Gamma-rays induced genome wide stable mutations in cowpea deciphered through whole genome sequencing.

PURPOSE: Gamma rays are the most widely exploited physical mutagen in plant mutation breeding. They are known to be involved in the development of more than 60% of global cowpea (Vigna unguiculata (L.) Walp.) mutant varieties. Nevertheless, the nature and type of genome-wide mutations induced by gamma rays have not been studied in cowpea and therefore, the present investigation was undertaken. MATERIALS AND METHODS: Genomic DNAs from three stable gamma rays-induced mutants (large seed size, small seed size and disease resistant mutant) of cowpea cultivar 'CPD103' in M6 generation along with its progenitor were used for Illumina-based whole-genome resequencing. RESULTS: Gamma rays induced a relatively higher frequency (88.9%) of single base substitutions (SBSs) with an average transition to transversion ratio (Ti/Tv) of 3.51 in M6 generation. A > G transitions, including its complementary T > C transitions, predominated the transition mutations, while all four types of transversion mutations were detected with frequencies over 6.5%. Indels (small insertions and deletions) constituted about 11% of the total induced variations, wherein small insertions (6.3%) were relatively more prominent than small deletions (4.8%). Among the indels, single-base indels and, in particular, those involving A/T bases showed a preponderance, albeit indels of up to three bases were detected in low proportions. Distributed across all 11 chromosomes, only a fraction of SBSs (19.45%) and indels (20.2%) potentially altered the encoded amino acids/peptides. The inherent mutation rate induced by gamma rays in cowpea was observed to be in the order of 1.4 × 10-7 per base pair in M6 generation. CONCLUSION: Gamma-rays with a greater tendency to induce SBSs and, to a lesser extent, indels could be efficiently and effectively exploited in cowpea mutation breeding.

Molecular and physiological characterization of early semi-dwarf mutants of rice and localization of SNP variants in Sd1 locus generated through gamma radiation.

PURPOSE: The 'Improved White Ponni' (IWP) rice variety, which is susceptible to lodging, leading to yield losses. Our primary goal is to develop new rice lines with non-lodging traits, enhancing stem strength and resistance to adverse conditions. Additionally, we aim to improve yield-contributing agronomic traits, benefiting farmers, food security, and the environment. Our work contributes to scientific knowledge and addresses a significant issue in Southern Indian rice cultivation. MATERIALS AND METHODS: In the present study, early and semi-dwarf early mutants of IWP were developed without altering the native grain quality traits using gamma ray-mediated mutagenesis. The seeds (500) were irradiated with γ-rays after fixing the Lethal Dose 50 (LD50), and selection for semi-dwarfism and earliness was imposed on a large M2 population. The selected traits were confirmed by evaluating the M3 lines at morpho-physiological, biochemical, and molecular levels. RESULTS: The response of mutants to gibberellic acid has been studied, which identified responsive mutants as well as slow-responding mutant lines including IWP-11-2, IWP-48-2, IWP-50-11, and IWP-33-2. Agar plate assay indicated low α- amylase content in IWP-50-11, IWP-33-2, IWP-43-1, IWP-47-2, and IWP-18-1. The scanning electron microscopy demonstrated that the mutants displayed an increased cellular dimension in comparison to the wild type. In dwarf mutants, null alleles were observed for the SD1 gene-specific primers which depicts gene undergone mutation. Further sequencing revealed the presence of single nucleotide polymorphisms in the SD1 gene resulting in semi-dwarfism in the mutant IWP-D-1. CONCLUSIONS: The impact of a defective gibberellic acid-mediated signaling pathway in mutants to produce a novel high-yielding and early maturing semi-dwarf rice variety.

Development and characterization of gamma ray and EMS induced mutants for powdery mildew resistance in blackgram.

PURPOSE: During post-rainy and rice fallow cropping seasons, popular blackgram varieties are severely affected by powdery mildew leading to severe yield loss. The lack of natural genetic variability for powdery mildew resistance in blackgram germplasm warrants mutation breeding. Hence, in this study, blackgram cultivar CO6 was mutagenized with gamma ray and ethyl methanesulphonate (EMS) to create variability for powdery mildew resistance. MATERIALS AND METHODS: Seeds of blackgram CO6 were irradiated with three doses of gamma ray (200 Gy, 300 Gy and 400 Gy) followed by two doses of ethyl methanesulphonate (EMS; 20 and 30 mM) to achieve six combination treatments. Selected resistant mutants of M2 generation were characterized for agronomic, histological, enzyme and biochemical traits along with powdery mildew resistant LBG 17 and susceptible CO6 checks. Molecular variability was studied using 72 simple sequence repeat (SSR) markers. RESULTS: In the M2 generation, 60 powdery mildew resistant mutants were identified and a total of 25 high yielding mutants were evaluated further to confirm powdery mildew resistance and yield. Nine resistant mutants (PM 13, PM 20, PM 21, PM 42, PM 53, PM 54, PM 56, PM 57 and PM 60) and the resistant check (LBG17) showed significantly higher values for leaf density, trichome density, SOD, CAT, POX, PPO, total phenols, phytic acid and silica content. SSR markers viz., CEDG154, CEDG290, CEDG139, CEDG259, CEDG191, CEDG024, CEDG 282, CEDG 166, CEDG 232 and CEDG 088 were found polymorphic between resistant mutants and the parent CO6. CONCLUSION: The study has demonstrated that sufficient variability was induced in the blackgram for powdery mildew resistance. The elevated levels of SOD, CAT, POX, PPO, total phenols, phytic acid, and silica content observed in selected mutants may be attributed to powdery mildew resistance. The superior mutants identified in this study may be used as donors for the development of powdery mildew resistant lines or released as a new variety.