PagMYB180 regulates adventitious rooting via a ROS/PCD-dependent pathway in poplar.

The formation of adventitious roots (AR) is an essential step in the vegetative propagation of economically woody species. Reactive oxygen species (ROS) function as signaling molecules in regulating root growth and development. Here, we identified an R2R3-MYB transcription factor PagMYB180 as a regulator of AR formation in hybrid poplar (Populus alba × Populus glandulosa). PagMYB180 was specifically expressed in the vascular tissues of poplar roots, stems and leaves, and its protein was localized in the nucleus and acted as a transcriptional repressor. Both dominant repression and overexpression of PagMYB180 resulted in a significant reduction of AR quantity, a substantial increase of AR length, and an elevation of both the quantity and length of lateral roots (LR) compared to the wild type (WT) plants. Furthermore, PagMYB180 regulates programmed cell death (PCD) in root cortex cells, which is associated with elevated levels of ROS. Transcriptome and reverse transcription-quantitative PCR (RT-qPCR) analyses revealed that a series of differentially expressed genes are related to ROS, PCD and ethylene synthesis. Taken together, these results suggest that PagMYB180 may regulate AR development via a ROS/PCD-dependent pathway in poplar.

Long non-coding RNA-mediated competing endogenous RNA regulatory network during flower development and color formation in Melastoma candidum.

M. candidum, an evergreen shrubby flower known for its superior adaptation ability in South China, has gained increased attention in garden applications. However, scant attention has been paid to its flower development and color formation process at the non-coding RNA level. To fill this gap, we conducted a comprehensive analysis based on long non-coding RNA sequencing (lncRNA-seq), RNA-seq, small RNA sequencing (sRNA-seq), and widely targeted metabolome detection of three different flower developmental stages of M. candidum. After differentially expressed lncRNAs (DElncRNAs), differentially expressed mRNAs (DEmRNAs), differentially expressed microRNAs (DEmiRNAs), and differentially synthesized metabolites (DSmets) analyses between the different flower developmental stages, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were conducted to identify some key genes and metabolites in flavonoid, flavone, anthocyanin, carotenoid, and alkaloid-related GO terms and biosynthetic pathways. Three direct-acting models, including antisense-acting, cis-acting, and trans-acting between lncRNAs and mRNAs, were detected to illustrate the direct function of lncRNAs on target genes during flower development and color formation. Based on the competitive endogenous RNA (ceRNA) regulatory theory, we constructed a lncRNA-mediated regulatory network composed of DElncRNAs, DEmiRNAs, DEmRNAs, and DSmets to elucidate the indirect role of lncRNAs in the flower development and color formation of M. candidum. By utilizing correlation analyses between DERNAs and DSmets within the ceRNA regulatory network, alongside verification trials of the ceRNA regulatory mechanism, the study successfully illustrated the significance of lncRNAs in flower development and color formation process. This research provides a foundation for improving and regulating flower color at the lncRNA level in M. candidum, and sheds light on the potential applications of non-coding RNA in studies of flower development.

Overexpression of a gibberellin 20-oxidase gene in poplar xylem led to an increase in the size of nanocellulose fibrils and improved paper properties.

Cellulose, the major component of secondary cell walls, is the most abundant renewable long-chain polymer on earth. Nanocellulose has become a prominent nano-reinforcement agent for polymer matrices in various industries. We report the generation of transgenic hybrid poplar overexpressing the Arabidopsis gibberellin 20-oxidase1 gene driven by a xylem-specific promoter to increase gibberellin (GA) biosynthesis in wood. X-ray diffraction (XRD) and sum frequency generation spectroscopic (SFG) analyses showed that cellulose in transgenic trees was less crystalline, but the crystal size was larger. The nanocellulose fibrils prepared from transgenic wood had an increased size compared to those from wild type. When such fibrils were used as a reinforcing agent in sheet paper preparation, the mechanical strength of the paper was significantly enhanced. Engineering the GA pathway can therefore affect nanocellulose properties, providing a new strategy for expanding nanocellulose applications.

Chloroplast (Cp) Transcriptome of P. davidiana Dode×P. bolleana Lauch provides insight into the Cp drought response and Populus Cp phylogeny.

