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| Newsletter 3 | Agosto 2021 |
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 VqBGH40a isolated from Chinese wild Vitis quinquangularis degrades trans-piceid and enhances trans-resveratrol
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Resveratrol is a phytoalexin that can prevent plants from pathogen attacks. Piceid is the glycosylation product of resveratrol and the main storage form of stilbenes in grapevines. Here, we reported the function of a β-glycoside hydrolase gene, VqBGH40a, from the Chinese wild grapevine Vitis quinquangularis accession Danfeng-2 in the regulation of plant resistance to powdery mildew (Uncinula necator). VqBGH40a was transiently overexpressed in Danfeng-2 leaves and then artificially inoculated with powdery mildew showed that VqBGH40a protein could hydrolyze trans-piceid in vivo. These results demonstrate that VqBGH40a can hydrolyze trans-piceid, enhance trans-resveratrol content, and participate in the defense mechanism of grapevine against powdery mildew.
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Rapid developments in the field of plant genome editing using CRISPR-Cas systems necessitate more detailed consideration of the delivery of the CRISPR system into plants. Successful and safe editing of plant genomes is partly based on efficient delivery of the CRISPR system. Along with the use of plasmids and viral vectors as cargo material for genome editing, non-viral vectors have also been considered for delivery purposes. These non-viral vectors can be made of a variety of materials, including inorganic nanoparticles, carbon nanotubes, liposomes, and protein- and peptide-based nanoparticles, as well as nanoscale polymeric materials. They have a decreased immune response, an advantage over viral vectors, and offer additional flexibility in their design, allowing them to be functionalized and targeted to specific sites in a biological system with low cytotoxicity.
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Application of CRISPR/Cas9 technology in wild apple (Malus sieverii) for paired sites gene editing
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Xinjiang wild apple is an important tree of the Tianshan Mountains, and in recent years, it has undergone destruction by many biotic and abiotic stress and human activities. It is necessary to use new technologies to research its genomic function and molecular improvement. The CRISPR/Cas system has been successfully applied to genetic improvement in many crops, but its editing capability varies depending on the different combinations of the synthetic guide RNA (sgRNA) and Cas9 protein expression devices. In this study, we used 2 systems of vectors with paired sgRNAs targeting to MsPDS. We successfully induced the albino phenotype of calli and buds in both systems.
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The CRISPR/Cas9 system has been used for genome editing in several plant species; however, there are few reports on its use in trees. Here, CRISPR/Cas9 was used to mutate a target gene in Populus alba × Populus glandulosa hybrid poplars. The hybrid poplar is routinely used in molecular biological studies due to the well-established Agrobacterium-mediated transformation method. A single guide RNA (sgRNA) with reported high mutation efficiency in other popular species was designed with a protospacer adjacent motif sequence for the phytoene desaturase 1 (PagPDS1) gene. This technique will be useful to improve tree quality in hybrid poplars (P. alba × P. glandulosa); for example, by enhancing biomass or stress tolerance.
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Muscat Flavor in Grapevine: A Digital PCR Assay to Track Allelic Variation in VvDXS Gene
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The aroma of grapes and derived wines has long been one of the major traits considered in the selection of grapevine varieties through the centuries. In particular, Muscat aromatic grapes have been highly appreciated and widespread since ancient times. Monoterpenes are the key compounds responsible for the Muscat flavor. A major QTL affecting monoterpene level has been found to co-localize with the 1-deoxy-D-xylulose 5-phosphate synthase (VvDXS) gene, encoding for the 1-deoxy-D-xylulose 5-phosphate synthase enzyme involved in the plastidial pathway of terpene biosynthesis. In more detail, a single nucleotide polymorphism (SNP 1822) in the coding region of the gene causes a "gain of function" mutation, which is involved in Muscat flavor. We have developed a digital PCR-based assay to target allelic variations in the VvDXS gene, SNP1822, with the aim to propose a fast and sensitive analytical tool for targeting Muscat-flavored grapevine genotypes.
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Grape growers use rootstocks to provide protection against pests and pathogens and to modulate viticulture performance such as shoot growth. Our study examined two grapevine scion varieties ('Chardonnay' and 'Cabernet Sauvignon') grafted to 15 different rootstocks and determined the effect of rootstocks on eight traits important to viticulture. We assessed the vines across five years and identified both year and variety as contributing strongly to trait variation. The effect of rootstock was relatively consistent across years and varieties, explaining between 8.99% and 9.78% of the variation in growth-related traits including yield, pruning weight, berry weight and Ravaz index (yield to pruning weight ratio).
