Biochemical studies of comparative haemolymph constituents in fourth instar larvae of Daba trivoltine ecorace of topical tasar silkworm Antheraea mylitta D.
Authors
-
,IN
P. K. Mishra -
,INL. Jaiswal
-
,IND. Kumar
-
,INA. Kumar
-
,INJ. P. Pandey
-
,INS. K. Sharan
-
,INB. M. K. Singh
-
,INB. C. Prasad
DOI:
https://doi.org/10.18311/jeoh/2010/18281Keywords:
Haemolymph, Amino acids. Protein, Glycogen, Glycerol, Trehalose, Antheraea mylitta, NDDAbstract
Diapause is endocrine mediated dormancy and the preparatory stages are fourth and fifth instar lan/ae of tasar silkworm, Antheraea mylitta Drury. Different haemolymph biochemical constituents like total protein, free amino acids, trehalose, glycerol and glycogen were estimated in fourth instar lan/ae of Daba trivoltine ecorace of tropical tasar silkworm so as to differentiate between non-diapause destined (NDD- the first and second crops) and diapause destined (DD- third crop) generations and also to relate them as preparatory for undergoing diapause. There was significant difference in the presence of haemolymph glycogen in female larvae (1.9361+0.9605mg/ml) than male larvae (1.0859±0.5393) in TV third crop. No significant different was observed in remaining constituents. The concentration of trehalose was significantly higher in female lan/ae (0.9501 ±0.3784 mg/ml) than male lan/ae (0.3811±0.2420) in TV second crop. A comparison made between the female and male lan/ae of second crop shows the higher level presence of amino acids and proteins in male larvae. Rest of the constituents was higher in female larvae than male in second crop. During third crop, except for total free amino acids all other biochemical constituents were higher in male lan/ae and the difference was observed to be significant in the concentration of glycerol and amino acids A comparison between the non-diapause destined (2"" crop) and diapause destined (3"^ crop) generations indicated that the diapause destined larvae has significantly higher level of amino acids trehalose (6.5873 + 3.2437 mg/ml in male, 8.1217 ± 3.0710 mg/ml in female) and glycerol (in female). However, a comparison between NDD first crop and third crop indicated that all the haemolymph constituents were significantly higher in diapause destined fourth instar lan/ae, like amino acids protein (11.3999 ± 2.5290mg/ml in male, 11.3264 ± 2.5672mg/ml in female), trehalose glycogen and glycerol. The findings indicate that fourth instar lan/ae of Daba trivoltine of tasar silkworm accumulate energy reserves to skip forthcoming unfavorable conditions in pupal stage. SDS PAGE studies did not indicate any major difference in protein profile but a sharp increase in band intensity of 85kDa, probably a major storage protein and a 43kDa diapause specific band in DD larvae.
Downloads
Metrics
Downloads
Published
How to Cite
Issue
Section
Accepted 2017-11-30
Published 2017-11-30
References
Adedokun.T. A. and Denlinger, D. L. (1985) Metabolic reserves associated with pupal diapause in flesh fly Sarcophaga crassipalpis. J. Insect physiol., 31, 229-233
Agrawal, S. C; Jolly, M. S. and Banerjee, N. D. (1981) Lipid content in silkworm, Antheraea mylitta D. Indian J.Entomol.. 43, 294-296.
Beadle, L. C. and Shaw, J. (1950) The retention of salt and regulation of the non-protein fraction in the blood of aquatic larva, Sialis lutaria. J. Exp. Biol., 27, 96-100.
Boctor, I. Z. (1981) Changes in the free amino acids of the haemolymph of diapause and non-diapause pupae of the cotton ball worm, Heliothis armigera Hbn. (Lepidoptera: Noctuidae). Expehntia, 37, 125-126.
Brown, J J. (1980). Haemolymph protein reserves of diapausing and non-diapausing codling moth larvae, Cydia pomonella (L) (Lepidoptera: Tortricidae). J. Insect Physiol., 26, 487491.
Brown, J. J. and Chippendale, G. M. (1978). Juvenile hormone and protein associated with the larval diapause of the south western corn borer, Diatraea grandiosella. Insect Biochem. 8, 359-367.
Buck, T. B. (1953) Physiological properties and chemical composition of insect blood. In Insect Physiology (K. D. Roeder, ed.). John Wiley, NewYork pp 147-190.
Chen, P. S. (1966)Amino acid and protein metabolism in insect development. In Advances In insect Physiology, (J. W. L. Beament, J. E Trehen and V. B. Wigglesworth - eds.) 3, 53-132
Chen, P. S. and Levenbook, L. (1966a) Studies on haemolymph proteins of the blow fly Phormia regina. I. Changes in ontogenetic pattern. J. Insect. Physiol., 12, 1595-1609.
