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MUTATION BREEDING IN SOLANACEOUS AND LEGUME VEGETABLES

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University of Horticultural Sciences, Bagalkot Kittur Rani Channamma College of Horticulture, Arabhavi Seminar-I By, MAHBOOB UHS12PGM240 Dept. of Vegetable Science ON Mutation breeding in solanaceous and legume vegetable crops

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Preamble Topic division Mutation

preamble :

preamble Mutation Is a sudden heritable change in the characteristics of an organism. Mutagenesis Treating of a biological material with a mutagen in order to induce mutation. Mutagens Agents used for induction of mutations . Irradiation Exposure of a biological material to one of the radiations. Mutation breeding Is induction and isolation of mutants. (Sharma, 2009)

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Historical event

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Physical mutagens Chemical mutagens Type of Mutation (Sharma, 2009)

Result of change :

Result of change Gene mutation: genes of nuclear or plasma, chloroplast mytochondria etc. Change in base sequence. 1) Hypomorphs - less efficient than normal allele 2) Hypermorphs - more efficient 3) Amorphs - lost the function of normal allele 4) Neomorphs - allele acquired a new function 5) Antimorphs - antagonising the action of normal allele Chromosome mutations: 1) Chromosome number 2) Translocation 3) Inversion 4) Deletions 5) Duplications (Sharma, 2009)

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MUTATION CAN BE GOOD BAD OR NEUTRAL

Mutagens:

Mutagens A) Physical mutagens (radiations) i ) Ionizing radiation a) Particulate radiation α- ray (DI) – Densely ionizing β- ray (SI) – Sparsely ionizing Fast neutron (DI)* Thermal neutron (DI) b) Non particulate radiation (electromagnetic radiations) X-rays (SI)* γ - rays (SI)* ii) Non ionizing radiation or Ultraviolet radiations (Sharma, 2009)

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B) Chemical mutagens Problems: Poor solubility, toxicity and reactivity. 1) Alkylating agents a) Sulphur mustards b) Nitrogen mustards c) Epoxides d) Ethylene imine (EI) * e) Sulphates and sulphonates Ethyl methane sulphonate (EMS)* Methyl methane sulphonate (MMS)* f) Diazoalkanes – Nitroso compounds: N-methyl-N-nitro-N- nitroso – guadinine (MNNG)

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2. Acridine dyes Acriflavine Proflavine Acridine orange Ethidium bromide* 3. Base analogues 5-Bromouracil 5-Chlorouracil 4. Others Nitrous acid Hydroxyl amine Sodium azide* (Sharma, 2009)

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Table 1: Different mutagen source, Organs suitable and Mutation produced. Plant mutation report ., 2010 Category Gamma rays x- rays β -rays Neutrons source Gamma garden (radio active decay of 14 C, 60 CO) x- ray machine Decay radio active Isotopes ( 3 H, 32 P, 32 S) Nuclear reactor (rad heavier element) Organs suitable Seeds and seedlings Seeds seeds Seeds Mutation produced Chromosome and point mutation Point mutation

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Effects of mutagens (Sharma, 2009)

Table 2: Number of officially released mutant cultivars developed with different types of radiation. :

Table 2: Number of officially released mutant cultivars developed with different types of radiation. Type of mutagen Number of released mutant cultivar Percentage of total Radiation* 1411 100.00 X Rays * 311 22.04 Gamma chronic * 61 4.32 Fast neutrons ** 48 3.40 Thermal neutrons 22 1.56 Gamma rays 910 64.49 Other 24 1.70 http://mvgs.iaea.org/AboutMRPs.aspx *including various treatment ** including neutrons

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S.N. Type of radiation Main properties Mode of action or changes caused 1. X-rays S.I., penetrating and non-particulate. Induce mutations by forming free radicals and ions. Cause addition, deletion, transitions and transversions. 2. Gamma- rays S.I., very penetrating and non particulate. Induce mutations by ejecting atoms from the tissues. Cause addition, deletion, transitions and transversions. 3. Alpha-particles D.I., particulate, less penetrating and positively charged. Act by ionization and excitation. Cause chromosomal and gene mutations. 5. Fast & thermal neutrons D.I., particulate, neutral particles, highly penetrating. Cause chromosomal breakage and gene mutations. 6. Ultra violet rays Non- ionizing, low penetrating. Cause chromosomal breakage and gene mutations. Table 3: Mode of action of different physical and chemical mutagens . Janardhan et al ., 2008, Gujrath

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Chemical agents Examples Mode of action or changes caused 1) Alkylating agents 2) Base analogues 3) Acridin dyes 4) Others EMS,EI, MMS, etc. 5Bromo uracil, 2 Amino purine. Acriflavin, proflavin Nitrous acid, Hydroxylamine Sodium azide Base substitution AT↔GC Transition Base substitution AT↔GC Transition Frame shift mutation (addition, deletion of nucleotides) GC↔AT Transitions Cont. …….

Table 4: Mutagens used and attribute improved in mutant cultivars released in India. Mahla et al., 2010, Jodhpur:

Table 4: Mutagens used and attribute improved in mutant cultivars released in India. Mahla et al ., 2010 , Jodhpur Mutagen No. of mutants Main attribute % of occurrence Gamma rays 169 High yield 86 X-rays 26 Early maturity 65 Neutrons 7 Disease resistance 57 Ethyl methane sulphonate 15 Quality characters 39 Dimethyl sulphonate (DMS) 4 Grain quality 67 Ethylene Imine (EI) 2 A biotic stress resistance 65 Sodium azide (NaN 3 ) 2 Improved plant type 81 Other mutagens 29 Other 9 Cross bred 47 Natural mutants 12

Fig 1: Number of cultivars developed by mutation breeding in each 5-year period from 1961-2010 :

Fig 1: Number of cultivars developed by mutation breeding in each 5-year period from 1961-2010 2714 Plant mutation report ., 2010

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Fig 2: Cumulative number of officially released mutant varieties in various regions of the world. Plant mutation report.,2010

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Country Common name and no. of released varieties Total Australia Soybean(1) 6(1) Austria Faba bean (1) 17(1) Bangladesh Tomato (3) 23(1) Belgium Potato (1) 22(1) Brazil Common bean (3) 9(3) Bulgaria Soybean (3) 30(3) Canada Common bean (12) 35(12) China Common bean(1) Soybean (58) 605 (59) CSFR/ CzechREP . Common bean (1) Soybean (1) 36(2) Egypt Common bean (1) 4(1) Estonia Potato (1) 5(1) Germany Faba bean(1) common bean (2) Soybean (1) 138(4) Hungary Soy bean(1) 7(1) India Common bean (1), cowpea(10), eggplant (1), pea(1), tomato(4), Soybean (7), cluster bean (1), chilli (1) 343 (26) Indonesia Soybean 3, 11(3) Iraq Faba bean 2, 23(2) Italy Common bean 2, eggplant3, pea 6, potato 1, 35(12) Japan Soybean 6, tomato 4, 120(10) Korea Soybean 2 11(2) Poland Faba bean 5, pea 14, 30(19) Russia Pea 1, 6(1) Sweden Pea1, 26(1) Thailand Soybean 1 9(1) USA Common bean (26), 125 (26) USSR Common bean (4), faba bean (4), pea (8),soybean (9), tomato(2), cowpea (8), 204 (35) Table 5: Number of variety released in legumes and solanaceous vegetables by various country Plant mutation report., 2010