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Dynamic changes in transposable element and gene methylation in mulberry (Morus notabilis) in response to Botrytis cinerea
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DNA methylation has been proposed to regulate plant stress resistance. However, the dynamic changes in DNA methylation in woody plants and their correlations with pathogenic responses are not fully understood. Here, we present single-base maps of the DNA methylomes of mulberry (Morus notabilis) leaves that were subjected to a mock treatment or inoculation with Botrytis cinerea. Compared with the former, the latter showed decreased mCG and mCHG levels and increased mCHH levels. DNA methylation inhibitors reduced resistance gene methylation levels and enhanced mulberry resistance, suggesting that the hypomethylation of resistance genes affects mulberry resistance to B. cinerea.
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Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most devastating citrus diseases worldwide. Generating disease-resistant citrus varieties is considered one of the most efficient and environmentally friendly measures for controlling canker. Xcc causes canker symptoms by inducing the expression of canker susceptibility gene LOB1 via PthA4, a transcription activator-like (TAL) effector, by binding to the effector binding element (EBE) in the promoter region. Two different sgRNAs were used to target both EBE types. Both 35S promoter and Yao promoter were used to drive the expression of SpCas9p to modify EBEPthA4-LOBP in grapefruit. Using 'Duncan' grapefruit epicotyls as explants, 19 genome-edited grapefruit plants were generated with one biallelic mutant line (#DunYao7). Xcc caused canker symptoms on wild-type and non-biallelic mutant plants but not on #DunYao7. This study represents the first time that CRISPR-mediated genome editing has been successfully used to generate disease-resistant plants for 'Duncan' grapefruit, paving the way for utilizing disease-resistant varieties to control canker.
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VvXYLP02 confers gray mold resistance by amplifying jasmonate signaling pathway in Vitis vinifera
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Xylogen-like proteins (XYLPs) are essential for plant growth, development, and stress responses. However, little is known about the XYLP gene family in grape and its protective effects against gray mold a destructive disease caused by Botrytis cinerea. We identified and characterized six common XYLPs in the Vitis vinifera genome (VvXYLPs). VvXYLP expression pattern analyses with B. cinerea infection showed that VvXYLP02 was significantly up-regulated in the resistant genotype but down-regulated or only slightly up-regulated in the susceptible genotype. VvXYLP02 overexpression in Arabidopsis thaliana significantly increased resistance to B. cinerea, indicating that the candidate gene has functional importance. Furthermore, JA treatment significantly up-regulated VvXYLP02 expression in V. vinifera. JA-responsive genes were also up-regulated in VvXYLP02 overexpression lines in A. thaliana under B. cinerea inoculation. These findings suggest that VvXYLP02, which is induced by JA upon the pathogen infection, enhances JA dependent response to enforce plant resistance against gray mold disease.
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The present invention provides the identification and use of EGTom1 and/or EGTom2, homologs of EGTom1 and/or EGTom2, orthologs of EGTom1 and/or EGTom2, paralogs of EGTom1 and/or EGTom2, and fragments and variations thereof for altering salt tolerance, drought tolerance and/or sugar content of fruit (sweetness) in plants.
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Genome editing including the introducing of precise gene edits is well established in diploid plants. Methods well established in the art introduce double strand DNA breaks in the genome of a plant applying technologies such as Zn-finger nucleases, homing endonucleases, TALEN or RNA guided nuclease e.g. Cas9 or Cas12a.
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The invention provides an RNA bacteriostatic agent phasiRNA5. The RNA bacteriostatic agent phasiRNA5 has an RNA sequence as shown in SEQ ID NO.1. The invention also provides a crop pathogen inhibitor. The inhibitor at least comprises phasiRNA5. Specifically, the crop pathogen inhibitor can be a spray containing the phasiRNA5, and can effectively prevent and treat gray mold through spraying. According to the invention, an applied phasiRNA5 solution can significantly inhibit the germination of botrytis cinerea spores and the infection toxicity of botrytis cinerea to plants, and can be used for preventing and treating gray mold of vegetable crops.
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Dirección de Innovación y Transferencia Tecnológica
Vicerrectoría de Investigación y Doctorado - Universidad Andrés Bello ®
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IDENTIFICATION AND USE OF GRAPE GENES CONTROLLING SALT/DROUGHT TOLERANCE AND FRUIT SWEETNESS