Project description:Purpose: To study pre-meiotic (21-nt) and meiotic (24-nt) phasiRNA pathways in non-grass monocots Methods: Anthers were dissected using a 2 mm stage micrometer in a stereo microscope, and immediately frozen in liquid nitrogen until total RNA isolation was performed. Small RNA, mRNA libraries were generated using short-read (Illumina) and Single Molecule Real Time SMRT (PacBio) sequencing approaches. Stages were assigned based on the morphology of archesporial (AR) and tapetal cells of A. officinalis (Asparagus) anthers.
Project description:Sex chromosomes evolved from autosomes many times across the eukaryote phylogeny. Several models have been proposed to explain this transition, some involving male and female sterility mutations linked in a region of suppressed recombination between X and Y (or Z/W, U/V) chromosomes. Comparative and experimental analysis of a reference genome assembly for a double haploid YY male garden asparagus (Asparagus officinalis L.) individual implicates separate but linked genes as responsible for sex determination. Dioecy has evolved recently within Asparagus and sex chromosomes are cytogenetically identical with the Y, harboring a megabase segment that is missing from the X. We show that deletion of this entire region results in a male-to-female conversion, whereas loss of a single suppressor of female development drives male-to-hermaphrodite conversion. A single copy anther-specific gene with a male sterile Arabidopsis knockout phenotype is also in the Y-specific region, supporting a two-gene model for sex chromosome evolution. Additionally, we test for the presence of Y-specific small RNA loci in several XX, XY, and YY genotypes that may be acting as sex determination loci.
Project description:Nicotiana benthamiana plants were infected with Asparagus Virus 2 and its mutant version. Upper non-inoculated leaves were collected at various time points and used for sample preparation. RNA-seq was performed on the WT infected, mutant infected and mock uninfected samples. Ribo-seq was performed on the WT infected and mutant infected samples.
Project description:Most physiological and molecular mechanisms of salinity stress are researched based on salt shock conditions. However, salt shock doesn’t occur in agricultural practice or natural ecosystem. In the fields, salts accumulate gradually through high salt or sodium irrigation water and poor managements that allow ground water to rise to soil surface. Therefore, it is more reasonable to research salinity stress in a mild way (stepwise salt addition). The objective of this study is to select marker genes to differentiate between salt shock (Phase 0) and salt stress (Phase 1). Three replicates were used for all RNA-Seq experiments conducted on control, Phase 0 and Phase 1 samples in Arabidopsis thaliana. Phase 0 samples (rosette leaves) were harvested 1 hour later from 105 mM NaCl salt shock treatment; Phase 1 samples (rosette leaves) were harvested 2 days later after reaching 90 mM NaCl by a stepwise addition (15 mM NaCl per day).
Project description:Purpose:Identification of genes and miRNAs responsible for salt tolerance in upland cotton (Gossypium hirsutum L.) would help reveal the molecular mechanisms of salt tolerance. We performed physiological experiments and transcriptome sequencing (mRNA-seq and small RNA-seq) of cotton leaves under salt stress using Illumina sequencing technology. And quantitative reverse transcription polymerase chain reaction (qRT–PCR) methods and to evaluate protocols for optimal high-throughput data analysis Methods:We investigated two distinct salt stress phases—dehydration (4 h) and ionic stress (osmotic restoration; 24 h)—that were identified by physiological changes of 14-day-old seedlings of two cotton genotypes, one salt tolerant and the other salt sensitive, during a 72-h NaCl exposure. A comparative transcriptomics approach was used to monitor gene and miRNA differential expression at two time points (4 and 24 h) in leaves of the two cotton genotypes under salinity conditions. Results:During a 24-h salt exposure, 819 transcription factor unigenes were differentially expressed in both genotypes, with 129 unigenes specifically expressed in the salt-tolerant genotype. Under salt stress, 108 conserved miRNAs from known families were differentially expressed at two time points in the salt-tolerant genotype. Conclusions:Our comprehensive transcriptome analysis has provided new insights into salt-stress response of upland cotton. The results should contribute to the development of genetically modified cotton with salt tolerance.
Project description:In the present study, we utilized RNA-SEQ and TMT techniques to identify differentially expressed genes and differentially expressed proteins in R. soongorica seedling leaves treated with 0, 200, and 500 mM NaCl for 72 hours. These analyses were based on morphological and physiological responses to salt stress. Potential salt tolerance-related genes were identified, which laid the foundation for revealing the molecular regulatory mechanisms of moso bamboo adaptation to salt-stressed environments.