A systematic genome-wide analysis and screening of the NAC family genes related to wood formation in Cinnamomum camphora.

Many processes, such as growth, aging, and adaptation to abiotic stress, are regulated in plants by NAC transcription factors. In woody plants, NAC transcription factors acts as a primary switch that regulates secondary xylem development by activating various downstream transcription factors and modulating expression levels of genes involved in the synthesis of the secondary cell wall. Our team had previously sequenced the whole genome of the camphor tree (Cinnamomum camphora). Here, we performed a detailed analysis of the NAC gene family of C. camphora and examined its evolutionary history. The genomic sequences of 121 NAC genes of C. camphora were identified and classified into 20 subfamilies in 2 major classes based on the phylogenetic analysis and structural features. Expansion of the CcNAC gene family occurred mainly by fragment replication and was influenced by the purifying selection. By analyzing predicted interactions of the homologous AtNAC proteins, we identified five CcNACs that potentially regulate xylem development in C. camphora. RNA sequencing revealed distinct expression profiles of CcNACs in seven different plant tissues. Subcellular localization prediction revealed that 120, 3, and 2 CcNACs have biological functions in the nucleus, cytoplasm, and chloroplast, respectively. Furthermore, we verified expression patterns of five CcNACs (CcNAC012, CcNAC028, CcNAC055, CcNAC080, and CcNAC119) in various tissue types using qRT-PCR. Our results will facilitate further in-depth studies of the molecular mechanisms by which CcNAC transcription factors regulate wood formation and other processes in C. camphora.

Transcriptome comparison analyses in UV-B induced AsA accumulation of Lactuca sativa L.

BACKGROUND: Lettuce (Lactuca sativa L.) cultivated in facilities display low vitamin C (L-ascorbic acid (AsA)) contents which require augmentation. Although UV-B irradiation increases the accumulation of AsA in crops, processes underlying the biosynthesis as well as metabolism of AsA induced by UV-B in lettuce remain unclear. RESULTS: UV-B treatment increased the AsA content in lettuce, compared with that in the untreated control. UV-B treatment significantly increased AsA accumulation in a dose-dependent manner up until a certain dose.. Based on optimization experiments, three UV-B dose treatments, no UV-B (C), medium dose 7.2 KJ·m- 2·d- 1 (U1), and high dose 12.96 KJ·m- 2·d- 1 (U2), were selected for transcriptome sequencing (RNA-Seq) in this study. The results showed that C and U1 clustered in one category while U2 clustered in another, suggesting that the effect exerted on AsA by UV-B was dose dependent. MIOX gene in the myo-inositol pathway and APX gene in the recycling pathway in U2 were significantly different from the other two treatments, which was consistent with AsA changes seen in the three treatments, indicating that AsA accumulation caused by UV-B may be associated with these two genes in lettuce. UVR8 and HY5 were not significantly different expressed under UV-B irradiation, however, the genes involved in plant growth hormones and defence hormones significantly decreased and increased in U2, respectively, suggesting that high UV-B dose may regulate photomorphogenesis and response to stress via hormone regulatory pathways, although such regulation was independent of the UVR8 pathway. CONCLUSIONS: Our results demonstrated that studying the application of UV-B irradiation may enhance our understanding of the response of plant growth and AsA metabolism-related genes to UV-B stress, with particular reference to lettuce.

An Acoustic Localization Sensor Based on MEMS Microphone Array for Partial Discharge.

Partial discharge (PD) localization is important for monitoring and maintaining high-voltage equipment, which can help to prevent accidents. In this work, an acoustic localization sensor based on microelectromechanical system (MEMS) microphone array is proposed, which can detect and locate the partial discharge through a beam-forming algorithm. The MEMS microphone array consists of eight commercial MEMS microphones (SPV08A0LR5H-1, Knowles Electronics, IL, USA) with an aperture size of about 0.1 m × 0.1 m, allowing for a small hardware size and low cost. In order to optimize the acoustic performance of the array, a random array topology is designed. The simulation analysis indicates that the designed random topology is superior to several commonly used topologies. In terms of the localization algorithm, a deconvolution method called Fourier-based fast iterative shrinkage thresholding algorithm (FFT-FISTA) is applied. Simulation and experiment results demonstrate that FFT-FISTA used in the proposed acoustic localization sensor has significant advantages over the conventional beam-forming algorithm on spatial resolution and sidelobe suppression. Experimental results also show that the average localization error of the proposed scheme is about 0.04 m, which can meet the demands of practical application.

