Mutation breeding ppt

Mutagenesis is the process by which the genetic information
of an organism is changed in a stable manner.

The term ‘mutation breeding’ has become popular as it
draws attention to deliberate efforts of breeders and
the specific techniques they have used in creating and
harnessing desired variation in developing elite breeding
lines and cultivated varieties.

1. Mutation Breeding By- SHRIKANT YANKANCHI

2. Introduction Micro- and Macro-mutations Procedures for micro mutations breeding/polygenic Handling of segregating population Screening/selection Breeding fo r A Biotic and Biotic stress Methods for Validation of mutants Achievements of mutation breeding Advantages and limitations of breeding Contents...... 2

3. Introduction The term mutation breeding (‘ Mutationszüchtung ’) was first coined by Freisleben and Lein (1944) He referred mutation breeding as the deliberate induction and development of mutant lines for crop improvement Milestones in mutation breeding 300BC- The ancient Chinese book “ Lulan ” provides the first documentation of mutant selection in plant breeding: maturity and other trait in cereals in China (Huang and Liang, 1980) 1590 - The first verifiable (spontaneous) plant mutant described, ‘ incisa ’ mutant of greater celandine 3

4. 1667 -The first known description of a graft-chimera; Bizarria -orange, Florence, Italy 1901-1904 - de Vries suggests and promotes radiation to induce mutations in plants and animals 1907-Cramer publishes extensive examples of spontaneous mutants in crop plants 1927 - First proof of induced mutations in plants; radium ray treatment of Datura stramonium ( Gager and Blakeslee) 1927 - Muller working with Drosphila provides proof of mutation induction by X-rays Muller champions induced mutation for animal and plant breeding and opens a new era in genetics and breeding 1928 Stadler publishes the first results of mutation induction in crop plants, barley, maize, wheat and oat, but is sceptical about the use of induced mutation for crop improvement 4

5. 1936 The first induced mutant variety is released, tobacco var. ‘ Chlorina ’ using X-rays in Indonesia (then the Dutch East Indes ) 1942 First report of induced disease resistance in a crop plant; X-ray induced mildew resistance in barley ( Freisleben and Lein , 1942) 1944/46 First reports of chemical induced mutation ( Auerbach and Robson, 1944) 1949 First plant mutation experiments using 60Co gamma ray installations 1954 The first release of a mutant variety in a vegetatively propagated crop: tulip var. Faraday with an improved flower colour and pattern (see van Harten and Broertjes , 1989) 2000-2009 Development of high-throughput genotyping and phenotyping using automated, robotic and computerised systems 2000 onwards Development of TILLING populations 5

6. Mutations called micro- and macro-mutations depending on the magnitude of phenotypic effect produced by them Macromutation produces a large phenotypic effect easily recognizable on individual plant basis Oligogenic in nature Can be easily selected in M2 generation Micro- and Macro-mutations 6

7. Micromutations Produces Small phenotypic effect Cannot be recognizable on individual plant basis Detected only in group of plants and need treatment of statistical data Polygenic nature and selection delays till M3

8. Procedures for micro mutations breeding/polygenic

9. M1 M2 M3 M4 M5 M6-M8 M9 i . Mutagen -treated seeds space planted ii. Seeds from individual plants harvested separately i . Individual plant progenies are grown ii. Fertile, vigorous, normal looking plants harvested separately i . Individual plant progenies from selected plants grown ii. Superior plants selected from superior progenies showing segregation iii. Homogeneous mutant progenies may be harvested in bulk i . Individual plant progenies from selected plants grown ii. Superior homogeneous lines harvested in bulk i . Preliminary yield trial with a suitable check ii. Superior lines selected i. Multi-location yield trial ii. Outstanding line released as new variety i. Seed multiplication for distribution 9

10. 1.Selection of the variety for Mutagen treatment It should be the best variety available in crop Seed s hold be pure Oladosu et al , 2016 10

11. 2. Part of the plant to be treated Seeds Pollen grains Vegetative propagules Corns bulbs complete plants

12. Mutagen Treatments reduces germination growth rate vigour & fertility Mutagens generally induce a high frequency of chromosomal changes and meiotic and mitotic irregularities 3. Dose of mutagen 12

13. Optimum mutagen dose is one, which produces maximum frequency of mutations and causes the minimum killing Close to LD 50 dose is optimum Varies with crops eg :- 46 krad for Vicia faba , 120-140 krad for Brassica napus varies with mutagens eg : EMS – 0.3-1.5 %, for 2-6 hours 13

14. Optimum dose rate of physical and chemical mutagens for seed treatment of cereals 14

15. Handling of segregating population M1 generation Seeds treated with chemical mutagens should be washed thoroughly and be planted as soon as possible Large M1 generation is raised from treated seeds (Wider spacing) E.g :- 25,000 plants are to be grown to obtain a useful mutation in M1 generation Mutagens with high mutation frequency - M1 generation size can be reduced R. Roychowdhury and J. Tah ., 2013

16. Continued...... The M1 plants should not be allowed to cross pollinate M1 population should be planted 75-100 m apart from the parental or other genotypes of the same crop species Mechanical isolation M1 generation Dominant mutations are selected each plant selfed and harvested separately for M2

17. M2 generation Two methods of sowing M2 generation can be followed M1 plant to row where all seeds produced from a single plant are grown in row M1 spike or branch to row , Oligogenic mutants with distinct features are identified and selected

18. Screening/selection Mainly three types screening/selection techniques in M2 and subsequent generation i) Visual ii) Mechanical/Physical iii) Other methods

19. i ) Visual screening -most effective and efficient method for identifying mutant phenotypes -Visual selection often is the prime basis for selecting for disease resistance , earliness, plant height, colour changes, ion-shattering, adaptation to soil, climate, growing period etc.

