Advanced Mutation Breeding by Ion Beams. Atsushi TANAKA Quantum Beam Science Directorate, Japan Atomic Energy Agency 20 March 2014

Transcription

1 Advanced Mutation Breeding by Ion Beams Atsushi TANAKA Quantum Beam Science Directorate, Japan Atomic Energy Agency 2 March 214 1

2 TAKASAKI TSURUGA 2

3 Bird s Eye View of JAEA-Takasaki North Gate Ion Beam Irradiation Facility (TIARA) Co-6 Irradiation Facilities Laboratory Bldg. Administration Bldg. Electron Accelerator Bldg. Information Materials Bldg. Experiment Bldg. 1 Takasaki Venture Bldg. N Food Irradiation Facility International Lodging Main Gate 1 3

4 Irradiation Facilities in Takasaki Gamma-ray Irradiation Facility Co-6 gamma-ray source TIARA (Takasaki Ion Accelerators for Advanced Radiation Application ) Japan s First Large-scale Gamma-ray Irradiation Facilities Wide Range of Six-digit Adsorption Rate 8 Irradiation Rooms in 3 Facilities Electron Beam Irradiation Facility Cyclotron Rapid Beam Stabilize & Exchange High-energy Heavy Ion Microbeam Tandem Accelerator Heavy Ion Microbeam & Single Ion Hit Swift Cluster Ions Electron beam irradiation room Japan s First Electron Accelerator for Industrial Scale (2 MV, 3 ma) Ion Implanter Wide Variety of Ions Including Fullerene Combination Use with SEM Single-ended Accelerator High Spatial Resolution Light Ion Microbeam Micro-PIXE Analysis & Proton Beam Writing World s First Facility Specialized for R&D of Materials and Biotechnology More than 8 Joint Research Activities: collaborated with Private companies, Public research institutes, Universities, etc. 4 Irradiation service: 25 subjects (65 millions yen Revenue )

5 Recent Activities in Takasaki Observe Small Tumors Create(Modify) New features Cure Cancer therapy 1 µm Nobel RI-labeled compound Nitrogen fixation Imaging of plant functions using Positron Imaging Fuel cell electrolyte membrane Kusatsu Adsorbent for Scandium Hydrogen separation membrane Resistance evaluation to radiation Effects of heavy ion micro beams New mutants (low cadmium rice) 25 μm (Si K α ) White lung Detection of elements using Micro-PIXE Biodegradable dummy lens JAX A 宇宙用デバイス Semiconductors for aerospace Radiation grafting & cross-linking Evaluation of materials used in Accelerator Ion beam breeding 5

6 FNCA Mutation Breeding Project 4-7 March 214, Indonesia 6

7 IAEA Technical Meeting August 213, Mongolia 7

8 Mutation Breeding in Asia Researched by A. Tanaka (FAO/IAEA Database, October 211) Country No. of cv. Total Rice Barley Wheat Maize Soybean Chrysanthemum All countries China Japan India Russia Netherland Germany USA Bangladesh 44 Indonesia 29 Korea 35 Pakistan 53 Thailand 2 Viet Nam 55 Chemical 6.6% X-ray 9.5% Tissue Culture 15.7% Others 2.1% Ion beam 5.8% γ-ray 6.3% More than half varieties with ion beams created by using TIARA Mutagen (Japan) Nakagawa, TechnoInnovation (27) 8

9 Mutation induced by Ion Beams Energy deposition Irradiation to cell g-rays: low LET radiation 3nm spur (~3nm) Nucleus g-rays: 2, spurs/gy Can ion beams cause different mutation? Ion beams: high LET radiation Core (~3nm) Nucleus Carbon ions: 4 tracks/gy 9

10 Started from 1991 by using TIARA 1

11 Mutation frequency Arabidopsis tt and gl mutant phenotypes W T t t W T g l 11

12 Mutation frequency Mutation frequencies for mutant groups induced by carbon ions and electrons Mutagen (Dose) No. of M1seeds sown No. of M2 plants Mutant group (loci) No. of mutants in M2 Mutation frequency/ locus/ diploid cell/ dose(gy) ( 1-6 ) Carbon ions (15Gy) 26,2 14,88 tt3, tt4, tt5, tt6, tt7, tt18,tt19 gl1, gl2, gl3, ttg1, ttg Electrons (75Gy) c.a.15,6 8,827 tt3, tt4, tt5, tt6, tt7, tt18,tt19 gl1, gl2, gl3, ttg1, ttg times higher per dose, and 4 times higher per plant (seed) Shikazono et al. Genetics(23) Shikazono et al. JXB (25) 12