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Crop Total Soybean 170 Faba bean 20 Common bean 57 Cow pea 12 Pea 20 Potato 16 Egg plant 06 Tomato 21 Chilli and capsicum 01 (328) Table 6: Number of mutant variety released in legumes and solanaceous vegetables in world. Plant mutation report ., 2010

General properties of mutation:

General properties of mutation Imanish et al .,2007,Japan

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(Sharma, 2009)

Steps of Mutation breeding :

Steps of Mutation breeding Formulation of objectives of the programme . Selection of the plant part to be used for mutation. Selection of mutagen (physical or Chemical mutagen). The dosage of mutagen. Giving the mutagen treatment. Handling of mutagen treated population. (Sharma, 2006)

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Selection of the plant part to be treated

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Not (Sharma, 2006)

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Case studies

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solanaceous TOMATO

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Table 7: Germination percentage, survival and mutation frequency of gamma irradiated tomato seeds. Cv. CO1 Dose No. of seeds Germination percentage Survival % after 40 days Mutation frequency (M 1 V 2 ) In moist chamber after 7 days In soil After 7 days Control 100 83 72 92.80 - 10kR 100 7 4 6 5 93.32 0.38 20kR 100 60 39 91.5 0.37 30kR 100 53 39 88.6 0.46 40kR 100 30 28 76.7 0.49 50kR 100 30 28 76.7 0.49 Emmanuel et al ., 2012, Gujarat

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Table 8: Mutagenic effect of EMS and EI on quantitative characters in M 1 generation of tomato. George et al ., 2011, India Chemical mutagen Concentration (%) Germination % Days to flower Height of the plant Pollen fertility% Control - 53 66.66 36.33 88.00 EMS 0.05 0.10 0.20 0.40 60.0* 52.0 49.0 28.0 55.66* 70.66 64.66 72.00 30.33 24.66 28.00 33.66 90.65 79.32 80.78 89.28 EI 0.05 0.10 0.20 0.40 42.0 37.0 25.0 0.00 60.33 61.66 61.66 - 43.00 40.00 33.33 - 78.85 68.57 76.00 - EMS+EI 0.05 0.10 0.20 0.30 0.40 57.0 50.0 58.0 32.0 31.0 68.66 61.00 59.66 72.33 72.33 21.33 40.00 26.00 31.00 28.66 88.11 83.40 90.65 81.44 70.05 CD at 5% - 14.38 3.77 5.05 9.96 CD at 1% - 19.49 5.12 6.85 13.49

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(A) Divided leaves (B) The loci for hairy (high trichome number) and high pigmented fruits (C) Seedling dwarfism (D) Large number of fruits per plant is characteristic of MT plants (E) The Tm-2 gene (resistance to the TMV) Both trays containing 35-days old plants were sprayed with a TMV solution. (F) The Aubergine ( Abg ) gene was introgressed epidermis (G) 13 flowers per inflorescence which is twice of that observed in MT.

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Table 9: Analysis of abnormalities in M 2 generation irradiated with 30Gy gamma rays Micro-Tom a wild type of Tomato. Imanish et al ., 2007, Japan Studied Survived Candidate of morphological mutant Confirmed inheritance Fruit mutant No .of M 1 families 2,500 2,344 ( 93.7%) 435 17 12 No. of M 2 families 19,000 6,644 (34.96%) 326 18 13

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Phenotypes of gamma rays induced mutant lines in ‘Micro-Tom’ (A). The first screening was performed in a field-grown M 2 population (B) Based on morphological alterations, such as small and abundant fruits (C) Elongated fruits (D) Similar to ovate mutation, small plant size (E) Absence of abscission layer in pedicels and presence of leaves in inflorescences (F) Similar to the jointless mutation, short pedicels (G) High pigmented (dark green) fruits (H) Incomplete ripening fruits

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Line Leaf surface colour Total fruit carotenoid contents (μg/g fw) L a* b* Lycopene β -carotene Dark green Wt 49.1 -11.2 24.1 68.7 2.8 Pale leaf mutant 56.4 -10.0 34.3 68.8 3.4 Table 10: Characterization of a colour mutant in M 3 generation irradiated with 30Gy gamma rays Micro-Tom of Tomato. Imanish et al ., 2007,Japan Colour Minolta CR-10 and carotinoid by HPLC a* a green to red scale , b* a blue to yellow scale, L* total carotenoid content,

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Common name Name of the mutant released Country of release Year of release Mutagen used Parent variety Main character induced tomato Bahar Bangladesh 1992 Cross - Determinate tomato Binatomato-2 Bangladesh 1997 Gamma rays - Yield tomato Binatomato-3 Bangladesh 1997 Gamma rays - Yield tomato Co.3 India 1977 EMS CO 1 Compact growth tomato Kagyoku Japan 1986 Cross - Disease resistance tomato Kyoryoku-reikou Japan 1984 Gamma rays Shugyokuxl.peruvianum TMV resistance tomato Kyouryokuogatareiko Japan 1984 Cross - Diseases resistance tomato Lanka Cherry Sri Lanka 2010 - - Colour tomato Luch 1 Russian Federation 1965 Gamma rays Pushkinsky E arliness tomato Lufanqie 7 China 1997 Gamma rays - TMV resistance tomato Lufanqie 7 China 1997 X-rays - Yield tomato MAGINE Cuba 2007 Gamma rays - tolerance to drought and good quality. tomato Maybel Cuba 2007 X-rays - Yield tomato PKM-1 India 1980 Gamma rays Annanj Yield tomato Pusa Lal Meeruti India 1972 Gamma rays Meeruti Fruit ripening tomato Rannii Nuch Russian Federation 1991 EI jubilejnii Earliness tomato Ryugyoku Japan 1986 Cross - Disease resistance tomato S.12 India 1969 Gamma rays Sioux Dwarfness tomato Yufan 1 China 2000 Gamma rays - TMV resistance tomato Yufan 2 China 2005 Gamma rays - TMV resistance Achivements http://mvgs.iaea.org/AboutMRPs.aspx

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POTATO

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Explant production Shoots formed by the tubers were surface sterilized by placing them in 70% ethanol for 2 min and 5% hypochloride solution for 10 min . rinsed 3 times with distilled water, dried and planted in MS , medium containing 30g/L saccharose . Th e shoots were incubated for 10 days at 26 °C in growth chambers with 16 h light/8h dark periods, and the node explants used in the study were obtained. Potato tissue culture and irradiation of the explants Node explants from Marfona potato variety were planted in MS medium containing 0.5 mg/L ZR and 1.5mg/L IAA. To create the M 1 V 1 generation, explants

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Table 11: Physiological parameters of the 28 day-old cultures of control and mutated node explants of potato .cv . marfona O rkun and Sema ., 2012, Turky Irradiation level Number of explants Regeneration rate (%) Average plant height ( cm) Average leaf count Root formation (%) control 40 93 4.61±0.27a* 4.8±0.26a 60 5 Gy 40 92 3.91±0.26ab 5.3±0.35a 63 10 Gy 40 83 3.43±0.28ab 4.7±0.38a 47 15 Gy 40 73 3.05±0.32bc 4.6±0.48a 47 20 Gy 40 60 2.27±0.33cd 2.6±0.35b 37 25 Gy 40 50 1.95±0.3d 2.53±0.38bc 23 30 Gy 40 30 1.83±0.37d 1.6±0.33c 13 50 Gy 40 6.5 0.21±0.12a 0.23±0.13d 0

