Chemical Changes in Soybean Oil Due to Seed Borne Fungi

International Journal of Botany and Research (IJBR) ISSN (P): 2277–4815; ISSN (E): 2319–4456 Vol. 10, Issue 1, Jun 2020, 1–8 © TJPRC Pvt. Ltd.

CHEMICAL CHANGES IN SOYBEAN OIL DUE TO SEED BORNE FUNGI KESARE. U. T Department of Botany, Adarsh College, Omerga, Osmanabad, Maharashtra, India ABSTRACT Soybean oil was known for its contents, as it is commonly used for its consumption purpose. The oil extracted from artificially inoculated Soybean seeds shows biochemical changes. For the study purpose, changes were made in different parameters of oil, and five fungi were artificially inoculated to the three varieties of soybean seeds i.e. JS-335, JS-80 and Pooja. It is found that due to the association of Alternaria alternata, Aspergillus flavus, Aspergillus ustus, Fusarium oxysporum and Trichoderma viride, there is chemical changes in Soybean oil.

KEYWORDS: Soybean Oil, Chemical Changes & Soybean Varieties

Received: Oct 28, 2019; Accepted: Nov 18, 2019; Published: Jan 02, 2020; Paper Id.: IJBRJUN20201

INTRODUCTION

transmission. This takes place either in the field or in ill storage condition. Neergard (1977) reported that in the presence of seed-borne pathogens, several types of abnormalities like reduction in seed size, seed rotting, discoloration of seeds, seed necrosis, loss in germ inability, toxification and other physiological disorders.

According to Sandikar (1990), the species of fusarium are found to be significantly destructive and

Original Article

Seed plays a vital role for the production of healthy crop. These seeds are also responsible for disease

responsible to cause harmful effect on seed health resulting in to seed deterioration and poisoning of seeds.

During the process of bio-deterioration, fungi produce enzymes to degrade proteins, carbohydrates and oil.

Sharma(1987)and Agarwaland Gupta(1990) reported; Alternaria, Aspergillusflavs, Fusarium species, Trichothecium and Cladosporium, Collototrichum dematium and Macrophomina phaseolina from Soybean seeds. Laxmi et.al (1989) also observed some species of fungi from Soybean seeds during storage conditions.

Ouf et.al (1999) reported Rhzoctoniasolani, alternariatenuissima, Cercosporakikuchi and Colletotric humtruncatum on seeds of Soybean. Ramesh and Marihal(2006) observed the dominant fungi such as Alternaria alternate, Aspergillusniger, A.sydowi, A.ustus, Cercospora kikuchi and Fusarium moniliforme.

Therefore, the present study aimed to throw light on the changes in physical parameters of Soybean oil due to associated my coflora

MATERIALS AND METHODS Collection of Seed Samples and Detection of Seed Mycoflora For the collection of seed samples, the method described by Neergaard (1973) has been adopted. Accordingly from fields, store houses market places and seed companies. A composite sample of each variety was prepared by mixing the individual samples together. The seed mycoflora was isolated by using standard moist blotter paper method (SMB) and agar plate method (APM), as recommended by International seed testing association (ISTA 1966), De Tempe (1970) , Neergaard (1973) and Agarwal (1976).

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Identification of Seed Borne Fungi The fungi, occurring on each and every seed in the plates were identified preliminary on the basis of sporulation characters like sexual or asexual spores with the help of stereoscopic binocular microscope. The identification and further confirmation of seed-borne fungi was made by preparing slides of the fungal growth and observing them under compound microscope. The identification was made with the help of manuals as per Nelson, et.al. (1983),Singh, et.al(1991), Mukadam D.S.(1997) and Mukadam et.al (2006).

Calculations Estimation of Oil 10 gm of seed was taken and crushed using a mortar and pestle. Carefully mixed the ground sample and transferred the ground material to an extraction thimble, the thimble was placed in the extractor, previously fitted with a weighed flask.

