Original Article Determination of Physicochemical Properties and Morphological Observation in Syzygium Cumuni Leaf Galls P. Srilakshmi* Department of Biotechnology, Gokaraju Rangaraju institute of engineering and technology, Hyderabad, Andhra Pradesh, India Address for Correspondence Department of Biotechnology, Gokaraju Rangaraju institute of engineering and technology, Hyderabad, Andhra Pradesh, India Tel:+91-8885517198. E-mail: srilakshmi.penumatsa @yahoo.com ABSTRACT Galls are plant tissue which is controlled by the insect. Galls are outgrowths on the surface of leaf caused by invasion of various organisms like Bacteria, Fungi, Insects, Parasites and Mites. Galls may be found on leaves, stems, twigs, branches, trunks and roots. There are hundreds of unique galls caused by insects and mites. They are formed on a variety of plants and in a broad range of sizes, shapes, colors and textures. Some galls are common and abundant and easily noticed. Others are rare or less conspicuous. This paper reports physicochemical study in leaf galls of Syzygium cumini. The parameters assayed were Total ash, acid insolubility and water solubility values and extractive values (Alcohol, water and petroleum ether). Keywords: Galls, Physicochemical studies, Syzygium cumini. INTRODUCTION Plant galls are abnormal growths of plant cells formed as a response to the insects stimulus caused by egg laying insects or larvae or nymphs feeding. Gall production is believed to result when the cambium and other meristematic tissues react to stimuli produced by the larvae and cause the abnormal growths. The immature insects often can be found in a cell or cells within the developing gall. After a brief period of cell growth, all development stops. The insect becomes enclosed by the gall and feeds only on gall tissue during its development. Small holes on the outside of the gall indicate that the adult insects have emerged. After formation, it is impossible to eliminate the galls or the parts with insecticides because they are enclosed and well protected inside the gall. Those that occur on the leaves will drop off with the leaves, but those occurring on the trunk, roots and stems may persist for several years. Most galls do not adversely affect plant health. For insects/mites that act on the host plant, horticultural oil application can be made before insect/mite activity begins in the spring. Syzygium cumini belongs to American Journal of Phytomedicine and Clinical Therapeutics www.ajpct.org Srilakshmi _________________________________________________ ISSN 2321 – 2748 family Myrtaceae, in Hindi it is commonly known as Jamun. It is a medicinal plant. Common names are Java palm, Black plum, Jambul and Indian Blackberry1,2. The native home of jamun is India, distributed throughout India, in the forest up to 1800m generally along the bank and moist localities. The flowers are fragrant and small, about 5mm in diameter. The fruits develop by May or June and resemble large berries. Powdered seeds are used as a remedy in diabetes and in Metrorrhagia3,4. The seeds are sweet, astringent to bowels and good for diabetes. Jambul trees start flowering from March to April. The ash of the leaves is used for strengthening the teeth and gums. Vinegar prepared from the juice of the ripe fruit is an agreeable stomachic and carminative and used as diuretic5 and it is also useful in spleen enlargement and an efficient astringent in chronic diarrhea. In Unani medicine various parts of Jambul act as liver tonic, enrich blood, strengthen teeth and gums and form good lotion for removing infection of the head6. Other parts have been reported to possess anti-diabetic7. The fruits are berries and are oblong, 1.5-3.5 cms long, dark-purple or black, luscious, fleshly and edible; it contains a single large seeds8,9. Insect gall formation is uncommon and specific to only certain plants10. Syzygium cumini is one such plant. Galls are seen on leaves. Galls may also provide the insect with physical protection from predators11,12. There are few studies about the physiochemical parameters on Syzygium cumini leaves. However, such