Effective genome editing and identification of a regiospecific gallic acid 4-O-glycosyltransferase in pomegranate (Punica granatum L.).

Pomegranate (Punica granatum L.) trees are woody perennials that bear colorful and nutritious fruits rich in phenolic metabolites, e.g., hydrolyzable tannins (HTs) and flavonoids. We here report genome editing and gene discovery in pomegranate hairy roots using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) (CRISPR/Cas9), coupled with transcriptome and biochemical analyses. Single guide RNAs (sgRNAs) were designed to target two UDP-dependent glycosyltransferases (UGTs), PgUGT84A23 and PgUGT84A24, which possess overlapping activities in β-glucogallin (a galloylglucose ester; biosynthetic precursor of HTs) biosynthesis. A unique accumulation of gallic acid 3-O- and 4-O-glucosides (galloylglucose ethers) was observed in the PgUGT84A23 and PgUGT84A24 dual CRISPR/Cas9-edited lines (i.e., ugt84a23 ugt84a24) but not the control (empty vector) or PgUGT84A23/PgUGT84A24 single edited lines (ugt84a23 or ugt84a24). Transcriptome and real-time qPCR analyses identified 11 UGTs with increased expression in the ugt84a23 ugt84a24 hairy roots compared to the controls. Of the 11 candidate UGTs, only PgUGT72BD1 used gallic acid as substrate and produced a regiospecific product gallic acid 4-O-glucoside. This work demonstrates that the CRISPR/Cas9 method can facilitate functional genomics studies in pomegranate and shows promise for capitalizing on the metabolic potential of pomegranate for germplasm improvement

Effect of sea quarks on the single-spin asymmetries ALW±A^{W^{\pm}}_{L} in polarized pp collisions at RHIC

We calculate the single-spin asymmetries $A^{W^{\pm}}_{L}$ of $W^{\pm}$ bosons produced in polarized pp collisions with the valence part of the up and down quark helicity distributions modeled by the light-cone quark-spectator-diquark model while the sea part helicity distributions of the up and down quarks treated as parametrization. Comparing our results with those from experimental data at RHIC, we find that the helicity distributions of sea quarks play an important role in the determination of the shapes of $A^{W^{\pm}}_{L}$. It is shown that $A^{W^{-}}_{L}$ is sensitive to $\Delta \bar u$, while $A^{W^{+}}_{L}$ to $\Delta \bar d$ intuitively. The experimental data of the polarized structure functions and the sum of helicities are also important to constrain the sizes of quark helicity distributions both for the sea part and the valence part of the nucleon.Comment: 19 latex pages, 5 figures, final version for publicatio

Rigidity of noncompact complete Bach-flat manifolds

Let $(M,g)$ be a noncompact complete Bach-flat manifold with positive Yamabe constant. We prove that $(M,g)$ is flat if $(M, g)$ has zero scalar curvature and sufficiently small $L_{2}$ bound of curvature tensor. When $(M, g)$ has nonconstant scalar curvature, we prove that $(M, g)$ is conformal to the flat space if $(M, g)$ has sufficiently small $L_2$ bound of curvature tensor and $L_{4/3}$ bound of scalar curvature.Comment: 10 pages, To appear J. Geom. Physic

Exploring the Phytochemical Landscape of the Early-Diverging Flowering Plant Amborella trichopoda Baill.

Although the evolutionary significance of the early-diverging flowering plant Amborella (Amborella trichopoda Baill.) is widely recognized, its metabolic landscape, particularly specialized metabolites, is currently underexplored. In this work, we analyzed the metabolomes of Amborella tissues using liquid chromatography high-resolution electrospray ionization mass spectrometry (LC-HR-ESI-MS). By matching the mass spectra of Amborella metabolites with those of authentic phytochemical standards in the publicly accessible libraries, 63, 39, and 21 compounds were tentatively identified in leaves, stems, and roots, respectively. Free amino acids, organic acids, simple sugars, cofactors, as well as abundant glycosylated and/or methylated phenolic specialized metabolites were observed in Amborella leaves. Diverse metabolites were also detected in stems and roots, including those that were not identified in leaves. To understand the biosynthesis of specialized metabolites with glycosyl and methyl modifications, families of small molecule UDP-dependent glycosyltransferases (UGTs) and O-methyltransferases (OMTs) were identified in the Amborella genome and the InterPro database based on conserved functional domains. Of the 17 phylogenetic groups of plant UGTs (A-Q) defined to date, Amborella UGTs are absent from groups B, N, and P, but they are highly abundant in group L. Among the 25 Amborella OMTs, 7 cluster with caffeoyl-coenzyme A (CCoA) OMTs involved in lignin and phenolic metabolism, whereas 18 form a clade with plant OMTs that methylate hydroxycinnamic acids, flavonoids, or alkaloids. Overall, this first report of metabolomes and candidate metabolic genes in Amborella provides a starting point to a better understanding of specialized metabolites and biosynthetic enzymes in this basal lineage of flowering plants