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Radiation dosage M 1 V 3 NaCl concentration in the media Salt-tolerant mutants produced 50mM 100mM 125mM Amount % Number of explants (Control ) 40 40 40 0 0 5 Gy 40 40 40 0 0 10 Gy 30 30 30 0 0 15 Gy 40(5) 40(9) 40(6) 20 17 20 Gy 40(5) 40(9) 40(3) 17 14 25 Gy 30 30 30 0 0 30 Gy 40(5) 40(6) 40(3) 14 12 Table 12: Selection of salt-tolerant explants in the M 1 V 3 generation of Marfona potato. Orkun and Sema ., 2012, Turky

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Numbers of the salt tolerant mutant plants ≠1 ≠2 ≠3 ≠4 ≠5 ≠6 ≠7 ≠8 ≠9 ≠10 ≠11 ≠12 ≠13 ≠14 ≠15 ≠16 ≠17 ≠18 ≠19 Gamma irradiation dosage ( Gy ) 15 15 20 20 30 15 15 15 20 20 30 15 15 15 20 20 30 15 30 NaCl concentration ( mM ) 125 125 125 125 100 100 100 100 100 100 50 50 50 50 50 50 50 100 100 Table 13: Classification of the 19 salt-tolerant mutants of Marfona potato. Orkun and Sema ., 2012, Turky

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Fig 3: RAPD profiles obtained from the OPH-12 primer. Orkun and S ema ., 2012, Turky M: Marker, N: Negative, K: Control, average polymorphism ratio of the 6 primers used was estimated as 89.66%.

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Table 14 : Genetic distance between the mutants and control plants of marfona potato. Orkun and Sema ., 2012, Turky

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Fig 4: Dendrogram of the control and mutants based on distances produced by RAPD-PCR. Of marfona potato Orkun and Sema ., 2012, Turky

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Table 15: Effect of gamma radiation on in vitro cultured callus weight, root and shoot initiation of Gao-14 cv . Of potato . Zhen, 2008, China Gamma rays ( Gy ) Mean callus weight *(mg) Root weight (mg) Frequency of shoot initiation (%) 1 2.22 1.03 23 5 2.18 0.89 10 10 2.17 0.15 0 30 1.76 0.13 0 50 1.80 0.11 0 100 1.31 0.11 0 Control 2.04 0.97 25 *after 66 day of culture.

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Table16: Variation obtained in M 1 V 2 generation after irradiation with gamma rays of potato in lab condition . Zhen, 2008, China Character No. of Mutant plants Frequency (%) Leaf shape 6 0.8 Pigmented leaf 3 0.4 Pigmented stem 3 0.4 Tuber skin colour 1 0.1 Tuber flesh colour 1 0.1 High tuber number* 2 0.3 Increased tuber weight** 3 0.4 High yield plant *** 2 0.3 Poor growth # 18 2.3 *Number of tubers was 6 or more per plant. **Mean tuber weight exceeded 500g ***Tuber yield per plant exceeded 2000g. #Tuber yield per plant was under 250g.

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Table 17: Peroxidase and Polyphenol oxidase activity and phenol content in the variants of potato varieties Kufri Chandranmukhi and Kufri Jyothi isolated after gamma irradiation in M 1 V 3 generation for late blight. Das et al .,2002, Shimla

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Variety Treatment(dose ) Disease reaction (Late blight) Enzyme activity Phenol content (mg) Peroxidase (OD/min/gm ) Polyphenol oxidase (OD/min/gm ) Kufri Chandramukhi 20 Gy 1PLB20 R 0.072 0.035 48.2 2PLB20 R 0.084 0.030 46.50 3PLB20 MR 0.012 0.024 36.30 4PLB20 S 0.012 0.012 10.30 5PLB20 MR 0.091 0.017 33.80 6PLB20 S 0.010 0.014 14.60 40Gy 1PLB40 MR 0.056 0.016 32.30 1PLB40 R 0.028 0.033 41.40 1PLB40 S 0.014 0.015 15.50 1PLB40 R 0.091 0.030 41.40 Control S 0.020 0.012 10.80 Kufri Jyothi 20Gy 1PLB20J R 0.077 0.020 40.60 2PLB20J R 0.078 0.021 48.20 3PLB20J MR 0.046 0.019 39.80 4PLB20J R 0.082 0.030 41.20 5PLB20J S 0.019 0.012 14.62 6PLB20J S 0.025 0.012 11.80 40Gy 1PLB40J MR 0.086 0.016 33.60 2PLB40J R 0.086 0.019 35.90 3PLB40J R 0.082 0.019 51.30 4PLB40J R 0.086 0.018 56.80 5PLB40J R 0.055 0.019 39.20 Control S 0.023 0.013 14.20

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Common name Name of the mutant released Country of release Year of release Mutagen used Parent variety Main character induced Potato Desital Italy 1987 Gamma rays Desiree Skin colour KonkeiNo.45 Japan 1973 X-rays - Whit Skin colour Mariline 2 Belgium 1968 X-rays Mariline Yield Sarme Estonia 1993 Cross EMS - Late maturity, high yield, White Baron Japan 1997 Gamma rays Yellow tolerant to browning C Japan 1994 Gamma rays Local Yield Achivements http://mvgs.iaea.org/AboutMRPs.aspx

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Capsicum and chilli

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Table 18: Effect of different exposures of gamma radiations on reproductive parameters in capsicum annum. Jabeen et al ., 2004, Pakistan Treatments Days require for floral initiation Percent meiotic aberrations Percent pollen sterility Number of capsules per plant Control 100.0 0.00 3.60 100.00 2kR 88.20(12) 3.30 5.70 116.40(16) 4kR 92.20(8) 4.20 8.70 106.70(6) 8kR 96.10 7.60 10.60 97.50 12kR 100.0 10.90 11.80 81.90 16kR 104.90 14.80 25.90 76.80 20kR 105.90 21.80 32.80 59.50

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Table 19 : Mutagenic effectiveness and efficiency of mutagens in M 2 generation of chilli . Cv . K1, (pot) Devi and Mullainathan ., 2011, Tamil Nadu Mutagenic effectiveness Mutagenic efficiency Mutagenic dose Lethality % M 2 families M 2 plants M 2 families M 2 plants Gamma rays( kR ) 10 42(58) 0.71 0.32 0.07 0.076 20 45(55) 0.42 0.14 0.19 0.061 30 48(52) 0.36 0.12 0.21 0.068 40 33(67) 0.12 0.06 0.15 0.068 50 20(80) 0.50 0.05 0.124 0.112 EMS( mM ) 10 38(62) 0.21 0.052 0.22 0.054 20 42(58) 0.07 0.025 0.14 0.17 30 43(57) 0.05 0.013 0.12 0.032 40 32(68) 0.02 0.059 0.09 0.029 50 24(76) 0.006 0.005 0.05 0.040