The morter and pestle was washed with petroleum hydrocarbon solvent, transferring the washings to the thimble in the extractor.

The quantity of solvent required for efficient working of the extractor was added to this apparatus, the condenser has been fixed and allowed through it a cold water to flow, and heated the extractor so that the action was moderate and not violent. The extraction was continued to four hours. Then the extractor was removed from its bath, the extraction thimble was taken, after it has drained out of the extractor and allowed to evaporate from it in a current of air.

The thimble was dried in the oven at 105oC + 20oC for 30 minutes. The material was taken into a morter and added approximately 10 gm of fine sand or quartz and grinded the mixture as finely as possible.

The grinded material was transferred to the thimble again and placed the thimble in the extractor. The pestle and morter rinsed with solvent and transferred rinsings to the extractor. The extraction process was repeated further for two hours.

Again, the thimble was removed from the extractor, dried it with the contents as before, and grinded the contents again without the addition of more sand or quartz. Again, the ground material was transferred to the extractor; a second weighted flask was fitted to the extractor. Pestle and mortar was rinsed with solvent as before and extracted for two hours.

The greater part of the solvent was removed in the flasks by distillation. The flasks were heated in an oven at 103oC + 2oC for one hour to remove solvent.

The flask was cooled to room temperature and weighed. The heating process and cooling repeated, until the weight of the flasks was constant. If the weight of oil in the second flask does not exceed 10 mg, the extraction was considered to be completed. If the oil recovered as above is not clear, it may be dissolved in solvent and filtered though a filter paper. The paper should be washed thoroughly with solvent, an oil solution and the washing should be combined. Solvent should be evaporated and the flask containing should be dried and weighed as above to obtain the amount of oil.

Calculation Oil % = W1 x 100 ----------------

W Where, Impact Factor (JCC): 5.5837

NAAS Rating: 4.08

Chemical Changes in Soybean Oil Due to Seed Borne Fungi

W1=

Weight in gm of oil extraction thimble and

W=

Weight in gm of the sample taken for the test.

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Ref.: SOPA (2003)

RESULTS AND DISCUSSIONS In the present paper, seed mycoflora of only three varieties of Soybean are given in Table no.1.Actually, ten different varieties of Soybean from Marathwada region (MH) are selected for the study, and it is observed that total thirty species of fungi were isolated. Mycoflora mainly dominated by seven species of Aspergillus six species of fusarium followed by Curvularia lunata, Alternaria altenata, Pnenicillium chrysogenum ,Spicaria violacia, Trichoderma viride and Verticillium species, which occurs comparatively higher incidence on Soybean varieties studied. The qualitative and quantitative mycoflora varies in different varieties of Soybean.

Soybean oil is popularly used for consumption purpose by human being. Oil extracted from artificially inoculated seeds shows biochemical changes. In order to study the changes in physical parameters of oil, five dominant fungi were selected and are artificially inoculated on three popular varieties of Soybean, which are grown on large scale in Marathwada region (MH).

Table 1: Incidence of Fungi on Three Varieties of Soybean by Agar Plate Method Soybean Varieties Sr. No. Fungi Js-80 Js- 335 Pooja 1 Alternaria alternata 20 10 16 2 Aspergilluscandidus 18 20 20 3 Aspergillusflavus 80 13 50 4 Aspergillusfumigatus 40 12 35 5 Aspergillusglaucus 22 08 10 6 Aspergillusniger 26 16 16 7 Aspergillusrubrer 35 10 28 8 Aspergillusustus 35 12 25 9 Botrytis cinerea 09 10 13 10 Cladosporiumsps 06 06 06 11 Colletotrichumsps 04 08 06 12 Curvularialunata 09 10 13 13 Curvulariatetramera 06 12 10 14 Drechsleraoryzae 06 06 06 15 Fusariumchladosporium 10 08 00 16 Fusariumdimerum 28 25 32 17 Fusariumequiseti 20 35 30 18 Fusariummoniliformae 25 24 16 19 Fusariumoxysporum 17 24 16 20 Fusariumroseum 45 22 32 21 Helminthsporiumsps 06 22 32 22 Hormodendrumcladosporiodes 09 08 08 23 Macrophominaphaseolina 09 10 12 24 Moniliasitopila 10 12 13 25 Pencilliumchrysogenum 06 08 06 26 Rhizoctoniabataticola 10 10 06 27 Spicariaviolecia 20 12 06 28 Trichodermaviride 09 09 12 29 Tubercularia vulgaris 05 10 07 30 Verticilliumsps.

