RESUMO
Anthurium andraeanum is a tropical flower with high ornamental and economic value. Cold stress is one of the major abiotic stresses affecting the quality and value of A. andraeanum; thus, improving the cold tolerance of this species is an important breeding objective. MicroRNAs (miRNAs) have a critical role in plant abiotic stress responses, but their specific molecular regulatory mechanisms are largely unknown, including those related to the cold stress response in A. andraeanum. Here, we identified and cloned the precursor of miR158 from A. andraeanum (Aa-miR158). Both Aa-miR158 and its target gene (c48247) had higher expression levels in strong leaves than in other tissues or organs. Further study revealed that the transcript level of Aa-miR158 was increased by cold stress. Heterologous overexpression of Aa-miR158 improved cold stress tolerance in Arabidopsis, which was associated with decreases in the malondialdehyde (MDA) concentration and relative electrical conductivity (REC) as well as increases in peroxidase (POD) and catalase (CAT) activity. Moreover, overexpressing Aa-miR158 significantly increased the expression of endogenous genes related to cold stress tolerance and reactive oxygen species (ROS) levels in transgenic Arabidopsis under cold stress. Overall, our results demonstrate that Aa-miR158 is significantly involved in the cold stress response and provide a new strategy for cold tolerance breeding of A. andraeanum.
RESUMO
Anthurium andraeanum Lind. is a popular potted and cut-flower plant with an attractive spathe and foliage. It is native to tropical rainforest areas and is able to blossom throughout the year under suitable conditions. However, various abiotic stresses seriously restrict the ornamental value of A. andraeanum and increase the costs of cultivation. A dark green (dg) leaf color mutant of A. andraeanum 'Sonate', which accumulates high levels of anthocyanin, has shown increased vigor and tolerance to stresses during cultivation and is, thus, an ideal germplasm for studying stress tolerance in this species. Here, we show that the anthocyanin content in dg mutant plants at different stages of leaf development was higher than in wild-type (WT) plants, and the ability to tolerate under low-temperature (LT, 14 °C) stress was stronger in dg than in WT plants. RNA-Seq of cDNA libraries from young leaves of dg and WT identified AabHLH35 as a differentially expressed gene (DEG) that was significantly up-regulated in dg. Furthermore, heterologous expression of AabHLH35 improved tolerance to cold and drought stresses in Arabidopsis. These results have built an important molecular foundation for further study of stress tolerance in A. andraeanum.
RESUMO
MicroRNAs (miRNAs) are a class of non-coding small RNAs that play important roles in the regulation of gene expression. Although plant miRNAs have been extensively studied in model systems, less is known in other plants with limited genome sequence data, including Anthurium andraeanum. To identify miRNAs and their target genes in A. andraeanum and study their responses to abiotic stresses, we conducted deep-sequencing of two small RNA (sRNA) libraries prepared from young leaves of wild-type (WT) and dark green (dg) leaf color mutant plants of A. andraeanum 'Sonate'. A total of 53 novel miRNAs were identified, 32 of which have been annotated to 18 miRNA families. 10 putative miRNAs were found to be differentially expressed in WT and dg, among which two miRNAs were significantly up-regulated and eight down-regulated in dg relative to WT. One differentially expressed miRNA, Aa-miR408, was dramatically up-regulated in dg. qRT-PCR analysis and heterologous expression of Aa-miR408 in Arabidopsis under different stress treatments suggest that Aa-miR408 is involved in abiotic stress responses in A. andraeanum. Our results provide a foundation for further dissecting the roles of miRNAs and their targets in regulating abiotic stress tolerance in A. andraeanum.