, University School of Sciences, Gujarat University, Ahmedabad, India Abstract: The God’s gift to mankind is flower. They are integral part of human livings. In addition to serve as the reproductive organs of flowering plants, flowers have long been admired and used by humans mainly to make their environment beautiful with their pleasant aroma and diverse colours and even as a source of food. After flowering and during the production of seeds and fruits, annuals undergo senescence. Senescence refers to the final steps leading to the death of the organism. It comprises of a highly regulated series of cytological and biochemical events to coordinate the degradation of macromolecules and remobilization of nutrients from senescing tissue into reproductive and young organs as well as storage tissues. Protein plays an important role during senescence process. In these experiments it was observed that level of protein has been decreased significantly. Proteolysis plays an important role in protein degradation. As proteolysis become dominant, a decline in amount of protein has been found during senescence of Chrysanthemum coronarium L flower petals.

Introduction ’ Flowers are indeed nature s smile; they add color and beauty to life. Morphologically flower is considered as a highly condensed and modified shoot. Its function is reproduction. The branch of Horticulture dealing with the cultivation of flowering plants and use of flowers is known as floriculture. Research work on floriculture is being carried out at several research institutes under the Indian Council of Agriculture Research (ICAR) and Council of Scientific and Industrial Research (CSIR), in the horticulture/floriculture department of state. Agriculture University and under All India Co-ordination Floriculture Improvement Project (AICFP) with a network of about twenty centres. The crops which have received larger attention include Rose,Gladiolus, Chrysanthemum, Orchid, Jasmine, Tuberose, Aster, Marigold etc. In the present study, Chrysanthemum coronarium L. (Crown Daisy) plant is selected. It is a weed in cornfield in England.30-60 cm tall, with flowers having shades of yellow and ivory-white. The crown daisy is much branched plant with finely cut foliage reaching up to a height of 1m. Both single and double cultivars are available but single are common. Chrysanthemum coronarium L. is free flowering and grows in all types of soil in sunny situation, in beds and pots and is used as cut flowers. Flowering is controlled by the developmental age of the plant and environmental signals, including photoperiod, vernalization, light quality, and the availability of water and nutrients (Bernier, 1988; Bernier et al., 1993). Senescence is a dynamic process that occurs naturally concurrent with age or is prematurely induced by a variety of biotic and abiotic factors. Senescence can broadly be defined as the combination of events that leads to the death of cells, tissues, or organs (Patel et al., 2010). Plant senescence is known to consist of genetically driven degradative processes which lead to the death of cells, organs, and organisms (Patel et al., 2012) Senescence in plants is characterized by the breakdown of cell wall components and membrane, leading to the loss of tissue structure and cellular compartmentation and ultimately, homeostasis (Thimann, 1980; Thompson et al., 1987; Nooden, 1988). The degradative processes are accompanied by the decay of chlorophyll and the onset of proteolysis, which are also more rapid in darkness (Martin and Thimann, 1972; Trippi and DeLuca D’Oro, 1985; Kanazawa et al., 2000). Differential degradation of thylakoid proteins is responsible for the loss of photochemical activities during senescence (Roberts et al., 1987). Material and Method: In order to study the total protein status and the changes in it during the senescence period in uncut Chrysanthemum coronarium L. flower petals, biochemical estimation were done using dry flowers. The seeds were obtained from Anand Agricultural University (AAU), Anand. The plants grown in the experimental plots of the botanical garden of the department served as the source of the material. Various stages identified: It was observed that the uncut flowers of Chrysanthemum coronarium L. remained fresh on the th plant for 9 days with 10 day as the senescent day at which the petals started abscising. Data was recorded at the interval of 2 days. Thus, 5 stages were defined as follows: Stage 1: After 48 hours of flower opening (Day 2) Stage 2: After 96 hours (Day 4) Stage 3: After 144 hours (Day 6) Stage 4 After 192 hours (Day 8) Stage 5 (Senescent stage): After 240 hours (Day 10)

