Characterization of α-amylase in the midgut and the salivary glands of rice striped stem borer, Chilo suppressalis Walker (Lepidoptera: Pyralidae)

다국어 초록 (Multilingual Abstract)

The rice striped stem borer, Chilo supprressalis, is a destructive pest of rice that was introduced to Iran in
1973 and has since become widely distributed. Amylases are hydrolytic enzymes that catalyze the
hydrolysis of the α-D-(1,4)-glucan linkage in glycogen and other related carbohydrates. Laboratory-reared
4th stadium larvae were randomly selected; the midgut and the salivary glands were removed by
dissection under a dissecting microscope and α-amylase activity was assayed using the dinitrosalicylic acid
procedure. The activity of α-amylase in the midgut and salivary gland were 0.06 and 0.036 μmol/min/mg
protein, respectively. The optimal pH and temperature for α-amylase were 9 and 35–40 °C, which is
consistent with reports of other lepidopteran insects. The enzyme activity was inhibited by addition of
NaCl, KCl, MgCl2, Urea, EDTA, and SDS. CaCl2 enhanced enzyme activity. Ca2+ ions also had inhibitory effects
on glucosidase activity. Plant amylase inhibitors play important role against insect pests. Hence, the
characterization of digestive enzymes and the examination of inhibitors on enzyme activity could be useful
in tackling of insect pests.

다국어 초록 (Multilingual Abstract)

The rice striped stem borer, Chilo supprressalis, is a destructive pest of rice that was introduced to Iran in
1973 and has since become widely distributed. Amylases are hydrolytic enzymes that catalyze the
hydrolysis of the α-D-(1,4)-glucan linkage in glycogen and other related carbohydrates. Laboratory-reared
4th stadium larvae were randomly selected; the midgut and the salivary glands were removed by
dissection under a dissecting microscope and α-amylase activity was assayed using the dinitrosalicylic acid
procedure. The activity of α-amylase in the midgut and salivary gland were 0.06 and 0.036 μmol/min/mg
protein, respectively. The optimal pH and temperature for α-amylase were 9 and 35–40 °C, which is
consistent with reports of other lepidopteran insects. The enzyme activity was inhibited by addition of
NaCl, KCl, MgCl2, Urea, EDTA, and SDS. CaCl2 enhanced enzyme activity. Ca2+ ions also had inhibitory effects
on glucosidase activity. Plant amylase inhibitors play important role against insect pests. Hence, the
characterization of digestive enzymes and the examination of inhibitors on enzyme activity could be useful
in tackling of insect pests.

참고문헌 (Reference)

1 Chen, M.S., "α-Amylase from three species of stored grain coleopteran and their inhibition by wheat and corn proteinaceous inhibitors" 22 : 261-268, 1992

2 Estruch, J.J., "Transgenic plants: An emerging approach to pest control (1997)" 15 : 137-141, 1997

3 Chapman, R.F., "The insects: structure and function" Cambridge university press 770-, 1998

4 Podoler, H., "The alpha-amylase of the beetle Callosobruchus chinensis" 121 : 321-325, 1971

5 Kanekatysa, U., "Studies on further properties for an alkaline amylase in the digestive juice of the silk worm" 76 : 1-21, 1978

6 Barbosa Pereira, P.J., "Specific inhibition of insect α-amylases: yellow meal worm α-amylase in complex with the Amaranth α-amylase inhibitor at 2.0 Å resolution" 7 : 1079-1088, 1999

7 Hori, K., "Some properties of salivary amylases of Adelphocoris suturalis (Miridae), Dolycoris baccarum (Pentatomidae), and several other heteropteran species" 12 : 454-466, 1969

8 SAS institute, "SAS/STAT User's Guide for Personal Computers" SAS Institute 1997

9 Dezfoulian, A., "Rice striped stem borer" Agri. Org. Iran Pub 149-, 1972

10 Campos, F.A.P., "Resolution and partial characterization of proteinases andα-amylases from midgut of larvae of the bruchid beetle Callosobruchus maculates (F)" 92 : 51-57, 1989

11 Doani,W.W., "Quantification of amylases in Drosophila separated by acrylamide gel electrophoresis" 164 : 363-378, 1967

12 Baker, J.E., "Purification and partial characterization of α-amylase allozymes from the lesser grain borer Rhyzopertha dominica" 21 : 303-311, 1991

13 Baker, J.E., "Purification and partial characterization and postembryonic levels of amylases from Sitophilus oryzae and Sitophilus granaries" 2 : 415-428, 1985

14 Mohammed, M.A., "Purification and characterization of α-amylase from the infective juveniles of the nematode Heterorhabditis bacteriophaga" 139 : 1-9, 2004

15 Ishaaya, I., "Protease and amylase activity in larvae of the Egyptian cotton worm, Spodoptera littoralis" 17 : 945-953, 1971

16 Ferreira, C., "Properties of the digestive enzymes snd the permeability of the perithrophic membrane of Spodoptera frugiperda (Lepidoptera) larvae" 107 : 631-640, 1994

17 Ferreira, C., "Properties of digestive glycosidase and peptidase and the permeability of the peritrophic membranes of Abracris flavolineata (Orthoptera: Acrididae)" 123 : 241-250, 1999

18 Baker, J.E., "Properties of amylase from midgets of larvae of Sitophilus zeamais and Sitophilus granaries" 13 : 421-428, 1983

