Heat transcription factors (Hsfs) belong to a large gene family classified into A, B, and C groups, with classes A and B Hsfs being well-characterized and known for their roles in plant tolerance to abiotic stresses. The functions and roles of Class C Hsfs are not well-documented. The objectives of this study were to characterize a class C Hsf gene (FaHsfC1b) cloned from tall fescue (Festuca arundinacea), a perennial grass species, and to determine the physiological functions of FaHsfC1b in regulating heat tolerance by overexpressing FaHsfC1b in Arabidopsis thaliana. Full length cDNA of FaHsfC1b was cloned and the sequence alignment showed that it had high similarity to OsHsfC1b with typical DNA binding domain, hydrophobic oligomerization domain, and a nucleus localization signal. Transient expression with FaHsfC1b-eGFP in protoplasts of Arabidopsis leaves indicated its nucleus localization. qRT-PCR analysis showed that FaHsfC1b responded to heat, osmotic, salt, and cold stress in leaves and roots during 48-h treatment. Physiological analysis showed that FaHsfC1b overexpression enhanced plant survival rate, chlorophyll content, and photochemical efficiency, while it resulted in decreases in electrolyte leakage, H₂O₂ and O²⁻ content under heat stress. qRT-PCR showed that endogenous HsfC1 was induced in transgenic plants and the expression levels of heat protection protein genes, including several HSPs, AtGalSyn1, AtRof1, and AtHSA32, as well as ABA-synthesizing gene (NCED3) were significantly upregulated in transgenic plants overexpressing FaHsfC1b under heat stress. Our results first demonstrate that HsfC1b plays positive roles in plant tolerance to heat stress in association with the induction and upregulation of heat-protective genes. HsfC1b may be used as a candidate gene for genetic modification of cool-season plant species for improving heat tolerance.

Tiller production in grass species is controlled by both axillary bud initiation and bud outgrowth, which may be regulated by plant hormones. However, how gibberellic acid (GA) affects tillering in perennial grass species is still unclear. This study aims to elucidate the roles and the underlying mechanisms of GA in regulating tiller development. Tall fescue seedlings were treated with different concentrations of GA3 by foliar application, dose-dependent inhibitory effects of GA on tiller production were observed. GA3 (25 μM) slowed down the transition from axillary buds to tillers by specifically inhibiting the outgrowth of axillary buds. GA-inhibition of tillering were not related to endogenous content for auxin or strigolactone, but was mainly due to the antagonistic interaction with cytokinins (CK), as shown by the decreased CK content and up-regulation expression of CK degradation genes in GA3-treated plants. Furthermore, GA could act through regulating the expression of FaTB1 specifically expressed in axillary buds to repress bud outgrowth. These results provide insights for the regulatory mechanisms of GA for tiller bud outgrowth through crosstalks with CK and signaling of FaTB1 expression.

Background: Explored the molecular science of anther development is important for improving productivity and overall yield of crops. Although the role of regulatory RNAs, including long non-coding RNAs (lncRNAs) and microRNAs (miRNAs), in regulating anther development has been established, their identities and functions in Camellia oleifera, an important industrial crop, have yet not been clearly explored. Here, we report the identification and characterization of genes, lncRNAs and miRNAs during three stages of the tropical C. oleifera anther development by single-molecule real-time sequencing, RNA sequencing and small RNA sequencing, respectively. Results: These stages, viz. the pollen mother cells stage, tetrad stage and uninucleate pollen stage, were identified by analyzing paraffin sections of floral buds during rapid expansion periods. A total of 18,393 transcripts, 414 putative lncRNAs and 372 miRNAs were identified, of which 5,324 genes, 115 lncRNAs, and 44 miRNAs were differentially accumulated across three developmental stages. Of these, 44 and 92 genes were predicted be regulated by 37 and 30 differentially accumulated lncRNAs and miRNAs, respectively. Additionally, 42 differentially accumulated lncRNAs were predicted as targets of 27 miRNAs. Gene ontology enrichment indicated that potential target genes of lncRNAs were enriched in photosystem II, regulation of autophagy and carbohydrate phosphatase activity, which are essential for anther development. Functional annotation of genes targeted by miRNAs indicated that they are relevant to transcription and metabolic processes that play important roles in microspore development. An interaction network was built with 2 lncRNAs, 6 miRNAs and 10 mRNAs. Among these, miR396 and miR156 family were up-regulated, while their targets, genes (GROWTH REGULATING FACTORS and SQUAMOSA PROMOTER BINDING PROTEIN-LIKE genes) and lncRNAs, were down-regulated. Further, the trans-regulated targets of these lncRNAs, like wall-associated kinase2 and phosphomannose isomerase1, are involved in pollen wall formation during anther development. Conclusions: This study unravels lncRNAs, miRNAs and miRNA-lncRNA-mRNA networks involved in development of anthers of the tropical C. oleifera lays a theoretical foundation for further elucidation of regulatory roles of lncRNAs and miRNAs in anther development.