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Table 20: Mutation rate of various doses of gamma rays and EMS in M 2 generation in chilli Cv . K1, (pot) Devi and Mullainathan ., 2011, Tamil Nadu Chlorophyll mutants M 2 families bases M 2 plants bases Viable mutants Total mutants M 2 Family basis Mutagen N umber M 2 families Number M 2 plants No. % No. % No. % No. % Gamma rays(kR) 10 198 845 14 7.07 27 3.19 5 0.59 32 3.78 20 212 916 18 8.49 25 2.73 6 0.66 31 3.38 30 196 813 21 10.7 29 3.57 16 1.23 39 4.80 40 143 758 7 4.89 17 2.24 7 0.92 23 3.03 50 121 445 3 2.48 10 2.25 2 0.45 12 2.70 EMS ( mM ) 10 167 917 14 5.38 19 2.07 5 0.55 24 2.62 20 190 943 11 5.79 19 2.01 9 0.95 28 2.97 30 207 828 12 5.80 20 2.57 14 1.69 27 3.26 40 167 844 5 2.99 8 0.94 7 0.83 15 1.78 50 153 510 2 1.31 5 0.98 4 0.78 9 1.76

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Table 21: Spectrum and frequency of induced chlorophyll deficiency mutations in M 2 plants of C hilli . Cv . K1, (pot) Devi and Mullainathan ., 2011,Tamil Nadu Albino % Chlorine % Virescent % Mutagenic treatment No. % No. % No. % Gamma ray(kR) 10 5 0.59 7 0.82 11 1.54 20 6 0.66 5 0.55 7 0.76 30 6 0.98 15 1.85 11 1.35 40 4 0.53 19 2.50 11 1.45 50 7 1.57 20 4.49 13 2.92 EMS( mM ) 10 - - 7 0.76 3 0.32 20 6 0.64 10 1.06 10 1.06 30 6 0.72 11 1.33 9 1.09 40 11 1.30 18 2.13 11 1.30 50 8 1.57 10 1.96 12 2.35 Albino : Neither carotenoids nor chlorophyll is formed. Viridis : Young leaves are yellowish, but later change to normal green Virescens : Light green gradually changing to dark green and mostly viable. Chlorina : Greenish yellow variegation or chlorophyll deficiency

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Table 22: Induced viable mutations for various traits in M 2 generation of C hilli . Cv . K1, (pot) Devi and Mullainathan ., 2011, Tamil Nadu Gamma rays(kR) Total viable mutant EMS (mM) Total viable mutants Grand total viable mutants Trait 10 20 30 40 50 0 10 20 30 40 50 0 0 Tall mutant - 1 3 1 - 4 2 - 5 1 - 8 13 Profuse branching - 2 4 3 - 9 - 1 7 2 - 10 19 Curved leaf 1 - 4 - - 5 1 - 3 - - 4 9 Diminished morphology 1 - 2 2 4 9 - - 1 - - 1 10 Dwarf plant 1 - 4 2 2 9 1 1 1 2 4 9 18 Flower mutant - 2 6 1 - 9 - 2 12 4 - 18 27 Rosette leaf - - - - - - - - 2 - - 2 2 Pointed fruit apex - 1 1 1 - 3 - 1 5 - - 6 9 Male sterile 1 - 7 4 4 16 - 1 6 1 3 11 27 Long pods - 1 4 1 - 6 - 1 16 1 - 18 24 Dark green pods - - 6 2 2 10 - 2 14 1 - 17 27 Yellowish green pods - 1 4 - - 5 - - 1 - - 1 6 Total 4 8 47 17 12 85 4 9 67 8 7 105 190

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Tall Mutant : plant height, 65 to 80cm; few branches flowers and fruits and frequency was higher with chemical mutagens than gamma irradiation. Curved Leaf : leaves have curved leaf blades with long petioles, fewer branches, flowers and fruits, seed set poor. Diminished Morphology : extremely small (2cm in length 1cm in width) leaves with normal shape; main stem before the first cyme has 18 to 20 internodes. Dwarf Plant : 15 to 20cm in height, short internodes, thick, dark green leaves . Flower Mutant : generally Capsicum species has pentamerous flower but an abnormal behavior of trimerous , tetramerous, heptamerous nature. This is also reported mutants with change in number of sepals, petals and ovules than normal in Borago Rosette Leaf : the rosette plants were shorter, had smaller leaves which is dependent upon the length of the petiole and delayed floral development. Genic Male Sterility : Male sterility is utilized for economic production of hybrids on a large scale in various crops. In chilli , 30, 40 and 50 kR of gamma rays and 30 mM of EMS mainly induced genic male sterility ( androecium transformed into petaloid structures). Long Pods : Commonly, consumers prefer long pods. A total of 16 such mutants were isolated from 30mM of EMS. These mutants had a pod length of 8 to 10.5cm

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Common name mutants name Country of release Year of release Mutagen used Parent variety Main character induced Sweet pepper F 1 Orange Beauty Russian Federation 2011 EMS Yield Chilli MDU.1 India 1976 Gamma rays Capsaicin and yield Sweet pepper Nush-51 Russian Federation 1991 Gamma rays Yield Sweet pepper Orangeva Kapija Bulgaria 1991 EMS Yield Sweet pepper Pirin Bulgaria 1991 EMS (FAO/IAEA Database, 2010) Achivements

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CLUSTER BEAN

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Ethyl methane sulphonate Gamma rays Fig 5: Effect of Gamma rays and EMS on reduction in germination and survival of cluster bean seeds of RGC and HGS, Mahla et al ., 2010, Jodhpur

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Table 23: Frequency of morphological and chlorophyll mutation in cluster bean. (RGC and HGS ) Mahla et al ., 2010, Jodhpur Cultivars / treatment Morphological mutation % Chlorophyll deficiency mutation % Spreading Dwarf Mating ( turfing ) Vigorous Total Albino Chlorina Xantha Total RGC , Gamma rays control - - - - - - - - - 10 kR 0.87 - 0.43 0.40 1.70 1.14 0.57 0.85 2.56 20 kR 2.42 0.85 1.30 0.76 5.33 1.78 0.88 1.78 4.44 30 kR 3.65 4.56 5.87* 2.05 16.13 3.23 1.29 0.00 4.52 40 kR 16.72* 8.18* 3.09 4.21* 32.20 3.39 1.69 0.00 5.08 RGC , EMS 0.1% - 0.62 0.43 - 1.05 0.85 0.42 0.40 1.67 0.2% 2.15 0.62 1.40 0.25 3.42 1.03 0.68 0.65 2.36 0.3% 4.54 2.12 2.02 0.46 9.14 1.71 1.14 1.15 4.00 0.4% 6.77 13.42 5.84 1.21 17.24 2.75 1.38 0.00 4.13 HGS , Gamma rays Control - - - - - - - - - 10 kR 1.16 0.28 0.57 - 2.01 1.34 0.67 1.34 3.35 20 kR 2.39 1.92 1.14 1.58 7.03 2.70 1.08 1.08 4.86 30 kR 4.66 3.72 10.31 4.47 23.16 2.86 2.10 2.11 7.07 40 kR 15.18 8.96 7.34 16.76 48.24 5.88 0.00 2.35 8.28 HGS , EMS 0.1% 0.36 0.28 0.67 0.59 1.90 1.09 0.54 0.82 2.45 0.2% 1.71 0.80 1.26 2.23 6.00 2.50 1.00 1.50 5.00 0.3% 3.28 1.74 10.65 2.42 18.09 2.86 1.90 1.90 6.66 0.4% 4.76 16.35 8.98 4.39 34.48 5.13 0.00 2.56 7.69

