Abstract The type II clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 system (CRISPR/Cas9) has been successfully applied to edit target genes in multiple plant species. However, it remains unknown whether this can be used for genome editing grape. In study, we described and targeted gene mutation ‘Chardonnay’ suspension cells plants via the CRISPR/Cas9 system. Two single guide RNAs (sgRNAs) were designed distinct sites of L-idonate dehydrogenase ( IdnDH...

Abstract Cultivated grapevine (Vitis) is a highly valued horticultural crop, and cold stress affects its growth productivity. Wild Amur grape (Vitis amurensis) PAT1 (Phytochrome A signal transduction 1, VaPAT1) induced by low temperature, ectopic expression of VaPAT1 enhances tolerance in Arabidopsis (Arabidopsis thaliana). However, little known about the molecular mechanism during response grapevine. Here, we confirmed overexpression transformed calli enhanced tolerance. Yeast two-hybrid...

Sugar accumulation is a critical event during grape berry ripening that determines the market values. Berry cells are highly dependent on sugar transporters to mediate cross-membrane transport. However, role of in improving berries not well established grapes. Herein we report Sugars Will Eventually be Exported Transporter (SWEET), is, VvSWEET10, was strongly expressed at onset (véraison) and can improve content. VvSWEET10 encodes plasma membrane-localized transporter, heterologous...

E-cadherin is a well characterized adhesion molecule that plays major role in epithelial cell adhesion. Based on findings expression of frequently lost human cancers, it has been implicated as tumor suppressor carcinogenesis most cancers. However, ovarian cancer development, our data from the current study showed uniquely elevated 86.5% benign, borderline, and malignant carcinomas irrespective degree differentiation, whereas normal samples do not express E-cadherin. Thus, we hypothesize may...

Clustered regularly interspersed short palindromic repeats (CRISPR)/Cas system is an efficient targeted genome editing method. Although CRISPR/Cas9-mediated mutagenesis has been applied successfully in grape, few studies have examined the technique's efficiency. To optimize CRISPR/Cas9 efficiency Vitis vinifera, we surveyed three key parameters: GC content of single guide RNA (sgRNA), variety transformant cells used, and SpCas9 expression levels transgenic cell mass. Four sgRNAs with...

Shoot branching is an important trait of plants that allows them to adapt environment changes. Strigolactones (SLs) are newly identified plant hormones inhibit shoot in plants. The SL biosynthesis genes CCD7 (carotenoid cleavage dioxygenase 7) and CCD8 have been found regulate several herbaceous by taking advantage their loss-of-function mutants. However, the role for control grapevine still unknown due lack corresponding mutants.Here we employed CRISPR/Cas9 system edit VvCCD7 VvCCD8 grape...

Abscisic acid (ABA) plays a crucial role in abiotic stress responses. The pyrabactin resistance (PYR)/PYR-like (PYL)/regulatory component of ABA receptor (RCAR) proteins that have been characterized as receptors function the core components signaling pathway. However, functions grape PYL genes response to different stresses, particularly cold stress, remain less studied.In this study, we investigated expression profiles upon treatment and isolated VaPYL4 gene from Vitis amurensis, cold-hardy...

Abstract Overexpression and knockout (or knockdown) of gene interest are two commonly used strategies for functional study. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system-mediated has been applied in most plant species, including grapevine. However, CRISPR/dCas9 (deactivated Cas9)-based transcriptional activation is still unreported fruit crops, although a few studies have documented Arabidopsis rice. Here, we tested activators VP64 TV endogenous genes grape....

Nuclear factor Y (NF-Y) transcription is composed of three distinct subunits: NF-YA, NF-YB and NF-YC. Many members NF-Y family have been reported to be key regulators in plant development, phytohormone signaling drought tolerance. However, the function less known grape (Vitis vinifera L.). A total 34 genes that distributed unevenly on (V. vinifera) chromosomes were identified this study. Phylogenetic analysis was performed predict functional similarities between Arabidopsis thaliana genes....

CRISPR/Cas9 has been recently demonstrated as an effective and popular genome editing tool for modifying genomes of humans, animals, microorganisms, plants. Success such is highly dependent on the availability suitable target sites in to be edited. Many specific have computationally identified several annual model crop species, but not reported perennial, woody fruit species. In this study, we characterized five types widely cultivated grape species Vitis vinifera developed a user-friendly...