13 Mutation spectrum Mutation breeding system in chrysanthemum Acclimatization Callus induction from leaf or petal Original var. Taihei With pink-color petals Plantlet culture Gamma irradiation or Ion beam irradiation Regeneration 13

14 Mutation spectrum T a b l e F l o w e r m u t a t i o n o f r e g e n e r a t e d p l a n t s f r o m f l o r a l o r g a n c u l t u r e i n d u c e d b y c a r b o n i o n s a n d g r a y s M u t a g e n M u t a t i o n f r e q u e n c y o f m u t a t e d c h a r a c t e r ( % ) W h i t e L i g h t p i n k D a r k p i n k O r a n g e Y e l l o w C o m p l e x / S t r i p e U n i r r a d i a t e d g r a y s C a r b o n i o n s Original var. TAIHEI ION NO KOKI ION NO SEIKO ION NO MAHO First cultivation of new chrysanthemum varieties using ion beams (1998) 14

15 Mutation spectrum Flower mutation of carnation regenerated from leaf cultures treated by mutagen Mutagen Light pink Pink Dark pikn Mutation frequency( 1-1 %) Flower color Red Salmon Yellow Cream Stripe Complex Round petals Shape Dianthus type petals EMS Soft X-rays Gamma-rays Carbon ions Mutation spectrum is much broader Okamura et al. NIMB (23) 15

16 Mutation spectrum New varieties of carnation induced by ion beams Original var. Vital Commercialized in 22 EU & Japan ($ 13 million in 21) with KIRIN Brewery Co.Ltd. 16

17 Model plants Novel mutants and varieties Crops Ornamentals Trees & fruit Microorganism UV-B resistant Blast resistant rice Bacterial-Leaf-Blight resistant rice New flower-color chrysanthemum Ficus with high NO 2 assimilation Yeast producing higher amount of ethyl caproate Frilled flower Potato Virus Yellow resistant tobacco Carnation with new flower color/shape Wax rich conifer Protease high-producing Aspergillus Flavonoid accumulation Yellow Mosaic Virus resistant barley Chrysanthemum with reducing axillary buds Melon with good growth in low temperature High temperature resistant rizobium Hyper-nodulation Low-nitrogen-fertilizer grown rice Osteospermum with new pastel coloration Thornless mutant of Yuzu (Citrus junos) Oyster mushroom with new character 17

18 New Varieties with Kagoshima Biotechnology Institute Greatest goal in chrysanthemum: reducing axillary buds Jimba Ion beams Aladdin Ion beams Aladdin 2 Original Var. Reduced Axillary Buds Reduced Axillary Buds + Early Flowering Products: 22 Million flowers (212) Requested license from more than 35 marketing cooperatives Ueno et al. JJSHS (213) 18

19 New Varieties Shizuoka prefecture Saving energy: Netted melon with a large fruits under low temperature (cut down on heating expense) 19 New variety as Shizuiku No.1 ( 静育 1 号 ) was established in 211. Grown in 2 below & 2 % cost saving

20 NO2 absorbance(μg/g) New Variety Phytoremediation of air pollution by greening with Hiroshima University 7 Ficus pumila 6 5 WT KNOX WT KNOX 4-8% Up of the capability to assimilate atmospheric nitrogen dioxide (NO 2 ) Press release (Aug. 27) Technology transfer (29) & Commercialized (21) 2 Sep. 213 Tokyo Station Utilized for wall greening, Takahashi et al. J. Phytoremediation (212) green roof, etc. 2

21 New Variety A Solution to Eutrophication - Rice breeding suitable for low nitrogen input - Biwako Lake with Shiga Prefecture Original var. Akino-uta Low-nitrogen-fertilizer grown rice mutant var Press Announcement 21

22 Food safely & security: development of low-cadmium Koshihikari rice Grain Cd New Variety WT: コシヒカリ Koshihikari Low-cadmium mutant lcd-kmt1 variety WT: コシヒカリ Koshihikari Low-cadmium mutant lcd-kmt1 variety Press release Press release Conclusive evidences WT: Koshihikari 1. The same yield and good taste as Low-cadmium original Koshihikari mutant variety 2. Three independent mutants on the same gene: transporter responsible for manganese Ion beam Field breeding A Field B is definitely Field C (High Cd) (High Cd) (High Cd) suitable for staple grain! Maximum allowed Cd concent. Ishikawa et al. Proc. Natl. Acad. Sci.USA (212) 22