reports in Syzygium cumini leaf galls have not been recorded. Hence we have conducted physicochemical studies in leaf galls of Syzygium cumini. The results were compared. AJPCT[2][2][2014]174-179 MATERIALS AND METHODS Collection of Plant Material Young galled leaves of equal size were collected from the local areas of Hyderabad, Andhra Pradesh, India during August and September 2013. Fresh leaves were collected in bulk, washed, shade dried and pulverized in a mechanical grinder to obtain coarse powder and their physicochemical study was done. Determination of Physicochemical Parameters Moisture content The percentage of active chemical constituents in crude drug is given in terms of air dried drugs. So the moisture content of a drug should be determined. 2g of powdered drug was transferred into a china dish and the contents were distributed evenly to a depth not exceeding 10mm. The loaded plate was heated at 105°C in hot air oven and weighed at different time intervals until a constant weight was obtained. The experiment was repeated six times for precision and percentage of moisture of the sample was calculated by taking the difference in weight after drying and initial weight of the drug13. Total ash value About 2g of powdered drug was weighed accurately into a crucible and incinerated at a temperature of 450°C in muffle furnace until free from carbon. Then the crucible was cooled and weighed. Percentage of ash was calculated with reference to air dried plant material13.14. Acid insoluble ash Ash obtained from total ash was boiled with 25ml of 2N HCl for few minutes and filtered through an ash less filter paper. The filter paper was transferred into a silica crucible and incinerated at 650°C in muffle furnace until free from carbon. Then the Srilakshmi _________________________________________________ ISSN 2321 – 2748 crucible was cooled and weighed. Percentage of acid insoluble ash was calculated with reference to air dried substance14. Water soluble ash Ash obtained from total ash was boiled with 25ml of distilled water for few minutes and filtered through an ash less filter paper. The filter paper was transferred into a silica crucible and incinerated at 450°C in muffle furnace until free from carbon. Then the crucible was cooled and weighed. Percentage of water soluble ash was calculated with reference to air dried substance13,14. Determination of ethanol soluble extractive value Accurately weighed 5g of air dried powdered plant material was macerated with 100ml of ethanol in a closed flask for 24hrs, shaking frequently during first 6hrs and allowed to stand for 18hrs. It was then filtered rapidly, taking precautions against loss of the solvent and 25ml of the filtrate were evaporate to dryness in a flat-bottomed shallow dish and dried at 100°C. The % w/w of ethanol soluble extractive value was calculated with reference to the air dried plant material13-15. Determination of water-soluble extractive value Procedure was the same as alcohol soluble extractive using chloroform and water (chloroform: water-1:399) instead of alcohol. Petroleum ether extractive values 5g drug was refluxed with 100ml of petroleum ether for 24hrs and filtered through Whatman filter paper. 10ml of the filtrate was evaporated in a tarred dish at 105°C and weighed. Then ether soluble extractive values were calculated13. AJPCT[2][2][2014]174-179 pH determination pH of 1% and 5% was determined by using pH meter 16 . pH 1%: 1g of the accurately weighed powder of leaves of Syzygium cumini was dissolved in water and filtered. pH of filtrate was determined by using pH meter. pH 5%: 5g of the accurately weighed powder of leaves of Syzygium cumini was dissolved in water and filtered. pH of filtrate was determined by using pH meter. Morphological Features (See table 1 and 2) RESULT AND DISCUSSION There was a substantial increase in total ash and slight increase in acid insolubility and water solubility when compared to normal leaf. But there was decrease in loss on drying and low alcoholic extractive value and high extractive values of petroleum ether and water. Gall removed leaf is also