Sequencing Greenlip Abalone - We have used the greenlip abalone draft genome (PRJNA433241) to generate basic genetic knowledge of the reproductive process with a specific focus on the key interactive processes of maturation, neural signalling, communication and the stress response.
Ontology highlight
ABSTRACT:
INSTRUMENT(S): TripleTOF 5600
ORGANISM(S): Haliotis Laevigata
SUBMITTER: 
Tianfang Wang
LAB HEAD: Tianfang Wang
PROVIDER: PXD009218 | Pride | 2019-07-16
REPOSITORIES: Pride
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Project description:Switchgrass plants were grown in a Sandwich tube system to induce gradual drought stress by withholding watering. After 29 days, leaf photosynthetic rate decreased significantly, compared to the control plants which were watered regularly. The drought-treated plants recovered to the same leaf water content after three days of re-watering. Root tip (1cm basal fragment, designated as RT1 hereafter) and the elongation/maturation zone (the next upper 1 cm tissue, designated as RT2 hereafter) tissues were collected at the 29th day of drought stress treatment, (named SDT for severe drought treated), one (D1W) and three days (D3W) of re-watering. The tandem mass tags mass spectrometry-based quantitative proteomics analysis was performed to identify the proteomes, and drought-induced differentially expressed proteins (DEPs). From RT1 tissues, 6,156, 7,687 and 7,699 proteins were quantified, and 296, 535 and 384 DEPs were identified in the SDT, D1W and D3W samples, respectively. From RT2 tissues, 7,382, 7,255 and 6,883 proteins were quantified, and 393, 587 and 321 proteins DEPs were identified in the SDT, D1W and D3W samples. Between RT1 and RT2 tissues, very few DEPs overlapped at SDT, but the number of such proteins increased during the recovery phase. A large number of hydrophilic proteins and stress-responsive proteins were induced during SDT and remained at a higher level during the recovery stages. A large number of DEPs in RT1 tissues maintained the same expression pattern throughout drought treatment and the recovery phases. The DEPs in RT1 tissues were classified in cell proliferation, mitotic cell division, and chromatin modification, and those in RT2 were placed in cell wall remodeling and cell expansion processes. This study provided information pertaining to root zone-specific proteome changes during drought and recover phases, which will allow us to choose the proteins (genes) as better defined targets for developing drought tolerant plants.
2020-06-04 | PXD017441 | Pride
Project description:Environmental stresses such as drought, salinity and both high and low temperature are frequently faced by crops all over the world and can be considered major limiting factors for plant geographical distribution and productivity. Breeding has allowed creation of crops more adapted to some of the adverse environmental conditions and to overcome the geographical limitation without major consequences to productivity. However, due to the climate changes observed in the last few decades, some agricultural areas have experienced the âgreen seed problemâ characterized by chlorophyll retention in mature seeds. This is problematic in oil seed crops such as soybean and canola since it is related to lower seed and oil quality, resulting in serious financial losses. Besides the environmental factors, there are also genetic components controlling the susceptibility of different cultivars to green seed production. Understanding the molecular mechanisms controlling chlorophyll retention in seeds is crucial to allow advanced molecular breeding techniques and genetic engineering as a way to increase tolerance to this growing problem. We have used maturing soybean seeds of the cultivar MG/BR 46, harvested in R6, R7 and R8, produced under non-stressed and stressed environmental conditions, to understand he molecular basis of chlorophyll degradation and, consequently, its retention during soybean seed maturation. Soybean seeds were produced under stressed and non-stressed conditions and harvested in 3 different stages of maturation: R6 non-stressed, R7 non-stressed, R8 non-stressed, R6 stressed, R7 stressed, R8 stressed.
2016-01-21 | E-GEOD-70999 | biostudies-arrayexpress
Project description:The winged pearl oyster, Pteria penguin, is a widely distributed and economically important bivalve with unique and primitive byssus. The byssogensis of this species is largely unknown. Here we report the lable free quantification analysis on different tissues of P. penguin.Through the analysis, we found that the process of byssogenesis requires ATP to provide energy among the differential proteins; and seven (Tenascin-X, laminin, papilin, fibrillin-1, spondin, protease inhibitors and Mucins) of the 31 proteins identified in the byssus were derived from ECM. Functions of extracellular matrix proteins can be correlated with the formation and the nature of the byssus.
