Biochemical studies on the amylase of Mango Mealybug (Drosicha stebbingi Green)

Yıl 2014, , 307 - 322, 01.06.2014

Mubasshir Sohaıl Muhammad Anjum Aqueel Muhammad Afzal Abubakar Muhammad Raza Muhammad Karman Muhammad Sajjad Khalıl

https://doi.org/10.16970/ted.82875

Öz

Mealybug (Drosicha stebbingi Green) is voracious pest of the mango orchards. It causes heavy quality losses and low production of mango fruit (Luhanga & Gwinner 1993). The research was conducted to control the pest. Isolation, purification and characterization of amylase from mango mealybug were performed. Amylolytic activity was 365U/ml in crude homogenate of D. stebbingi whereas in acetone precipitation sample activity was 680 U/ml. The optimum pH and temperature for amylase activity was 6.4 and 50-60 ºC, respectively. However, amylase activity was maximum at 3% of substrate (starch) concentration. Enzymatic activity was inhibited by Pb2+, Na1+, Hg2+, Ni2+and Cu2+ but Ca2+ enhanced the enzymatic activity. Natural plant extract like Datura (Datura alba), Neem (Azadirachta indica) and citrus (Citrus reticulata) inhibited the amylolytic activity. Molecular weight of amylase was 26 kDa measured by sodiumdodisylsulphate-polyacrylamide gel electrophorasis (SDS-PAGE). Purpose of the research was to evaluate the isolation, purification and biochemical characterization of the digestive enzyme of mango mealybug. The experiment provides better understanding about digestive physiology of mealybug for its efficient control. Biochemical characteristics of digestive amylolytic activity in adults of mango mealybug may open new horizons for its control

Anahtar Kelimeler

Mango Mealybug, amylase, biochemical characterization

Mango unlubitlerinde (Drosicha stebbingi Green) amilaz üzerinde biyokimyasal çalışmalar

Öz

Mango unlubiti (Drosicha stebbingi Green) mango bahçelerinde oburca beslenen bir zararlıdır. Mango meyvesinde ağır kalite kaybı ve düşük üretime sebep olmaktadır (Luhanga & Gwinner 1993). Bu çalışma zararlının kontrolü için yapılmıştır. İzolasyon, saflaştırma ve amilaz karakterizasyonu Mango unlubitinden yapılmıştır. Amilolitik aktivite D. stebbingi ham homojenat içinde 365U / ml iken aseton içinde çökeltme örnek aktivitesi 680 U / ml olmuştur. Amilaz aktivitesi için optimum pH ve sıcaklık sırasıyla, 6.4 ve 50-60 ºC olmuş, bununla birlikte, amilaz aktivitesi, maksimum alt-tabaka (nişasta) konsantrasyonunun% 3’ü olmuştur. Enzimatik aktivite Pb2 +, nA1- +, Hg2 +, Ni + 2 ve Cu + 2 tarafından önlenmiş, fakat Ca + 2 enzimatik aktiviteyi arttırmıştır. Datura (Datura alba), Neem (Azadirachta indica) ve narenciye (Citrus reticulata) gibi doğal bitki ekstraktları amilolitik aktiviteyi engellemiştir. Sodiumdodisylsulphatepoliakrilamid jel elektroforez (SDS-PAGE) ile ölçülen amilaz molekül ağırlığı 26 kDa olmuştur. Araştırmanın amacı izolasyonu, saflaştırmayı ve mango unlubitlerin sindirim enziminin biyokimyasal karakterizasyonunu değerlendirmektir. Bu çalışma unlubitlerin etkin kontrolü için, unlubitlerin sindirim fizyolojisinin daha iyi anlaşılmasını sağlar. Mango unlubitlerin erginlerindeki sindirim amilolitik aktivitesinin biyokimyasal özellikleri zararlının kontrolü için yeni ufuklar açabilir

