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Arvind Kumar,Antony Ginny,Rajesh M.K.,Josephrajkumar A.,Grace Tony 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.2
Oryctes rhinoceros L. (Coleoptera: Scarabaeidae), the coconut rhinoceros beetle (CRB), is a primary pest of co conut in South and Southeast Asia and the Pacific Islands. The beetle has the potential to severely affect the economies of local communities, many of whom are marginal and small farmers who rely on coconut as the main source of livelihood. Reverse transcription-quantitative real-time PCR (qRT-PCR) based targeted gene expression analysis has emerged as a powerful tool due to its sensitivity and reproducibility. However, calculating the relative expression of target genes requires normalization with reference genes across specific experimental conditions. To identify suitable reference gene(s) possessing stability, we selected six prospective genes (viz., NADH, ACTIN, EF1A, RPL3, SDHA, and ARF6) and evaluated them for their potential use as reference gene(s) across different developmental stages of O. rhinoceros. A comprehensive approach based on five statistical models viz., GeNorm, BestKeeper, NormFinder, RefFinder and the ΔCt value, was utilized, and based on the obtained stability values of candidate genes, a consensus ranking was generated. The expression levels of NADH, EF1A and RPL3 were observed to be the most stable across the developmental stages with significant statistical reliability. Further, this study identified NADH/EF1A as the most reliable reference gene combination which could provide robust normalization of RT-qPCR data in gene expression studies in O. rhinoceros. This is the first report iden tifying the suitable reference genes for normalizing gene expression in O. rhinoceros across different develop mental stages, facilitating future elucidation of gene expressions in this species.
Regulation of coconut somatic embryogenesis: decoding the role of long non-coding RNAs
Sabana A. A.,Antony Ginny,Gangaraj K. P.,Grace Tony,Rajesh M.K. 한국식물생명공학회 2024 Plant biotechnology reports Vol.18 No.1
Long non-coding RNAs (lncRNAs) are transcripts longer than 200 nucleotides that lack significant protein coding potential and have been shown to regulate various biological processes. This study was designed to identify lncRNAs in coconut and their role in the process of somatic embryogenesis in coconut, a crop with high recalcitrance to in vitro culture. RNA-Seq data of coconut embryogenic calli of the West Coast Tall cultivar was exploited for in silico prediction of lncRNA. From a total of 6328 transcripts, which were annotated as uncharacterised or with no homology hits with the existing database, 5110 putative lncRNAs are identified. We also studied the relationship between lncRNAs, microRNAs (miRNAs) and mRNAs and found that some of the lncRNAs act as miRNA precursors, some as potential miRNA targets and some function as endogenous target mimics (eTMs) for miRNAs. Real-time quantitative PCR confirmed that 10 selected lncRNAs showed significant differences in the expression pattern in different stages of coconut somatic embryogenesis. Our results suggest the existence of diverse lncRNAs in coconut embryogenic calli, some of which are differentially expressed. The information generated in this study could be of great value in understanding the molecular mechanisms governing somatic embryogenesis in coconut.