The plant GATA family is one of the most important transcription factors involved in light-responsive development, nitrogen metabolism, phytohormone signaling, and source/sink balance. However, function gene less known grape (Vitis vinifera L.). In this study, we comprehensively analyzed grape, particularly phylogenetic evolution, duplication patterns, conserved motifs, structures, cis-elements, tissue expression predicted VvGATA genes response to abiotic stress. potential roles berry...

Cold tolerance is regulated by a variety of transcription factors (TFs) and their target genes. Except for the well-characterized C-repeat binding (CBFs)-dependent transcriptional cascade, mechanisms cold mediated other regulatory networks are still largely unknown. Here, we used assay transposase-accessible chromatin with sequencing (ATAC-seq) RNA-seq to identify responsive TFs in Vitis amurensis, grape species high hardiness. Nine TFs, including CBF4, RAV1 ERF104, were identified after...

The development of clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) system has revolutionized genome editing and plant breeding. Applications CRISPR/Cas9 technology in fruit crops, including grapevine, enable precise improvement agronomically important traits. In this review, we first describe based on the mostly used newly developed CRISPR technologies. We then focus applications disease resistance, optimization system, multiplex off-target...

Clustered regularly interspaced short palindromic repeats (CRISPR) /Cas12a system, also known as CRISPR/Cpf1, has been successfully harnessed for genome engineering in many plants, but not grapevine yet. Here we developed and demonstrated the efficacy of CRISPR/Cas12a from Lachnospiraceae bacterium ND2006 (LbCas12a) inducing targeted mutagenesis by targeting tonoplastic monosaccharide transporter1 (TMT1) dihydroflavonol-4-reductase 1 (DFR1) genes 41B cells. Knockout DFR1 gene altered...

Anthocyanins are essential for the quality of perennial horticultural crops, such as grapes. In grapes, ELONGATED HYPOCOTYL 5 (HY5) and MYBA1 two critical transcription factors that regulate anthocyanin biosynthesis. Our previous work has shown Vitis vinifera B-box protein 44 (VvBBX44) inhibits synthesis represses VvHY5 expression in grape calli. However, regulatory mechanism underlying this regulation was unclear. study, we found loss VvBBX44 function resulted increased accumulation...

Wood vinegar (WV), a byproduct from the charcoal production process, has been reported to have excellent antioxidant capability by chemical examination. However, biological effect of WV in living animals is still unknown. In this study, simple model organism, nematode Caenorhabditis elegans, was used as an vivo system assess effects wood through development, lifespan, brood size, germline cell apoptosis and superoxide dismutase (SOD) level.Wood extract (WVE) promoted prolonged lifespan...

Abstract Various studies demonstrated that bone morphogenetic proteins (BMPs) and their antagonists contribute to the development of cancers. Chordin‐like 2 (CHRDL2) is a member BMP antagonists. However, role its relative mechanism CHRDL2 in osteosarcoma remains unclear. In present study, we expression was significantly upregulated tissues cell lines compared with adjacent human normal osteoblast. Inhibition decreased proliferation colony formation cells vitro, as well migration invasion....

The cure rate for patients with osteosarcoma (OS) has stagnated over the past few decades. Penfluridol, a first-generation antipsychotic, demonstrated to prevent lung and esophageal malignancies from proliferation metastasis. However, effect of penfluridol on OS its underlying molecular mechanism remains unclear. This study revealed that effectively inhibited cell migration, induced G2/M phase arrest in cells. In addition, treatment was found increased reactive oxygen species (ROS) levels...

A motile, Gram-stain-negative, non-pigmented bacterial strain, designated MGL06T, was isolated from seawater of the South China Sea on selection medium containing 0.1 % (w/v) malachite green. Strain MGL06T showed highest 16S rRNA gene sequence similarity to Rhizobium vignae CCBAU 05176T (97.2 %), and shared 93.2-96.9 with type strains other recognized species. Phylogenetic analyses based housekeeping sequences that strain belonged genus Rhizobium. Mean levels DNA-DNA relatedness between R....

Many investigations have shown that nickel exposure can induce micronuclei generation, inhibit DNA repair and cell apoptosis, both in cells tissues. However, there is a lack of appropriate vivo animal models to study the underlying mechanisms nickel-induced apoptosis. The model organism, Caenorhabditis elegans, has been be good for investigating many biological processes. In present study, we detected 0.01 mM induced significantly germline apoptosis after treatment 12 hours, which...