Anahtar Kelimeler

Mango unlubiti, amilaz, biyokimyasal karakterizasyon

Kaynakça

• FAO, Production Yearbook (2010) Food and agricultural organization of the United Nations, Rome.
• Abraham EG, Nagaragu J, K. DR (1992) Biochemical study of amylase in the silkworm,Bombyx mori L.: comparative analysis in diapausing and nondiapausing strain. Insect Biochemistry and Molecular Bioliology:22:867-873.
• Agounkè D, Agricola U, Bokonon-Ganta HA (1988) Rastrococcus invadens Williams (Hemiptera: Pseudococcidae), a serious pest of fruit trees and other plants in West Africa. Bulleton of Entomological Research:78:695–702.
• Akhtar M, Anwar M, Pasha MK, Alam GS (2000) Geological Road Log along the Islamabad-Lahore Motorway. Geological Survey of Pakistan.
• Azevedo PV, Silva BBd, da Silva VPR (2003) Water requirements of irrigated mango orchards in northeast Brazil. . Agricultural Water Management:58 241-254.
• Bokonon-Ganta AH, Groote HD, Neuenschwander P (2002) Socio-economic impact of biological control of mango mealybug in Benin. Agriculture, Ecosystem and Environment:93:367-378.
• Bradford MM (1976) A dye binding assay for protein. Analytical Biochemistry:72:248-254.
• Burhan A, Nisa U, Gokhan C, Ashabil A, Osman G (2003) Enzymatic properties of novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. Process Biochemistry:38:1397-1403.
• Chary SJ, Reddy SM (1985) Starch degrading enzymes of two speciesof Fusarium folia. microbiology:30:452.
• Dow JA (1984) Extremely high pH in biological systems: a model for carbonate transport. American Journal of physiology:246: 633-635.
• Kadziola A, Abe J, Svensson B, Haser R (1993) Crystal and molecular structure of barley α-amylase. Journal of molecular biology 250:195–202.
• Khan MA (2001) Control of insect pests of mango. Proc. Inter. Mango workshop 27 Feb.-1 Mar. Direct. Agric-Multan Region, Punjab:224.
• Luhanga WW, Gwinner J (1993) Mango mealybug (Rastrococcus iceryoides) on Mangifera indica in Malawi. FAO Plant Protection Bulletin:41:125-126.
• Mehrabadi M, Ali RB (2009) Study on Salivary Glands α-amylase In Wheat Bug Eurygaster maura (Hemiptera: Scutelleridae). American Journal of Applied Sciences:6:555-560.
• Mendiola-Olaya E, Valencia-Jiménez A, Valdés-Rodríguez S, Délano-Frier J, Blanco-Labra A (2000) Digestive amylase from the larger grain borer, Prostephanus truncatus Horn. Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology:126:425-433.
• MINFAL (2010) Agriculture Statistics of Pakistan. Govt. of Pakistan, Min.of Food, Agri. and Livestock Economic Wing, Islamabad.
• Mohammed MA (2004) Purification and characterization of alpha-amylase from the infective juveniles of the nematode Heterorhbditis bacteriophaga. Comparative Biochemistry and Physiology part B:139:1-9.
• Narula S (2003) Nip mango mealybug in the bud now Spectrum, Sunday 14, 2003. Tribune (GARDEN LIFE) India.
• Oliveira-Neto O, Batista JAN, Rigden DJ, Franco OL, Falcao R, Fragoso RR, Mello LV, Santos RCD, Grossi-de-sa MF (2003) Molecular cloning of amylase from cotton boll weevil, Anthonomus grandis and structural relations to plant inhibitors: An approach to insect resistance. Journal of protein chemistry:22:77-87.
• Prasad V, Singh RK (1976) Prevalence and control of mango mealy bug Drosicha stebbingii (Green) in Bihar. Indian Journal of Entomology:38:2144-224.
• Rao MS, Reddy NS, Rao GV, Sambasiva Rao KRS (2005) Studies on the extraction and characterization of thermostable a-amylase from pericarp of Borassus indica. African Journal of Biotechnology 4:289-291.
• Rehman FU, Aslam M, Taiq MI, Shaheen MA, Amtul JS, Naveed NH, Iram BA (2009) Isolation of cellulolytic activities from (red flour beetle) Tribolium castaneum. African Journal of Biotechnology:8:6710-6715.
• Sami AJ, Akhtar MW (1990) Purification and characterization of two native extracellular carboxymethyle cellulose of carboxymethylcellulase of Cellulomonas flavigena. Biochemical Society Transactions:18:651.
• Shalom G, Pratten J (2008) Cloning, heterologous gene expression and biochemical characterization of the alpha- 1,3-glucanase from the filamentous fungus Penicillium purpurogenum. Protein Expression Purification:60:170- 175.
• Sivakumar D, Jiang Y, Yahia EM (2011) Maintaining mango (Mangifera indica L.) fruit quality during the export chain. Food Research International:44:1254-1263. DOI: http://dx.doi.org/10.1016/j.foodres.2010.11.022.
• Tandon PL, Lal B (1978) The mango coccid, Rastrococcus iceryoides Green (Homoptera: Coccidae) and its natural enemies. Current Science:13:46-48.
• Tandon PL, Verghese A (1985) World List of Insect, Mite and other Pests of Mango. Technical Document No. 5, IIHR, Banglore:22.
• Watanabe H, Tokudat G (2001) Animal cellulases. Cellular and Molecular Life Sciences:58:1167-1178.
• Yousuf M (1993) Mango mealybug control with polyethylene bands. Pakistan Entomologist:15:129.
• Zeng F, Cohen AC (2000) Comparison of α-amylase and protease activities of a zoophytophagus and two phytophagous Heteroptera. Comparative Biochemistry and Physiology:126:101-106.
• Zibaee A, Bandani AR, Kafil M, Ramzi S (2008) Characterization of α-amylase in the midgut and the salivary glands of rice striped stem borer, Chilo suppressalis Walker (Lepidoptera: Pyralidae). Journal of Asia-Pacific Entomology:11:201-205. DOI: http://dx.doi.org/10.1016/j.aspen.2008.09.003.
• Zoltowska K (2001) Purification and characterization of alpha-amylases from the intestine and muscle of ascaris suum (Nematoda. Acta Biochimica Polonica:48:763-774.