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Sl.no Agronomic traits (Number of superior mutant) RGC HGS Mutants Parent Mutants Parent 1 Early maturity (days ) (20) 66 days (13 plants) 72 65 days (7plants) 73 2 High yielding (g/plant ) (20) 6 (10 ) 5.23 6.4(10 ) 5.58 3 Plant height (cm ) (5) 45 (3 ) 38.0 50 (2) 42.0 4 Number of pods per plant (9) 24 (5 ) 18.0 29 (4) 19.0 5 Number of seeds per pod (10) 9 (6) 8.2 9 (4 ) 8.4 (27) (37) Table 24: Selected progenies for agronomic traits in M 3 generation of cluster bean RGC and HGS Mahla et al ., 2010, Jodhpur

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Mutants Yield per plant(g ) Days to maturity Plant height (cm) Branches per plant Clusters per plant Pods per plant Seeds per pod Mutant-1 6.89 67.5 42.7 5.7 12.3 25.5 9.2 Mutant-2 8.02* 68.0 46.0 5.9 12.9 27.1 9.7 Mutant-3 7.93 65.5 43.7 5.9 12.9 26.5 9 .8 Mutant-4 6.81 66.5 43.4 5.6 11.8 22.6 8.5 Mutant-5 7.54 66.5 44.8 5.8 12.2 23.8 9.0 Mutant-6 7.48 68.5 41.9 5.7 11.9 22.2 8.9 Mutant-7 7.90 65.0 43.0 5.5 12.6 22.9 9.7 RGC 936 Control 4.26 71.5 38.1 4.5 9.2 18.1 7.9 Mutant-1 7.82 67.5 44.6 5.4 11.9 24.6 8.8 Mutant-2 7.35 68.0 44.3 5.4 12.0 24.6 9.0 Mutant-3 7.74 69.0 50.0 5.5 11.8 25.1 8.8 Mutant-4 7.84 65.0 49.3 4.8 10.2 25.8 8 .1 Mutant-5 9.95* 61.0 48.8 6.3 13.7 34.8 9.1 Mutant-6 9.50* 60.5 45.9 6.2 13.4 30.6 9 .8 HGS 365 Control 4.55 72.3 36.3 4.4 9.2 18.3 8.3 Table 25: Agronomic performance of promising M 4 progenies of cluster bean. RGC and HGS. Mahla et al ., 2010, Jodhpur

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FRENCH BEAN

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Table 26 : Frequency of EMS induced seed coat colour mutants in the M 2 generation of French bean cv. BAT 65, Swapnil , and Rajendra , 2011, Brazil EMS concentration (%) No. of treated seeds No. of surviving M 1 plants plants with mutant seed coat colour in M 2 Mutant characters N0. % 0 control 6 00 5 65 0 0 Black colour 0.25 6 00 259 16 0.20 10 different hues of beige, 6 red brown 0.5 6 00 260 23 0.57 3 yellow brown, 4red brown, 2 black with brown strips, 6different hues of beige, 5grayish yellow, 1.0 6 00 150 18 0.22 8red brown, 5yellow brown, 5different hues of beige, 1.5 6 00 93 16 0.20 6red brown, 8yellow brown, 2different hues of beige,

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Table 27 : Frequency of Gamma rays induced seed coat colour mutants in the M 2 generation of French bean cv . BAT 65. Swapnil , and Rajendra , 2011, Brazil Radiation dose ( kR ) No. of treated seeds No. of surviving M 1 plants plants with mutant seed coat colour in M 2 Mutant characters N0. % control 6 00 5 76 0 0 Black colour 6 6 00 265 6 0.95 3different hues of beige ,2red brown, and 1yellow 12 6 00 263 20 1.05 5 yellow brown, 3red brown, 2black with brown strips, 18 6 00 251 5 0.75 3red brown, 1yellow brown, 1different hues of beige, 14 6 00 121 6 0.98 2red brown, 2yellow brown, 2different hues of beige, 24 6 00 23 4 0.84 2red brown, 1yellow brown, 1different hues of beige,

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Table 28: Mutagenic effectiveness and efficiency of Gamma rays and EMS in French bean. Mehata et al ., 2001, India Treatment Sterility % Mutated progeny (%) Mutagenic effectiveness Mutagenic efficiency Gamma rays (kR) 5 10 15 24.50 29.30 30.30 6.30 8.40 9.20 1.52 0.82 0.47 0.26 0.29 0.32 Mean 28.03 7.96 0.93 0.24 EMS (%) 0.3 0.5 0.7 25.30 28.40 32.70 7.90 12.30 8.60 5.36 3.90 2.13 0.26 0.59 0.87 Mean 28.80 9.60 3.46 0.67

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Table 29: Seed germination in M 1 , M 2 and M 3 generations treated with X-rays in French bean. Gill and Vans, 2011, China X-ray Treatments Generation Number of seeds sown Number of seeds germinated Germination percentage 7kR M 1 M 2 M 3 90 1196 500 69 895 334 76.67 74.83 66.80 M 75.76 14kR M 1 M 2 M 3 90 1124 500 64 910 315 71.11 68.04 63.00 M 69.33 21kR M 1 M 2 M 3 90 436 500 43 200 190 47.78 66.87 39.00 Control - - - 90 436 500 58 303 333 64.44 69.81 66.67 66.02

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Fig 6 : Effect of mutagens on the frequency of flower colour mutations in M 2 generation of French bean ( cv . Varun pink) Avinash and More, 2010, Pune Doses of radiation by gamma rays(kR) percentage of total number of plants 0.954 white colour flowers

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Fig 7: Effect of mutagens on the frequency of flower colour mutations in M 2 generation of French bean ( cv . V arun pink) Avinash and More., 2010, Pune percentage of total number of plants Doses of EMS 0 0.1 0.15 0.2 0.25 3.725

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Common name Name of the mutant released Country of release Year of release Mutagen used Parent variety Main character induced common bean AC Hensall Canada 1997 EMS cross - Earliness common bean AC Skipper Canada 1996 cross - Earliness common bean Black Magic United States 1987 EMS cross - Seed colour common bean Blackhawk United States 1990 EMS cross - Seed colour common bean C-20 United States 1982 EMS cross - Seed colour common bean Campeiro Brazil 2003 Gamma rays cross - - common bean CAP-1070 Brazil 1986 Gamma rays Carioca Bushy type common bean Centralia Canada 1988 EMS cross - Earliness common bean CIAT 899 Tunisia 2007 EMS cross - - common bean Domino United States 1987 EMS cross - Seed colour common bean Dresden Canada EMS cross - Earliness common bean Fleetwood Canada 1977 X-ray cross - Earliness common bean Frontier United States 1998 EMS cross - Seed colour common bean FT- Paulistinha Brazil 1992 EMS cross - Yield common bean Giza 80 Egypt 1980 Gamma rays Fin de Villeneuve Rust resistant common bean Gratiot United States 1962 X-rays Michelite Stiffness common bean Harkovskaya 8 Russian Federation 1985 Gamma rays - Seed colour common bean IAPAR 57 Brazil 1992 X-rays Crosss - GMVD Resistance common bean Pusa parvati India 1970 X-rays Wax P Earliness common bean Svetlaya USSR 1992 MNH Shchedraya Yield common bean Unima FRG 1957 Cross EMS Disease Resistant (FAO/IAEA Database, 2010) Achivements

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Cow pea Figure 1 Root distribution patterns of the cowpea drought tolerant control line IT96D-602and the more susceptible line TVu7778.

