BIOTEKNOLOGI DALAM BIDANG PERLINDUNGAN TANAMAN · 2013-04-17 · Bioteknologi bidang perlindungan...
Transcript of BIOTEKNOLOGI DALAM BIDANG PERLINDUNGAN TANAMAN · 2013-04-17 · Bioteknologi bidang perlindungan...
BIOTEKNOLOGI DALAM BIDANG PERLINDUNGAN
TANAMAN
Dr. Noor Istifadah, Ir., MSc. Prof. Dr. Hj. Hersanti, Ir., MP.
Dr. Danar Dono, Ir., MSi. Endah Yulia, SP., MSc., PhD.
Fitri Widiantini, SP., MBtS., PhD. Rika Meliansyah, SP., MSi.
Kompetensi Dasar
Setelah mengikuti perkuliahan ini, mahasiswa mampu menjelaskan tentang:
Perlunya perlindungan tanaman
Penggunaan bioteknologi dalam bidang perlindungan tanaman
Metode untuk meningkatkan kinerja biokontrol
Feeding the hungry world
What we are hoping
The reality
Yield losses
Plant pests and diseases: 20-40% (FAO, 2013).
Weeds
Natural disaster Drought
Excessive rain
Hurricane
Earthquake
Volcano eruption
Fires
etc
Pemanfaatan bioteknologi dalam perlindungan tanaman
Bioteknologi bidang perlindungan
tanaman
Peningkatan kinerja agen biokontrol
Deteksi dan identifikasi patogen
Pembuatan tanaman resisten OPT
Peningkatan Kinerja Agen Biokontrol
Mutasi gen
Kultur jaringan
Modifikasi gen dengan rekayasa genetik
http://evolution.berkeley.edu/evolibrary/article/0_0_0/mutations_04
Peningkatan Kinerja Agen Biokontrol-Mutasi
Substitution Insertion Deletion
Apakah mutasi itu?
Perubahan secara permanen pada susunan DNA yang menyusun suatu gen
Peningkatan Kinerja Agen Biokontrol-Mutasi
Peningkatan Kinerja Agen Biokontrol-Mutasi
www.nature.com
Somatic mutation
Germ-line mutation
Peningkatan Kinerja ABK-Mutasi
Bagaimana mutasi bisa terjadi?
Inherited
Acquired
Penyebab mutasi:
Random mistakes
Environmental
Radiation
Chemical
Transposon insertion
Type of Radiation
Non-ionizing radiation
Ionizing radiation
http://www.epa.gov/rpdweb00/understand/
Mutation-Radiation
Radiation
Mutation-Chemical
Bahan kimia penyebab terjadinya mutasi-mutagen, umumnya carcinogenic
Golongan utama penyebab mutasi: DNA Reactive chemicals-directly reactive
with DNA or when involved in metabolism process in cells is able to produce mutagens Alkylating agents-chemicals that add an alkyl
group (CnH2n+1) to another molecule. Alkylation of a base may change the normal base pairing
Base analogs-mengganti susunan basa pada susunan DNA
Intercalating agent-able to insert itself into the stacking of DNA bases EtBr
Mutation-Transposon element
First found by Barbara McClintock 1940an-colour pattern on corn
Small piece of DNA
Capable to move and to integrate into different location within genome
http://www.broadinstitute.org/education/glossary/transposable-elements
Seleksi Mutant-positive selection
Seleksi Mutant-negative selection
RODAC-replicate organism direct agar contact
Peningkatan Kinerja Agen Biokontrol-Mutasi Cheng et al., 2003
Pseudozyma flocculosa
BCA pada powdery mildew
WT mode of action-antibiosis (A)
Insertion mutagenesis
Some loss their activity
Some have higher activity compared to WT
MeOH extraction P. flocculosa Phomopsis sp.
Pseudomonas fluorescens F113 Mutant with Enhanced Competitive Colonization Ability and Improved Biocontrol Activity
against Fungal Root Pathogens
Barahona et al., 2011
Directed mutagenesis, insertion of target gene fragment
Controlling pest insect
Three strategies:
Manipulating the genes of entomopathogenic bacteria, viruses or fungi to produce more efficient bio-pesticides
Cloning the gene encoding insecticidal protein and incorporate it into plants, resulting transgenic plant producing its own bio-pesticides
Genetic modification of insect pest to target it for biocontrol
Sterile Insect Technique
British Colombia Sterile Insect Release Program
Launch in 1992
Sterilised by radiation
Successful to keep the moth population low
http://pmtp.wsu.edu/newslettersV4I7.html
Sterile Insect Technique by TE
Mycoherbicides-Gressell et al. 2007
Peningkatan Kinerja ABK-Kultur Jaringan
Tissue culture can be used to increase the BCA agent through the following:
The use of fusion protoplast to enhance the biological control activities
Maintaining obligate biological control agent
Production of secondary metabolite
Fusion Protoplast
Type of genetic modification in plant as a resulted from fusion of two distinct species of plants with the characteristics of both
http://www.eplantscience.com/
Fusion Protoplast
Type of genetic modification in plant as a resulted from fusion of two distinct species of plants with the characteristics of both
http://www.eplantscience.com/
Gene transfer Trichoderma and Aspegillus through protoplast fusion to increase production of citric acid and cellulase
El-bondkly A.M. (2006)
To convert ground rice straw into citric acid and cellulose
T. reesei NRRL 18670, Trichoderma harzianum NRRL 13879, and T. viride strain as cellulases producers
A. niger NRRL 599 as citric acid producer
Protoplasts were prepared through enzymatic hydrolysis of mycelium suspension
Protoplasts prepared from each of the two strains were mixed
Gene transfer Trichoderma and Aspegillus through protoplast fusion to increase production of citric acid and cellulase
Fusants produced citric acid three fold more than A. niger
Transfer of tuber soft rot and early blight resistances from Solanum brevidens into cultivated potato
Tek et al., 2004
Non cultivated S. brevidens shows significant resistance to tuber soft rot (Erwinia carotovora) and early blight (Alternaria solani)
A somatic hybrid between potato (S. tuberosum) and S. brevidens was developed by protoplast fusion
Clone C75-5+297 has consistently out-yielded common varieties under disease stress
Using both molecular and cytogenetic approaches demonstrated that a single copy of chromosome 8 from S. brevidens replaced a potato chromosome 8 in C75-+297.
