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    Mutagenesis Intro

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    krishnamruthatsuresh
    The document discusses mutagenesis methods for enhancing desirable traits in crop plants. It describes using chemical mutagens like EMS to induce mutations and generate genetic variability in rice varieties. Traits that were improved in one variety through EMS mutagenesis include complete panicle exertion, strong culm, improved root architecture, and resistance to bacterial blight, brown plant hopper, and yellow stem borer. It also provides details on the methodology used, including maintaining a large mutagenized population and next-generation sequencing to identify causal mutations.

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    Mutagenesis Intro

    Uploaded by

    krishnamruthatsuresh
    13 views11 pages
    The document discusses mutagenesis methods for enhancing desirable traits in crop plants. It describes using chemical mutagens like EMS to induce mutations and generate genetic variability in rice varieties. Traits that were improved in one variety through EMS mutagenesis include complete panicle exertion, strong culm, improved root architecture, and resistance to bacterial blight, brown plant hopper, and yellow stem borer. It also provides details on the methodology used, including maintaining a large mutagenized population and next-generation sequencing to identify causal mutations.

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    The document discusses mutagenesis methods for enhancing desirable traits in crop plants. It describes using chemical mutagens like EMS to induce mutations and generate genetic variability in rice varieties. Traits that were improved in one variety through EMS mutagenesis include complete panicle exertion, strong culm, improved root architecture, and resistance to bacterial blight, brown plant hopper, and yellow stem borer. It also provides details on the methodology used, including maintaining a large mutagenized population and next-generation sequencing to identify causal mutations.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    13 views11 pages

    Mutagenesis Intro

    Uploaded by

    krishnamruthatsuresh
    The document discusses mutagenesis methods for enhancing desirable traits in crop plants. It describes using chemical mutagens like EMS to induce mutations and generate genetic variability in rice varieties. Traits that were improved in one variety through EMS mutagenesis include complete panicle exertion, strong culm, improved root architecture, and resistance to bacterial blight, brown plant hopper, and yellow stem borer. It also provides details on the methodology used, including maintaining a large mutagenized population and next-generation sequencing to identify causal mutations.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 11

    Mutagenesis: An approach for enhancing desirable

    traits in crop plants


    • Introduction
    • Mutagenesis methods
    • Traits improved by EMS mutation of BPT5204
    • Complete Panicle Exertion
    • Strong culm
    • Root system architecture
    • Bacterial blight resistance
    • Brown Plant Hopper
    • Yellow Stem Borer resistance
    • Methodology
    • Implications
    1 03/10/2024 Add a footer
    Mutagenesis: Basics and methods
    • A tool for gene functional studies and to create genetic variability
    • Novel variants for morphological, physiological, and biotic stress
    tolerance traits, induced mutations
    Spontaneous mutation rates in higher plants are low, ranging from 10 −5 to 10−8

    Chemicals
    Physical mutagens Sodium azide, ethyl
    Gamma rays, alpha methanesulphonate
    TILLING particles, UV and (EMS), methyl
    radioactive decays
    methanesulphonate
    (MMS), hydrogen
    fluoride (HF), diethyl
    sulphate,
    hydroxylamine and N-
    Insertional
    mutagenesis methyl-N-nitrosourea
    T DNA tagging Error prone PCR (MNU)
    RNAi
    VIGS EMS preferentially alkylates
    Transposon AC/Ds G residues, inducing preponderantly
    C-to-T and G-to-A transitions (CG > TA) Scheme of developing mutant lines in crops
    2 03/10/2024 Add a footer
    Mutagenesis in crops: Status

    Major Rice mutant projects in India

    ICAR-IIRR and CSIR-CCMB : BPT 5204 Samba Mahsuri


    Multi institutional Network project - N22
    IGKV Raipur and Bhabha Atomic Centre Trombay

    3 03/10/2024 Add a footer


    Mutation mapping and identification of causal SNP: CRITICAL points
    • Selection of the appropriate mutagen
    • Concentration and time of mutagen
    • Maintenance of a large population
    • Phenotyping
    • Record maintenance
    • Selection of a method for analyzing causal SNP Phenotyping
    Bulking on the basis of phenotyping
    • Methodology Equimolar concentration of DNA/ RNA
    RNA-bulk Seq
    • Generation of MutMap population
    • Generation of bi-parental mapping population
    • Generation of back cross population
    • Application of Next-Generation sequencing
    • Bioinformatics data analysis
    • Selection of pipeline and parameters

