Project description:Arbuscular mycorrhizal symbiosis improves water and nutrient uptake by plants and provides them other ecosystem services. Grapevine is one of the major crops in the world. V. vinifera scions are generally grafted onto a variety of rootstocks that confer different levels of resistance against different pests, tolerance to environmental stress, and influence the physiology of the scions. Arbuscular mycorrhizal fungi are involved in the root architecture and in the immune response to soil-borne pathogens. However, the fine-tuned regulation and the transcriptomic plasticity of rootstocks in response to mycorrhization are still unknown. We compared the responses of 10 different grapevine rootstocks to arbuscular mycorrhizal symbiosis (AMS) formed with Rhizophagus irregularis DAOM197198 using RNA sequencing-based transcriptome profiling. We have highlighted a few shared regulation mechanisms, but also specific rootstock responses to R. irregularis colonization. A set of 353 genes was regulated by AMS in all ten rootstocks. We also compared the expression level of this set of genes to more than 2,000 transcriptome profiles from various grapevine varieties and tissues to identify a class of transcripts related to mycorrhizal associations in these 10 rootstocks. Then, we compared the response of the 351 genes upregulated by mycorrhiza in grapevine to their Medicago truncatula homologs in response to mycorrhizal colonization based on available transcriptomic studies. More than 97% of these homologs were expressed in at least one transcriptome profile, and 64% in all profiles. At the intra-specific level, we described for the first time shared and specific grapevine rootstock genes in response to R. irregularis symbiosis. At the inter-specific level, we defined a shared subset of mycorrhiza-responsive genes
Project description:Several pathogens infect grapevine, including viruses and viroids. Considering that there are no effective plant protection treatments against these pathogens and vineyards are cultivated through decades usage of high quality and pathogen-free propagation material (rootstocks and scions) is essential. Although presence of regulated pests is routinely checked using ELISA or rarely RT-PCR, these diagnostics methods can detect only particular pathogens moreover can fail to detect variant strains. High-throughput sequencing of small RNAs can be an effective, alternative method to avoid these disadvantages. Since for production of grafts, pathogen free cultivars and rootstocks must be used, 17 grapevine rootstock plantations and 2 rootstock variety collections were selected for characterisation of their virom by high throughput sequencing of virus derived small RNAs.
Project description:Since the roots of grapevine rootstocks have a direct contact with drying soil and has an important role in abiotic stimuli, any plasticity on the architecture of the rootstocks would enable grapevine varieties to a better respond to drought stress. However, genomics evidences behind the physiological responses of rootstocks under prolonged drought stress are poorly documented in the literature. In the current study, eight widely used hybrid grapevine rootstocks in viticulture were firstly grafted with sultana seedless and subjected to drought stress to test their physiological and biochemical responses. The results of experiment indicated that the roots of V.rupestris X V.berlandieri (110 R, 1103P, 140 Ru) rootstocks possessed much higher water content as well as non-structural carbohydrate and nitrogen concentrations compared to V.riparia X V.berlandieri (SO4, 5BB, 420A, 8B) and V.vinifera X V.berlandieri (41B) hybrids under drought. V.rupestris X V.berlandieri hybrids were also performed much higher root elongation performance under drought compared to other rootstock hybrids. Three rootstock varieties (110R, 5BB and 41B) having different pedigrees and root architectural responses to drought were also investigated at transcriptome level to find out gene regulation network behind differential physiological responses to drought. Transcriptome analysis revealed 2795, 1196 and 1612 differentially expressed transcripts for the roots of 110R, 5BB and 41B, respectively. The highest expression increases in 110R compared to other rootstocks were recorded for the transcripts functional in carbohydrate (SWEET14, CWINV) and nitrate/peptide (NRT1/ PTR FAMILY) transportation as well as osmoregulation (dehydrins, osmotins, LEAs and proline-glycine rich proteins) during drought. Higher induction of these genes in the roots of tolerant 110R genotype indicated importance of efficient uptake of carbohydrate and nitrogen source released from canopy under drought and preservation of water with osmotic regulation on the root elongation and drought tolerance of grapevines. Expression increases in several other pathogenesis related proteins, regulation of cell wall modification enzymes and activity of several secondary metabolites have been also associated to altered root architecture and drought tolerance in the grapevine rootstocks for the first time with the current study.
Project description:Differences in root transcriptional profile between two grapevine rootstocks (1103P and SO4) showing different tolerance to Mg deficiency were analyzed after 4 and 14 days of growth with (+Mg) or without (-Mg).
