Oligo-dT anchored cDNA-SRAP and cDNA-SCoT aided identification of transcripts differentially expressed during the early stages of recovery of resurrection plant Haberlea rhodopensis Friv. from freezing-induced desiccation

Abstract

Resurrection plants are a unique group of flora showing an extraordinary ability to survive extreme water shortages. Among them, the Bulgarian endemic plant Haberlea rhodopensis Friv. has been used as a model to study the mechanisms underlying desiccation tolerance induced by water and low temperature stress. Although extensive transcriptome data, including two RNAseq experiments, are available for drought induced desiccation and recovery, the gene expression during recovery after freezing-induced desiccation (RAF) remains largely unexplored. In this study we performed a differential screening using a new approach, based on oligo-dT anchored cDNA-SRAP (copy DNA sequence-related amplified polymorphism) and a modified oligo-dT anchored cDNA-SCoT (copy DNA start codon targeted) assays to provide new information on genes involved in the tolerance to low temperature stress and the recovery of H. rhodopensis. The putative functions of 19 identified transcript derived fragments were established by searching for homology with known sequences in public databases and the expression of 10 of them during the early stages of recovery was confirmed by quantitative reverse transcription polymerase chain reaction. Some of the identified genes have known functions in the response to abiotic stress in other plant species including resurrection plants, which confirms the effectiveness of the approach used in the present study. The results of this study will contribute to obtaining new information on the defence mechanisms in the recovery process of H. rhodopensis, especially at the early stages of recovery, which is the most vulnerable period for plants.

Keywords

• Resurrection plants
• differential screening
• recovery after freezing-induced desiccation
• cDNA-SRAP
• cDNA-SCoT
• qRT-PCR