Genome-Wide Characterization and Analysis of bHLH Transcription Factors Related to Anthocyanin Biosynthesis in Cinnamomum camphora ('Gantong 1').

Cinnamomum camphora is one of the most commonly used tree species in landscaping. Improving its ornamental traits, particularly bark and leaf colors, is one of the key breeding goals. The basic helix-loop-helix (bHLH) transcription factors (TFs) are crucial in controlling anthocyanin biosynthesis in many plants. However, their role in C. camphora remains largely unknown. In this study, we identified 150 bHLH TFs (CcbHLHs) using natural mutant C. camphora 'Gantong 1', which has unusual bark and leaf colors. Phylogenetic analysis revealed that 150 CcbHLHs were divided into 26 subfamilies which shared similar gene structures and conserved motifs. According to the protein homology analysis, we identified four candidate CcbHLHs that were highly conserved compared to the TT8 protein in A. thaliana. These TFs are potentially involved in anthocyanin biosynthesis in C. camphora. RNA-seq analysis revealed specific expression patterns of CcbHLHs in different tissue types. Furthermore, we verified expression patterns of seven CcbHLHs (CcbHLH001, CcbHLH015, CcbHLH017, CcbHLH022, CcbHLH101, CcbHLH118, and CcbHLH134) in various tissue types at different growth stages using qRT-PCR. This study opens a new avenue for subsequent research on anthocyanin biosynthesis regulated by CcbHLH TFs in C. camphora.

The MKK2a Gene Involved in the MAPK Signaling Cascades Enhances Populus Salt Tolerance.

Mitogen-activated protein kinase (MAPK) cascades are highly conserved signal transduction modules, which transmit environmental signals in plant cells through stepwise phosphorylation and play indispensable roles in a wide range of physiological and biochemical processes. Here, we isolated and characterized a gene encoding MKK2 protein from poplar through the rapid amplification of cDNA ends (RACE). The full-length PeMKK2a gene was 1571 bp, including a 1068 bp open reading frame (ORF) encoding 355 amino acids, and the putative PeMKK2a protein belongs to the PKc_like (protein kinase domain) family (70-336 amino acids) in the PKc_MAPKK_plant subfamily and contains 62 sites of possible phosphorylation and two conserved domains, DLK and S/T-xxxxx-S/T. Detailed information about its gene structure, sequence similarities, subcellular localization, and transcript profiles under salt-stress conditions was revealed. Transgenic poplar lines overexpressing PeMKK2a exhibited higher activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) than non-transgenic poplar under salt stress conditions. These results will provide insight into the roles of MAPK signaling cascades in poplar response to salt stress.

Integrated SMRT and Illumina Sequencing Provide New Insights into Crocin Biosynthesis of Gardenia jasminoides.

Crocins are valuable bioactive components of gardenia fruit, and their biosynthesis and accumulation have attracted widespread interest. Studies have investigated the biosynthesis and accumulation of crocin based on Illumina sequencing, but there is a lack of reports based on full-length transcriptome sequencing. Utilising SMRT sequencing and high-performance liquid chromatography (HPLC), we explored crocin biosynthesis and accumulation in the fruit of Gardenia jasminoides. HPLC analysis showed that crocins specifically exist in fruit and that the content of crocins increases gradually during fruit development. SMRT sequencing generated 46,715 high-quality full-length isoforms, including 5230 novel isoforms that are not present in the G. jasminoides genome. Furthermore, a total of 46 genes and 91 lncRNAs were involved in the biosynthesis and accumulation of crocin. The qRT-PCR indicated that genes involved in crocin biosynthesis reached a peak in the NOV stage. These findings contributed to our understanding of crocin biosynthesis and accumulation.

Genome-Scale Identification, Classification, and Expression Profiling of MYB Transcription Factor Genes in Cinnamomum camphora.