Chen, P. S. and Levenbook, L. (1966b) Studies on haemolymph of proteins of the blow fly Phormia regina. II: Synthesis and breakdown as revealed by isotopic labelling. J. Insect Physiol., 12, 1611-1625.
Denlinger, D. L. (1985) Hormonal control of diapause, pp. 353412, In, Comprenhensive insect physiology, biochemistry and pharmacology, Vol, 8. (Eds, G, A. Kerkut and L. I, Gilbert), Pergamon, New York.
Denlinger, D, L. (2002) Regulation of Diapause, Ann. Rev. Entomol., 47, 93-122,
Doira, H. (1968) Development and sexual differences of blood proteins in the silkworm, Bombyx mori . Sci, Bull, Fac, Agric, Kyushu Univ,, 23, 205-214,
Evans, P. D, and Crossley, A. C. (1974) Free amino acids in the haemocytes and plasma of the larva of Calliphora vicinia. J. Exp. Biol., 61, 463-472,
Firling, C, (1977) Amino acid and protein changes in the haemolymph of developing fourth instar Chironomous tentans. J. Insect Physiol., 23, 17-22,
Florkin, M, and Jeuniaux, C. (1974) Haemolymph composition. In Physiology of Insects (M.Rockstein, ed.) Vol. V., 2nd edition, pp. 255-307.
Fukuda, T, and Matuda, M. (1953) Nippon Sanshigaka Zasshi (In Japanese, English abstract), 22, 243,
Goldewski, J. and Kludekiewicz, B,, Grazelak, K, and Cymborowski, B, (2001), Expression of lan/al haemolymph proteins (Lhp) genes and protein synthesis in the fat body of greaterwax moth {Galleria mellonella) larvae during diapause, J. Insect Physiol., 47, 759-766,
Hagen, J. H, and Hagen, P. B, (1962) An enzymatic method for the estimation of glycerol in blood and its use to determine the effect of noraderanaline on the concentration of glycerol in blood. Can. J. Biohem. Physiol., 40, 11291139.
Hayakawa, Y and Chino, H. (1982a) Phosphofructokinase as a possible key enzyme regulating glycerol or trehalose accumulation in diapausing insects. Insect Biochem., 12, 639-642,
Hayakawa, Y and Chino, H. (1982b) Temperature dependent activation of glycogen phosphorylase and synthatase of fat body of silkworm Philosamia cynthia. The possible mechanism of the temperature-dependent interconversion between glycogen and trehalose. Insect Biochem., 12, 361-368.
Hirano, M. and Yamashita, 0. (1983) Developmental changes in trehalose synthesis in fat body of the silkworm, Bombyx mori: Trehalose synthatase related to regulation of haemolymph trehalose during metamorphosis. Insect Biochem., 13, 593-599.
Horie, Y. (1961) Physiological studies on the alimentary canal of the silkworm, bombyx mori III, Absorption and utilization of carbohydrates, Bull. Seric. Exp. Jpn., 16, 287-309.
Jo Hyun-ii and Kim Yonggyyun (2001) Relationship between cold hardiness and diapause in the smaller fruit Tortix, Adoxophyes orana (Fischer von Roslerstamm), J. Asia Pacific Entomol, 491, 1-9.
Jolly, M. S.; Sinha.A. K. and Agrawal, S. C. (1972) Free amino acids in the larval and pupal haemolymph of Antheraea mylitta D. reared on Terminalia tomentosa. Indian J. Sen., 11, 63-67.
Karaki, M. (1969) Electrophoretic studies on blood proteins of Samia cynthia ricini during development, Biol. J 18, 32 34.
Koeppe, J. K. and Gilbert, L. I. (1973) Immunological evidence for the transport of haemolymph protein into the cuticle of Manduca sexta. Biol.Rev., 19, 615-624.
Koopmanschap.A. B., Lammer, J. H. M. and de Korte, C. A. D.(1995). The structure of the green coding diapause proteins 1 of the Colorado potato beetle (Leptinotarsa decem/ineata) J. Insect Physiol., 41, 509-518.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond.), 227, 690-685
Leather, S. R., Walters, K. F. A and Bales, J. S. (1993) The ecology of Insect Overwintering.Cambridge University Press, Cambridge.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. S.(1951) Protein measurement with the Folin Phenol reagent. J. Biol. Chem., I. 93, 265-275.
Mishra, P. K., Kumar D., Jaiswal, L., Ashutosh Kumar, Singh B. M. K., Sharan S. K., Pandey J. R and Prasad B. C. (2010a) Biochemical aspects of diapause preparation in ultimate instar of tropical tasar silkworm Antheraea mylitta Drury. Eco-physiolo. Occupati. HItt)., 9, 253-60.