19 Hosseinkhani, S., "Partial unfolding of carbonic anhydrase provides a method for its immobilization on hydrophobic adsorbents and protects it against irreversible thermoinactivation" 33 : 179-184, 2003

20 Da Silva, M.C.M., "Mutants of common bean alpha-amylase inhibitor-2 as an approach to investigate binding specificity to alpha-amylases" 39 : 201-208, 2004

21 Oliveira-Neto, O., "Molecular cloning of α-amylase from the cotton ball weevil, Anthonomus grandis and structural relations to plant inhibitors: an approach to insect resistance" 22 : 77-87, 2003

22 Terra, W.R., "Midgut amylase lysozyme, aminopeptidase, and trehalase from larvae and adults of Musca domestica" 9 : 283-297, 1988

23 Ghoshal, D., "Introduction and expression of cowpea trypsin inhibitor (CpTI) gene in transgenic tobacco" 10 : 19-25, 2001

24 Slansky, F., "Insect nutrition: an adaptationist's perspective" 65 : 45-71, 1982

25 Dow, J.A.T., "Insect midgut function" 19 : 187-328, 1986

26 Majidi, F., "Identification of some entomopathogei fungi on Chilo suppressalis Walker in Guilan province" 23 : 37-49, 2002

27 Dow, J.A., "Extremely high pH in biological systems: a model for carbonate transport" 246 : 633-635, 1984

28 Pusztai, A., "Expression of the insecticidal bean alpha-amylase inhibitor transgene has minimal detrimental effect on the nutritional value of peas fed to rats at 30% of the diet" 129 : 1597-1603, 1999

29 Maqbool, S.B., "Expression of multiple insecticidal genes confers broad resistance against a range of different rice pests" 7 : 85-93, 2001

30 Bandani, A.R., "Effects of efrapeptin and destruxin, metabolites of entomogenous fungi, on the hydrolytic activity of a vacuolar type ATPase identified on the brush border membrane vesicles of Galleria mellonella midgut and on plant membrane bound hydrolytic enzymes" 1510 : 367-377, 2001

31 Somadder, K., "Digestive enzymes in the gut and salivary gland of the larvae of Chilo auricilius Ddgn" 36 : 218-223, 1980

32 Mendiola-Olaya, E., "Digestive amylase from the larger grain borer" 126 : 425-433, 2000

33 Cohen, A.C., "Demonstration of alpha-amylase in the sweetpotato whitefly, Bemisia tabaci (Heteroptera: Aleyrodidae)" 109 : 593-601, 1994

34 Khanjani, M., "Crop pests of Iran" Boali Sina university press 717-, 2006

35 Applebaum, S.W., "Comprehensive Insect Physiology, Biochemistry and Pharmacology" Pergamon Press 279-307, 1985

36 Zeng, F., "Comparison of α-amylase and protease activities of a zoophytophagus and two phytophagous Heteroptera" 126 : 101-106, 2000

37 Lammli, U.K., "Cleavage of structural proteins during the assembly of bacteriophage T4" 227 : 680-685, 1970

38 Kommano, S., "Chilo suppressalis Walker rearing" handbook of insect rearing. Elsvier 219-226, 1985

39 Agblor, A., "Characterization of alpha-amylase and polygalacturonase from Lygus spp" 27 : 321-326, 1994

40 Terra, W.R., "Biology of the insect midgut" Chapman and Hall 207-235, 1996

41 Terra, W.R., "Biology of the Insect Midgut" Chapman and Hall 153-193, 1996

42 Kfir, R., "Biology and management of economically important lepidopteran cereal stem borers in Africa" 47 : 701-731, 2002

43 Abraham, E.G., "Biochemical study of amylase in the silkworm,Bombyx mori L.: comparative analysis in diapausing and nondiapausing strain" 22 : 867-873, 1992

44 Kazzazi, M., "Biochemical characteristics of a-amylase of the sunn pest" 8 : 371-377, 2005

45 Morton, R.L., "Bean α-amylase inhibitor 1 in transgenic peas (Pisum sativum) provides complete protection from peaweevil (Bruchus pisorum) under field conditions" 3820-3825, 2000

46 Muralidharan, K., "Assessments of crop losses in rice ecosystems due to stem borer damage (Lepidoptera: Pyralidae)" 25 : 409-417, 2006

47 Bernfeld,P., "Amylases, α and β" 1 : 149-158, 1955

48 Gutierrez, C., "Alpha-amylase activities of agricultural insect pests are specifically affected by different inhibitor preparations from wheat and barely endosperms" 72 : 37-44, 1990

49 Franco, O.L., "Activity of wheat α-amylase inhibitors towards bruchid α-amylases and structural explanation of observed specificities" 267 : 2166-2173, 2000

50 Terra, W.R., "Action pattern, kinetical properties and electrophoretical studies on an alpha-amylase present in midgut homogenates from Rynchosciara americana (Diptera) larvae" 56 : 201-209, 1977

51 Zibaee, A., "A study on biochemical differences among five different groups of rice striped stem borer Chilo suppressalis Walker (Lepidoptera: Pyralidae)" 5 : 20-29, 2008

52 Bradford, M., "A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding" 72 : 248-254, 1976

53 Strobl, S., "A novel strategy for inhibition of α-amylases: yellow meal worm α-amylase in complex with Ragi bifunctional inhibitor at 2.5 Å resolution" 6 : 911-921, 1998