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Change in Chemical Parameters of Oil in Soybean Variety JS-335 Soybean oil was known for its content, as it is commonly used for its consumption purpose. In order to see the effect of fungi for change in chemical composition of oil, the experiment was set and the results are given in table no.2.

It is observed that free fatty acid increases considerably in the presence of Fusariumroseum. The Iodine values did not show much variation, whereas peroxide value increases due to Fusariumroseum, Aspergillusustus, Alternariaalternata and Trichodermaviride. Saponification and unsaponification value did not show much variation due to association of fungi.

Change in Chemical Parameters of Oil in Soybean Variety JS-80 Soybean Variety JS-80 and its change in the chemical composition of oil is studied and their results are given in Table. 3.

It was observed that percent free fatty acid increased due to association of Fusariumroseum, Trichodermaviride, Aspergillusustus and Alternariaalternata similarly peroxide value also increased due to F. roseum followed by Austus, T. viride, A. alternataand A. flavus. There is no much variation was reported in saponification and unsaponification value.

Change in Chemical Parameters of Oil in Soybean Variety Pooja Soybean variety Pooja and its change in its chemical composition of oil were studied and their results are given in Table no.4.

It is observed that percent free fatty acid increase due to association of Fusariumroseum. Trichodermaviride, Aspergillusustus and Alternariaalternata. Similarly peroxide value also increased due to F. roseum followed by A. ustus, T. viride, Aalternata and A flavus. There was not much variation was reported in saponification and unsaponification value.

In case of Iodine value also, there was not much variation was reported.

Fatty Acid Composition of Oil in Soybean Variety JS-335 Fatty acid composition is one of the important phenomena of edible oil; therefore detail investigation of change in different parameters of fatty acid was studied. The results of soybean variety JS-335 was shown in Table No. 5.

It is observed that myristic acid is reduced due to Alternariaalternata and increased due to Fusariumroseum.

Plamitic acid content was increased due to A. alternata, Aspergillusustus and F. roseum. Palmitooleic acid content show slight variation in its content stearic acid content increased due to A. alternataand F. roseum. Oleic acid content reduced due to F. roseum. Similarly linoeic acid reduced due to A. alternata and spergillusflavus. Arachidic acid content was increased due to A. ustus, whereas beheneic acid content is increased due to all test fungi.

Fatty Acid Composition of Oil in Soybean Variety JS-80 Change in fatty acid composition due to associated fungi is shown in table no.6.

It was observed that myristic acid is reduced due to Alternariaalternata, whereas palmitio acid content was increased due to A. alternata, Aspergillusustus and Fusariumroseum. Palmite oleic acid content show slight variation in its content stearic acid content increased due to Alternariaalternata and Fusariumroseum. Oleic acid content reduced due to F. roseum. Similarly,Linoeic acid was reduced due to Alternariaalternata and Aspergillusflavus. Arachidic acid content was increased due to Aspergullusustus, Trichodermaviride. Whereas, Beheheic acid content was increased due to all test fungi.

Impact Factor (JCC): 5.5837

NAAS Rating: 4.08

Chemical Changes in Soybean Oil Due to Seed Borne Fungi

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Fatty Acid Composition of Oil in Soybean Variety Pooja Change in fatty acid composition due to associated fungi is shown in Table No. 7.