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ISBN:978-81-909047-9-7, p-ISSN:2249-2569, e-ISSN:2320-2955

Collection and preparation of material for biochemical estimations In order to carry out the estimation from dry material, the ray florets were collected from every stage of flower starting from the day it opened till its senescence. Every day the field was surveyed in the morning and the flowers which had just opened were tagged. These flowers were considered as stage 0 (0 hr.) flowers. Ray florets from some of the stage 1 (after 48 hours) flowers were collected and packed separately with proper labels. Similarly, ray florets for stage 2 (96 hrs.), stage 3 (144 hrs.), stage 4 (192 hrs.), stage 5 (240 hrs.) (senescent stage) flowers were also collected. These ray florets were then placed in the oven at 80° C for drying. In order to study the changes in the proteins, the biochemical estimations of total protein was done from the 100 mg dry material of all stages of Chrysanthemum coronarium L. Total protein activity was estimated by the method of Lowry et al., (1951). The results were expressed as total protein mg/ g dry ray florets. For statistical analysis, means were based on ten replicates for each stage and the standard error was computed. It was also statistically examined by One-way ANOVA calculated at 0.05% level of significance (Table-1).

Source of Variation

Sum of Squares (SS)

Degree of Freedom (DF)

Mean Square (MS)

Between Groups

6.434

4

1.609

Within Groups

.029

10

.003

Total

6.464

14

F ratio

Table value F*

551.167

3.5

Result and Discussion: Protein plays an important role during senescence process. In these experiments level of protein decreased. There was no significant difference between in stage-4 and stage-6. After that, level of total protein decreased till senescent stage (fig-1). Protein degradation may occur during senescence process to make free amino acids. By the process of proteolysis amount of free amino acids increases which may use in the respiration process. During the course of petal aging there is a drop in the level of macromolecules like proteins (Borochov et al., 1976; Parups, 1971; Paulin, 1977). Carfantan (1970) reported that in tulip petals, accumulation of proteins is rapid in initial stages and proteolysis becomes dominant even before opening of the flower. Decreasing trend of total protein in Tithonia rotundifolia Blake was reported (Patel et al., 2012). Lay-Yee et al. (1992) reported that flower senescence was accompanied by a decline in the petal protein content as well as breakdown of specific proteins in petals. Treatment with compounds that inhibit translation or transcription delayed the visible symptoms of petal senescence, showing that protein synthesis is required (Xu et al., 2007). Van Doorn and Woltering (2007) have reported that a decrease in over all protein levels can be due to a decrease in synthesis as well as increase in degradation. Proteins are degraded to a mixture of smaller polypeptides and amino acids (Parups, 1971) and a significant increase in the level of ammonia was noted in senescing roses. A decrease in overall protein levels can be due to a decrease in synthesis as well as an increase in degradation. In Ipomoea, no upsurge of in vitro protease activity was detected during petal senescence (Winkenbach, 1970), but in other species, such as Hemerocallis (Stephenson and Rubinstein, 1998), Iris (Pak and van Doorn, 2005), and Petunia (Jones et al., 2005), a massive increase in protease activity was found. Fig 1: Total Protein (mg/g dry petals) in uncut flower petals of Chrysanthemum coronarium L.

Total Protein 6

mg /g Total protein

5 4 3

Total Protein

2 1 0 Stage 2

Stage 4

Stage 6

Stage 8


Stage 10

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ISBN:978-81-909047-9-7, p-ISSN:2249-2569, e-ISSN:2320-2955

Conclusion: The reduction in protein content involved degradation to a mixure of smaller polypeptides and amino acids. The total protein content of the corolla was declining indicating the requirement of protein synthesis for the onset of senescence. The onset of hydrolysis of structural cell components like proteins is initiated in response to depletion of the free sugars used as respirable metabolites in order to supply alternative substrates for respiration such as the carbon skeleton of amino acids. This supposition is supported by the fact that an exogenous supply of sugars delays the onset of excessive protein degradation. References 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21.

22.

