Molecular tagging of seed size using MITE markers in an induced large seed mutant with higher cotyledon cell size in groundnut.

A large seed mutant, TG 89 having a 76.7% increment in hundred kernel weight in comparison to its parent TG 26, was isolated from an electron beam-induced mutagenized population. Studies based on environmental scanning electron microscopy of both parent and mutant revealed that the mutant seed cotyledon had significantly bigger cell size than parent. A mapping population with 122 F2 plants derived from the mutant and a distant normal seed genotype (ICGV 15007) was utilized to map the QTL associated with higher HKW. Bulk segregant analysis revealed putative association of three markers with this mutant large seed trait. Further, genotyping of F2 individuals with polymorphic markers detected 14 linkage groups with a map distance of 1053 cM. QTL analysis revealed a significant additive major QTL for the mutant large seed trait on linkage group A05 explaining 12.7% phenotypic variation for the seed size. This QTL was located between flanking markers AhTE333 and AhTE810 having a map interval of 4.7 cM which corresponds to 90.65 to 107.24 Mbp in A05 chromosome, respectively. Within this genomic fragment, an ortholog of the BIG SEEDS 1 gene was found at 102,476,137 bp. Real-time PCR revealed down-regulation of this BIG SEEDS 1 gene in the mutant indicating a loss of function mutation giving rise to a large seed phenotype. This QTL was validated in 11 advanced breeding lines having large seed size from this mutant but with varied genetic backgrounds. This validation showcased a highly promising selection accuracy of 90.9% for the marker-assisted selection. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03909-0.

Development of candidate gene-based markers and map-based cloning of a dominant rust resistance gene in cultivated groundnut (Arachis hypogaea L.).

A dominant rust resistance gene, VG 9514-Rgene was isolated through map-based cloning. Sequence analysis revealed non-synonymous mutations in the TIR, NBS and LRR region of the R-protein. Candidate gene-based markers from these SNPs revealed complete co-segregation of the isolated VG 9514-Rgene with rust resistance in a RIL population and confirmed their map position in between FRS 72 and SSR_GO340445 markers in arahy03 chromosome. Blastp search of VG 9514-Rprotein detected Arahy.T6DCA5 with >80.0% identity that localized at 142,544,745.0.142,549,184 in arahy03 chromosome. Ka/Ks calculation revealed that VG 9514-Rgene had undergone positive selection compared to four homologous genes in the groundnut genome. Homology based structure modelling of this R-protein revealed a typical consensus three-dimensional folding of TIR-NBS-LRR protein. Non-synonymous mutations in susceptible version of R-protein were mapped and found E268Q mutation in hhGRExE motif, Y309F in RNBS-A motif and I579T in MHD motif of NB-ARC domain are probable candidates for loss of function.

Identification of major consensus QTLs for seed size and minor QTLs for pod traits in cultivated groundnut (Arachis hypogaea L.).

Hundred kernel weight is an important indicator for large-seeded genotype selection. A recombinant inbred line population was used to decipher the genetic architecture of seed size and three pod traits in cultivated groundnut based on the phenotypic data from six and three environments, respectively. The study revealed a consensus major QTL for HKW in B07 group that explained 10.5-23.9% phenotypic variation due to seed size. Further, two other minor QTLs were identified in B03 and B08 group for the seed size. Two minor QTLs for pod beak were positioned in B03 and A08. A minor QTL for pod reticulation was also mapped in the same map interval with the pod beak QTL in A08. Another minor QTL for pod constriction was co-mapped with the minor QTL for HKW in B08. The other minor QTL for pod constriction was placed in the neighboring map interval with the consensus QTL for seed size in B07 that suggests linkage of pod constriction with large seed trait. Analysis of the flanking markers profile in 71 cultivated groundnut genotypes revealed a strong association of pPGPseq_2E06 marker with large seed trait.

Mapping of a dominant rust resistance gene revealed two R genes around the major Rust_QTL in cultivated peanut (Arachis hypogaea L.).

KEY MESSAGE: A consensus rust QTL was identified within a 1.25 cM map interval of A03 chromosome in cultivated peanut. This map interval contains a TIR-NB-LRR R gene and four pathogenesis-related genes. Disease resistance in plants is manifested due to the specific interaction between the R gene product and its cognate avirulence gene product (AVR) in the pathogen. Puccinia arachidis Speg. causes rust disease and inflicts economic damages to peanut. Till now, no experimental evidence is known for the action of R gene in peanut for rust resistance. A fine mapping approach towards the development of closely linked markers for rust resistance gene was undertaken in this study. Phenotyping of an RIL population at five environments for field rust score and subsequent QTL analysis has identified a 1.25 cM map interval that harbored a consensus major Rust_QTL in A03 chromosome. This Rust_QTL is flanked by two SSR markers: FRS72 and SSR_GO340445. Both the markers clearly identified strong association of the mapped region with rust reaction in both resistant and susceptible genotypes from a collection of 95 cultivated peanut germplasm. This 1.25 cM map interval contained 331.7 kb in the physical map of A. duranensis and had a TIR-NB-LRR category R gene (Aradu.Z87JB) and four glucan endo-1,3 ß glucosidase genes (Aradu.RKA6 M, Aradu.T44NR, Aradu.IWV86 and Aradu.VG51Q). Another resistance gene analog was also found in the vicinity of mapped Rust_QTL. The sequence between SSR markers, FRS72 and FRS49, contains an LRR-PK (Aradu.JG217) which is equivalent to RHG4 in soybean. Probably, the protein kinase domain in AhRHG4 acts as an integrated decoy for the cognate AVR from Puccinia arachidis and helps the TIR-NB-LRR R-protein to initiate a controlled program cell death in resistant peanut plants.

Electron beam irradiation revealed genetic differences in radio-sensitivity and generated mutants in groundnut (Arachis hypogaea L.).

Electron beam accelerators are being used for many industrial applications including food and agriculture. A 10MeV linear accelerator facility was standardized for low dose application 0.1-1 kGy) in pulse mode using unscanned scattered beam for irradiation of groundnut seeds for mutation breeding. Using this facility, 50% growth reduction (GR50) dose was standardized in five groundnut genotypes. There were significant differences for radio-sensitivity among these genotypes. Seed mutagenesis of two groundnut genotypes, TG 26 and TG 68 with electron beam has generated one large seeded and four high yielding mutants in preliminary field trials.

Inheritance and molecular mapping of a gibberellin-insensitive dwarf mutant in groundnut (Arachis hypogaea L.).

A dark green dwarf mutant, TGM 167, was isolated from a gamma ray + sodium azide mutagenized population of cultivated groundnut breeding line, TFDRG 5. The mutant had a 45.8% reduction in height due to its shorter internodal length. Further, it was found to be insensitive towards exogenous GA(3) application, although it had nearly the same level of endogenous GA(3) as the parent. Genetic analysis revealed that the dwarfism is under the control of a single dominant gene. This dominant dwarfing gene was mapped with an SSR marker TC3H02 at a distance of 9.7 cM.