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Table 30: Efficiency and effectiveness of Gamma rays, EMS and combination of both in cow pea cv. Gomati. Dube et al ., 2011, Wardha Dose/ Treatment ( kR ) % Chromosomal aberrations in mitosis (CA) % Lithality in germination ( L) % Injury in seedling height (I) Mutagenic Efficiency Mutagenic Effectiveness , 10kR 11.3 28.6 27.8 0.4 1.4 0.9 20kR 17.6 43.2 25.0 0.3 0.5 1.1 30kR 18.0 50.6 6 1.0 0.3 0.3 0.6 40kR 18.6 52.4 72.0 0.3 0.2 0.4 0.1%18h EMS 12.0 15 21.0 0.8 0.5 5 .6 0.2% 16.0 19.0 32.6 0.8 0.4 2.9 0.3% 19.6* 14.2 55.2 0.9* 0.7 6.0* 0.4% 18.0 59.5 66.3 0.3 0.3 2.5 10kR+18h 0.2 % EMS 10.0 64.2 30.0 0.2 0.3 0.7 20kR+0.2 % 7.0 79.9 60.0 0.08 0.1 0.3 30kR+0.2 % 5.3 80.9 78.9 0.06 0.06 0.1 40kR+0.2 % 3.3 85.7 97.3 0.04 0.03 0.07

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Developmental problem

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Table 31: Mean nutritive values of cowpea mutants treated with gamma rays of IT84S 2246D obtained by proximate analysis. (potted) Olabisi et al ., 2007, Nigeria Means with different letters within column are significantly different at 0.05 level of probability..

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Table 32: Means of various parameters evaluated on gamma induced mutants infested with adult females of maruca vitrata in a free choice test. Olabisi et al ., 2008, Nigeria Means followed by different letters in the same column are significantly different at probability of 0.05 Mutant No. of larvae recovered/plant No . of pods/plant No . of damaged pods/plant % pod damage No . of seed/plant No . of damaged seeds/plant Seed yield/plant(g) % yield loss % seed damage Mutant 1 9.3· bcd 11.7 cdef 7.7 cd 50.6 de 75.0 cde 14.3 bcd 14.4 8b 39.2 efg 21.2 ed Mutant 2 9.7 bcd 12.0 cdel 7.7 ed 72.0 abcde 74.0 cde 11.7 bcd 13.3 8bcd 19.7 g 14.9 d Mutant 3 10.3 bcd 4.3 f 4.3 d 100.0 8 20.7 fg 4.7 cd 1.1 f 78.5 abc 22.6 ed Mutant 4 7.0 ed 13.3 bcdef 7.3 ed 55.9 ede 98.7 abcd 8.7 bed 15.8 a 46. 1 edefg 7.8 d Mutant 5 9.3 bcd 15.7 bcde 11.0 abcd 69.2 abcde 87.3 bcd 10.7 bcd 15.6 a 54.4 cdef 12.2 d Mutant 6 19.3 a 21.7 ab 4.0 a 48.8 ab 98.3 abcd 3.3 ab 13.6 abcd 19.3 abcde 14.9 ed Mutant 7 12.7 bc 12.3 bcdef 8.7 bcd 77.3 abcd 103.3 abc 17.7 ab 13.3 8bcd 47. 1 cdefg 17.4 ed Mutant 8 12.7 bc 19.3 abc 15.7 8b 80.3 abc 130.7 ab 25.7 a 13.3 abcd 47.3 edelg 19.9 cd Mutant 11 11.7 bed 5.0 f 4.4 d 91.7 a 5.0 g 3.67 d 0.7 f 88.5 ab 91.1 a Mutant 14 11.7 bed 12.7 bcdef 12.0 abcd 95.6 a 63.0 cdel 9.3 bed 7.6 cde 24.6 fg 14.9 d Mutant 15 8.0 ed 15.3 bcdef 12.7 abc 84.3 abc 23.7 efg 5.0 ed 3.6 f 71.6 abcde 20.5 ed 16 (parent) 15.7 ab 11.3 cdef 10.7 abcd 94.3 a 46.7 defg 14.0 bed 5.4 def 75.1 abcd 32.1 bcd ( Tvu 946) 5.7 d 12.7 bcdef 5.5 d 44.3 e 144.0 a 7.7 bed 8.5 bcde 37.1 fg 5.3 d S.E 2.0 2.9 2.3 9.1 16.1 3.3 2.0 10.3 9.2

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Plant type Larval Index Pod damage Yield loss Seed damage Overall susceptibility Tolerance** Index Index Index Index Rating Mutant 1 0.59 0.54 0.52 0.66 2.31 MS Mutant 2 . 0.62 0.76 0.26 0.46 2.10 MS Mutant 3 0.66 0 .06 1.05 0.70 3.47 S Mutant 4 0.45 0.59 0.16 0.18 1.89 MT Mutant 5 0.59 0.73 0.72 0.38 2.42 MS Mutant 6 0.48 0.62 0.15 0.16 1 .71 MT Mutant 7 0.81 0.82 0.63 0.54 2.80 MS Mutant 8 0.81 0.85 0.62 0.62 2.90 MS Mutant 9 0.76 0.65 0.57 0.58 2.56 MS Mutant 10 0.75 1.06 0.77 0.63 3.21 S Mutant 13 0.51 1.02 1.27 1.47 4.27 HS Mutant 14 0.75 1.01 0.33 0.46 2.55 MS Mutant 15 0.51 0.89 0.95 0.65 3.00 S (parent)16 1.00 1.00 1.00 1.00 4.00 S (Tvu946)17 0.36 0.47 0.49 0.17 1.49 MT Table 33: Susceptibility indices and resistance ratings of Maruca infested gamma induced mutants in a free choice test Olabisi et al ., 2008, Nigeria

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Table 35: Yield of M 2 Vita 7 exposed to different doses of the three mutagens to cowpea seeds. Odeigah et al ., 2009, Nigeria Treatment Yield Parameters 100R gamma rays 200R gamma rays 10mMx6hr . EMS 1.0NaN 3 x2hr Control Peduncles/Plant 19.19±5.63* (45.60) 16.49±4.46 (25.11) 42.60±12.99* (223.22) 35.88±11.78* (172.23) 13.18±3.9 Pods/plant 22.51±7.38* (47.32) 20.59±7.90* (34.16) 36.98±15.10* (142.02) 34.74±14.71* (125.59) 15.28±4.8 Plant height (cm) 25.26±10.07 (14.20) 28.47±10.53* (28.71) 21.15±4.55 (-4.39) 138.74±48.3 (527.22 ) 22.12±1.6 100 seeds weight (g) 13.23±0.43 (0.23) 13.20±0.48 (0.00) 18.83±0.29* (42.65) 12.15±0.41 (-7.95) 13.20±0.4 Seeds/Pod 11.59±3.61 (-0.7) 10.14±2.99 (-12.59 12.76±2.05* (10.00) 10.46±2.48 (-8.28) 11.60±2.1 Germination per cent 57.30 (-10.47) 31.00 (-51.56) 67.708* (5.78) 50.45 (-21.17) 64.00 50% flowering 39* (8.75) 41* (0.0) 39* (8.75) 45 (2.86) 49 1 Values are sample means ± S.D. * Better than control, p=0.05, ** Better than control, p=0.01

