Project description:A grape-bud-oriented genomic platform was produced for a large-scale comparative analysis of bud responses to two stimuli of grape-bud dormancy release, hydrogen cyanamide (HC) and heat shock (HS). The results suggested considerable similarity in bud response to the stimuli, both in the repertoire of responding genes and in the temporary nature of the transcriptome reprogramming. Nevertheless, the bud response to HC was slower, more condensed and stronger, as reflected by a higher number of regulated genes and a higher intensity of regulation compared to the response to HS.
Project description:Bud endodormancy induction response of two genotypes (Seyval, a hybrid white wine grape and Vitis riparia, PI588259, a native North American grape species) was compared under long (15 h) and short (13 h) photoperiods. Proteins were extracted from both genotypes for all time points and experimental conditions. The proteins were separaed by 2D-PAGE, trypsin digested, and the peptides identified with a MALDI-TOF-TOF mass spectrometer. A master gel was made and mapped with all proteins from both genotypes. The proteins were identified by matching the peptide sequences against the 8X Vitis vinifera grape genome in NCBI. This study was funded by NSF grant DBI064755 and is the result of a collaboration between Dr. Anne Fennell at South Dakota State University and Dr. Grant R. Cramer at the University of Nevada, Reno.
Project description:This study investigated changes in gene expression of controlled environment chilled (4C) grape overwintering buds as they accumulated from 0 to 2000 chilling hours. Keywords: time course, chilling, endodormancy release, axillary bud, grape
Project description:A grape-bud-oriented genomic platform was produced for a large-scale comparative analysis of bud responses to two stimuli of grape-bud dormancy release, hydrogen cyanamide (HC) and heat shock (HS). The results suggested considerable similarity in bud response to the stimuli, both in the repertoire of responding genes and in the temporary nature of the transcriptome reprogramming. Nevertheless, the bud response to HC was slower, more condensed and stronger, as reflected by a higher number of regulated genes and a higher intensity of regulation compared to the response to HS. To facilitate large-scale comparative analysis of early changes in the bud transcriptome by cDNA microarray, HC and HS were applied to canes collected from three vineyards, located in different regions, in three different years. This experimental scheme resulted in two true biological replicates for each treatment, differing in both timing and location, and loop design of technical replicates within time series. Consistent with our previous studies, both application of 5% Dormex (HC) and incubation for 1 h in 50oC water (HS) resulted in increased bud-break rates compared to respective controls. Bud break of HS-treated and HC-treated buds started 10 to 12 days after treatment. Three weeks after treatment, HS-treated buds exhibited 100% bud break while HC-treated buds had reached 80% bud break. The control showed significantly lower levels of bud break during this period. Bud material was collected from control, HC- and HS-treated cuttings at six time points (3, 6, 12, 24, 48 and 96 h) after the treatments and used to prepare total RNA samples.
Project description:This study investigated changes in gene expression of controlled environment chilled (4C) grape overwintering buds as they accumulated from 0 to 2000 chilling hours. Keywords: time course, chilling, endodormancy release, axillary bud, grape A loop design with 3 biological replicates (RNA from buds collected at 0, 500, 1000, 1500, and 2000 hr of chilling in 2002, 2004, and 2005).
Project description:Bud dormancy in perennials in boreal and temperate ecosystems is crucial for survival in harsh winter. Dormancy is released by prolonged exposure to low temperatures and is followed by reactive growth in the spring. Lysine acetylation (Kac) is one of the major post-translational modifications (PTMs) involved in plant response to environment signals. However, little information is available on the effects of Kac modification on bud dormancy release. Here, we report the dynamics of lysine acetylome in hybrid poplar (Populus tremula x alba) dormant buds. A total number of 7,594 acetyl sites from 3,281 acetyl proteins were identified, representing the largest to date dataset of lysine acetylome in plants. Of them, 229 proteins were differentially acetylated during bud dormancy release and were involved mainly in the primary metabolism. Site-directed mutagenesis enzymatic assays showed that acetylation strongly modified the activities of two key enzymes of primary metabolism, pyruvate dehydrogenase (PDH) and isocitrate dehydrogenase (IDH). We thus propose that Kac of enzymes could be an important strategy for reconfiguration of metabolic processes during bud dormancy release. In all, our results reveal the importance of Kac in bud dormancy release and give a new perspective to understand the molecular mechanisms of tree’s seasonal growth.
Project description:Solar ultraviolet C(UV-C)radiation reaching the Earth’s surface is little due to the filtering effects of the stratospheric ozone layer. At present, artificial UV-C irradiation is utilized for different biological processes. Grape is a major fruit crop around the world. Research has shown that UV-C irradiation induced the biosynthesis of phenols. However, changes at the molecular level in response to UV-C and leading to these effects are poorly understood. To elucidate the effect of UV-C on expression of genes in grape and the response mechanism, transcript abundance of grape (Vitis vinifera L.) leaves was quantified using the Affymetrix Grape Genome oligonucleotide microarray (15,700 transcripts)
Project description:Solar ultraviolet C(UV-C)radiation reaching the Earth’s surface is little due to the filtering effects of the stratospheric ozone layer. At present, artificial UV-C irradiation is utilized for different biological processes. Grape is a major fruit crop around the world. Research has shown that UV-C irradiation induced the biosynthesis of phenols. However, changes at the molecular level in response to UV-C and leading to these effects are poorly understood. To elucidate the effect of UV-C on expression of genes in grape and the response mechanism, transcript abundance of grape (Vitis vinifera L.) leaves was quantified using the Affymetrix Grape Genome oligonucleotide microarray (15,700 transcripts) Grape leaves were exposed to UV-C irradiation at 6W/m2 for 10 min. LCK-0-1, LCK-0-2 and LCK-0-3 are 0 h after the initiation of treatment and as the controls; LTR-6-1, LTR-6-2 and LTR-6-3 are for 6 h after the initiation of treatment; LTR-12-1, LTR-12-2 and LTR-12-3 are for 12 h after the initiation of treatment. Three replicates for each time point. 9 samples in all.