2023-12-20 | PXD031701 | Pride
Project description:A genomic overview of in vivo binding of a transcription factor FoxA-a in the ascidian 64-cell stage embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged FoxA-a. After fixation, Chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
2010-03-08 | E-GEOD-17687 | biostudies-arrayexpress
Project description:⢠Herbivore-induced plant volatiles (HIPVs), in addition to attracting natural enemies of herbivores, can serve a signaling function within plants by acting as wound signals that induce or prime defenses. However, particularly in woody plants, which compounds within HIPV blends are capable of acting as signaling molecules are largely unknown. ⢠Leaves of hybrid poplar (Populus deltoides x nigra) saplings were exposed in vivo to naturally wound-emitted concentrations of the green leaf volatile (GLV) cis-3-hexenyl acetate (z3HAC) and then subsequently fed upon by gypsy moth larvae (Lymantria dispar L.). Volatiles were collected throughout the experiments, and leaf tissue was collected to measure phytohormone levels and expression of defense-related genes. ⢠Relative to controls, z3HAC-exposed leaves had higher levels of jasmonic acid and ï¡-linolenic acid following gypsy moth feeding. Further, z3HAC primed transcripts of phytohormone signaling (lipoxygenase 1) and direct defense (a Kunitz proteinase inhibitor) genes. These qRT-PCR results were supported by microarray analysis using the AspenDB 7K EST microarray containing ~5400 unique gene models. Moreover, z3HAC also primed the release of herbivore-induced terpene volatiles. ⢠The widespread priming response suggests an adaptive benefit to detecting z3HAC as a wound signal. Thus, woody plants can detect and use z3HAC as a signaling cue to prime defenses before actually experiencing damage. GLVs may therefore have important ecological functions in arboreal ecosystems. Plants and Gypsy Moths Hybrid poplar (Populus deltoides x nigra, clone âOGYâ: Magnoliopsida; Malpighiales; Salicaceae; Aigeiros section of Populus) saplings were grown from cuttings in a walk-in growth chamber maintained at 25oC with a 16:8 photoperiod. Cuttings were planted in five-gallon pots in commercial potting soil (MetroMix 250, SunGro, Bellevue, WA, USA) and watered as necessary. Due to supplements in the soil, fertilization was not required and poplars grew to heights of ~1.5 m in 12-15 weeks. Newly molted 4th instar L. dispar larvae were used for all experiments. Egg masses were obtained from APHIS (Otis Plant Protection Lab, Cape Cod, MA, USA) and reared through 3rd instar on artificial diet. Newly molted 4th instar larvae cleared their gut contents prior to molting. Experimental Manipulations All experiments were conducted from March-April 2007 in a single walk-in growth chamber maintained as described above. Poplars were equilibrated to the chamber for 24 hours prior to manipulations. Two fully-expanded source leaves (leaf plastochron index LPI 8 and 9) were isolated per sapling using individual 15x15x1 cm Teflon/glass chambers (for photo see Appendix in Frost et al., 2007). These two leaves are non-orthostichous, which means that they do not readily share assimilates or wound signals (Davis et al., 1991). Thus, for the purposes of these experiments, treatment leaves did not interact with their respective control leaves. However, the leaves are of slightly different ages and, to control for this potential effect, we randomized which leaf would receive the z3HAC treatment. Flow into each chamber was set at 2.0 L/min, which was sufficient to avoid condensation within the chamber. The experiments were designed to have two consecutive albeit separate treatments: exposure to z3HAC and subsequent application of L. dispar larvae. We used commercially available z3HAC (Sigma-Aldrich), and prepared concentrations to infuse into each septum such that it delivered ~40 ng z3HAC per application. After a pretreatment volatile collection (Day 1 and night starting Day 2), z3HAC-infused rubber septa were placed into the leaf chambers with each treatment leaf (morning of Day 2). Control leaves each received a septum infused with just the dichloromethane solvent. Septa were replaced twice, once before the afternoon collection on Day 2, and before the morning collection on Day 3. All septa were removed during the night of Day 3, since most volatile production stops at night in this hybrid (Fig 5a). On the morning of Day 4, two newly molted 3rd or 4th instar gypsy moth larvae were added to each leaf, regardless of whether the leaf received the treatment or control septa on Days 2 and 3. The herbivores remained on the leaves for ~24 hours. On the day of collection, each leaf was excised at the petiole, photographed, and then placed in a labeled coin envelope and submerged directly into liquid N2. Each photograph also contained a standard of known area, from which leaf area and % damage calculations were determined (Sigma Scan 500, Systat Software Inc.). Larvae consumed similar amounts on the treatment and control leaves (24 hours: z3HAC-treated leaves: 4.7 ± 1.8 cm2; controls: 4.3 ± 1.7 cm2). The snap-frozen leaves were stored at -80oC until grinding. Each leaf was ground in an individual mortar and pestle under liquid N2. Ground tissue was weighed into vials for RNA extraction and JA analysis (see below). Care was taken to not allow the ground tissue to thaw at any point prior to extraction. This method therefore allowed us to measure gene expression and JA from the same tissue. Microarray Analysis To explore a transcriptome-level response of poplar leaves to z3HAC and herbivory, we