Department of Life Sciences

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Browsing Department of Life Sciences by Author "Dur-E -Aeman"

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    DEVELOPMENT OF GENETIC TRANSFORMATION SYSTEM IN OKRA (ABELMOSCHUS ESCULENTUS (L). MOENCH)
    (University of Management and Technology, 2017) Dur-E -Aeman
    This study appraises the development of genetic transformation system in okra (Abelmoschusesculentus L. Moench). Okra plays an important role in human diet and is an excellent source of carbohydrates, proteins, fats and minerals. Okra is affiliated with Malvaceae ancestors and is highly susceptible towards insects and viruses. Reported begomoviruses of okra reported throughout the world are Bhendi yellow vein mosaic virus, Okra yellow vein mosaic virus, Okra yellow crinkle virus,Okra yellow mosaic Mexico virus and Cotton leaf curl Alabad virus. Conditions were optimized by establishing a simple regeneration system of okra. 193 embryos were isolated and 69% total regeneration frequency was calculated. Best regeneration frequency was calculated when embryos were grown on MS medium containing 1g GA/L and B5 vitamin. 342 embryos were isolated for development of transgenic plants by using embryo as explant. The transformation efficiency was 2.92 %. Presence of transgene was confirmed by Green fluorescent protein (GFP) expression and PCR analysis. Expression of GFP were effortlessly and rapidly distinguished when examined under a fluorescence microscope at 40X- 100X resolving power. Microscopic analysis was done by using fluorescence stereo microscope to check GFP expression. A tissue sample of okra plant leaf was placed on glass slide. Expression of Green fluorescent protein genes was quickly distinguished at 40-100X resolution power. PCR analysis confirmed the presence of RNAi constructs in transgenic plants of okra when amplified PCR product of size 417 bp was visualized on 1 % agarose gel stained with ethidium bromide.