Lithuanian university of agriculture

LITHUANIAN UNIVERSITY OF AGRICULTURE
Department of Crop Science and Animal Husbandry APPLIED BIOTECHNOLOGY

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, 10 000 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 • have good knowledge about application of plant tissue culture techniques in • obtain certain practical training in methods used in plant biotechnology. 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 15 hours
Test 15 hours
Examination 30 hours
LITERATURE
1. Plants, genes, and crop biotechnology (2nd edition). Marten J. Chrispeels, David E. Sadava, Martin J. Chrispeels. Jones & Bartlett Pub., 2002, p.562. 2. In vitro plant breeding. Acram Taji, Prakash P. Kumar, Prakash Lakshmanan. 3. In vitro embryogenesis in plants. Edited by Trevor A. Thorpe. Kluwer Academic 4. Morphogenesis in plant tissue cultures. Edited by Woong – Young Soh and Sant S. Bhojwani. Kluwer academic publishers, 1999, p. 520. 5. Cloning Agricultural Plants via in vitro techniques. Edited by B. V. Conger.CRC Study programme prepared by Acting assoc.prof. dr. Natalija Burbulis Department of Crop Science and Animal Husbandry, Faculty of Agronomy, Lithuanian University of Agriculture Studentu 11, Akademija, Kaunas distr., LT-53361 Lithuania Phone: +370 37 752314 Fax: +370 37 752271 e-mail: natalija.burbulis@lzuu.lt CONTENT OF STUDY PROGRAMME
LESSONS (15 hours)
1. Introduction (5%)
• Applications of plant tissue culture.
2. Basics medium (10%)
• Macro-, micro nutrient, source of carbohydrate, vitamins. • Growth • 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 • 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%)
• 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. • Factors affecting androgenesis: genetic potential, physiological status of the donor plant, stage of pollen development. • External factors; culture medium, culture density, colchicines treatment,
6. Cell culture and selection of desirable traits (20%)
• 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%)
• 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
microspores. Regeneration of haploid plants and their transplanting. Evaluation of ploidy level. Colchicine treatment. Results evaluation and interpretation.

Source: http://intern.asu.lt/socrates/Moduliai/Burbulis%20-%20Applied%20Biotechnology%20EN.pdf