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| Cloning of Cold Tolerance-related Genes RsMYBS3 and RsRCI2B of Rhododendron sphaeroblastum var. wumengense and Their Expression Analysis |
| PENG Wan-Ming, TAN Yi, ZHANG Xiao-Li, LUO Liang, LI Xin-Yi, HUANG Hai-Quan*, HUANG Mei-Juan* |
| College of Landscape Architecture and Horticulture Sciences, Southwest Research Center for Engineering Technology of Landscape Architecture (State Forestry and Grassland Administration)/Yunnan Engineering Research Center for Functional Flower Resources and Industrialization/Research and Development Center of Landscape Plants and Horticulture Flowers, Southwest Forestry University, Kunming 650224, China |
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Abstract Rhododendron sphaeroblastum var. wumengense is a high-altitude and cold-tolerant plant. MYBS3 and rare cold inducible 2 (RCI2B) genes were found to play a significant role in enhancing cold stress tolerance in plants based on the pre-screening from R. sphaeroblastum transcriptome. In this study, the key transcription factors RsMYBS3 (GenBank No. OR178946) and RsRCI2B (GenBank No. OR178945) genes that regulated cold tolerance of R. sphaeroblastum were cloned, whose cDNA lengths were 795 and 165 bp, encoding 264 and 54 aa, respectively. Bioinformatics analysis showed that RsMYBS3 was a hydrophilic unstable protein with a typical SANT conserved structural domain of the SANT superfamilies; RsRCI2B was a hydrophobic stable protein with a highly conservative structural domain typical PMP3. The results of gene homology comparison showed that both RsMYBS3 and RsRCI2B proteins were highly homologous to MYBS3 and RCI2B proteins in red horse psyllium (Rhododendron vialii). The similarities were 93.73% and 94.44%, respectively. Phylogenetic analysis showed that RsMYBS3 and RsRCI2B were clustered in a clade with RvMYBS3 and RvRCI2B from R. vialii, which suggested that 4 genes were orthologous. qPCR analysis showed that both genes were significantly expressed in R. sphaeroblastum under the low-temperature stress, but had different expression patterns under different stress temperatures and times. The RsMYBS3 gene had the highest expression level at -6 ℃ treatment on 1 d, and then gradually decreased on 4 d with the prolonging of stress time, and expression gradually up-regulated on 7 d. The expression level gradually increased and reached a peak from 1 d to 4 d, and gradually decreased on 7 d under the -12 ℃ treatment. While the expression level of RsRCI2B gene had gradually increased from 1 d to the 7 day at -6 ℃ treatment, and at -12 ℃ treatment, except for the slight decrease expression on 1 d, the expression level gradually increased and reached a peak from 4 d to 7 d as the stress time prolonged. The above mentioned results suggested that both RsMYBS3 and RsRCI2B genes might be involved in the cold-tolerance-response physiological process of R. sphaeroblastum. This study provides some certain basic data and theoretical basis for exploring their cold tolerance molecular mechanism in the future.
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Received: 25 July 2023
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Corresponding Authors:
*xmhhq2001@163.com; haiquanl@163.com
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