It was observed from the table that myristic acid was reduced due to Alternariaalternata Palmitic acid is reduced due to Alternariaalternata Palmitic acid content was increased due to Fusariumroseum, Aspergillusustas and Alternariaalternata. Palmitooleic acid show slight variation due to associated fungi. Steric acid content was increased due to Fusariumroseum and Alternariaalternata. Oleic acid content is reduced due to F. roseum.

Linoeic acid content was reduced due to Alternariaalternata and Aspergillusflavus.linoleheic acid content do not show much variation due to associated fungi. Arachidic acid was increased due to Aspergillusustus, however beheneic acid was increased due to all fungi.

Table 2: Change in Chemical Parameters of Oil in Soybean Variety Js-335 Parameters Control Aal Asf Asu Fur Triv Free fatty acid % 0.58 5.14 3.14 5.76 12.78 6.78 Iodine Value % 126.70 127.06 126.70 126.34 124.52 125.26 Peroxide Value 4.22 7.04 5.14 8.44 12.28 6.28 Soap (ppm) 00 00 00 00 00 00 Saponification Value 191.21 187.7 191.06 190.46 190.0 190.68 Un saponification Value 1.84 1.92 1.468 1.9 1.9 1.85 AalAlternariaalternata AsfAspergillusflavus AsuAspergillusustus FurFusariumroseum TrivTrichodermaviride

Table 3: Chemical Parameters of Oil in Soybean Variety JS-80 Parameters Control Aal Asf Asu Fur Free fatty acid % 0.58 5.13 3.15 5.73 12.78 Iodine Value % 126.70 127.08 126.7 126.30 124.41 Peroxide Value 4.22 7.04 5.12 8.43 12.27 Soap (ppm) 00 00 00 00 00 Saponification Value 191.21 187.7 191.06 191.06 190.46 Unsaponification Value 1.84 1.93 1.469 1.9 1.9 AalAlternariaalternata AsfAspergillusflavus AsuAspergillusustus FurFusariumroseum TrivTrichodermaviride

Triv 6.78 125.19 8.28 00 190.68 1.87

Table 4: Change in Chemical Parameters of Oil in Soybean Variety Pooja Parameters Control Aal Asf Asu Fur Triv Free fatty acid % 0.58 5.14 3.14 5.76 12.78 6.78 Iodine Value % 126.08 127.07 126.70 126.32 124.42 125.26 Peroxide Value 4.22 7.03 5.14 8.44 12.26 8.26 Soap (ppm) 00 00 00 00 00 00 Saponification Value 191.21 187.6 190.05 190.46 190.00 190.00 Unsaponification Value 1.84 1.93 1.468 1.9 1.9 1.88 AalAlternariaalternata AsfAspergillusflavus www.tjprc.org

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Kesare. U. T

AsuFurTriv-

Aspergillusustus Fusariumroseum Trichodermaviride

Table 5: Fatty Acid Composition of Oil in Soybean Variety JS-335 Parameters Control Aal Asf Asu Fur Triv Myristic acid 0.06 0.04 0.06 0.05 1.05 0.05 Palmitic acid 11.74 12.10 11.62 12.04 12.64 11.71 Palmito oleic acid 1.43 1.58 1.54 1.82 1.71 1.41 Stearic acid 2.84 3.61 2.84 2.93 3.82 2.84 Oleic acid 25.72 25.61 26.84 25.47 24.76 25.14 Linoeic acid 51.01 49.81 50.05 50.65 50.45 51.01 Linoleneic acid 6.82 6.64 6.74 6.84 6.42 6.79 Arachidic acid 0.20 0.29 0.22 0.33 0.28 0.32 Beheneic acid 0.15 0.21 0.25 0.25 0.18 0.20 AalAlternariaalternata AsfAspergillusflavus AsuAspergillusustus FurFusariumroseum TrivTrichodermaviride

