katedra speciÁlnÍ produkce rostlinnÉ cultivationof

KATEDRA SPECIÁLNÍ PRODUKCE ROSTLINNÉ září 2014

prof. Ing. Vladislav Čurn, Ph.D.University of South BohemiaFaculty of Agriculture, Biotechnological CentreNa Sádkách 1780, 370 05 České Budějovice, CZ

CULTIVATION OF GM PLANTS/CROPS

GMO technology, GMO cultivation

• GMO = organism whose genetic material has been altered bygenetic modification

• GMO represents deliberate change of the genetic material in away that is not possible to achieve by natural recombination

• GMO is the incorporation of foreign genetic material intorecipient organism or deletion of part of the heritable materialusing recombinant DNA techniques and transgenesis

recombinant DNA technology - gene splicing, molecular cloning, plasmid engineering

GMO/GMP - Genetically modified organisms/plants


• recombinant DNA technology– joining together DNA molecules from two different

organisms– construction recombinant DNA molecule (chimeric gene)– insertion new DNA molecule into a host organism to

produce new genetic combinations• any DNA sequence may be created and introduced into any

organisms

Recombinant DNA technology


• construction recombinant DNA molecule



• selection of bacterial cells with inserted recombinant DNA


Plant genomics and GMO/GMP

• Methods used for introduction of foreign genes intoplants:

• Indirect introduction of cloned genes into the plants usingAgrobacterium

• Direct transformation




tobacco plant transformed with the insect luciferase gene

THE PROCESS OF GENETIC MODIFICATION


• Plant tumor – A. tumefaciens, Gram-negative soil bacteriacauses crown-gall disease in plants



• Insertion of the cloned genes into plants using bacteria Agrobacterium tumefaciens



• Indirect introduction of cloned genes into the plants using Agrobacterium



• leaf disk method for Agrobacterium mediated transformation of higher plants(Horsch et al. 1985):

– precultivation of plants in vitro– precultivation of bacteria with Ti plasmid with inserted gene of interest– preparation of leaf discs or nodal segments– cocultivation of plant tissue and Agrobacterium– transfer disks to MS selection medium– cultivation of calli or regenerated shoots, rooting plants and transplantation

into soil



• Indirect introduction of cloned genes into the plants using Agrobacterium



• Direct transformation – electroporation or „gene gun“



• Direct transformation – biolistic method



• Direct introduction of cloned genes using biolistic method



Most frequently used transgenes:

• Transgenes for tolerance to herbicides• Transgenes for resistance to insect pests• Transgenes male-sterility• Transgenes for resistance to viruses• Transgenes for prolonging ripening• Transgenes changing fatty acid synthesis• Transgenes changing storage proteins• Transgenes for industrially important enzymes• Transgenes for antibodies• Genes for biosynthesis of β-carotene in rice ("yellow rice") ...

GMO - transgenes useful for crop improving


GMO - global status of biotech crops


GMO – why to grow GMO crops?

non-GMO x GMO maize

corn borerinfestation ofmaize


GMO – why to grow GMO crops?

• Comparison of yield of GM andconventional maize hybrids

• Estimation of the economic lossesdue to non-cultivation of GM maize

• Health risks – toxins in non-GM(non-resistant) maize


• principles of coexistence• co-existence of multiple cultivation/growing systems:

– conventional without GMO– conventional with GMO– organic (ecological) agriculture

GMO growing in different agricultural systems


• principles of coexistence– liability to notify GMO cultivation– isolation distances– evidence / GMO cultivation records

GMO growing in different agricultural systems

minimum isolation distance

GMO maize non-GMO maize

minimum isolation distance in organic agriculture

GMO maize non-GMO maize


• Case study in rape

• Occurrence of GM rape plants on plots where they fieldexperiments with GM rape were carried out

• Molecular analysis of all cruciferous plants with the aim todetermine whether there is hybridization of rape and otherrelated species

Monitoring of the occurrence of GM plants


• Case study in rape



• Case study in potatoTransgene detection in potato using PCR and RT-PCRtransgene = GN lectin, endogene - UDP-glucose pyrophorylase (UGPase)


GN lectin

UGPase


• DNA microarray detection

Detection of GMO in food and agricultural products


Possible risks associated with the cultivation of GM crops

• Comparison of yield of GM andconventional maize hybrids

• Estimation of the economic lossesdue to non-cultivation of GM maize

• Health risks – toxins in non-GM(non-resistant) maize


• contamination of water and soil residues of herbicides• but the standard herbicides contaminate surface/underground

water and soil twenty-times more• resistance of pests to transgene• not while maintaining the refuges and growing non-GMO• negative impact of GMO to pollinators, parasitoids, predators• not reported• negative impact of GMO to environment• reported in the case of large concentration of GMO and

lowering agrobiodiversity



• Cisgenic plants - cisgenic plants are similar to traditionally bred plants and cisgenesis is equivalent to traditional breeding

• Into cisgenic plant are transfered complete plant genes, genesfrom the same plant species or from closely related species, which can hybridize with recipient plant.

New trends in plant biotechnology and plant breeding


• cisgenesis



• GMOs in pharmacy• Technologies of „molecular pharming“

• applications in pharmacy have a long history• 1979 - first insulin and somatotropin production in E. coli• 1982 – commercial production of insulin - the first clinical use of recombinant

protein


Děkuji za pozornost

Vladislav ČurnJihočeská univerzita v Českých BudějovicíchZemědělská fakulta, Katedra speciální produkce rostlinnéBiotechnologické centrum

Na Sádkách 1780, 370 05 České Budějovicewww.zf.jcu.czhttp://biocentrum.zf.jcu.cz

GENOMICS AND GMO CULTIVATION OF GM PLANTS/CROPS

katedra speciÁlnÍ produkce rostlinnÉ cultivationof

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