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APPLICATION OF MOLECULAR BIOLOGY IN AGRICULTURE GROUP II

MPAKA PETER MUHWEZI OBED KALEMBE SOLOME

DEFINITIONS
Molecular biology: The branch

Agriculture: The science, art, or occupation concerned with cultivating land, raising crops, and feeding, breeding, and raising livestock.

of biology that deals with the


formation, structure, and function of macromolecules

essential to life, such as


nucleic acids and proteins, and especially with their role in cell replication and the transmission of genetic information.

INTRODUCTION
Application of molecular biology in agriculture dates way back to at

least 10000years.
Humans have constantly searched for improved varieties of their crop plants and animals. Gene cloning in animals has been on going, knowingly or unknowingly The rate of evolution of new species through natural selection was slow, but now molecular methods like gene cloning have been discovered that can help fasten the process

Plant agriculture
Objectives

To have varieties with better nutritional qualities


Higher yields Crops that aid cultivation and harvesting Crops which are pest resistant To understand plant evolution

GENE CLONING
This is the insertion of a fragment

of DNA, carrying a gene, into a


cloning vector and subsequent propagation of the

recombinant DNA molecule in


the host organism. Ways

Gene addition
Gene subtraction

GENE ADDITION
Alters the characteristics of a plant by provision of one or more new genes Applications : Plants that make their own insecticides Herbicide resistant crops

Modified fruit ripening Drought tolerance Improved sweetness Improved flower colour etc

Plants that make their own insecticide


Plants can be attacked by bacteria, viruses, fungi and animals: large scale destruction is by insects. Insecticides are normally used to reduce crop damage but they are non specific, toxic and insects can evade them

The -endotoxin of Bacillus thuringiensis


Bacteria are prey to insects and have a defense mechanism against insects. e.g. B. thuringiensis produces -endotoxin that is highly poisonous than insecticides Molecular biologists have taken advantage of this toxin to help in self crop protection.

By introduction of the gene that codes for the -endotoxin into a


plant.

Cloning of -endotoxin gene in maize

FINDINGS
The factors assessed were damage to foliage and length of tunnels produced by the larvae boring into the plant. Transformed plants showed better results than the non modified. Eg length of tunnels reduced from 40.7cm to 6.3cm

Other applications of gene addition

Gene subtraction
The unwanted gene is inactivated Strategies are several but the best is use of antisense RNA This has been used to delay tomato ripening The gene that codes for polygalacturonase enzyme,responsible for ripening is inactivated.

GRAPH
Expression in a normal plant
Comparison: expression in normal and modified plant

Other Applications Of Gene Subtraction

Genetic modification in animals


Why ? Scientific and medical research Treatment of human disease Production of modified food producing animals

Genetic modification of animals ctd


How?

Four steps involved in gene transfer


I. II. Identification of a gene with a significant and desired effects Introducing the DNA fragment coding for the desired gene

III.

Regulating the expression of the introduced gene

IV. Confirming transmission of the transferred gene to the next generation of animals

Limitations
Escape genes Poor produce due to target pests resistance Possible harmful effect of the marker genes used with plant cloning vectors Terminator technology Disruption of the farmland biodiversity ethical issues arise:-moral, religious etc

It is expensive
Un expected yet undetected

mutations can alter the


future in unpleasant ways

Future of molecular biology in agriculture


Transgenic plants are being used in pharmaceutical industries to produce many therapeutic proteins, including antibodies, blood products, cytokines, growth factors, hormones, recombinant enzymes and human and veterinary vaccines (Twyman et al, 2005). Several PDP products for the treatment of human diseases are approaching commercialization, including recombinant gastric lipase for the treatment of cystic fibrosis, and antibodies for the prevention of dental caries and the treatment of non-Hodgkin's lymphoma (Ma et al, 2003). There are also several veterinary vaccines in the pipeline.

Future of molecular biology in agriculture


Product Various singlechain Fv antibody fragments CaroRx E.coli heatlabile toxin Gastric lipase Hepatitis B Virus surface antigen Class Antibody Indication Non-Hodgkin's lymphoma Dental caries Diarrhoea Company/Organiza Crop tion Large Scale Biology Viral vectors in Corp tobacco Planet Biotechnology Transgenic tobacco Inc. Prodigene Inc. Meristem Therapeutics Transgenic maize Transgenic maize Status Phase I

Antibody Vaccine

Phase II Phase I Phase II Phase I Phase I

Therapeutic enzyme Cystic fibrosis, pancreatitis Vaccine Hepatitis B

Arntzen group Transgenic potato (Richter et al, 2000) Thomas Jefferson Transgenic lettuce University/Polish Academy of Sciences

Human intrinsic factor

Dietary

Vitamin B12 deficiency

Cobento Biotech AS Transgenic Arabidopsis

Phase II

Lactoferrin

Dietary

Gastrointestinal infections
Norwalk virus infection Rabies

Meristem Therapeutics
Arntzen group (Tacket et al, 2000)

Transgenic maize
Transgenic potato

Phase I
Phase I Phase I

Norwalk virus capsid Vaccine protein Rabies glycoprotein Vaccine

Yusibov et al (2002) Viral vectors in

References
http://www.ncbi.nlm.nih.gov/pmc/articles/P MC26347/ http://findarticles.com/p/articles/mi_hb1366/ is_1_15/ai_n29297863/ T.A brown () gene cloning and DNA analysis 6th edn