Mishra, P. K., Kumar D., Jaiswal, L., Ashutosh Kumar, Singh B. M. K., Sharan S. K., Pandey J. P and Prasad B. C. (2010b) Role of haemolymph biochemical constituents and brain Proteins in maintenance of pupal diapause in Daba bivoltine ecorace of tropical tasar silkworm Antheraea mylitta Drury, Eco-physiolo. Occupati. HIth., 9, 243-252.
Moore, S. and Stein, W. H. (1948) In Method in Enzymology. Vol.III. (Colowick, S. P. and Kaplan, N. O., eds). Academic Press. New York.
Mullin, D. E. (1985) Chemistry and physiology of the haemolymph. In Comprehensive Insect Physiology and Pharmacology (G. A. Kerkut 8. L. I. Gilbert eds.), Pergamon Press. 3,355400.
Nordin, H. J., Zheng Cul and Chih-Ming Yin (1984) Cold-induced glycerol accumulation by Ostrinia nubilalis larvae is developmentally regulated. J. Insect. Physiol., 30, 563566.
Osir, E. O., Labongo, L. V. and Unithan, G. C. (1989). A high molecular weight diapause associated protein from the stem borer Busseola fuses, purification and properties./Arc/)./nsecf Biochem. Physiol., 11, 173-187.
Palli,, S. R., Ladd, T. R., Ricci, A. R., Primavera, M., Mungrue, I. N., Pang, A. S. D. and Retnakaran, A. (1998) Synthesis of the same two proteins prior to larval diapause and pupation in spruce bud worm Choristoneura fumiferana. J. Insect Physiol., 44, 509-524.
Pawar, V. M. and Ramakrishna, N. (1977) Biochemical changes in larval haemolymph of Spodoptera litura Fabricius due to nuclear polyhedrosis virus infection. Indi J. Exp. Biol., 15, 755-758.
Sactor, B. (1965) The physiology of insects Vol.11 Academic Press, New York.
Sakamoto, E. and Horie, Y. (1979) Qualitative changes of phosphorous compound inhaemolymph during development of silkworm Bombyx mori. J. Seri. Sci. Jpn., 49, 509-511.
Salama, M. and Miller, T. (1992). A diapause associated protein of the pink bollworm Pectinophora gossypiellas. Arch. Insect Biochem. Physiol., 21, 1-11.
Sinha, A.K., Chaudhary, S.K. and Sengupta, K. (1988) Changes in free amino acids in the larval and pupal haemolymph of Antheraea mylitta Drury reared on Terminalia arjuna and Terminalia tomentosa. Indi J. Seri., 27, 95-108.
Sinha, U.S.P. and Sinha, A.K. (1994) Changes in concentrationof proteins and carbohydrates in developing embryos and larval haemolymph of temperate tasar silkworm, Antheraea proylei. Jolly Indi J. Seri., 33, 84-85.
Sowri D. M. K. and Sarangi, S. K. (2002) A comparative study on the trehalose level in different varieties of the silkworm, Bombyx mori, during fifth instar larval development. Entomon, 27, 57-61.
Sula, J. Kodrik, D. and Socha, R. (1995) Hexameric haemolymph proteins related to adult diapause in the red firebug, Pyrrochoris apterus (L.) (Heteroptera). J. Insect Physiol.. 41,793-800.
Tauber, M. J.; Tauber, C. A. and Masaki, M. (1986) Seasonal adaptations of insects, Oxford University Press, London.
Wyatt, G. R. (1961) The Biochemistry of insect haemolymph. Ann. Rev. Entomol., 6, 75-102.
Wyatt, G. R. (1967) The biochemistry of sugars and polysaccharides in insects. Advances in Insect Physiology (J. W. L. Beament, J. E. Treherne and V. B. Wigglesworth, (eds). Vol 4. Academic Press, Londn. pp. 287-360.
Wyatt, G. R. (1980) The fat body as a protein factory In Insect Biology in Future. (M. Locke and D. S. Smith, eds ). Academic Press, New York. pp. 201-225.
Wyatt, G. R. and Kalf, G. F. (1956) Trehalose in insects. Fed. Proc, 15, 188.
Wyatt, G. R. and Kalf, G. F. (1957) The chemistry of insect haemolymph. I. Trehalose and other carbohydrates. J. Gen. Physiol., 40, 833-847.
Wyatt, G. R. and Pan, M. L. (1978). Insect Plasma Proteins, Ann. Rev. Biochem., 47, 799-817.
