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BIOTEKNOLOGI

TANAMAN
PEMULIAAN TANAMAN
KONVENSIONAL :
PEMULIAAN TANAMAN YANG MELIBATKAN
AKTIVITAS PERSILANGAN DAN SELEKSI PADA
TINGKAT INDIVIDU TANAMAN.

PEMULIAAN TANAMAN
BIOTEKNOLOGI (REKAYASA GENETIKA):
PEMULIAAN PADA TATARAN SEL/MOLEKULER
Plant transformation technologies

Genetic transformation creating transgenic


organisms
Agrobacterium
Gene gun
Gene transfer to protoplasts
Transgenic plant technologies also require
tissue culture
Modern plant biotech = recombinant DNA Start with bacterial DNA and your gene
technology (from bacteria, goats, fungus, maize)
involves gene transfer in a much more
precise manner than traditional breeding
but still involves manipulation of
biochemistry, physiology and development
DNA recombination involves taking DNA
from one organism and moving it to another
The resulting transgenic plants can be called
genetically modified organisms (GMOs)
How do you move DNA around?
STEP 1: Get your gene
Locate and remove DNA of interest using
restriction enzymes that recognize specific
target DNA sequences
Place into a plasmid for amplification in a Grow lots
bacterium (E. coli)
of bacteria,
make lots
of DNA!
Transfecting target cells requires gene gun (biolistics) or Agrobacteria

STEP 2: Prepare your receiving


tissue
Involves tissue culture
techniques
Often use sterile young leaf
segments as targets
STEP 3: Get your DNA into the
target plant
Gene gun and technique
Method 1: gene gun
Use naked DNA (linear) DNA
Coat DNA onto beads
(tungsten or gold)
Use air pressure to fire into
tissue
Invented at Agracetus in
Wisconsin
Agrobacteria allow controlled DNA insertion
Method 2: Agrobacteria
Use engineered instead of wild-type A.
tumefasciens Ti plasmid Original T-DNA coding
Still possesses virulence genes (allow for PGR, opine genes
removed
transfer of T-DNA to target cell) but
lacks opine and PGR synthesis genes
Wounded tissue (cut) attracts
Agrobacteria that can infiltrate through
wounds into apoplasm
Transfers T-DNA to genome
Can do in high throughput in immersion
culture

Both methods integrate their DNA


randomly into the genome
Not really desirable: would like to
target transgene to appropriate
segment of genome for expression at Agrobacteria were
correct developmental stage
first isolated from
Agrobacterium transformation tends to
give lower copy numbers better for crown galls
controlling silencing in long term
Selectable markers aid greatly in identifying positive
transformants
STEP 4: Regenerate transgenic plants
Transformation is not 100% efficient not
every targeted plant cell will be transgenic!
Just as in bacteria, use a selectable marker to
find positives
Antibiotic, herbicide resistance common
Transformed explants taken through a
dedifferentiating callus stage
Then manipulate auxin and cytokinin ratios to
regenerate shoots and roots
Thus, tissue culture is an integral part of
making transgenic plants
Antisense technology
Used to produce the Flavr-Savr tomato in 1994.
Enzyme polygalacturonase breaks down
structural polysaccharide pectin in wall of a plant.
This is part of the natural decay process in a plant
Monsanto identified the gene than encodes the
enzyme and made another gene that blocked the
production of the enzyme.
Antisense molecules
Pertanian GM Food

Bioteknologi dapat dimanfaatkan dalam:

Bt Corn
Peningkatan kualitas biji-bijian
Peningkatan kadar protein
Pembentukan tanaman resisten hama, penyakit,
dan herbisida
Pembentukan tanaman toleran kekeringan, tanah
masam, suhu ektsrim
Pembentukan tanaman yang lebih bernilai nutrisi
tinggi, seperti vit C, E dan -karoten

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Bt Cotton

Bt cotton Testing : bollworms resistance


PERAKITAN TEBU PRG TOLERAN KEKERINGAN
DI LABORATORIUM BIOTEKNOLOGI, PT.
PERKEBUNAN NUSANTARA XI (PERSERO)
Dilepas sebagai varietas bina melalui
Kepmentan no. : 4571/Kpts/SR.120/8/2013
Definisi dan istilah
Produk Rekayasa Genetik (PRG) :
adalah organisme hidup , bagian-
bagiannya dan/atau hasil olahannya
yang mempunyai susunan genetik baru
dari hasil penerapan bioteknologi
modern.
Tanaman PRG : tanaman yang
dihasilkan dari penerapan teknik
rekayasa genetik.
Tebu PRG toleran kekeringan menggunakan gen betA,
mengkode choline dehydrogenase
Sintesis glicine betaine (GB)
dari choline pada bacteria

Sintesis choline and GB pada


tanaman. Tdk semua tanaman
mensintesis GB.

Bacteria Choline dehy- Betaine aldehyde


drogenase (CDH) dehydrogenase (BADH)

Choline mono-
Plants oxygenase (CMO) Compatible solute
Tebu Toleran Kekeringan NXI-4T
1. Kajian Keamanan Lingkungan

1. Molecular testing (PCR,


Genetic Stability Southern Blot)
2. Biochemistry testing
(using HPLC)
Genetic expression 3. Morphology character
4. Physiological character
test (NaCl test)
Plant performance

Pollen
Study of gene flow and Seed

Soil Parameter
Soil
Weedyness/invasive character Bacteria
Diversity of Soil
Non target Microbe
organism
2. Kajian Keamanan Pangan

Genetic Stability
Comparative study Wild
Substance Equivalence Type and Reference

Animal testing using mice


Acute Toxicity
Homology / Similarity (software
program )
Alergenicity Alergene Reaction (animal
testing using rat)

Gastric enzyme
Digestive Tract Intestinal enzyme
Protein analysis (SDS PAGE)
TERIMA KASIH
TERIMA KASIH

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