Bernier G, (1988). The control of floral evocation and morphogenesis. Annual Review of Plant Physiology and Plant Molecular Biology. 39: 175–219. Bernier G, Havelange A, Houssa C, Petitjean A, Lejeune P, (1993). Physiological signals that induce flowering. The Plant Cell. 5: 1147–1155. Borochov, A.; Mayak S. and Halevy, A. H. (1976) : Combined effect of acid and sucrose on growth and senescence of rose flowers, Physiol. Plant. 36: 221-224. Carfantan N, (1970). Observation preliminary studies in tulip protein during senescence, Acta Hort. 41: 31-43. Jones M L, Chaffin G S, Eason J R, Clark D G, (2005). Ethylene-sensitivity regulates proteolytic activity and cysteine protease gene expression in Petuniacorollas. Journal of Experimental Botany. 56: 2733–2744. Kanazawa S, Sano S, Koshiba T, Ushhimaru T, (2000). Changes in antioxidative enzymes in cucumber cotyledons during natural senescence: comparison with those during dark-induced senescence. Physiol. Plant. 109: 211–216. Lay-Yee M, Stead A D and Reid M S, (1992). Flower senescence in Dayliliy (Hemerocallis). Physiol. Plant. 86: 308-314. Lowry O H, Rosebrough N J, Farr A L, Randall R J, (1951), Protein measurement with the folin phenol reagent, J. Biol. Chem. 193: 265-275. Martin C, Thimann K V, (1972). The role of protein synthesis in the senescence of leaves. I. The formation of protease. Plant Physiol. 49: 64–71. Nooden L D, (1988). The phenomenon of senescence and aging. In: Senescence and Aging in Plants. Eds. L. D. Nooden and A. C. Leopold, Acad. Press, San Diego. pp-2–50. Pak C, Van Doorn W G, (2005). Delay of Iris flower senescence by protease inhibitors. New Phytologist. 165: 473–480. Parups E V, (1971) Disc electrophoresis of proteins of senescing and fresh leaves and petals of certain ornamental plants. J.Am. Soc. Hortic. Sci. 55: 775-781. Patel R G, Desai R J, and Mankad A U, (2012). Sugar metabolism during ray florets Senescence in uncut flowers of Tithonia rotundifolia Blake, Bioscience Guardian, 2(2):191-195. Paulin A., (1977). Metabolism glucidique et proteique sur les fleur d’oecillet alimenteeou nin avec une solution de saccharose, ActaHort., 71 : 241-257. Roberts D R, Thompson J E, Dumbroff E B, Gepstein S and Mattoo A K, (1987). Differential changes in the synthesis and steady state levels of during thylakoid proteins during bean senescence. Plant Mol. Biol. 19: 343-353. Stephenson P, Rubinstein B, (1998). Characterization of proteolytic activity during senescence in daylilies. Physiologia Plantarum. 104: 463–473. Thimann K V, (1980). The senescence of leaves. In: Senescence of Plants. Ed. K. V. Thimann, CRC Press Inc., Boca Raton, Florida, 85–115. Thompson J E, Legge R L, Barber R F, (1987). The role of free radicals in senescence and wounding. New Phytol. 105: 317–344. Trippi V S, M De Luca d’Oro, (1985). The senescence process in oat leaves and its regulation by oxygen concentration and light irradiance. Plant Cell Physiol. 26: 1303–1311. Van Doorn W G and Woltering E J, (2007) Physiology and molecular biology of petal senescence. J. of Expt. Bot. 59(3): 453-480. Winkenbach F, (1970a). Zum Stoffwechsel der aufbluhenden und welkenden korolle der Prunkwinde Ipomoeapurpurea I. Beziehungen zwischer Getaltwande 1, Stofftransport, Atmung und Invertaseaktivitat. BerichtederschweizerischenbotanischenGesellschaft 80 : 391 - 406. Xu X, Gookin T, Jiang C, Reid M S, (2007). Genes associated with opening and senescence of the ephemeral flowers of Mirabilis jalapa. Journal of Experimental Botany. 58: 2193–2201.


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