Tyagi, WrichaVerma, Satish Kumar M.2023-01-272023-01-272017-12https://krishikosh.egranth.ac.in/handle/1/5810192813Pigeon pea belongs to legume family which has a specific physiology including organs like nodule and pod. The understanding of genes/transcripts involved in development of various organs of pigeon pea is limited. The present study was conducted to identify tissue specific transcripts from pigeon pea by ortholog-based approach and to isolate and characterize 5'UTR region of tissue specific transcript(s) from pigeon pea. An in silico search was conducted using phytozyme database and plant transcription factor database (PTFDB) separately for four tissues viz, seed, flower, leaf and pod. A total numberof 39 putative transcription factors (TFs) belonging to 18 different TFs families were identified for pod. The highest numbers of transcripts were observed for GATA and C2H2 (Cys (2) His (2)) zinc finger family members, and similarly, 54 TFs belonging to16 different TFs families were identified for flower tissue. Higher transcript numbers were observed for MYB (Myelobastosis), MIKC (MADS Intervening K-box Cterminal) family. Likewise, a total number of 92 TFs belonging to 28 different TFs families were identified for leaf tissues. In this case the highest number of TFs belonged to C2H2 and NAC (No Apical Meristem CUC domain) containing families. Similarly, a total number of 23 TFs with 12 different TFs families were identified expressing in seed were like MYB, NAC. Based on in silico analysis 10 transcription factors for each of the four tissues were identified. Out of the 10 transcription families (36280, 13238, 20528, 25279, 13634, 00220, 11572, 11926, 13354, 19191) identified for seed specific expression, five (00220, 11572, 11926, 13354, 19191) were targeted for RT-PCR using five different tissues (seed, flower, leaf, pod and stem). Simultaneously, members of auxin and TCP transcription family were also targeted for validation. Internal gene for studying tissue specificity in pigeon pea was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). This gene was selected based on RT-PCR of ß-actin (ACT), (GAPDH), ubiquitin family (UBI), elongation factor 1-A (EF1A) and ß-tubulin (TUB) on five different tissues (seed, flower, leaf, pod and stem). Our RT-PCR data led to conformation of 2 transcripts, 00220 and 11572 as seed specific pigeon pea transcripts. While 00220 is a member of MADS (MCM1/AGAMOUS/ DEFIEIENS/SRF) transcription family,11572 is a member of TALE (Three Amino acid Loop Extension) transcription family.Thermal asymmetric interlaced PCR (TAIL PCR) approach led to identification of 229 bp 5' UTR (Untranslated Region) of 00220 transcript. Upon sequencing and analysis using Plant CARE database presence of cis-acting elements like ARE,CAAT-box, G-box, GT1-motif, Skn-1 motif, and TATA-box was detected, three unnamed motifs were also identified at -10 and -66 position. The transcripts identified and putative promoter isolated in this study after further characterization might generate useful information regarding development in pigeon pea.EnglishTransciptionPigeonpeasThesis