23 New Varieties Microbial Ion Beam Breeding in Yeast Aims Development of a new variety superior to the Gunma prefecture original yeast Gunma KAZE Yeast Target phenotype: Collaborative research with the Gunma Industrial Technolgy Center Improvement of alcohol fermentation property High production of good flavor component (Increase of ethyl caproate Fruity flavor) Current status Press release Success in obtaining a new mutant variety that produces a higher amount of ethyl caproate than Gunma KAZE Yeast New variety No.277 Now on sale! (from April 213) 23

24 Chromosomal DNA A gene Mutation Analysis Intragenic point-like mutation TT4 locus (395 a.a.) Electron TC AA Ser 196 Asn 196 Carbon ion A deletion at 259 a.a Carbon ion 8 base deletion A at 14 a.a Intron Exon Ion beams induce deletion, leading to null mutation Electron T A Ileu 252 Asn 252 Carbon ion A deletion at 259 a.a. Carbon ion 22 base deletion at 344 a.a. 24

25 Mutation Analysis Large DNA alteration A gene Chromosomal DNA T T 4 b r e a k d e l e t i o n I n v e r s i o n t t 4 ( C 1 ) C h r o m o s o m e 5 b r e a k r e j o i n r e j o i n d e l e t i o n Ion beams induce large DNA alteration with deletion t t g C h r o m o s o m e 5 C h r o m o s o m e 3 C h r o m o s o m e 3 g l 1-3 C h r o m o s o m e 2 T T G 1 b r e a k b r e a k A t p k 7 b r e a k G L 1 b r e a k b r e a k b r e a k d e l e t i o n d e l e t i o n d e l e t i o n d e l e t i o n d e l e t i o n Reciprocal translocation Inversion Insertion r e j o i n r e j o i n r e j o i n r e j o i n 25

26 Effect of ion beams for mutation induction Energy deposition Irradiation to cell Mutation Gamma rays 3 nm Spur (~3 nm) Cell nucleus Gamma rays: 2, spurs/gy Point mutation: base substitution, frame shift A gene: ~2, nm Ion beams Core (~3 nm) Cell nucleus Carbon ions: 4 tracks/gy Large DNA alteration: deletion, translocation, inversion Ion-beam induced mutation for plant breeding 1. High mutation rate :Small samples & spaces for screening 2. Broad mutation spectrum :Producing new varieties and mutants 3. Minimum No. of DNA damage :Pinpoint-breeding without bad characters Tanaka et al. JRR (212) 26

27 TIARA Users for Ion Beam Breeding in Japan ARTI, Korean Atomic Energy Research Institute (KAERI) Forum for Nuclear Cooperation in Asia (FNCA) 群馬県農業技術センター群馬県林業試験場群馬県産業技術センターサッポロビール森産業カネコ種苗東洋大学前橋工科大学 青森県農林総合研究センター弘前大学 秋田県農業試験場 新潟農業総合研究所 北海道大学 東北大学宮城県農業 園芸総合研究所 Malaysian Nuclear Agency 福岡県農業総合試験場福岡農業協同組合 佐賀県果樹試験場 大分県農林水産研究センター大分ゼネラルサービス 熊本県立大学 鹿児島県バイオテクノロジー研究所 高知県農業技術センター 京都府農業資源研究センター京都府立大学 アースノート 広島大学景設計 島根大学 岡山大学 埼玉県農林総合研究センター埼玉大学朝日工業 金沢大学 兵庫県農林水産技術総合センター 住化農業資材 ミヨシ 岐阜県生物工学研究所 大阪大学石原産業 滋賀県農業技術振興センタータキイ研究農場 東京大学東京農工大学東京農業大学名糖産業国際基督教大学 愛知県農業総合試験場名城大学 栃木県農業試験場キリンアグリバイオ JT 葉タバコ研究所上都賀農業振興事務所 放射線育種場森林総合研究所筑波大学 千葉大学東京理科大学千葉県農業総合研究センターキッコーマンヤマサ醤油 静岡県農林技術研究所静岡大学石井育種場横浜植木 27

28 Malaysia:gamma-rays, ion beams D. jayakarta Original Variety Mutant (Gamma-ray) D. mirbellianum Original Variety 2 mutant lines resistant to thrips Mutant (Ion-beam) 1 mutant line resistant to both mites and thrips technology transferred to commercial laboratory at Nuclear Malaysia Economic Effect: USD 16,/year (214-) 28

29 Vietnam 28 Sep. 21 Original var. Khang dan Lodging-resistant Lodging-resistant mutant (Gamma-rays) mutant (Ion beams) High yielding 29

30 Bangladesh Salt tolerance & High yielding 3