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Common name Name of the mutant released Country of release Year of release Mutagen used Parent variety Main character induced cowpea Co 5 India 1984 Gamma rays Co-1 Nutritional cowpea COCP 702 India 2002 - - - cowpea Cowpea-88 India 1990 Radiation Yield cowpea Gujarat cowpea-1 India 1984 - - - cowpea ICV 11 Kenya 1985 Gamma rays ICV 1 Aphid resistant Semi-erect cowpea ICV 12 Kenya 1985 Gamma rays ICV 1 Aphid resistant, Yield cowpea ( Kalleshwari ) India 2007 - - Nutritional cowpea Uneca -Gama Costa Rica 1986 Gamma rays Centa Yield cowpea V16 ( Amba ) India 1981 DMS Pusa Phalguni Yield cowpea V240 India 1984 DMS Pusa Phalguni Yield cowpea V37 ( Shreshtha ) India 1981 DMS Pusa Phalguni Yield cowpea V38 ( Swarna ) India 1981 DMS Pusa Phalguni Yield (FAO/IAEA Database, 2010) Achivements

IRB IBARAKI JAPAN:

IRB IBARAKI JAPAN GAMMA ROOM is shielded building equipped with a 44.4TBq Co-60 source. The facility is used for the acute irradiation of seed, bulbs, tubers and scions. Gamma room GAMMA FIELD is a circular field of 100m radius with 88.8TBq Co-60 source at the center. The field is surrounded by a shielding dike of 8m in height. gamma greenhouse is an octagonal greenhouse of 7m radius for the chronic irradiation of subtropical plants. Irradiation tower of gamma field Gamma room Shield box of 60 co

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Building of gamma room for acute irradiation Irradiation to seeds Irradiation to in vitro materials Irradiation to plants

Malaysia :

Malaysia

Institutes handling Mutation work in India.:

Institutes handling Mutation work in India. Bose Research I nstitute Calcutta (BRI) IARI, New Delhi Bhabha Atomic Research Centre (BARC) Mumbai, Tamil Nadu Agricultural University (TNAU) Coimbatore, National Botanical Research Institute (NBRI) Lucknow .

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Space mutation breeding is characterized by inducing a high frequency and large amplitude variation, with good heredity and beneficial modifications. (270000 hectares).400 spp RAPD patterns of control tomato (youfan 1) plants and plants from seeds carried in the space station generated by primers S13, S14 and S15. Note: CK means the ground control. 1, 2, 3, 4 and 5 are five plants from seeds carried in the space. Space Medicine & Medical Engineering 2000,

China Aerospace Breeding Research Center (CABRC):

TF873 China Aerospace Breeding Research Center (CABRC)

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CONCLUSION Induce mutation can rapidly create variability in quantitatively and qualitatively inherited traits. Mutation breeding is one of such tool used to broaden the genetic base, to identify, and develop useful genotypes. Modern mutation breeding with modern technologies in combination with refined selection methods has great potential for the improvement of both sexually and asexually propagated crop plants .

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(Sharma, 2006)

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Mutation breeding is a potentially powerful tool for legume vegetable improvement, since it has very limited exploitable and useful genetic variation. The creation of variability through hybridization is very difficult and cumbersome owing to very small and delicate flower structures , which often result in very poor seeds setting in the manually hybridized buds and higher frequency of flower drop during and after crossing

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Table 34 : Yield of M 2 IT84E-124 exposed to different doses of the three mutagens . Odeigah et al ., 2009, Nigeria Treatment /Yield Parameters 100R gamma rays 200R gamma rays 10mMx6hr . EMS 1.0NaN 3 x2hr 1.0NaN 3 x2hr Control Peduncles/Plant 11.79±4.59 (-7.53) 13.97±4.13 (9.56) 10.22±2.17 (-19.84) 0.60±2.82 (-16.86) 21.29±10.36* (66.98) 12.75±2.77 Pods/plant 17.27±8.36 (-3.21) 18.59±5.64 (4.26) 11.68±3.42 (-34.49) 27.81±15.70* (55.97) 34.86±17.09* (95.51) 17.83±4.16 Plant height (cm) 16.60±3.59 (-3.21) 17.73±3.15 (3.38) 11.31±1.85 (-34.05) 36.49±15.40* (112.77) 29.26±36.17* (70.49) 17.15±1.41 100 seeds weight (g) 13.35±0.36 (-0.37) 13.85±0.55* (3.36) 12.70±0.71 (-5.22) 11.75±0.35 (-12.31) 13.80±0.34 (3.30)* 13.40±0.21 Seeds/Pod 9.11±3.43 (8-72) 9.00±2.38 (-9.82) 5.78±2.29 (42.08) 10.65±2.91 (6.71) 11.73±3.51* (17.53) 9.98±2.46 Germination percent 70.60 (-39.44) 48.70 (-32.36) 62.10 (-13.75) 45.80 (-36.39) 72.80 (1.11) 72.00 50% flowering 36 (2.86) 41 (-17.14) 35 (0.0) 36 (2.86) 32 (8.75) 35 1 Values are sample means ± S.D. * Better than control, p=0.05, ** Better than control, p=0.01

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Table 7: Mutagenic effect of EMS and EI on mutation frequency in tomato. Sagar et al .,2011, India Chemical mutagen and concentration Mutation frequency in M 2 Control - EMS 0.05 % 0.01 % 0.02% 0.04% Mean 6.0 63.65* 14.75 10.34 31.17 EI 0.05% 0.10% 0.20% Mean 8.34 3.00 28.57 13.30 EMS+EI 0.05% 0.10% 0.2% 0.3% 0.4% Mean 12.50 8.33 15.36 12.90 17.39 13.29

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Brinjal

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Treatment /dose Effectiveness M/Kr Pusa Purple Long Bhagumathi Local Round Gamma rays ( kR ) 5 10 15 20 25 30 35 40 0.054 0.056 0.031 0.026 0.018 0.044 0.045 0.028 0.084 0.034 0.024 0.021 0.034 0.039 0.041 0.044 0.058 0.038 0.026 0.019 0.032 0.048 0.038 0.029 EMS (%) 0.05 0.1 0.15 0.2 0.25 0.30 0.35 0.40 2.000 1.350 0.608 0.800 0.940 0.808 1.039 0.900 1.875 1.075 0.708 0.850 1.015 1.054 1.032 1.037 2.800 1.575 1.353 0.775 1.440 1.158 0.917 0.806 Gamma rays +EMS 20kR+0.1 % 20kR+0.2 % 20kR+0.3 % 20kR+0.4 % 1.904 1.353 1.341 1.585 1.684 1.175 1.999 0.748 1.737 0.910 0.532 0.522 Table 18: Mutagenic effect of gamma rays and EMS in brinjal. Shivaputra et al., 2008,India

