Isolation of oleosin and determination of oil body size in Brassica juncea
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Authors
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Ibandalin Mawlong
ICAR-Directorate of Rapeseed Mustard Research, Bharatpur-321 303, Rajasthan, India
Author
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MS Sujith Kumar
ICAR-Directorate of Rapeseed Mustard Research, Bharatpur-321303, Rajasthan, India
Author
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PD Meena
ICAR-Directorate of Rapeseed Mustard Research, Bharatpur-321303, Rajasthan, India
Author
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Devesh Pathak
Department of Plant Protection, Faculty of Agricultural Sciences, AMU, Aligarh-202002, Uttar Pradesh (UP), India
Author
Keywords:
Brassica juncea, oil bodies, oleosinAbstract
Oil bodies are structural entities that store triacylglycerols (TAG) in oilseeds. Each oil body contains a matrix of
triacylglycerol (TAG) surrounded by a layer of phospholipids and its associated oil body proteins. Description of oil
body and its associated proteins has not been well documented in Brassica juncea unlike Arabidopsis which is a related
species. In this study extraction of oil bodies (OB) was done by flotation method following which the presence and
integrity of oil bodies was confirmed using compound light microscope. The size of OB ranged from 0.58 (Maya) to 1.05
μm (HB9902). The major phospholipid as profiled by TLC was phospotidylcholine. Purification of structural proteins
was done by altering the time of urea (8M) treatment. The proteins isolated from the oil bodies showed multiple bands
in SDS-PAGE that included oleosin protein. Downstream purification of oil body proteins was standardized to remove
further unspecific bands by additional urea treatment with time of incubation 30 min, 1h and overnight at room
temperature.Out of the three, overnight incubation showed less appearance of unspecific bands. A comparative study
among selected genotypes was also done showing oil content to be inversely related to oil body size and positively
correlated to oleosin content. Hence by manipulating the oleosin proteins, it may be possible to increase the oil content.
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