Maintaining obligate biological control agent
vasicular arbuscular mycorrhiza pada akar inangnya atau dg teknik transformasi Agrobacterium rhizogenes membuat akar rambut (hairy roots)
Compiled by Noor Istifadah-Danar Dono
Production of secondary metabolite
Medicinal plants are most exclusive life-saving drug sources Taxol-taxus tree-an anticancer
Morphine and codein-opium poppy-analgesic
Hairy root culture for secondary metabolite production
Based on the inoculation with Agrobacterium rhizogenes
Production of secondary metabolite
No System Mechanism of action Compound Plant source
1 Cholinergic system
Inhibition of acetylecholinestrase (AChE)
Essential oils Azadirachtina indica, Mentha spp., Lavendula spp.
Cholinergic acetylcholine nicotinic receptor agonist/antagonist
Nicotine Nicotiana spp., Delphinium spp., Haloxylon salicornicum, Stemona japonicum
2 GABA system GABA-gated chloride channel Thymol, Silphinenes
Thymus vulgaris
3 Mitochondrial system
Sodium and potassium ion exchange disruption
Pyrethrin Crysanthemum cinerariaefolium
Inhibitor of cellular respiration (mitochondrial complex I electron transport inhibitor or METI)
Rotenone Lonchocarpus spp.
Affect calcium channels Ryanodine Ryania spp.
Affect nerve cell membrane action Sabadilla Schoenocaulon officinale
4 Octopaminergic system
Octopaminergic receptors, Essential oils Cedrus spp., Pinus spp., Citronella spp., Eucalyptus spp.
Block octopamine receptors by working through tyramine receptors cascade
Thymol Thymus vulgaris
Miscellaneous Hormonal balance disruption Azadirachtin Azadiractina indica
Rattan, 2010
Peningkatan Kinerja ABK-Rekayasa Genetik
Identifying and characterizing the gene of interest Entomopathogenic nematodes
Fungal biocontrol for insect pest
Improving efficacy by genetic engineering (gene modification) Fitness, persistence, host range, enhance pathogenicity,
improve environmental tolerance, and alter host range
Characterization of the gene expression
Gene insertion, either on the original organism or into other organism such as plant to develop transgenic plant
Peningkatan Kinerja ABK-Rekayasa Genetik
Ryder et al., 2012 – genetic modification of chitinase-encode gene in Trichodema hamatum, impaired saprotrophic competitiveness against Rhizoctonia solani and promote the plant growth.
Segal D., and Glazer I, 2000 – genetic engineering entomopathogenic nematode (EPN) Heterorhabditis bacteriophora towards environmental extremes , selection for heat tolerance and resistance to nematicides
Reference
El-bondkly A.M. (2006) Gene transfer between different Trichoderma species and Aspergillus niger through intergeneric protoplast fusion to convert ground rice straw to citric acid and cellulases. Applied Biochemistry and Biotechnology 135:117-32.
Tek A.L., Stevenson W.R., Helgeson J.P., Jiang J. (2004) Transfer of tuber soft rot and early blight resistances from Solanum brevidens into cultivated potato. Theoretical and Applied Genetics 109:249-54.
Barahona E., Navazo A., Martinez-Granero F., Zea-Bonilla T., Perez-Jimenez R.M., Martin M., Rivila R. (2011) Pseudomonas fluorescens F113 mutant with enhanced competitive colonization ability and improved biocontrol activity against fungal root pathogens. Appl. Environ. Microbiol. 77:5412-5419.
Gressel J., Meir S., Herschkovitz Y., Al-Ahmad H., Greenspoon I., Babalola O., Amsellem Z. (2007) Approaches to and successes in developing transgenically enhanced mycoherbicides, in: M. Vurro and J. Gressel (Eds.), Novel Biotechnologies for Biocontrol Agent Enhancement and Management, Spinger, Dordrecht.
Grigliatti T.A., Meister G., Pfeifer T.A. (2007) TAC-TICS: transposon-based biological pest management systems, in: M. Vurro and J. Gressel (Eds.), Novel Biotechnologies for Biocontrol Agent Enhancement and Management, Spinger, Dordrecht.
Cheng Y., McNally D.J., Labbe C., Voyer N., Belzile F., Belanger R.R. (2003) Insertional mutagenesis of a fungal biocontrol agent let to discovery of a rare cellobiose lipid with antifungal activity. Applied and Environmental Microbiology 69:2595-2602.