Sugarcane Biotechnology: Genetic Engineering of ... - Philsurin

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Dec 11, 2012 (4 years and 8 months ago)

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Sugarcane Biotechnology:
Genetic Engineering
of Sugarcane
Antonio C. Laurena, PhD
Research Professor, Institute of Plant Breeding
Crop Science Cluster, College of Agriculture
UP Los Banos
Outline of Topics
Introduction/Background
The Process of Genetic
Engineering in Plants
GM Sugarcane:
Philippine Perspectives
Genetic Engineering of Sugarcane
Summary
Biology + technology
= Biotechnology
Biology is the study of living things
(plants, animals and microorganisms)
Technology is a set/system of methods
and instruments
Biotechnology is the application of
scientific techniques to improve
living things for man’s needs and the
environment.
Traditional Conventional Modern
Gene(s) Trait
GregorMendel (1860s) James Watson &
Francis Crick (1960s)
idea of a gene identity of a gene
(DNA)
IPB Plant Biotechnology Program
1975
DNA
(gene)
mRNA
protein
TRAIT
gene
Plasmid
vector
Gene
construct
DNA
Transfer
What is a genetic engineering?
Transgenic
Plant
molecular
switches
ONOFF
gene-of-interestselectable marker
“Gene Gun” Technique
“Gene Gun” Technique
DNA coated
golden particles
Gene gun
Cell division
A plant cell with
the new gene
Transgenic plant
Plant cell
Cell’s DNA
GM Corn: YieldGard, Bt Corn
IR Corn
1.This particular corn is resistant to the
Asiatic corn borer.
2. The bacterial gene Bt CryIabwas
transferred to corn by genetic
engineering
Golden Rice
Capacity of rice plant to produce
pro-VitaminA (beta-carotene)
Involves the insertion of three
genesin the rice plant
Institute of Plant Breeding
Crop Science Cluster
UP Los Banos
Genetic engineering
of plants
Genetically modified papaya
Delayed ripening (DR) trait
Gene
Agrobacterium
Particle
bombardment
Gene transfer to
plant tissue
Selection
Regeneration
Greenhouse
selection/testing
Field testing
Variety
Vector
Marker
gene
Cloning sites
Gene
construct
Gold bullets
shooting
infection
The Process of Transgenic Papaya Technology
Research and Development
Lab
Greenhouse
Single/multi-location field testsVariety
NCBP
DA/BPI
NCBP –National Committee on Biosafety of the Philippines
DA/BPI –Department of Agriculture/Bureau of Plant Industry
For GM Crops, there are two DA/BPI-related agencies:
1. BPI Biotech Core Team
2. BPI Plant Quarantine Service
19972002200720102011
Transgenic Sugarcane
or GM Sugarcane
Anyone?
Indonesia and India
Countries with GM sugarcane
Ref. William Lee Burnquistpowerpoint(CTC, Brazil)
Embryogeniccalli
Transformation by
Gene Gun
Plant Tissue Culture
Greenhouse/Field Test
Australia, Brazil and USAOther traits
Brazil and MauritiusAbioticstress resistance
Australia, Brazil, USA, S Africa,
and Colombia
Disease resistance
Brazil and S AfricaPest resistance
Australia, Brazil, USA, S Africa,
and Mauritius
Herbicide resistance
CountriesTransgenic Trait
GM sugarcane development in different countries
Source: Maldonado A, M Melgarand P Lamport. 2008. Worldwide
advances in sugarcane transgenesis. Sugar Journal pp.11-16
(April 2008) www.SugarJournal.com
Rosanne Casupowerpoint
Prof. Robert Birch
GM sugarcane “Sugarbooster”
This produce isomaltulosewithout affecting
sucrose yield resulting in a two-fold increase
in total sugar yield in the harvested juice.
The sucrose isomerasegene from Pantoea
dispersawas inserted into sugarcane
Q117N3.2N3.2H
Sugarcane plants were 16 months old with 59 internodes
in the third vegetative generation.
GM sugarcane
Ref. Wu L and RG Birch.2007. Doubled sugar content in sugarcane plants
modified to produce a sucrose isomer. Plant Biotech J 5:109-117
Why Isomaltulose?
10x the value of sucrose
Low glycemicindex
Anti-acariogenic
Potential raw material
for other chemicals
Drought Tolerant Genes
stomatalsensitivity
to ABA
protein regulator
SNAC1
osmotolerancedue to
prolineaccumulation
Enzyme –pyrroline5
carboxylatesynthase
P5CS
Enzyme
trehalosesynthase
otsAand
otsB
regulator of transpiration
stress signal protein
ERECTA
drought, cold and salt
tolerance
stress-induced
regulators
DREBs/CBF
photosynthesis, starch
and tolerance to photo-
oxidative damage
Ref. Cattivelliet al. 2008 Drought tolerance improvement in crop plants: an integrated
view from breeding to genomics. Field Crop Res105:1-14
Development of a Drought-Tolerant Sugarcane
PTPN XI: Government Enterprise for Estate Crops, Indonesia
Genes for drought resistance
1.pyrroline-5-carboxylate synthase
2.Na
+/H+
antiporter(yeast and Arabidopsis)
The actual cost of R&D is US$ 1,548,000
and at present the GM sugarcane is under
confined field trials in East Java.
???
Gene flow, non-target species, food
safety, feed safety and data evaluation
2006 to
present
US$ 1,567,753.00
Subtotal
US$ 12,416.00
Confined field trials (East Java)
2004-2005
US$ 7,337.00
Contained facility testing2003
US$ 1,548,000.00Research and Development
1999-2002
Total Actual Cost
ActivitiesYear
Actual Cost of Developing the Drought-Tolerant
Sugarcane in Indonesia from 1999 to 2005
Regulatory Cost US$ 99,870.00
Ref. FrikkieBotha’spowerpointMay 2005 “Redefining R&D
p
ro
g
rammesfor a su
g
arcane industr
y
”.
Ref. FrikkieBotha’spowerpointMay 2005 “Redefining R&D
programmesfor a sugarcane industry”.
Ref. FrikkieBotha’spowerpointMay 2005 “Redefining R&D
p
ro
g
rammesfor a su
g
arcane industr
y
”.
To develop a GM sugarcane:
1.Genes and Expression Cassettes
2.Transformation System
a. Gene gun
b. Tissue culture protocol
3.Molecular Analytical Methods
a. PCR
b. Northernsand Southerns
4.Contained facility (BL2 greenhouse)
5.Open field test site
Can the Philippines develop its
own GM sugarcane?