Pengantar

Bioteknologi Hutan
Kuliah Pengantar Bioteknologi Kehutanan
Pokok Bahasan 1 Pendahuluan - Pengertian dan Sejarah
- Prospek dan Tantangan
Pokok Bahasan 2 Genetika Molekuler Tanaman
- Materi Genetik
- Regulasi dan Ekspresi Gen
- Perubahan Materi Genetik
Pokok Bahasan 3 Rekayasan Gen
- Kloning-transformasi
- Metode-transformasi
Pokok Bahasan 4 Sistem Silvikultur Jaringan
- Mikropropagasi dan Kultur Sel
- Kultur Haploid dan Variasi Somaklonal
Pokok Bahasan 5 Penerapan Bioteknologi
- Identifikasi dan Klasifikasi Molekuler
(Isoenzym, RFLP, RAPD, sidik jari)
- Perlindungan terhadap Hama & Penyakit
- Organisme Pemacu Pertumbuhan Tanaman
- Organisme Transgenik
Bioteknologi :
Teknologi yang menggunakan sistem hayati
(proses-proses biologi) untuk mendapatkan
barang dan jasa yang berguna bagi kesejahteraan
manusia
Pengertian dan Sejarah
Sejarah
Minuman alkohol anggur
(pengawetan daging)

Khamir roti
Keju
Yogurt
Susu masam
kecap
1857 Pasteur menemukan
bahwa fermentasi merupakan
proses yang dilakukan oleh
organisme hidup
1920 fermentasi aseton,
etanol, butanol, gliserin

PD II Penicillium natatum
Antibiotik
Vitamin, steroid, enzim
Amerika Serikat mengijinkan digunakannya sel tubuh manusia untuk kegiatan penelitian mengenai terapi gen. 1990
Metode polymerase chain reaction (PCR) pertama kali dipublikasikan 1988
Badan paten Amerika Serikat mengumkan bahwa tikus yang direkayasa secara genetis peka terhadap kanker 1988
Ti plasmid yang telah direkayasa digunakan untuk transformasi pada tanaman 1983
Pertama kalinya vaksin hewan diproduksi menggunakan teknologi DNA rekombinan di Eropa 1982
Pertama kalinya kit diagnostik yang dibuat dari antibodi monoklonal digunakan secara luas di Amerika Serikat 1981
Pertama kalinya alat otomatis untuk sintesis DNA dijual secara komersial 1981
Badan pengadilan Amerika Serikat menetapkan peraturan dalam kasus Diamond VS Chakrabarty yang melakukan
manipulasi mikroorganisme yang dipatenkan.
1980
Perusahaan Genentech memproduksi insulin manusia pada sel bakteri Escherichia coli 1978
Pengembangan teknik untuk mengetahui sekuen DNA 1976
Pertama kali dikeluarkan pedoman untuk penelitian dalam bidang DNA rekombinan 1976
Kohler dan Milstein mendeskripsikan produksi antibodi monoklonal 1975
Boyer dan Cohen mengembangkan teknologi DNA rekombinan 1973
Khorana dan rekan-rekannya dapat mensintesis keseluruhan gen yang mengkode tRNA 1972
Isolasi pertama kali enzim endonuklease restriksi 1970
Keseluruhan kode genetik dapat diketahui 1961-1966
Watson dan Crick menemukan struktur DNA 1953
Avery, MacLeod dan McCarty membuktikan bahwa DNA merupakan materi genetik. 1944
Antibiotik penisilin diproduksi besar-besaran dalam skala industri 1943
Karl Ereky pertama kali menyatakan istilah bioteknologi 1917
Peristiwa Tahun
Lactid Acid Bacteri
Yeast
Fermented
Alcohol
Ether
Amino acid
Protein
Sugar
Fermentasi
Bioteknologi tradisional :
Seleksi bahan, mikrobia yaang digunakan dan modifikasi
lingkungan untuk memperoleh produk optimal.