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Table 19: Frequency of plants with translocations and range and frequency of different types of translocations in brinjal after mutagenic treatments of seeds. Zeerak , 2000, Tokyo Mutagen Dose % of plants with translocation Translocations/100 cells R ing Chain Range Frequency Range frequency Control 0 0 0 0 0 0 Gamma rays (Gy) 10.0 20.0 30.0 9 15 30 0-1 0-3 0-4 28.24 34.60 46.25 0.1 0-3 0-3 71.76 65.40 53-75 EMS 0.8% 1% 4 7 0-1 0-2 27.69 36.23 0-3 0-2 72.31 63.77 Gamma rays +EMS 10.0+0.8% 10.0+1% 20.0+0.8% 20.0+1% 12 14 18 21 0-2 0-2 0-5 0-4 36.23 30.12 45.50 39.33 0-2 0-4 0-3 0-2 63.77 69.88 54.50 60.67

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Table 20: Percentage of pollen and seed sterility in brinjal induced by mutagenic treatments. Zeerak , 2000, Tokyo Mutagen Dose Percentage sterility Pollen sterility Seed sterility Control 0 1.95 3.86 Gamma rays (Gy) 10.0 20.0 30.0 8.35 15.21 22.30 14.28 24.54 36.75 EMS 0.8% 1% 20.55 28.02 28.16 38.75 Gamma rays +EMS 10.0+0.8% 10.0+1% 20.0+0.8% 20.0+1% 25.72 36.65* 29.95 34.62 36.46 52.53* 51.68 45.72

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A-I, Meiotic irregularities in S. melongena. A, First metaphase showing early and late separation of bivalents. B, First metaphase showing stickiness of Chromosomes. C, First anaphase with four chromatin bridges. D, First telophase with laggards. E, First anaphase showing 10-14 chromosome distribution. F, Second telophase showing non- disjunction of chromo somes . G, Second telophase with laggards H, Second telophase showing eliminated chromo somes . I, Second telophase showing eliminated chromosomes forming micronucleus .

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Table 21: Mean Performance of Mutagens on different characters in M 1 generation. Aruna et al .,2010,Tamilnadu

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Common name Name of the mutant released Country of release Year of release Mutagen used Parent variety Main character induced eggplant Daijiro Japan 2004 EMS - - eggplant Floralba Italy 1985 EMS Florida Market shortness eggplant Macla Italy 1983 EMS Florida Market shortness eggplant Picentia Italy 1983 EMS Lunga violetta shortness eggplant PKM 1 India 1985 Gamma rays puzhuthikathiri yield (FAO/IAEA Database, 2010) Achivements

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Table: Mutagenic effectiveness and efficiency in chlorophyll and viable mutation in cow pea var.CO-6 Dhanavel,2008,Tamilnadu Mutagens (conc.) Total No. of plant studied Total No. of plant mutant Mutation frequency % (M) Lethality % (L) Injury (%) (I) Effectiveness Efficiency EMS 0.3% 310 8 2.58 45.24 13.81 143.33 5.70 18.68 0.4% 305 7 2.29 49.57 18.37 95.41 4.61 12.46 0.5% 395 6 2.03 54.88 27.56 67.66 3.69 7.36 DES0.05% 312 9 2.88 40.20 18.71 115.20 5.24 15.39 0.06% 306 7 2.28 44.20 25.52 96.00 5.15 8.93 0.07% 298 7 2.34 64.88 31.81 55.71 3.60 7.35 SA0.03% 275 6 2.18 27.38 17.63 121.11 7.96 12.36 0.04% 304 8 2.63 34.55 26.81 109.58 7.61 9.80 0.05 279 7 2.50 46.09 34.71 88.33 5.42 7.20

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Table: chlorophyll mutations in the M 1 and M 2 Generations of cowpea treated with EMS and MMS Narasinghani and sudhirkumar., 2009, India Sl. No. Treatment Generation Chlorophyll mutations (%) Albina Xantha Chlorina Striata Vifidis total 1 EMS 0.25% 0.25% M 1 M 2 - 4 - 8 - 7 - 2 - 1 00 22 2 MMS 0.025% 0.025% M 1 M 2 - 1 - 2 - 4 - - - - 00 7

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Table: chemical induced mutations in Dwarf French bean variety contender. Pande and Seth, 2009, India Treatment Total M 2 plant No. of mutant plants Mutant character Days to flower Seed yield per plant (gm) EMS 0.15% 97 9 Straw brownish pods with greenish background 43.40 139.42 0.3% 83 7 Meaty pods, fascinated stem and early flowering. 31.70 143.25 0.45% 51 4 Rust brown seeds 36.13 136.29 0.6% 49 6 Dwarf plants high yield 39.16 124.72 MMS 0.15% 117 13 Dark green pods with brown spot at maturity 40.21 139.27 0.3% 89 16 Seed small, pods normal 36.51 141.25 0.45% 52 8 - 40.12 136.52 0.60% 36 4 - 41.25 130.25 Control - - - 49.90 139.27

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Table: Segregation of male sterile plants in F 2 , F 3 and test cross in Chilli . Meshram and Narkhede , 2008, India Cross Generation No. of plants segregating Expected frequencies Goodness of fit (P) Male sterile Male fertile Male sterile X CA 452-1 fertile F 1 42 0 - - - F 2 208 69 3:1 0.95 0.99 F 3 260 85 3:1 0.90 0.95 Test cross Male sterile X F 1 F 1 116 110 1:1 0.90 0.95

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Table 3: Induced viable mutations for various traits in M 2 generation of C hilli . Meshram and Narkhede , 2008, India Gamma rays(kR) Total viable mutants EMS (mM) Total viable mutants Grand total viable mutants Trait 10 20 30 40 50 10 20 30 40 50 Tall mutant - 1 3 1 - 4 2 - 5 1 - 8 13 Profuse branching - 2 4 3 - 9 - 1 7 2 - 10 19 Curved leaf 1 - 4 - - 5 1 - 3 - - 4 9 Diminished morphology 1 - 2 2 4 9 - - 1 - - 1 10 Dwarf plan 1 - 4 2 2 9 1 1 1 2 4 9 17 Flower mutant - 2 6 1 - 9 - 2 12 4 - 18 27 Rosette leaf - - - - - - - - 2 - - 2 2 Pointed fruit apex - 1 1 1 - 3 - 1 5 - - 6 9 Male sterile 1 - 7 4 4 16 - 1 6 1 3 11 27 Long pods - 1 4 1 - 6 - 1 16 1 - 18 24 Dark green pods - - 6 2 2 10 - 2 14 1 - 17 27 Yellowish green pods - 1 4 - - 5 - - 1 - - 1 6 Total 4 8 47 17 12 4 9 67 8 7 190

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Table : Mutagenic effectiveness and efficiency of mutagens in M 2 generation of chilli. Meshram and Narkhede , 2008, India Mutagenic effectiveness Mutagenic efficiency Mutagenic dose Lethality % M 2 families M 2 plants M 2 families M 2 plants Gamma rays(kR) 10 42 0.71 0.32 0.07 0.076 20 45 0.42 0.14 0.19 0.061 30 52 0.36 0.12 0.21 0.068 40 33 0.12 0.06 0.15 0.068 50 20 0.50 0.05 0.124 0.112 EMS( mM ) 10 38 0.21 0.052 0.22 0.054 20 42 0.07 0.025 0.14 0.17 30 49 0.05 0.013 0.12 0.032 40 32 0.02 0.059 0.09 0.029 50 24 0.006 0.005 0.05 0.040

Breeding for polygenic traits:

Breeding for polygenic traits

Breeding for oligogenic traits:

Breeding for oligogenic traits

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