In silico characterisation and functional validation of chilling tolerant divergence 1 (COLD1) gene in monocots during abiotic stress

Abstract

The G protein-coupled receptor is one of the major transmembrane proteins in plants. It consists of an α subunit, a β subunit and three γ subunits. Chilling tolerant divergence 1 (COLD1) includes a Golgi pH receptor (GPHR) domain, which maintains cell membrane organisation and dynamics, along with abscisic acid linked G protein-coupled receptor (ABA_GPCR) that regulates the signalling pathways during cold stress. In the present study, we performed characterisation of a homologous COLD1 from the economically important monocot species Oryza sativa L., Zea mays L., Sorghum bicolor (L.)Moench and Erianthus arundinaceus (L.) Beauv. IK 76-81, a wild relative of Saccharum. COLD1 was isolated from E. arundinaceus IK 76-81, analysed for its evolution, domain, membrane topology, followed by prediction of secondary, tertiary structures and functionally validated in all four different monocots. Gene expression studies of COLD1 revealed differential expression under heat, drought, salinity and cold stresses in selected monocots. This is the first study on regulation of native COLD1 during abiotic stress in monocots, which has opened up new leads for trait improvement strategies in this economically important crop species.

Comparative de novo transcriptome analysis identifies salinity stress responsive genes and metabolic pathways in sugarcane and its wild relative Erianthus arundinaceus [Retzius] Jeswiet

“…Sugarcane wild relative Erianthus arundinaceus is a potential crop for the production of high biomass and reservoir of many genes related to agronomic, biotic and abiotic stresses. We have isolated and characterized many genes from E. arundinaceus 14 , 15 , 19 – 22 and many of genes are still unexplored. Besides, transcriptome studies were reported for many traits such as cold transcriptome 23 , sucrose 24 , agronomic traits 25 , lignin 28 , red stripe disease 26 , smut disease 27 except for salinity stress.…”

“…Introgression of many genes from E. arundinaceus into sugarcane through conventional and biotechnological approaches has improved the agronomic performance of sugarcane varieties 17 , 18 . We have isolated, characterized, and overexpressed many genes from E. arundinaceus such as heat shock protein 70 14 , 19 , DREB2 20 , glyoxalase gene 15 , α-expansin 1 21 and chilling tolerant divergence 1 (COLD1) gene 22 , which shows the genetic importance of E. arundinaceus in improving the tolerances to biotic and abiotic stresses in sugarcane . There were several RNASeq studies in sugarcane for many traits such as agronomic traits, cold stress, sucrose, lignin, red stripe and smut disease except for salinity tolerance 23 – 28 .…”

In Silico Dissection of Regulatory Regions of PHT Genes from Saccharum spp. Hybrid and Sorghum bicolor and Expression Analysis of PHT Promoters under Osmotic Stress Conditions in Tobacco

“…The quantitative real-time PCR (qRT-PCR) analysis was conducted using the StepOne real-time PCR system (Applied Biosystems, Burlington, ON, Canada). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an endogenous reference control [30][31][32][33][34][35][36][37]. The qRT-PCR profile was as follows: 10 min of denaturation at 95 • C, followed by 40 cycles 15 s of denaturation at 95 • C, 1 min of annealing and extension at 60 • C in a final volume of 25 µL reaction [26,29,30].…”

Isolation, characterization and expression analysis of novel water deficit stress-responsive DEEPER ROOTING 1 (DRO1) gene from drought-tolerant Erianthus arundinaceus

“…To perform qRT-PCR experiments, the cDNA converted using total RNA isolated from root samples of drought-stressed and control plants were used as a template. qRT-PCR was performed on a Step-One Plus Real-Time PCR system (Applied Biosystems, Canada) by following the method explained elsewhere (Anunanthini et al 2019;Manoj et al 2019;AswinNarayan et al 2019;Dharshini et al 2020aDharshini et al , 2020bSwathik Clarancia et al 2020). Each reaction was carried out in triplicates.…”