Table 6: Fatty Acid Composition of Oil in Soyaben Variety JS-80 Parameters Control Aal Asf Asu Fur Triv Myristic Acid 0.06 0.04 0.66 0.05 0.055 0.05 Palmitic acid 11.74 12.10 11.62 12.04 12.60 11.71 Palmito oleic acid 1.43 1.58 1.54 1.82 1.71 1.41 Stearic acid 2.84 3.61 2.85 2.93 3.82 2.82 Oleic acid 25.72 25.66 26.85 25.48 24.77 25.12 Linoeic acid 51.01 49.80 50.05 50.66 50.45 51.01 Linoleneic acid 6.82 6.62 6.74 6.83 6.44 6.78 Arachidic acid 0.20 0.29 0.22 0.33 0.28 0.31 Beheneic acid 0.15 0.21 0.25 0.25 0.19 0.20 AalAlternariaalternata AsfAspergillusflavus AsuAspergillusustus FurFusariumroseum TrivTrichodermaviride

Table 7: Fatty Acid Composition of Oil in Soybean Variety Pooja Parameters Control Aal Asf Asu Fur Triv Myristic Acid 0.06 0.04 0.06 0.05 1.05 0.05 Palmitic acid 11.74 12.10 11.62 12.03 12.66 11.71 Palmito oleic acid 1.43 1.58 1.54 1.81 1.71 1.41 Stearic acid 2.84 3.61 2.82 2.92 3.80 2.80 Oleic acid 25.72 25.64 26.82 25.44 24.74 25.10 Linoeic acid 51.01 49.78 50.05 50.62 50.42 51.01 Linoleneic acid 6.82 6.61 6.71 6.82 6.42 6.74 Arachidic acid 0.20 0.29 0.22 0.33 0.28 0.31 Beheneic acid 0.15 0.21 0.25 0.25 0.19 0.20 AalAlternariaalternata AsfAspergillusflavus Impact Factor (JCC): 5.5837

NAAS Rating: 4.08

Chemical Changes in Soybean Oil Due to Seed Borne Fungi

AsuFurTriv-

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Aspergillusustus Fusariumroseum Trichodermaviride

CONCLUSIONS Biodeterioration of soybean seeds of variety JS-335, JS-80 and pooja were carried out by artificially inoculating fungi at the laboratory condition. It was found that most of the fungi deteriorated the chemical content of the seed. Soybean oil is commercially used for consumption purpose by the human being and also used for pharmaceutical industries. Experiments were carried out to study the role of fungi in biodeterioration of oil in three different varieties. The chemical composition is also found to be changed from soybean oil due to associated fungi. Free fatty acid of soybean variety JS-335 increased considerably due to Fusarium roseum. Similarly peroxide value also increased due to Fusarium roseum, Aspergillus ustus, Alternaria alternata and Trichoderma viride. Saponification value do not show much variation. Chemical changes of soybean oil variety JS-80 also showed increase in fatty acid due to Fusarium roseum, Trichoderma viride, Aspergillus ustus and Alternaria alternata . The chemical composition of oil of pooja variety showed increase in fatty acid composition due to Fusarium roseum, Trichoderma viride, Aspergillus ustus and Alternaria alternata.

No much variation is reported in saponification and unsaponification value. Fatty acid composition is essential parameter to study in biodeterioration. In JS-335 variety myristic acid is reduced due to Alternaria alternata and increased due to Fusarium roseum.

Whereas, stearic acid content increased due to Alternaria alternata and Fusarium roseum. Similarly, linoeic acid reduced due to Alternaria alternata and Aspergillus flavus. Changes in fatty acid composition of soybean JS-80 variety also showed decrease and increase ratio in the parameter of fatty acid composition due to associated fungi. similarly fatty acid composition also changed in pooja variety . whereas myristic acid, palmitic acid reduced due to Alternaria alternata, while palmitic acid, palmito oleic acid and stearic acid increased due to associated fungi. Alternaria alternata and Aspergillus flavus also responsible for reduction in linoeic acid and Aspergillus ustus increase arachidic acid.