Misal : pembuatan tempe, tape, roti, pengomposan sampah
Bioteknologi modern :
Memanfaatkan ketrampilan manusia dalam melakukan
manipulasi makhluk hidup agar dapat digunakan untuk
menghasilkan suatu barang yang diinginkan. Misal rekayasa
genetik
Pengertian & Sejarah
Bioteknologi merupakan penerapan prinsip ilmiah dan
rekayasa pengolahan bahan oleh agen biologi untuk
menyediakan barang dan jasa
Rekayasa genetik :
Teknik untuk menghasilkan molekul DNA yang berisi gen
baru yang diinginkan atau kombinasi gen-gen baru atau
dapat dikatakan sebagai manipulasi organisme
Pengertian & Sejarah
Hasil-hasil rekayasa genetika tanaman yang
digambarkan dalam National Enquier, c. 1981.
Prospek & Tantangan
Pertimbangan Pemanfaatan Hasil Bioteknologi

1) Tinjauan ekologik
a) GMO mengalahkan spesies alami, menggantikan spesias
alami, mengurangi keanekargaman hayati.
b) GMO berubah prilaku dibandingkan dengan aslinya.
c) Gen yang disisipkan dapat pindah ke kerabat dekat
spesies GMO, termasuk karakteristik gen yang dimaksud.
Penambahan gen karakter baru, kemampuan baru,
kemampuan hayati baru.
Kesetabilan gen asing atau dapat berubah dan
diekspresikan berbeda oleh organisme yang tertular gen
asing mutasi
Penarikan gen asing jika masuk dalam organisme non
target
Terganggunya rantai makanan'
Terganggunya organisme non target

2) Tinjauan kesehatan
a) Gangguan kesehatan karena produk gen asing
(dalam waktu panjang)
b) Bahaya atau tidaknya gen-gen penyerta

1) Tinjauan sosial dan budaya
a) Etika, moral
b) Agama

2) Perdagangan global

The History of
Biotechnology
http://www.i-s-b.org/wissen/timeline/englisch/
1859
Charles Darwin publishes his
book (On the Origin of
Species by Means of
Natural Selection).
According to this book
evolution is lifes motor.
The interplay of mutation
and selection endows living
beings with optimized traits
in order to survive. These
principles are also valid for
the so-called 'chemical
evolution' of biomolecules
and are being used in
laboratories for in vitro
optimizing of wanted
qualities in molecules
1865
Gregor Mendel finds that
independent 'factors' are
responsible for the heredity
of traits from one
generation to the next
according to a set of
(Mendelian) priciples.
1869
Friedrich Miescher discovers
an acidic substance in the
nucleus of cells which he
names 'nuclein'. By
elemental analysis he finds
14 % nitrogen, 3 %
phosphorus, and 2 % sulfur
(from proteins). Since the
substance cannot be
cleaved by the proteolytic
enzyme pepsin Miescher
concludes that this
substance is not a protein.
1879
Walther Flemming observes
the separation of
chromosomes during
mitosis but fails to fully
understand its meaning. He
counts 24 pairs of
chromosomes, a number
which will be corrected to
23 later in 1956 by the
Indonesian scientist Joe-
Han Tijo.
1900
William Bateson introduces
'genetics' as a scientific
discipline
Hugo de Vries, Erich Tschermak
von Seysenegg and Carl Correns
independently rediscover the
Mendelian principles
Hugo de Vries
defines the
meaning of
'mutations'
1902
Walter Sutton observes in
grasshoppers' cells that
chromosomes carry the
Mendelian 'factors of
heredity', i.e. the genetic
information.
1909
Wilhelm Johansen coins the
terms 'gene', 'genotype',
and 'phenotype' referring to
Mendel's 'factors'.
Archibald Garrod confirms
the hereditary nature of four
metabolic diseases. He
publishes "Inborn Errors of
Metabolism"
1910
Thomas Hunt Morgan's
studies in the biology of
Drosophila melanogaster
confirm that certain traits
are inherited sex-
specifically. He also proves
that some phenotypes
result from several genes
located on different
chromosomes.
1927
Herman Muller finds that
energetic radiation causes
defects in the
chromosomes, i.e.
mutations.
1928
Fred Griffith demonstrates
that material from killed
bacteria of the pathogenic
strain Streptococcus
pneumoniae S ('smooth') is
taken up from living ones of
the non-pathogenic strain
Streptococcus pneumoniae
R ('rough') which are
subsequently 'transformed'
into the pathogenic S-strain.
