DNA barcoding and comparative RNA-Seq analysis provide new insights into leaf formation using a novel resource of high-yielding Epimedium koreanum

Epimedium koreanum Nakai, a well-known traditional Chinese medicinal herb, has been widely used to treat osteoporosis and sexual dysfunction for thousands of years. However, due to the decreasing population of East Asian natural resources, yearly output of Epimedium crude herb has been in low supply year by year. In this study, an unusual variety of E. koreanum was discovered in Dunhua, Jilin Province, the northernmost area where this variety was found containing 6 individuals, with three branches that had 27 leaflets, which is much more than the typical leaflet number of 9. Firstly, the novel E. koreanum varety was identified using DNA barcodes. Then, 1171 differentially expressed genes (DEGs) were discovered through parallel RNA-seq analysis between the newly discovered variety and wild type (WT) E. koreanum plant. Furthermore, the results of bioinformatics investigation revealed that 914 positively and 619 negatively correlated genes associated with the number of leaflets. Additionally, based on RNA-Seq and qRT-PCR analysis, two homologous hub TCP genes, which were commonly implicated in plant leaf development, and shown to be up regulated and down regulated in the discovered newly variety, respectively. Thus, our study discovered a novel wild resource for leaf yield rewarding medicinal Epimedium plant breeding, provided insights into the relationship between plant compound leaf formation and gene expression of TCPs transcription factors and other gene candidates, providing bases for creating high yield cultivated Epimedium variety by using further molecular selection and breeding techniques in the future

Effects of Combined Elicitors on Tanshinone Metabolic Profiling and SmCPS Expression in Salvia miltiorrhiza Hairy Root Cultures

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Molecular cloning and functional identification of a cDNA encoding 4-hydroxy-3-methylbut-2-enyl diphosphate reductase from Tripterygium wilfordii

The 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR) is the last step key enzyme of the methylerythritol phosphate (MEP) pathway, synthesizing isopentenyl diphosphate and its allyl isomer dimethylallyl diphosphate, which is important for regulation of isoprenoid biosynthesis. Here the full-length cDNA of HDR, designated TwHDR (GenBank Accession No. KJ933412.1), was isolated from Tripterygium wilfordii for the first time. TwHDR has an open reading frame (ORF) of 1386 bp encoding 461 amino acids. TwHDR exhibits high homology with HDRs of other plants, with an N-terminal conserved domain and three conserved cysteine residues. TwHDR cDNA was cloned into an expression vector and transformed into an Escherichia coli hdr mutant. Since loss-of-function E.coli hdr mutant is lethal, the result showed that transformation of TwHDR cDNA rescued the E.coli hdr mutant. This complementation assay suggests that the TwHDR cDNA encodes a functional HDR enzyme. The expression of TwHDR was induced by methyl-jasmonate (MJ) in T. wilfordii suspension cells. The expression of TwHDR reached the highest level after 1 h of MJ treatment. These results indicate that we have identified a functional TwHDR enzyme, which may play a pivotal role in the biosynthesis of diterpenoid triptolide in T. wilfordii

Characterization and phylogenetic analysis of the complete chloroplast genome of Cercis canadensis ‘Forest Pansy’

Cercis canadensis ‘Forest Pansy’ is a tree species with high ornamental value, which complete chloroplast (cp) genome was sequenced, assembled, and annotated. The genome size is 158,960 bp with a total GC content of 36.17%. The cp genome is made up of a large single-copy region (88,114 bp), a small single-copy region (19,590 bp), and two inverted repeat regions (25,628 bp each). It contains 128 genes, including 84 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. Eighteen genes were duplicated in IRs. The maximum-likelihood (ML) phylogenetic analysis indicated that the Leguminosae species are grouped together, and C. canadensis ‘Forest Pansy’ is closely related to C. canadensis. The result would provide valuable information for genetic studies on Cercis genus