REFRENCES 1.

A.O.C.S. 1976. Official methods of analysis XI Ed. Association of official Analytical Chemist, Washington ,D.C. cd1-25,cd-758,cd3-25 and ca 6a-40.

2.

Agarwl V. K. 1976 .Technique for the detection of seed borne fungi. Seed research.4:24-31.

3.

Agaewal S. C., R. K. Gupta (1989). Fungal pathogens detected on Soybean seeds grwn in different localities. Seed research 17(2):208-210.

4.

De, Tempe, J 1970. Tesing cereal seeds for Fusarium infection in the Netherlands. Prof. Int. Seed. Test. Ass. 33: 193-2006.

5.

ISTA 1966. Proc. International seed testing Association, 31: 1-52.

6.

Laxmi B. S., J. P. Gupta and M. S. Prasad (1998). Influence of fungicidal seed treatment on germination of mycoflora succession of Soybean seeds during storage. Annals of Plant protection-Sciences. 6(2): 136-141.

7.

Mukadam, D. S. 1997. The illustrated kingdom of fungi, Aksharganga Prakashan Aurangabad.(M.S.),India .

8.

Mukadam D. S., Ashok M. Chavan, Patil M. S. and Anjali R. Patil 2006. The illustrated fungi, Saraswati Printing Press, Aurangabad.(M.S.). India

9.

Neupane, S., Dhakal, K. H., & Darai, R. Variations in Seed Oil and Fatty Acid Content of Different Soybean genotypes and its Coorelation with Seed Oil.

10. Neergaard Paul 1973. Detection of Seed borne pathogens by culture tests. Seed Sci. And Tech.1:217-254.

www.tjprc.org

editor@tjprc.org

8

Kesare. U. T 11. Neergaard Paul 1977 .Seed Pathology, Vol. I: John Villy and sons N.Y. 12. Nelson P. E., Toussoun, T. A. and Marassas, W. F. O. 1983. Fusarim species an illustrated manual for identification.

Pennsylvania State 13. Ouf S. A., Abdel, N. F. Hady (1999).Influence of He-Ne laser irradiation of Soybean seed mycoflora, growth, nodulation and resistance to Fusarium solani. Folia-Microbiologica (Czech Republic).44(4):388-396.

14. Ramesh C. H and A. K. Marihal (2006).National seminar on frontiers in Plant pathology.28-30 sep.2006:42.

15. Sandikar 1990.Seed-borne safflower and sesame. Ph. D. thesis, Marathwada University, Aurangabad.

16. Sharma A.K. (1997).Seed mycycoflora of Soybean and its pathology in the north- western hills. Seed res. 15(1): 66-70.

17. Singh, D., & Sharma, N. (2017). Effect of chromium on seed germination and seedling growth of green garm (Phaseols Aureus L) and chickpea (Cicer Arietinum L). Int J App Nat Sci, 6, 37-46.

18. Singh, K., Frisvad, J. C., Thrane U and Mathur, S. B. 1991.An illustrated manual on identification of some seed borne Aspergilli, Fusaria, Penicillia and their mycotrins. Dan Govt. Inst. Beedpath for Devel Contries. pp. 133.

19. SOPA (2003). Soybean Processors Association, Indore.

20. Vijayakumar, H. P., & Vijayakumar, A. (2015). Standardization of accelerated ageing duration to evaluate seed storability of soybean cultivars. International Journal of Agricultural Science and Research (IJASR), 5(4), 93-98.

AUTHOR PROFILE

KESARE UMESH TUKARAM: Associate Professor, Department of Botany, Adarsh College Omerga, Osmanabad, Maharashtra, India. He Published 11 Papers in national and International Journals. Memberships are : Member in Board of Studies, Dr. BAMU, Aurangabad and Member in Mrathwada Botanical Association.

Impact Factor (JCC): 5.5837

NAAS Rating: 4.08