Presentation on theme: "Nanoparticle Mediated Genetic Transformation in Plants"— Presentation transcript:
1 Nanoparticle Mediated Genetic Transformation in Plants PBIO 4500/5500 BIOTECHNOLOGY AND GENETIC ENGINEERINGPresented by :ISHA SHRIVASTAVA
2 How small is a nano? A nanometer is one billionth of a meter The thickness of an individual page is 100,000 nmsA fine human hair is 10,000 nmsOur finger nails grow at the rate of 1nm per secondSource:
3 Why do we need to combine nanotechnology and genetic engineering? Nano FarmingSyngenta, BASF, Bayer, Monsanto using nano-scale materials to produce nanopesticides “gutbuster”Recent breakthrough includes replacing the genetic material of one bacteria from anotherNano Food“Smart Foods” which respond to allergies, dietary needs and food preferencesAlter the properties and traits of food including its nutrition, flavor, texture, heat tolerance & shelf life
4 Vehicles for Nuclear Transformation in Plants Agrobacterium mediated:Most extensively used, wide host range (mostly dicotyledonous)Incompatibility between tissues of plant speciesMicroparticle Bombardment:Capable of delivering DNA into nucleus, mitochondriaCell Damage, high copy number of transgene, expensive equipmentElectroporation:Generate transgenic plants by protoplast transformationCell damage by electric pulses of wrong length, ion imbalance and cell death
5 Comparative study of different delivery systems Source: Rai,M., Deshmukh, S. , Gade, A., Elsalam, K.A Current Nanoscience .8 :
6 Nanoparticle Mediated Genetic Transformation Nanoparticles combined with chemical compounds deliver genes into target cellsDecreasing the particle size from micro to nano scale, hindrance due to cell wall can be removedCell Damage can be minimizedThe particle can reach the chloroplast and mitochondria easilyDifferent NPs used are calcium phosphate, Carbon materials, silica, gold magnetite, strontium phosphate.Enable controlled release conditionsFigure: Synthesis of mesoporous silicaSource:
7 Experiment with MSNs – Genetic Transformation Purpose: To investigate interaction of MSN with plant cellsSynthesize series of MSNs with different surface functional groups/capsInvestigation of MSN in protoplasts (plant cells with cell wall removed)Protoplasts incubated with Type-I MSN didn’t take up nanoparticles, Type-II MSN ( Type-I functionalized with triethylene glycol) entered the protoplastsFigure : Type-I and Type-II MSNsConclusion:MSN system can serve as a new and versatile tool for plant endocytosis and cell biology studies
8 Purpose: To prove MSNs can function as DNA delivery agents Plasmid containing a green fluorescent protein (GFP) gene under the control of constitutive promoter is usedOptimal coating ratio for DNA/Type-II MSNs was 1/10Type-II MSN bound DNA not digested by restriction enzymeTransient GFP expression observed 36 hr after protoplasts were incubated with DNA coated Type-II MSNConclusion:Type-II MSN system can serve as efficient delivery system for protoplasts and make DNA accessible to transcription machinery
9 Representation of Nanoparticle Mediated Gene Transfer Source: Rai,M., Deshmukh, S. , Gade, A., Elsalam, K.A Current Nanoscience .8 :
10 Figure: Gene gun system for bombarding micro/nano particles Purpose: To introduce MSN into plants with gene gun systemAttempts to bombard Type-I and Type –II MSNs didn’t lead to successful transformationUse of Type-III MSNs, where mesopores are capped by surface functionalized gold nanoparticlesIn comparison to traditional system, allows to load biogenic moeities-including chemicals that are membrane impermeable or incompatible with cell growth media into poresFigure: Gene gun system for bombarding micro/nano particlesSource:
11 Purpose : To deliver different biogenic species simultaneously Release the encapsulated chemicals in a controlled fashionGenerate transgenic tobacco containing inducible promoter controlled GFP geneExpression of GFP observed only when chemical β-oestradiol ispresentTransgenic plantlets bombarded with Type-IV MSNs ( filled with β-oestradiol) , and pores capped with gold nanopartclesRelease of β-oestradiol is triggered by DTT ( Dithiotheritol)
12 Mesoporous silica Nanoparticles for plant cell internalization
13 ConclusionNanobiotechnology could take the genetic engineering of agriculture to the next level down – atomic engineeringFurther developments such as pore enlargement and multifunctionalization of these MSNs may offer new possibilites in target specific delivery of proteins, nucleotides and chemicals.Opposition is mounting from civil society, unions and world leading scientists who point to ecological, health and socio-economic risks associated with nanogenetics.
14 References:Husaini,A.M., Abdin, M.Z. , Parray, G.A. , Sanghera, G. S. , Murtaza, I. , Alam, T. , Srivastava, D. K. , Farooqi, H. , and Khan, H. N Vehicles and ways for efficient nuclear transformation in plants. Landes Bioscience . 1(5) :Nair, R. , Varghese, S. H. , Nair, B. G. , Maekawa , T. , Yoshida , Y. , and Kumar , D. S Nanoparticulate material delivery to plants. Plant Science. 179 :Rai,M., Deshmukh, S. , Gade, A., Elsalam, K.A Current Nanoscience .8 :Torney, F. , Trewyn, B. G. , Lin, V.S. , and Wang, K Mesoporous silica nanoparticles deliver DNA and chemicals into plants. Nature Nanotechnology . 2 :
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