Research Projects

Genesis: The current proposal is built on the leads from the National Medicinal Plant Board-funded grant on ‘Development of high-yielding pellitorine-rich Akarkara variety and its region-specific field trials'. Akarkara Anacyclus pyrethrum (Asteraceae) is an indispensable medicinal plant in Ayurveda, Unani, Siddha, Traditional Arabic and Islamic Medicine (TAIM). Currently, it is listed as globally vulnerable on the IUCN Red List of threatened/endangered species. PI, at CSIR-CIMAP, initiated from germplasm collection across Northern India with limited (0.1mg/g d.w. basis) Pellitorine content in roots (2015). An intensive chemical mutagenesis breeding programme was undertaken, which led to a valuable forward genetic screen resource of the functional mutants, to be used for systematic hybridizations and genetic characterization to develop high-yielding varieties, to meet the demand-supply gap through the Akarkara metabolite-targeted breeding program. Various agronomically desired stabilized functional mutants currently in M7 were retrieved and further used as parents in the genetic improvement of Akarakara (Anacyclus pyrethrum var depressus). Scientific Rationale: Anacyclus pyrethrum (Anthemideae; 2n=2x=18; 2C=13.1Gbp) is a large genome, self-incompatible, cross-pollinated species. Developing mapping populations such as Recombinant Inbred Lines (RILs) to understand genetic inheritance of a trait, MutMap population where the two parents only differ for the mutated loci (99.9% similar) regions and validation by classical gene complementation using double mutants and gene segregation analysis in F2 MutMap populations through targeted KASP panels for candidate causal SNPs mapping is most economic, precise, and reliable strategy, to fine map mutations. In this proposal, we aim to map the Anacyclus loss of function leaf mutant phenotype that has a thickened, condensed leaf pinna, short internodes, compact plant type, delayed flowering, and reduced number of flowers, and the root with high N-alkylamide (Pellitorine) content. The delayed flowering led to long vegetative growth, higher photosynthate accumulation being channelized to the underground sink, the alkylmide-rich (Pellitorine) roots. Possibly a major meristem regulator like homologues of TCP family KNOTTED 1-like homeobox, SHOOT MEISTEMLESS (STM), CLAVATA1(CLV1), knotted1(kn1) etc., controlling the balance between shoot apical meristem maintenance and organogenesis, affecting the development of both leaf and flower, stands mutated. Specific Aims: The aim is to identify mutation-causal candidate genes and SNPs in the exonic regions that lead to probable amino acid changes or frameshift changes in promoters that lead to probable transcriptional changes through DGE and MutMAP. The genetics of the phenotype/trait will be studied from its gene segregation analysis in RILS. The integrative characterization of the leaf mutants based on gene expression, anatomy, photoperiod, and carbon metabolism interaction with metabolic flux, metabolite profiling at various developmental stages, and ROS as a signal molecule will be studied. The validated SNPs with a confidence level 95-99% will be used to map the mutations in the F2 MutMap population through KASP assays. Novelty: Such forward genetic screens and mapping populations are not reported from the genus Anacyclus, which is a prerequisite to study the genetics of the trait and mapping of the mutation hence will be a novel study. Candidate gene/pathways deciphered for this trait will generate a genomic resource that could be utilized for genetic improvement and breeding of Anacyclus pyrethrum for various other economic traits. Long coding RNA dataset from the transcriptome data will help to better understand the gene interaction. Such valuable characterized mutants can be commercially utilized for basic as well as applied studies in Analcyclus and metabolite-targeted breeding programme.