Osteology of <i>Huabeisaurus allocotus</i> (Sauropoda: Titanosauriformes) from the Upper Cretaceous of China

<div><p>Background</p><p>The Late Cretaceous titanosauriform sauropod <i>Huabeisaurus allocotus</i> Pang and Cheng is known from teeth and much of the postcranial skeleton. Its completeness makes it an important taxon for integrating and interpreting anatomical observations from more fragmentary Cretaceous East Asian sauropods and for understanding titanosauriform evolution in general.</p><p>Methodology/Principal Findings</p><p>We present a detailed redescription of <i>Huabeisaurus allocotus</i> and a suite of anatomical comparisons with other titanosauriforms that demonstrate its validity via autapomorphies (e.g., division of some presacral vertebral laminae, reduced development of caudal ribs, the development of fossae relative to one another in caudal vertebral neural arches, high tibia-to-femur ratio). <i>Huabeisaurus</i> shares many features with other Cretaceous East Asian sauropods (e.g., pendant cervical ribs, anterior-middle caudal vertebrae with a nearly flat anterior centrum face and a concave posterior centrum face) that are absent in sauropods from other landmasses and strata, suggesting a close relationship among many of these forms within the clade Somphospondyli.</p><p>Conclusions/Significance</p><p>Restudy of <i>Huabeisaurus</i> provides further evidence for the existence of a clade of somphospondylans – Euhelopodidae – mainly found in the Cretaceous of East Asia. Euhelopodidae represents a fourth example of the evolution of narrow crowns within Sauropoda, along with diplodocoids, brachiosaurids, and advanced titanosaurs (lithostrotians). Despite being known from fewer species than Diplodocoidea, Brachiosauridae, or Lithostrotia, euhelopodids possessed a broader range of tooth shapes than any of these clades, suggesting that euhelopodids exemplified a comparably broad range of feeding strategies and perhaps diets.</p></div

Holotypic left femur of <i>H. allocotus</i> (HBV-20001) from the Upper Cretaceous of Shanxi, China.

<p>Photographed during original preparation in (A) posterior, (B) medial, (C) anterior, and (D) distal views. Silhouette between (B) and (C) shows schematic cross section of the diaphysis. <i>Abbreviations: fic, fibular condyle; ftr, fourth trochanter; tic, tibial condyle.</i> Striped pattern indicates broken surface, dashed lines indicate broken bone margins.</p

Measurements of the left humerus (HBV-20002) from the Upper Cretaceous of China previously referred to as the paratype of <i>Huabeisaurus allocotus</i>.

<p>Measurements of the left humerus (HBV-20002) from the Upper Cretaceous of China previously referred to as the paratype of <i>Huabeisaurus allocotus</i>.</p

Holotypic middle caudal vertebrae of <i>H. allocotus</i> (HBV-20001) from the Upper Cretaceous of Shanxi, China.

<p>In (A) anterior; (B) posterior, (C) left lateral, (D) right lateral, (E) dorsal, and (F) ventral views. Ninth, eleventh, twelfth, seventeenth, eighteenth, and nineteenth caudal vertebrae are depicted from left to right in each row. <i>Abbreviations: cf, chevron facet; pocdf; postzygapophyseal centrodiapophyseal fossa; posdf, postzygapophyseal spinodiapophyseal fossa, ri, ridge.</i> Striped pattern indicates broken surface; dashed lines indicate broken bone margins.</p

Holotypic left radius of <i>H. allocotus</i> (HBV-20001) from the Upper Cretaceous of Shanxi, China.

<p>In (A) proximal, (B) anterior, (C) lateral, and (D) posterior views. Silhouette between (C) and (D) shows schematic cross section of the diaphysis. (A) photographed in 2012; (B–D) photographed during original preparation. <i>Abbreviations: gl, glenoid; igl, infraglenoid lip; tu, tubercle.</i> Dashed lines indicate broken bone margins.</p