• To validate of chloroplast genome-based markers
• To identify cross species orthologos marker transferability in red sanders
• To identify metabolic biomarkers specific to red sanders.

Location & agro climatic sub region

:

Gujarat/ Karnataka/Andhra Pradesh etc.

Name of Research scheme and B.H.

BH: 12099

Year and Season

:

2022

Organisms

:

Red Sanders (Pterocarpus santalinus. L.)

Statistical Design

-

Experimental detail

:

• Sampling site: Various location of Gujarat as well as from Karnataka/ Telangana/ Andhra Pradesh (As per available resources) .

• Sample collection time: Round the year  
• Molecular work :
  • • Identification of cytoplasmic genome markers set for  red sanders specific polymorphism

Methodology /Procedure:

• DNA extraction  followed by Quantitative and qualitative analysis of extracted genomic DNA using  electrophoratic and Nanodrop analysis.
• Rbcl, Matk, ITS etc marker will be used for cpDNA region amplification
• DNA amplification will be carried out by termalcycler machine followed by electrophoratic analysis.
• Gel snap will be documented under UV gel documentation system.
• Amplicone size polymorphism (bp)
• Nucleotide level polymorphism of DNA sequence
• Phylogenetic analysis of species.

• • • Identification of the molecular markers forms the Conserved Ortholog Set Markers (COS) from genome database of Fabaceae family and related tree species to study cross species transferability.

Methodology /Procedure:

• • • • Database search for DNA based marker from Fabaceae/ Leguminosae  tree Genome database/resources
      • Synthesis of primers/markers
      • PCR amplification
      • Agarose Gel electrophoresis
      • Documentation of Gel profiling
      • Identification of polymorphic markers specific to red sanders.
    • Chromatography based metabolite profiling of  P.santalinus  for identification of biomarkers specific to red sanders.

Methodology /Procedure:

• Extraction of total metabolite from bark/leaf samples of more than one year old seedlings of P.santalinus, P. marsupium and etc related species.
• Derivatization in organic solvent
• Chromatagraphy separation of total metabolite
•  Identification of P.santelinus specific metabolites .

Observation to be recorded

:

• DNA quantification and quality analysis
• Polymorphic bands
• Monomorphic bands
• PIC value
• DNA Sequence data acquisition by  Sanger’s/ NGS platform (As per available resources )
• Data analysis on various Bioinformatics platforms e.g. NCBI, BioEDIT, SnapGEne, Metaboanalyst etc.

Expected Outcome

:

The data set developed through above said investigation will be useful for authentication of Red sanders planting material/samples and would be useful for future molecular investigation of the different aspects of spatial and temporal population genetic structure, identification of diversity hotspots and tree improvement in order to address the major concern of farmers of Gujarat related to quality planting material and authentication of species.

Statement of progress  

The sample collection of Red sanders and Bijsal from  the various location of Gujarat as well as from Karnataka/ Telangana/ Andhra Pradesh is under process. Seedlings have been grown from the seed sources which are already been collected. Molecular work will be initiated as soon as total samples will be collected.