conducted microarray analysis of the samples collected after 24 hours of herbivore damage. This allowed a comparison of transcriptome-level responses to herbivory in z3HAC-treated versus untreated leaves (i.e., the set of genes that were primed by z3HAC exposure). Total RNA was labeled using Ambion's Amino Allyl Message Amp II aRNA Amplification Kit (#AM1753) according to the manufacturer's instructions. Briefly, 1 ug of each sample was reversed transcribed using T7-oligo(dT) priming and subsequently second strand cDNA synthesis was performed. The resulting double stranded cDNA was purified and then used as a template for in vitro transcription during which amino allyl-modified UTP was incorporated. The resulting amino allyl-modified cRNA was purified and coupled with Cy3 or Cy5 (GE Amersham, #RPN5661) as appropriate. We used dye swapping to account for potential dye effects. Dye coupled cRNA was purified and 4 ug was fragmented and subsequently dissolved in 60ul of hybridization solution. We used an aspen 7K EST array containing replicate subarrays of 6,705 elements in a 4Ã4 grid with a spot diameter of ~150 µm and a spacing of 200 µm (Ranjan et al., 2004). The elements represented ~5,400 unique JGI Populus gene models, and included positive and negative controls (Lucidea Universal ScoreCard, Amersham) to monitor target labeling and hybridization efficiency, as well as many genes of known function. Clone information can be downloaded from the aspenDB website (www.aspenDB.mtu.edu). Arrays were hybridized following previously published methods (Xiang et al., 2002; Hughes et al., 2001). Chang S, Puryear J, Cairney J. 1993. A simple and efficient method for isolating RNA from pine trees. Plant Molecular Biology Reporter 11: 113-116. Davis JM, Gordon MP, Smit BA. 1991. Assimilate movement dictates remote sites of wound-induced gene expression in poplar leaves. Proceedings of the National Academy of Sciences (USA) 88: 2393-2396. Frost CJ, Appel HM, Carlson JE, De Moraes CM, Mescher MC, Schultz JC. 2007. Within-plant signalling via volatiles overcomes vascular constraints on systemic signalling and primes responses against herbivores. Ecology Letters 10: 490-498. Hughes TR, Mao M, Jones AR, Burchard J, Marton MJ, Shannon KW, Lefkowitz SM, Ziman M, Schelter JM, Meyer MR, Kobayashi S, Davis C, Dai HY, He YDD, Stephaniants SB, Cavet G, Walker WL, West A, Coffey E, Shoemaker DD, Stoughton R, Blanchard AP, Friend SH, Linsley PS. 2001. Expression profiling using microarrays fabricated by an ink-jet oligonucleotide synthesizer. Nature Biotechnology 19: 342-347. Xiang CC, Kozhich OA, Chen M, Inman JM, Phan QN, Chen YD, Brownstein MJ. 2002. Amine-modified random primers to label probes for DNA microarrays. Nature Biotechnology 20: 738-742.
2010-05-25 | E-GEOD-11955 | biostudies-arrayexpress
Project description:Recently, we have reported on a highly drug-resistant carbapenemase-producing isolate of Enterobacter cloacae (Nepal et al., Virulence. 2018; 9: 1377-1389). In the present study, we asked the question whether and, if so, how this isolate responds to a sub-inhibitory challenge with the antibiotic imipenem. To answer this question, we applied a SILAC proteomics approach that allowed the quantification of changes in the relative abundance of bacterial protein in response to imipenem. The results show that the investigated E. cloacae isolate mounts a highly specific response to counteract the detrimental effects of imipenem.
2021-08-10 | PXD013412 | Pride
Project description:Cardiovascular disease (CVD) is the leading cause of mortality, with sex and age being strong risk factors. Mitochondria are vital for normal cardiac function, and regulation of mitochondrial structure and function may impact susceptibility to CVD. We analyzed the basal expression levels of mitochondria-related genes in the hearts of male and female untreated control rats at different ages to identify potential role of mitochondria in differential susceptibility to CVD between the sexes. Hearts from 5 male and 5 female rats at three different ages (young (8-week), adult (21-week), and old (78-week)) were used for investigation of sex-related differences in expression levels of genes. At each age, rats were humanely euthanized by CO2 asphyxiation and whole hearts were removed quickly, flash frozen in liquid nitrogen, and stored at -80ËC until further investigation. The frozen hearts were individually ground into powder in liquid nitrogen using a mortar and pestle chilled on dry ice. Total RNAs were extracted from heart tissue powder and gene expression was measured using Agilent Rat Whole Genome Microarrays (4 x 44k format; Agilent Technologies, Inc.).
2014-09-30 | E-GEOD-58204 | biostudies-arrayexpress
Project description:A genomic overview of in vivo binding of a transcription factor Snail in the ascidian early gastrula embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged Snail. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
2010-05-06 | E-GEOD-17691 | biostudies-arrayexpress
Project description:A genomic overview of in vivo binding of a transcription factor Neurogenin in the ascidian 64-cell stage embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged Neurogenin. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
2010-05-06 | E-GEOD-17689 | biostudies-arrayexpress
Project description:A genomic overview of in vivo binding of a transcription factor Tbx6b in the ascidian early gastrula embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged Tbx6b. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
2010-05-06 | E-GEOD-17694 | biostudies-arrayexpress