used to observe the changes and compared with the gall and healthy leaf. The results are shown in Table-3. There was a morphological difference in the leaf the results were showed in Table-1 and 2. CONCLUSION Syzygium cumini leaves are used in Ayurveda Medicine preparation. Therefore, from the present investigation we can conclude galls cause a change in physicochemical parameters (Loss on drying, total ash, acid insolubility, water solubility and extractive values). Physicochemical studies are carried out to confirm the identity of plant and ascertain the quality and purity of the drug material. So these parameters may be useful for the future investigations. Srilakshmi _________________________________________________ ISSN 2321 – 2748 REFERENCES 9. 1. 10. 2. 3. 4. 5. 6. 7. 8. D. Kubmarawa, G.A. Ajoku, N.M. Enwerem and D.A. Okorie. Afr. J. Biotechnol, 2007, 6, 1690-1696. Edeoga H.A, Okwu DE, and Mbaebie BO Phytochemical constisstuent of some Nigerian Medicinal Plants, African Journal of Biotech academic journals 2005, 4:685688 Kritikar KR and Basu BD. Indian Medicinal Plants. Periodical Experts, vol II, 1975:1052-53. Nadkarni KM: Indian Materiamedica. Popular book Bombay, vol. I, 1954:516-518. Sharma P and Mehta PM: In pravygunavignayan (The Chowk hambavidya bhawan, Varanasi) Part II and III, 1969:586. Sagrawat H, Mann AS, Khavya MD. Pharmacological potential of Eugenia Jambolana: a review. Pharmacogn Mag 2006; 2: 96-104. Chakraborty P, Mahapatra PK, Chaudhuri AK. 986: A neuro psychopharmacological study of Syzyguim cuminiplanta Med; 139143. Gamble JS. The flora of the Predidency of Madras London: Adlard and Son ltd; 1935. AJPCT[2][2][2014]174-179 11. 12. 13. 14. 15. 16. Hooker JD. The flora of British India. London: Nabu press; 1879, p, 499 Dreger-JauffretF. And Short house J.D. (1992): Diversity of galls-Inducing insects and their galls. In: Biology of insect- induced galls, J.D. Short house and O. Rohfritsch (Eds), Oxford university press, New York. Pp.8-33. Weis, A. E., and A. Kapelinski. 1994. Variable selection on Eurosta’s gall size. II. A path analysis of the ecological factors behind selection. Evolution 48, P.734 – 745. Graham N. Stone and Karsten Schonrogge (2003). The adaptive significance of insect gall morphology. TRENDS in Ecology and Evolution 18(10): 512-522. World health organization: WHO guideline for the Assessment of herbal medicines, WHO expert committee on specification for pharmaceutical preparation. Technical Report series No. 863. Geneva, 1996. Practical pharmacognosy by Kandelwal. K.R., 1998 Pharmacopeia of India, Ministry of Health and Family Welfare, vol. II, Appendix 3, Pp A-53-54. Government of India New Delhi 1996. Dr. Ansari SH, Essential of Pharmacognosy III edi 2008-09, Birla Publication pvt ltd Delhi.591-592. Srilakshmi _________________________________________________ ISSN 2321 – 2748 Table 1: Morphological examination of Syzygium cumini leaf S. No. 1 2 Feature Color (Upper surface) Color (Lower surface) Observation Leaf of Syzygium cumini family Myrtaceae Glossy dark green Dull green 3 Odor Characteristic 4 Shape Ovate or elliptical 5 Size 5-25 cm long, 1 to 4 in 2.5-10 cm wide 6 Arrangement Alternate 7 Texture Smooth Table 2: Morphological examination of galls of Syzygium cumini S. No. Feature 3 Color (Outer surface) Color (Inner surface) Odor 4 Shape 5 Size 6 Arrangement 7 Texture 1 2 AJPCT[2][2][2014]174-179 Observation Green Light green Characteristic Hemispherical to globose 10 mm high, 1 – 2 mm thick at base and 5 mm thick apically Usually simple and free Smooth, but hard galls are greenish or yellowish Galls of Syzygium cumini family Myrtaceae Srilakshmi _________________________________________________ ISSN 2321 – 2748 Table 3: Physicochemical parameters of Syzygium cumini Parameter LOD Total ash value Acid insoluble Water soluble Petroleum ether Alcoholic Water/Aqueous PH (1%) PH (5%) PH (10%) AJPCT[2][2][2014]174-179 Gall 1.86% Ash values 9% 2.75% 3.5% Extractive values 44% 48% 68% 8.2 8.3 8.7 1.88% Gall removed leaf 1.82% 5.5% 2% 2.9% 6.2% 1.8% 3.1% 40% 57.6% 32% 8.1 7.9 7.4 42% 54.8% 39% 7.9 7.7 8.3 Healthy Leaf