Pneumococcus pneumoniae
1944
Oswald Theodore Avery, Colin McLeod, and Maclyn
McCarty find by careful analysis of Griffith' experiments
that desoxynucleic acid (DNA) is the carrier of the
'transforming principle'.
1945
Erwin Schrdinger proposes
in his famous book 'What is
life?' that genes must be
'aperiodic crystals'
consisting of a succession
of a small number of
isomeric elements whose
precise sequence
constitutes the heredity
code. Although these ideas
do nothing to identify the
responsible molecular
structures, they attract
many newcomers to the
field.
1950
Erwin Chargaff observes the
1:1 ratio of the nucleic
bases adenine/thymine and
guanine/cytosine. This will
be a decisive hint for
Watson & Crick to the
structure of the DNA
molecule.
1952
Alfred Day Hershey's and Martha
Chase's studies in bacteriophages
prove that only the nucleic acids carry
the genetic information; proteins are
definitively excluded from playing this
role.
Joshua Lederberg finds plasmids in
bacteria, small rings of
extrachromosomal DNA which can be
replicated autonomously.
1953
James D. Watson and Francis H. C. Crick elucidate the
structure of the DNA molecule. It consists of two strands
which are bound by hydrogen bonds bewtween opposite
basepairs of adenine-thymine and cytosine-guanine. Their
model is based on the results of many colleages, namely
Rosalind Franklin who discovered the helical structure and
the outward position of the phosphate-sugar backbone.
1955
Frederick Sanger finds a
method to sequence
proteins. With this he
determines the sequence of
the amino acids of insulin.
1956..
Arthur Kornberg reports the first in vitro
synthesis of a DNA molecule.
George Emile Palade
locates protein
biosynthesis to the
ribosomes, the
'factories of the cell'.
1956
Joe Han Tijo and Albert Levan correct
the number of the chromosome pairs
in human cells to 23. Since the days
of Walther Flemming 24 was believed
to be the true number.
Chromosomen einer
menschlichen Zelle
protein chains are built at the ribosomes
1959
T. Akiba, T. Koyama, Y.
Isshiki, S. Kimua, T.
Fukushima, T. Watanabe
and T. Fukusawa describe
the transfer of multiple
resistance to antibiotics by
bacterial plasmids.
Plasmids turn out to be
efficient vectors for
trafficking genetic
information between
bacteria of different strains.
bacterial plasmid (arrow)
1960
Francois Jacob and Jaques Monod recognize the function
of the messenger RNA (mRNA). They develop the operon
model of gene regulation in procaryotes.
1961
Sydney Brenner and Francis Crick
claim that every amino acid
corresponds to a triplett of
nucleotides, called 'codons'.
Marshall Nirenberg and
Heinrich Mathaei can
prove that the codon UUU
in mRNA codes for the
amino acid phenyl alanine
Sydney Brenner
Marshall Nirenberg
1962
John Gurdon claims to have
reproduced frogs from the
epithelium cells of Xenopus
laevis. J. B. S. Haldane
coins 'cloning' when
describing these debated
experiments.
Xenopus laevis
South African
clawed frogs
John Gurdon
1966
Mainly due to the work of Har
Gobind Khorana the
'Genetic Code' is
completely known.
1968
Werner Arber discovers
enzymes, so-called
nucleases, which digest
DNA double strands from
the ends.
1969
Jonathan Beckwith is
the first to isolate a
complete gene, in this
case a gene from the
bacterial sugar
metabolism.
1970
Hamilton Smith discovers an enzyme (HindII)
which specifically cleaves DNA double
strands
Har Gobind Khorana synthesizes
a complete gene (that of an
alanine t-RNA) in vitro.
David Baltimore and Howard Temin discover the
viral enzyme reverse transcriptase. It translates
RNA sequences to DNA sequences and thus
tumbles the age old 'Central Dogma of Genetics'
which defined the direction of the flow of the
genetic information to start from DNA via RNA to
the proteins.
1971
Paul Berg, Peter Loban, and
Dale Kaiser find enzymes
which attach short single
strands to the blunt ends of
double stranded DNA. The
single strands consist of
only one sort of nucleotides.
Complementary single
strands (sticky ends) allow
the coupling of ds DNA
fragments; the remaining
gaps can be closed by the
use of ligases.