The camphor tree (Cinnamomum camphora (L.) Presl.) is the representative species of subtropical evergreen broadleaved forests in eastern Asia and an important raw material for essential oil production worldwide. Although MYBs have been comprehensively characterized and their functions have been partially resolved in many plants, it has not been explored in C. camphora. In this study, 121 CcMYBs were identified on 12 chromosomes in the whole genome of C. camphora and found that CcMYBs were mainly expanded by segmental duplication. They were divided into 28 subgroups based on phylogenetic analysis and gene structural characteristics. In the promoter regions, numerous cis-acting elements were related to biological processes. Analysis of RNA sequencing data from seven tissues showed that CcMYBs exhibited different expression profiles, suggesting that they have various roles in camphor tree development. In addition, combined with the correlation analysis of structural genes in the flavonoid synthesis pathway, we identified CcMYBs from three subgroups that might be related to the flavonoid biosynthesis pathway. This study systematically analyzed CcMYBs in C. camphora, which will set the stage for subsequent research on the functions of CcMYBs during their lifetime and provide valuable insights for the genetic improvement of camphor trees.

Transcriptome analysis and identification of genes related to terpenoid biosynthesis in Cinnamomum camphora.

BACKGROUND: Cinnamomum camphora has been cultivated as an economically important tree for its medicinal and aromatic properties. Selective breeding has produced Cinnamomum plants for special uses, including spice strains with characteristic flavors and aromas and high-potency medicinal cultivars. The molecular biology underlying terpenoid biosynthesis is still unexplored. RESULTS: Gas chromatography-mass spectrometry was used to analyze the differences in contents and compositions of essential oil terpenoids in linalool- and borneol-type chemotypes of C. camphora. The data revealed that the essential oils consist primarily of monoterpenes with only very minor quantities of sesquiterpenes and diterpenes and that the essential oil differs in different chemotypes of C. camphora, with higher yields of (-)-borneol from the borneol-type than from the linalool-type. To study the terpenoid biosynthesis of signature compounds of the major monoterpenes, we performed RNA sequencing to profile the leaf transcriptomes of the two chemotypes of C. camphora. A total of 23.76 Gb clean data was generated from two chemotypes and assembled into 156,184 unigenes. The total length, average length, N50 and GC content of unigenes were 155,645,929 bp, 997 bp, 1430 bp, and 46.5%, respectively. Among them, 76,421 unigenes were annotated by publicly available databases, of which 67 candidate unigenes were identified to be involved in terpenoid biosynthesis in C. camphora. A total of 2863 unigenes were identified to be differentially expression between borneol-type and linalool-type, including 1714 up-regulated and 1149 down-regulated unigenes. Most genes encoding proteins involved in terpenoid precursor MVA and MEP pathways were expressed in similar levels in both chemotypes of C. camphora. In addition, 10 and 17 DEGs were significantly enriched in the terpene synthase activity and oxidoreductase activity terms of their directed acyclic graphs (DAG), respectively. Three monoterpene synthase genes, TPS14-like1, TPS14-like2 and TPS14-like3 were up-regulated in the borneol-type compared to the linalool-type, and their expression levels were further verified using quantitative real-time PCR. CONCLUSIONS: This study provides a global overview of gene expression patterns related to terpenoid biosynthesis in C. camphora, and could contribute to a better understanding of the differential accumulation of terpenoids in different C. camphora chemotypes.

A High Sensitivity AlN-Based MEMS Hydrophone for Pipeline Leak Monitoring.

In this work, a miniaturized, low-cost, low-power and high-sensitivity AlN-based micro-electro-mechanical system (MEMS) hydrophone is proposed for monitoring water pipeline leaks. The proposed MEMS Hydrophone consists of a piezoelectric micromachined ultrasonic transducer (PMUT) array, an acoustic matching layer and a pre-amplifier amplifier circuit. The array has 4 (2 × 2) PMUT elements with a first-order resonant frequency of 41.58 kHz. Due to impedance matching of the acoustic matching layer and the 40 dB gain of the pre-amplifier amplifier circuit, the packaged MEMS Hydrophone has a high sound pressure sensitivity of -170 ± 2 dB (re: 1 V/µPa). The performance with respect to detecting pipeline leaks and locating leak points is demonstrated on a 31 m stainless leaking pipeline platform. The standard deviation (STD) of the hydroacoustic signal and Monitoring Index Efficiency (MIE) are extracted as features of the pipeline leak. A random forest model is trained for accurately classifying the leak and no-leak cases using the above features, and the accuracy of the model is about 97.69%. The cross-correlation method is used to locate the leak point, and the localization relative error is about 10.84% for a small leak of 12 L/min.

