APPLIED BIOTECHNOLOGY

Transcription

1 LITHUANIAN UNIVERSITY OF AGRICULTURE FACULTY OF AGRONOMY Department of Crop Science and Animal Husbandry STUDY SUBJECT DESCRIPTION APPLIED BIOTECHNOLOGY Study level: BSc Number of ECTS credit points: 4,5 Teaching method: lectures, supported by PowerPoint presentation and slides Prerequisites: basic knowledge of plant biology and physiology. Teaching aids: scripts referring to the actual topics are distributed during lectures. Examination method: oral examination, upon appointment. Registration for course: two weeks before the beginning of the course. Registration for examination: with lecturer, personally or by phone. INTRODUCTION The invention of agriculture, years ago, heralded the dawn of civilization. The constant improvements in technology and crop productivity through selection and breeding of plants have contributed to the growth of human civilization. Bearing in mind that there are natural limits to increased productivity by environmental manipulations, one way to improve and maintain a sustainable level of crop productivity is through the exploitation of biotechnology. Indeed, one major aim of biotechnology is to increase yield, while maintaining stable human ecosystem. Plant biotechnology and molecular breeding have already proved their impact in enhancing the productivity of some of the major agricultural crops. They will continue to contribute to the production of plants with novel traits that are otherwise difficult or impossible to develop by conventional breeding. In an age in which the dynamic field of plant biotechnology advances at an accelerating rate, there is a growing need to explain how that science is applied. After completion of the course the students will: have a basic knowledge of biotechnology; be able to perform biotechnological experiments on plants and evaluate the results; have good knowledge about application of plant tissue culture techniques in basic research and plant breeding; obtain certain practical training in methods used in plant biotechnology.

2 Syllabus Theory (15 hours): Introduction. Basics medium. Micropropagation. Applications of micropropagation. Haploid plant production in vitro. Cell culture and selection of desirable traits. In vitro mutagenesis. The origin, nature, and significance of variation in tissue culture. Laboratory practices (45hours) Laboratory equipment and material (10%) Micropropagation technology (40%) Double haploid technology (50%) Independent work (60 hours) Individual home work Test Examination 15 hours 15 hours 30 hours LITERATURE 1. Plants, genes, and crop biotechnology (2 nd edition). Marten J. Chrispeels, David E. Sadava, Martin J. Chrispeels. Jones & Bartlett Pub., 2002, p In vitro plant breeding. Acram Taji, Prakash P. Kumar, Prakash Lakshmanan. Food Products Press, 2002, p In vitro embryogenesis in plants. Edited by Trevor A. Thorpe. Kluwer Academic Publishers, 1995, p Morphogenesis in plant tissue cultures. Edited by Woong Young Soh and Sant S. Bhojwani. Kluwer academic publishers, 1999, p Cloning Agricultural Plants via in vitro techniques. Edited by B. V. Conger.CRC Press, Inc, 1981, p Study programme prepared by Acting assoc.prof. dr. Natalija Burbulis Signature Department of Crop Science and Animal Husbandry, Faculty of Agronomy, Lithuanian University of Agriculture Studentu 11, Akademija, Kaunas distr., LT Lithuania Phone: Fax:

3 ANNEX 1. Introduction (5%) Types of in vitro culture. Applications of plant tissue culture. CONTENT OF STUDY PROGRAMME LESSONS (15 hours) 2. Basics medium (10%) Macro-, micro nutrient, source of carbohydrate, vitamins. Growth regulators: Auxin and auxin-like compounds. Cytokinins and cytokinin-like compounds. Gibberellins and gibberellin-like compounds. Hormonal regulation of morphogenesis in tissue culture. 3. Micropropagation (20%) Stage 0: the preparative stage. Growing stock plants. Changing the physiological status of the stock plant, the source of explant. Stage 1: initiation stage. Factors determine the success rate in stage one. Hypersensitivity reaction. Stage 2: multiplication. Callus-forming cultures. Physiological and morphological aspects of somatic embryogenesis. Factors affecting direct organogenesis. Stage 3: rooting stage. Stage 4: transplanting. 4. Applications of micropropagation (10%) Pathogen elimination. Methods of virus elimination. Virus indexing. Tissue culture for maintenance of plant genetic resources (germplasm storage). Storage using slow growth techniques. Storage using cryopreservation. 5. Haploid plant production in vitro (15%) Origin of androgenic haploids. Molecular and cytological aspects of androgenesis induction.

4 Factors affecting androgenesis: genetic potential, physiological status of the donor plant, stage of pollen development. External factors; culture medium, culture density, colchicines treatment, temperature shock. 6. Cell culture and selection of desirable traits (20%) General selection strategies. Positive selection. Negative selection. Other selection strategies. Selection for herbicide tolerance. Selection for disease resistant lines. Selection for amino acids accumulators. Selection for variants for resistance to abiotic stresses. 7. In vitro mutagenesis (10%) Types of mutagens. Physical mutagens. Chemical mutagens. Precautions for handling the chemical mutagens. Determining the type and suitable concentration of mutagens. The choice of plant tissues for in vitro mutagenesis. 8. The origin, nature, and significance of variation in tissue culture (10%) The basis of somaclonal variation. Genetic variation arising from source plant. Genetic variation arising during culture. Causes of somaclonal variations. Use of somaclonal variation in breeding. LABORATORY PRACTICES (45hours) Laboratory equipment and material (10%) Work safety in growth room and sterile laminar flow hood. Care of donor plants and regenerants. Basic medium preparation. Micropropagation technology (40%) Induction, regeneration and rooting medium. Explant selection, sterilization and isolation. Direct and indirect organogenesis in vitro. Regenerants subcultivation and their transplanting. Results evaluation and interpretation. Double haploid technology (50%) Cytology of microspores: pre-culture events, post-culture events. Medium for embryo induction and plant regeneration. Bud selection and sterilization, extraction of

5 microspores. Regeneration of haploid plants and their transplanting. Evaluation of ploidy level. Colchicine treatment. Results evaluation and interpretation.