1972
Janet Mertz and Ron Davis paste DNA fragments of
different origin resulting from specific enzymatic
restriction. They close the gaps using ligases and thus
produce recombinant DNA.
1973
Stanley Cohen and Herbert Boyer paste
enzymatically restricted DNA fragments into
plasmids which have been cleaved by
restriction enzymes. The resulting
recombinant plasmids can serve as vectors
for the transfer of foreign DNA into bacteria.
These experiments are generally regarded as
the beginning of the era of genetic
engineering and modern biotechnology.
1975
The first conference on safety issues of the new technology
is held at Asilomar in California.
Georges Khler and Cesar Milstein develop the 'Hybridoma
Technology' for the production of monoclonal antibodies. An
antibody producing plasma cell (derived from a B
lymphocyte) is fused with a tumor cell resulting in an
immortal hybridoma cell which (as well as its daughters)
continues to produce one kind of antibodies.
1976
Herbert Boyer and Robert
Swanson found Genentech
the first biotech company.
1977
Walter Gilbert and Frederick Sanger independently
develop two different methods for DNA sequencing.
1978
Walter Gilbert discovers that
genes of eucaryotic
organisms are composed of
coding and non-coding
parts. The non-coding
sequences, named introns,
are cut from the mRNA
before translation leaving
the coding exons as the
source of information for
protein synthesis.
1979
Thomas Cech and Sydney Altman discover the
autocatalytic activity of some RNA molecules to
cleave themselves at well defined positions. They
coin 'ribozyme' for this type of RNA.
1980
Jozef Schell and Marc van Montagu transfer
foreign DNA into plant cells by employing T-
plasmids of Agrobacterium tumefaciens as
vectors.
1981
The first patent for a genetically modified organism is granted
to Ananda Chakrabarty of General Electric. It covers a P.
aeruginosa strain which is equipped with genes of certain
enzymes in order to metabolize crude oil.
1982
The U.S. Food and Drug
Agency (FDA) approves
recombinant insulin for
marketing.
1984
Alec Jeffreys develops 'genetic fingerprinting' allowing the
comparison of minute variations, the so-called restriction
length fragment polymorphisms (RLFPs), in the genomes
of two organsims.
1985
Karry Mullis develops the
polymerase chain reaction
(PCR). This powerful tool
allows to copy and
accumulate extremely low
amounts of DNA from
various sources until a level
sufficient for analysis is
reached.
1986
Neal First develops protocols for
clonig cattle from bovine
embryos by separating cells at
early embryonic stages. These
experiments resemble John
Gurdons earlier attempts at the
cloning of frogs. The first cloning
of an adult animal will be
reported in 1997 by Ian Wilmut
who succeeds in cloning a
sheep, Dolly.
1988
The first patent for a
transgenic
mammal is granted
to Harvard's Philip
Leder and Timothy
Stewart. It refers to
a mouse which is
susceptible to
tumors and serves
as model organism
for studies in
cancer.
1990
French Anderson successfully tries the
first somatic gene therapy of a human.
Patient
Ashanti DeSilva is cured
from an deficiency in the
enzyme ADA, which
causes severe
immunodeficiency.
German
'Genetic
Engineering
Act' passes
parliament
Official kick-off
of the Human
Genome Project
(HGP)
1993
The German Genetic
Engineering Act of 1990 is
revised, i.e. many
restrictions of the 1990
version are eased.
The Biotechnology
Industry Organization
BIO is created by
merging two smaller
trade associations
1994
Genetically modified tomatoes are on sale in the U.S. Cans
containing puree from transgenic tomatoes are on the
shelves of british supermarkets.
1995
The Institute for Genomic
Research (TIGR) publishes the
first complete sequence of the
genome of a free-living
organism, the bacterium
Haemophilus influenzae
Germany launches the BioRegio
contest. 17 regions compete in
presenting the best blueprint for
the development of commercial
biotechnology to an
international jury. The three
winning regions will share the
price of DM 150m.
1996
The complete sequencing of the baker's yeast'
genome is accomplished by close collaboration
of laboratories in Europe, Japan, and the U.S.
The Saccharomyces cervisiae genome ist the
first eucaryotic genome completely sequenced.
Patrick Browne of
Stanford University
presents the first
'gene chip'
containing 6116
different gene
specific sequences
of the baker's yeast
genome.
