KATEDRA SPECIÁLNÍ PRODUKCE ROSTLINNÉ CULTIVATIONOF ...
Transcript of 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
GMO technology, GMO cultivation
• 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
GMO technology, GMO cultivation
• selection of bacterial cells with inserted recombinant DNA
Recombinant DNA technology
Plant genomics and GMO/GMP
• Methods used for introduction of foreign genes intoplants:
• Indirect introduction of cloned genes into the plants usingAgrobacterium
• Direct transformation
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
GMO/GMP - Genetically modified organisms/plants
tobacco plant transformed with the insect luciferase gene
THE PROCESS OF GENETIC MODIFICATION
Plant genomics and GMO/GMP
• Plant tumor – A. tumefaciens, Gram-negative soil bacteriacauses crown-gall disease in plants
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
• Insertion of the cloned genes into plants using bacteria Agrobacterium tumefaciens
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
• Indirect introduction of cloned genes into the plants using Agrobacterium
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
• 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
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
• Indirect introduction of cloned genes into the plants using Agrobacterium
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
• Direct transformation – electroporation or „gene gun“
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
• Direct transformation – biolistic method
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
• Direct introduction of cloned genes using biolistic method
GMO/GMP - Genetically modified organisms/plants
Plant genomics and GMO/GMP
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
Plant genomics and GMO/GMP
GMO – why to grow GMO crops?
non-GMO x GMO maize
corn borerinfestation ofmaize
Plant genomics and GMO/GMP
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
Plant genomics and GMO/GMP
• 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
Plant genomics and GMO/GMP
• 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
Plant genomics and GMO/GMP
• 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
Plant genomics and GMO/GMP
• Case study in potatoTransgene detection in potato using PCR and RT-PCRtransgene = GN lectin, endogene - UDP-glucose pyrophorylase (UGPase)
Monitoring of the occurrence of GM plants
GN lectin
UGPase
Plant genomics and GMO/GMP
• DNA microarray detection
Detection of GMO in food and agricultural products
Plant genomics and GMO/GMP
• DNA microarray detection
Detection of GMO in food and agricultural products
Plant genomics and GMO/GMP
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
Plant genomics and GMO/GMP
• 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
Possible risks associated with the cultivation of GM crops
Plant genomics and GMO/GMP
• 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
Plant genomics and GMO/GMP
• 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
New trends in plant biotechnology and plant breeding