20. ii) Mechanical/Physical -Very efficient for seed size, shape, weight, density, etc., using appropriate sieving machinery Iii) Other methods chemical, biochemical, physiological etc. E.g.- Low alkaloid content mutants can be selected using colorimetric tests -chromatographic or electrophoresis techniques may be used to select isolate protein variants

21. Methods for Validation of mutants Genome-wide chips Difference screening Microarray PCR screening TILLING and ECO-TILLING 21

22. Mutation breeding for Biotic and Abiotic stress

23. Methods for generating mutant varieties mutagenesis Forward genetics -chemicals -radiation Reverse genetics Insertional mutagenesis - Agrobacterium mediated transformation - Virus induced gene silencing - RNA mediated interference - transposon tagging TILLING Next generation sequencing

24. Breeding fo r disease resistance Numerous mutants have been developed through mutation induction, showing enhanced resistance to various diseases (virus, bacterial, and to some extent fungi ) E.g.: locus (ml-o) - located on the short arm of chromosome 4H in barley Induced mutations at the locus confers resistance to powdery mildew and barley yellow mosaic virus Chen et al , 2014

25. Quality, nutrition and functionality starch, protein, fatty acid, vitamins, etc . Elimination of undesired substances such as anti-nutritional factors Raising or lowering the concentration of specific substances such as fatty acids Through mutation is by inducing knock-outs in genes involved in the metabolic pathways Eg : high quality edible oil of canola was achieved by lowering the levels of glucosinolates and the erucic acid by gene knock outs induced by gamma ray irradiation

26. Achievements of mutation breeding

27. Distribution of mutant crop varieties by continents IAEA mutant database, http://mvgs.iaea.org (2015)

28. Number of mutant varieties released in the world https://mvd.iaea.org/#!Search?page=1&size=15&sortby=Name&sort=ASC&Criteria[0][field]=Country&Criteria[0][val]=136 28

29. Officially released mutant varieties in the FAO/IAEA Mutant Varieties Database, July 2015

30. INDIA Mutant cultivars

31. popular mutant cultivars of chickpea developed in India Variety Release year Main improved attributes Pusa-408 ( Ajay ) 1985, Resistant to Ascochyta blight, high yield, profuse branching, semi erect, maturity 140-155 d Pusa-413 ( Atul ) 1985, Resistant to Fusarium wilt, stunt virus & foot rot, high yield, profuse branching, semi erect, maturity 130-140 days Pusa-417 ( Girnar ) 1985, High resistance to Fusarium wilt & moderate resistance to Ascochyta blight, stunt virus, high yield, profuse branching, maturity 110-130 days Kharkwal M. C . et al (1985 )

32. Leading rice varieties obtained by mutation breeding

33. Mutant rice varieties released in India for cultivation

34. Gamma radiation-induced rice mutants were released in India as high-yielding varieties under the series ‘PNR’. Two early ripening and aromatic mutation- derived rice varieties ‘PNR381’ and ‘PNR102 ’ currently popular in Haryana and UP Mutant Varieties database https://mvd.iaea.org/#!Home

35. The primary research centres and institutes in India that participated in the development and release of various mutants Indian Agricultural Research Institute (IARI)- New delhi Bhabha Atomic Research Centre- Mumbai Tamil Nadu Agricultural University –TN and National Botanical Research Institute – Lucknow , UP

36. Advantages Possible to achieve instant progress in elite material Single trait improvements can be made to an established variety preferred by producers, processors and/or consumers Limited breeding effort required Novel variation can be produced Single gene mutants with no negative pleiotropic effects are possible For some mutagenic treatments such as gamma and X-ray, there is neither residual radiation nor chemical contamination of the treated material. The treated material is safe to handle Specific genes/traits can be targeted Possible to calculate chances of success (mutation frequency)

37. Limitations... The process is generally random and unpredictable Useful mutants are rare and predominantly recessive Large population sizes and effective mass screening methods are required to select rare mutants Mutants can have strong negative pleiotropic effects on other traits

38. Continued ..... Health risks: handling, chemical mutagens ; radiations, fast neutrons treatments Most mutants are of no use to breeding even if a large number of mutants can be produced Field trialling and germplasm storage can be expensive and require a lot of space and careful management if large mutant populations are handled

39. References.... Text books Plant breeding – B. D. Singh Plant Mutation Breeding and Biotechnology - Edited by Q.Y. Shu , B. P. Forster, H. Nakagawa Review papers Principle and application of plant mutagenesis in crop improvement: a review – By Oladosu et al., 2015 Mutagenesis - By Rajnikant Mishra (2012)

40. Thank you