Overexpression of Ginkgo BBX25 enhances salt tolerance in Transgenic Populus.

B-box (BBX) genes play important roles in plant growth, light morphogenesis, and environmental stress responses. Ginkgo (Ginkgo biloba L.) is known as a living fossil species that has a strong ability to adapt to environmental changes and tolerate harsh conditions. In this study, we chose this species to investigate the function of the GbBBX25 gene. We isolated the BBX gene from ginkgo and named it GbBBX25; this gene consists of an 819 bp open reading frame (ORF) that encodes 273 amino acids with two B-box domains but no CCT domain. GbBBX25 was localized in only the nucleus. The expression of GbBBX25 transcripts was observed in the leaves and was significantly enhanced under salt stress conditions. To further verify its function, we overexpressed the GbBBX25 gene in Populus davidiana × Populus bolleana and found that the transgenic Populus had greater soluble sugar levels and higher peroxidase (POD) activity in response to salt stress than nontransgenic (NT) Populus. Five genes related to salt stress were induced in transgenic plants with significantly higher expression levels than those in NT plants. This finding suggests that GbBBX25 improves the salt adaptation abilities of transgenic Populus and provides a scientific basis for related research.

Overexpression of PeHKT1;1 Improves Salt Tolerance in Populus.

Soil salinization is an increasingly serious threat that limits plant growth and development. Class I transporters of the high-affinity K⁺ transporter (HKT) family have been demonstrated to be involved in salt tolerance by contributing to Na⁺ exclusion from roots and shoots. Here, we isolated the PeHKT1;1 gene from hybrid poplar based on the sequences of the Populus trichocarpa genome. The full-length PeHKT1;1 gene was 2173 bp, including a 1608 bp open reading frame (ORF) encoding 535 amino acids and containing eight distinct transmembrane domains. Multiple sequence alignment and phylogenetic analysis suggested that the PeHKT1;1 protein had a typical S⁻G⁻G⁻G signature for the P-loop domains and belonged to class I of HKT transporters. PeHKT1;1 transcripts were mainly detected in stem and root, and were remarkably induced by salt stress treatment. In further characterization of its functions, overexpression of PeHKT1;1 in Populus davidiana × Populus bolleana resulted in a better relative growth rate in phenotypic analysis, including root and plant height, and exhibited higher catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) activities than non-transgenic poplar under salt stress conditions. These observations indicated that PeHKT1;1 may enhance salt tolerance by improving the efficiency of antioxidant systems. Together, these data suggest that PeHKT1;1 plays an important role in response to salt stress in Populus.

The complete chloroplast genome of Cinnamomum camphora and its comparison with related Lauraceae species.

Cinnamomum camphora, a member of the Lauraceae family, is a valuable aromatic and timber tree that is indigenous to the south of China and Japan. All parts of Cinnamomum camphora have secretory cells containing different volatile chemical compounds that are utilized as herbal medicines and essential oils. Here, we reported the complete sequencing of the chloroplast genome of Cinnamomum camphora using illumina technology. The chloroplast genome of Cinnamomum camphora is 152,570 bp in length and characterized by a relatively conserved quadripartite structure containing a large single copy region of 93,705 bp, a small single copy region of 19,093 bp and two inverted repeat (IR) regions of 19,886 bp. Overall, the genome contained 123 coding regions, of which 15 were repeated in the IR regions. An analysis of chloroplast sequence divergence revealed that the small single copy region was highly variable among the different genera in the Lauraceae family. A total of 40 repeat structures and 83 simple sequence repeats were detected in both the coding and non-coding regions. A phylogenetic analysis indicated that Calycanthus is most closely related to Lauraceae, both being members of Laurales, which forms a sister group to Magnoliids. The complete sequence of the chloroplast of Cinnamomum camphora will aid in in-depth taxonomical studies of the Lauraceae family in the future. The genetic sequence information will also have valuable applications for chloroplast genetic engineering.