In the U.S.
transgenic plants
grow on more than
1.9 million hectares
1997
Ian Wilmut (right) visits Dolly (center) the first mammal
cloned from an adult cell.
1998
Two research teams report embryonic
stem cells differentiate to specialized
tissue cells. The picture shows an early
developmental stage, the blastocyst.
The first genome of a
multicellular organism
(Caenorhabditis elegans) is
sequenced.
Craig Mello und Andrew Fire find that small double-stranded
RNA molecules can selectively block gene expression in
Caenorhabditis elegans, a phenomenon later referred to as
RNA interference (RNAi)
1999
Pluripotent stem cells from tissues
can be reprogrammed to other cell
types.
The sequence of
chromosome 22
is published.
The U.S. biotech
company Celera
starts its own
human genome
sequencing
project
2000
There are 1300 biotech companies on each
side of the Atlantic. About 400 are listed at
stock exchanges.
The genome of
Drosophila
melanogaster is
sequenced. The fruit
fly was introduced by
Thomas H. Morgan
into genetics
research.
The Human Genome Organization and
Celera present the first working draft of the
human genome. The picture shows Celera's
CEO J. Craig Venter (left) and Francis
Collins, the speaker of the HGP.
2001
Less genes than expected
The international Human Genome Project
(HGP) and biotech company Celera
Genomics publish their versions of the
sequence of the human genome. The
Celera team estimates the number of
genes to range from 26,000 to 39,000, HGP
scientist' estimate is between 30,000 and
40,000.
Rice genome
Syngenta (Basel, Switzerland), in
collaboration with Myriad Genetics
(Salt Lake City, UT), announced the
completed sequencing of the
genome of Oryza sativa japonica
2001
RNA interference
Thomas Tuschl is
the first to
demonstrate that
RNA interference
(RNAi) is also
working with
mammalian cells.
2002
Genomes of the malaria agent and its carrier sequenced
Researchers of the Institute for Genomic Research (TIGR),
the Sanger Institute (UK), and Stanford University publish the
genomic sequence of Plasmodium falciparum and
Plasmodium yoelii. The protozoa P. falciparum causes
malaria.
The genome of its carrier Anopheles gambiae is also
sequenced and published.
2003
Genome of SARS virus
sequenced
Within three weeks after its
discovery scientists of the
Michael Smith Genome
Sciences Centre in British
Columbia (Vancouver, Canada)
succeeded in sequencing the
genome of the virus causing
severe acute respiratory
syndrome (SARS). The RNA
virus remotely resembles
corona virus species and
contains roughly 30,000 genes.
2003
Germ cells from stem cells
Karin Hbner and Hans Schler
of the University of
Pennnsylvania succeeded in
generating oocytes from murine
stem cells which were able to
grow into further embryonic
stages. The experiment
demonstrates the potential of
pluripotent stem cells to acquire
totipotency via a germ line cell
stage
2004
99.999% accuracy
Three years after the publication of
the first draft a far more precise
version of the sequence of the
human genome is published.
The number of
identified genes
decreases again
compared with previous
estimations.
19,599 have been
confirmed to encode
proteins, 2188 are
suspected to encode for
proteins. 1183 genes
have been generated
through duplication of
genes, 33 genes
appear to have been
degenerated into
disfunctional
pseudogenes.
2004
Fatherless mouse
Tomohiro Kono of the Tokyo
University of Agriculture presented
the first mouse which was obtained
from oocytes only. His team
combined the genomes of oocytes
from adult and
newborn mice and
eventually one of
460 experiments
resulted in a living
animal, being 14
month old at the
time of publication.
Critical for success
was the
suppression of a
gene which
normally controls
the imprinting of
chromosomal DNA
from mother and
father.
2004
Mice from cancer cells
Rudolph Jaenisch of MIT and Lynda Chin of the Dana
Farber Cancer Institute generated cloned mice from cancer
cells by stuffing the nuclei of melanoma cells into de-
nucleated mouse oocytes. The resulting blastocysts yielded
stem cells which were suitable for transplantation into
normal blastocysts which eventually developed into living
animals. Some genetic markers of the melanoma cells were
found again in the animal, but the epigentic patterns had
gone lost - showing that epigenetic modifcations are both
reversible and late stage in the transformation of normal
cells into melanoma cells.