    4 03/10/2024 Add a footer


    Methods and analyzing the data

    MutMap gap method


    MutMap method MutMap+ protocol
    5 03/10/2024 Add a footer
    DNA level
    MutMap

    Expression
    based
    RNA-bulk
    Seq

    Simplified modified MutMap


    method scheme QTL-seq framework
    MutMap+
    Zegeye et al., 2018
    6 03/10/2024 Add a footer
    EMS mutagenized population of Samba Mahsuri
    10,500 Samba
    Mahsuri seeds
    EMS (1.2% & 0.8%)

    M1 Generation
    (10,500)

    M2 Generation
    (10,500)
    155
    M3 Generation

    31
    M4 Generation

    22

    M5 Generation

    Distribution of mutants for various agro-morphological


    CSIR-PLOMICS 13Phase I traits in M2 generation
    CSIR-FBR-Phase-II
    Towards producttodevelopment
    Forwarded in rice using
    further generations
    mutants that have traits of agronomic importance
    Gopi et al., 2020 (https://doi.org/10.1371/journal.pone.0258816)
    2-4cm panicle
    choking

    Wild type panicle Mutant panicle

    Variants for different traits in the mutagenized population of Samba Mahsuri (A—M); W- Wild type Samba Mahsuri; M- Mutant; K1- Wild type with score 9; K2- Resistant mutant
    (SB 6) with score 1 and K3- Susceptible check TN 1 with score 9.

    8 03/10/2024 Add a footer


    (a) Basal Leaf sheath: colour (b) Leaf: Auricle (c) Leaf: Collar (d) Leaf: Shape of ligule (e) Spikelet: colour of Stigma (f) Stem: Thickness

    Semierect Horizontal
    Semi straight Deflexed Drooping
    (h) Flag Leaf: Attitude of blade (late observation) (i) Panicle: Curvature of main axis
    (g) Stem: Nodes & inter nodes

    (j) Lemma & palea colour in wild type & mutants with different grain types (k) Colour of tip of lemma Strong Clustered
    (l) Panicle: Secondary branching

    Suneel et al., 2020


    Next Generation Sequencing Sequence statistics of Samba Mahsuri (wild type) and mutant lines with
    different traits
    Mapped QC
    Type or trait of Number of Raw QC Passed Sequen ced Reference
    S. No. Line Cover age (X) passed Reads Mapp ed Reads (%)
    Line Reads (R1+R2) Reads (R1+R2) bases (Gb) used
    (R1+R2)
    1 SM-E Wildtype 112285084 102241208 15.34 39.22 100885182 98.67
    2 SM-G Wildtype 110937222 111916395 16.79 42.94 110657624 98.88 R498
    3 SM-C Wildtype 123573016 111003694 16.65 42.59 122513940 98.27
    Complete Panicle
    4 TI-109 114514030 114191395 17.13 43.81 103577844 98.73
    Exsertion
    Complete Panicle
    5 TI-110 107814208 96892528 14.53 37.17 96892528 98.77
    Exsertion
    High Yield; High
    6 TI-128 Spikelets 113249162 102101092 15.32 39.17 102101092 98.89
    per Panicle
    Elongated
    7 TI-35 Uppermost 108635160 108364345 16.25 41.57 97603326 98.76
    Internode Samba
    Mahsuri
    Complete Panicle
    8 TI-38 113152500 102614244 15.39 39.37 102614244 98.86
    Exsertion
    Bacterial Blight
    9 TI-42 106403198 78348650 11.75 30.06 78348650 98.62
    Tolerance

    High Yield; High


    10 TI-50 109924526 99290257 14.89 38.09 99290257 98.9
    Spikelets per
    Panicle
    Sheath
    11 SB-6 126418290 125999511 18.9 48.34 112679044 98.62
    Blight Tolerance
    Related analysis indicated a high similarity among the mutant lines which is seen as single cluster
    containing majority of the mutant lines

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