BACKGROUND: Raw second-generation (2G) lignocellulosic biomass materials have the potential for development into a sustainable and renewable source of energy. Poplar is regarded as a promising 2G material (P. davidiana Dode×P. bolleana Lauch, P. bolleana, P. davidiana, P. euphratica, et al). However, their large-scale commercialization still faces many obstacles. For example, drought prevents sufficient irrigation or rainfall, which can reduce soil moisture and eventually destroy the chloroplast, the plant photosynthetic organelle. Heterosis is widely used in the production of drought-tolerant materials, such as the superior clone "Shanxinyang" selected from the offspring of Populus davidiana Dode×Populus bolleana Lauch. Because it produces good wood and is easily genetically transformed, "Shanxinyang" has become a promising material for use in tree genetics. It is also one of the most abundant biofuel plants in northern China. Understanding the genetic features of chloroplasts, the cp transcriptome and physiology is crucial to elucidating the chloroplast drought-response model. RESULTS: In this study, the whole genome of "Shanxinyang" was sequenced. The chloroplast genome was assembled, and chloroplast structure was analysed and compared with that of other popular plants. Chloroplast transcriptome analysis was performed under drought conditions. The total length of the "Shanxinyang" chloroplast genome was 156,190 bp, the GC content was 36.75%, and the genome was composed of four typical areas (LSC, IRa, IRb, and SSC). A total of 114 simple repeats were detected in the chloroplast genome of "Shanxinyang". In cp transcriptome analysis, we found 161 up-regulated and 157 down-regulated genes under drought, and 9 cpDEGs was randomly selected to conduct reverse transcription (RT)-qPCR., in which the Log2 (fold change) was significantly consistent with the qPCR results. The analysis of chloroplast transcription under drought provided clues for understanding chloroplast function under drought. The phylogenetic position of "Shanxinyang" within Populus was analysed by using the chloroplast genome sequences of 23 Populus plants, showing that "Shanxinyang" belongs to Sect. Populus and is sister to Populus davidiana. Further, mVISTA analysis showed that the variation in non-coding (regulatory) regions was greater than that in coding regions, which suggests that further attention should be paid to the chloroplast in order to obtain new evolutionary or functional insights related to aspects of plant biology. CONCLUSIONS: Our findings indicate that complex prokaryotic genome regulation occurs when processing transcripts under drought stress. The results not only offer clues for understanding the chloroplast genome and transcription features in woody plants but also serve as a basis for future molecular studies on poplar species.

Expression analysis of the BpARF genes in Betula platyphylla under drought stress.

Auxin response factors (ARFs) play an important role in modulating plant growth and development processes by regulating the expression of auxin-responsive genes. However, the modes of action of ARFs in birch (Betula platyphylla) remain largely unknown. In this study, fifteen ARF genes were identified in the birch (B. platyphylla) genome. Bioinformatics analysis revealed that the 15 BpARF genes were unevenly distributed on 7 chromosomes. The 15 BpARF proteins clustered into 6 groups, and all of them contained ARF and B3 motifs. The cis-acting elements present within the promoters of the BpARF genes were mostly related to stress resistance. Expression analysis revealed that most of the BpARF genes were significantly upregulated or downregulated in response to drought treatment in at least one organ. In particular, the expression of BpARF1 was significantly induced by drought stress. The function of BpARF1 was further studied via a transient transformation system. Under drought stress conditions, compared with vector control plants, BpARF1 RNA interference (RNAi)-inhibited plants presented reduced reactive oxygen species (ROS) accumulation, enhanced peroxide (POD) and superoxide dismutase (SOD) activities, increased ascorbic acid (AsA) and proline contents, and reduced electrolyte leakage and water loss rates. Conversely, BpARF1 overexpression plants displayed the opposite physiological changes. These results suggest that the silencing of BpARF1 can improve the drought tolerance of B. platyphylla.

Expression of a populus histone deacetylase gene 84KHDA903 in tobacco enhances drought tolerance.

Histone deacetylases (HDACs) play a key role in regulating plant growth, development and stress responses. However, functions of HDACs in woody plants are largely unknown. In this study, a novel gene encoding a RPD3/HDA1-type histone deacetylase was cloned from 84K poplar (Populus alba×Populus glandulosa) and designated as 84KHDA903. The 84KHDA903 encodes a protein composed of 500 amino acid residues, which contains a conserved HDAC domain. Transient expression of 84KHDA903 in onion epidermal cells suggested that it was exclusively localized in nucleus. The 84KHDA903 exhibited different expression patterns under drought, salt and ABA treatments. The expression of 84KHDA903 was responsive to drought and ABA but not to salt. To understand the function of 84KHDA903 in stress responses, the 84KHDA903 gene was transformed into tobacco. The expression of 84KHDA903 in tobacco increased the tolerance of transgenic seeds to mannitol but not to salt. In adult stage, the 84KHDA903-expressing tobacco exhibited drought tolerance and showed strong capacity to recover after drought. During the recovery period, the stress-responsive genes including NtDREB4, NtDREB3 and NtLEA5 were induced to be highly expressed in the 84KHDA903 transgenic plants in contrast to wild-type plants. Taken together, for the first time, we reported a RPD3/HDA1-type histone deacetylase from poplar, 84KHDA903, which acted as a positive regulator in drought stress responses.