Project description:We have sequenced a wild Prunus mume and constructed a reference sequence for this genome. In order to improve quality of gene models, RNA samples of five tissues (bud, leaf, root, stem, fruit) were extracted from the Prunus mume. To investigate tissue specific expression using the reference genome assembly and annotated genes, we extracted RNA samples of different tissues and conducted transcriptome sequencing and DEG analysis. Five RNA pools were created corresponding to different tissues of the Prunus mume.
Project description:Stamen development is an important developmental process that directly affects the yield of Prunus sibirica. In this study, the male sterile flower buds and male fertile flower buds of Prunus sibirica were used as materials to performed RNA-Seq analyses to compare transcription differences. The results would provide a theoretical basis for further investigation of the formation mechanism of male sterile flower.
Project description:We have sequenced a wild Prunus mume and constructed a reference sequence for this genome. In order to improve quality of gene models, RNA samples of five tissues (bud, leaf, root, stem, fruit) were extracted from the Prunus mume. To investigate tissue specific expression using the reference genome assembly and annotated genes, we extracted RNA samples of different tissues and conducted transcriptome sequencing and DEG analysis.
Project description:Purpose: The State of Rio Grande do Sul is the largest producer of peaches from Brazil. However, it still has low values of productivity when compared to other States. One of the problems associated to it this is the occurrence of drainage soils problems, which can suffer flooding situations potentially hampering the development and productivity such culture. For studies to assist in the selection of flood tolerant genotypes, it is essential to understand the physiological and molecular changes of the plants in situations of oxygen deprivation. Using Illumina Hiseq2500 we performed transcriptome analysis of leaves from ‘Capdeboscq’ (Prunus persica) and ‘Julior’ (Prunus insititia x Prunus domestica) rootstocks under flooding for 48 hours. Methods: The mRNA of Prunus spp. plants cv. Capdeboscq e Julior was generated using deep sequencing, in triplicate, using Illumina Hi-Seq 2500, for the following treatments:I) control: plants received irrigation daily until field capacity; and II) plants exposed to flood stress, maintaining a water level of approximately 3 cm above the ground. The sequence reads that passed quality filters were analyzed at the transcript level using this method: Mapping using STAR and identification of differentially expressed genes (DEGs) was performed with the edgeR (false discovery rates - FDRs of <0.05). RT–qPCR validation was performed using SYBR Green assays. Results: Flooding stress causes important high transcriptional changes in the ‘Capdeboscq’ compared to 'Julior' and this is mainly due to their sensitivity/tolerance levels. ‘Capdeboscq’ had photosynthesis as the most affected physiological process at the molecular level, showing a large number of down-regulated enriched GOs, even though it activated cellular signaling pathways under flooding. 'Julior' was more efficient in defense responses, which include the activation of flavonoid biosynthesis pathways. Conclusions: The analysis of two Prunus spp. rootstocks contrasting to the level of tolerance / sensitivity provide new insights into the process of plant flood stress tolerance.
Project description:Pistil development is an important developmental process that directly affects the yield of Prunus sibirica. Through transcriptome sequencing analysis of clones with abortive pistil (No. 595) and normal pistil (No. 28) of Prunus sibirica, a total of 1950 significantly differentially expressed genes were obtained, among which 1000 genes were up-regulated and 950 genes were down-regulated. The results provide a theoretical basis for further investigation of the formation mechanism of pistil abortion.
Project description:In this study, we used vascular specific promoters and a translating ribosome affinity purification strategy to identify phloem associated translatomes in Prunus domestica L. Three different promoter:FLAG-RPL18 lines were used. These included two phloem specific promoters (pSUC2 and pSULTR2;2), as well as the more ubiquitously expressed cauliflower mosaic virus 35S promoter (p35S). Immunopurification of ribosome-mRNA complexes was accomplished by the method described in Reynoso et al. (Plant Functional Genomics: Methods and Protocols, 185-207; 2015). The dataset includes samples from plum leaves taken at 2, 4 and 6 weeks post vernalization.
Project description:Bud dormancy is a critical developmental process for perennial plant survival, and also an important physiological phase that affects the next seasonM-bM-^@M-^Ys growth of temperate fruit trees. Bud dormancy is regulated by multiple genetic factors, and affected by various environmental factors, tree age and vigor. To understand molecular mechanism of bud dormancy in Japanese apricot (Prunus mume Sieb. et Zucc.), we constructed a custom oligo DNA microarray covering the Japanese apricot dormant bud ESTs referring to peach (P. persica) genome sequence. Because endodormancy release is a chilling temperature-dependent physiological event, genes showing chilling-mediated differential expression patterns are candidates to control endodormancy release. Using the microarray constructed in this study, we monitored gene expression changes of dormant vegetative buds of Japanese apricot during prolonged artificial chilling exposure. In addition, we analyzed seasonal gene expression changes. M-bM-^@M-^XNankoM-bM-^@M-^Y vegetative buds collected in November, and those exposed to chilling for 40 or 60 days were used as microarray samples. Among the 58539 different unigene probes, 2345 and 1059 genes were identified as being more than two-fold up-regulated and down-regulated, respectively, following chilling exposure for 60 days (P value < 0.05). The down-regulated genes included P. mume DORMANCY-ASSOCIATED MADS-box genes, which supported the previous quantitative RT-PCR and EST analyses showing that these genes are repressed by prolonged chilling treatments. The genes encoding lipoxygenase were remarkably up-regulated by prolonged chilling. Cluster analysis suggested that the expression of the genes showing expression changes by artificial chilling exposure were coordinately regulated by seasonal changes. Our parametric analysis of gene set enrichment suggested that genes related to jasmonic acid (JA) and oxylipin biosynthesis and metabolic processes were significantly up-regulated by prolonged chilling, whereas genes related to circadian rhythm were significantly down-regulated. The results obtained from the microarray analyses were verified by quantitative RT-PCR analysis of selected genes. Taken together, this study raised the possibility that the microarray platform constructed in this study is applicable for deeper understanding of molecular network related to agronomically important bud phisiologies including dormancy release. In this study, we used chilling exposed bud samples (0, 40, 60 days starting at November) and seasonal monthly bud samples (June to March). For the samples in dataset 1 (three different time points during chilling treatment), three technical replicates (60K M-CM-^W 3 per sample) with three biological replicates were averaged, whereas three technical replicates were averaged for the samples in dataset 2 (10 different seasonal time points)