Project description:In order to investigate the putative roles of the VvPLCP genes in grapevine resistance, the leaves-specific expression patterns of VvPLCPs were analyzed according to transcriptome data in two cultivars including V. vinifera cv. ‘Zitian Seedless’ and Vitis rootstocks ‘Kober 5BB’ when infected with P. viticola
Project description:Grapevine red blotch is a recently identified viral disease that was first recognized in the Napa Valley of California. Infected plants showed foliar symptoms similar to leafroll, another grapevine viral disease, on vines testing negative for known grapevine leafroll-associated virus. Later, the Grapevine red blotch virus (GRBV) was independently discovered in the US states of California and New York and was demonstrated to be the causal agent of red blotch disease. Due to its wide occurrence in the US, vector transmission and impacts on grape industry, this virus has the potential to cause serious economic losses. Despite numerous attempts, it was not possible to isolate or visualize viral particles from GRBV infected plants. Consequently, this has hampered the development of a serological assay that would facilitate GRBV detection in grapevine. We therefore decided to explore mass spectrometry approaches in order to quantify GRBV in infected plants and to identify potential biomarkers for viral infection. We present for the first time the physical detection on the protein level of the two GRBV genes V1 (coat protein) and V2 in grapevine tissue lysates. The GRBV coat protein load in leaf petioles was determined to be in the range of 100 to 900 million copies per milligram wet weight by using three heavy isotope labeled reference peptides as internal standards. The V1 copy number per unit wet tissue weight in leaves appeared to be about six times lower, and about 200-times lower in terms of protein concentration in the extractable protein mass than in petioles. We found a consistent upregulation of several enzymes involved in flavonoid biosynthesis in leaf and petiole extracts of GRBV-infected plants by label-free shotgun proteomics, indicating the activation of a defense mechanism against GRBV, a plant response already described for grapevine leafroll associated virus infection on the transcriptome level. Last but not least, we identified some other microorganisms belonging to the grapevine leaf microbiota, two bacterial species (Novosphingobium sp. Rr 2-17 and Methylobacterium) and one virus, Grapevine rupestris stem pitting associated virus.
Project description:This study aimed at investigating the effect in Grapevine of two different rootstocks (1103 Paulsen - P - and Mgt 101-14 - M) in comparison with not grafted plants - F - on the miRNome of berry skin in Pinot noir (clone Entav 115), to explore the influence of rootstock-scion interaction on grape quality. 7-year old grapevine plants were grown in 70-liters, in an open field arranged in a randomized block design with 9 replicates for each root system. The plants were maintained in the same agronomic conditions: all the pots were fertilized and were abundantly irrigated. Berry samples (15 per plant, 3 plants per replicate), were collected at two different developmental stage: veraison (1) and maturation (2), and dissected to separate skin tissues. Total RNA was extracted and DNase treated, small RNA libraries were prepared using the TruSeq Small RNA Sample Preparation Kit (Illumina®), following all manufacturers' instructions. Eighteen bar-coded small RNA libraries were constructed starting from 1 µg of total RNAs.
Project description:This study aimed at investigating the effect in Grapevine of two different rootstocks (1103 Paulsen - P - and Mgt 101-14 - M) in comparison with not grafted plants - F - on the transcriptome of berry skin in Pinot noir (clone Entav 115), to explore the influence of rootstock-scion interaction on grape quality. 7-year old grapevine plants were grown in 70-liters, in an open field arranged in a randomized block design with 9 replicates for each root system. The plants were maintained in the same agronomic conditions: all the pots were fertilized and were abundantly irrigated. Berry samples (15 per plant, 3 plants per replicate), were collected at two different developmental stage: veraison (1) and maturation (2), and dissected to separate skin tissues. Total RNA was extracted from berry skins and DNase treated. 18 mRNA seq libraries were prepared, starting from total RNA (1 µg), using TruSeq RNA sample preparation kit (Illumina), according to manufacturers’ instructions. Libraries were quantified through qRT-PCR, as recommended by the protocol, and single-end sequenced for 100 bases on an Illumina Genome Analyzer (GAIIx).
Project description:MicroRNAs (miRNAs) are a class of non-coding RNA molecules which have significant gene regulation roles in organisms. The advent of new high throughput sequencing technologies has enabled the revelation of novel miRNAs. Although there are two recent reports on high throughput sequencing analysis of small RNA libraries from different organs of two grapevine wine varieties, there were significant divergence in the number and kinds of miRNAs sequenced in these studies. More sequencing of small RNA libraries is still important for the discovery of novel miRNAs in grapevine. Here, we initially constructed a small RNA library of flower and fruit tissues of a table grapevine cultivar ‘Summer Black’ and performed sequencing and analysis of sRNAs using the Illumina Solexa platform, expecting to discover more miRNAs related to the development of grapevine flowers and berries and the formation of dessert quality in grapevine berries. Totally, 130 conserved grapevine miRNA (Vv-miRNA) belonging to 28 Vv-miRNA families were validated, and 92 novel potential grapevine-specific ones representing 80 unique ones were first discovered. Forty-two (48.84%) of the novel miRNAs possessed differential semi-quantitative PCR expression profiles in various grapevine tissues that could further confirm their existence in the grapevine, among which twenty were expressed only in grapevine berries, indicating some fruit-specificity. 130 target genes for 46 novel miRNAs could be predicted. The locations of these potential target genes on grapevine chromosomes and their complementary levels with the corresponding miRNAs were also analyzed.