RNA sequencing of Nipponbare control and OsCNGC4, OsCNGC5 double knockout mutant in mature anthers
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ABSTRACT: We created a double loss-of-function/knockout mutant targeting two rice genes simultaneously. The selected genes are as follows: OsCNGC4(LOC_Os03g44440) and OsCNGC5(LOC_Os12g28260). These two CNGCs are strongly transcriptional expressed in the rice mature anthers (stages 13-14). The mutant of these OsCNGC4/5 displayed a low seed-setting rate. This data refers to the transcriptome of mature anthers from the double mutant of OsCNGC4 and OsCNGC5. We sampled mature anther for the analysis.
Project description:We generated single loss-of-function knockout mutant targeting a specific rice gene, GTrD5; LOC_Os11g20384), a member of the Suppressor of Actin 1 (Sac1) domain phosphoinositide phosphatase subfamily. GTrD5 is strongly expressed in mature anthers and pollen, and its knockout results in germination defects. Consequently, GTrD5 mutants have reduced self-fertilized seed production. This dataset comprises mature anther transcriptome data from GTrD5 single mutants, with samples collected from mature anthers for analysis.
Project description:We created a triple loss-of-function/knockout mutant targeting three rice genes simultaneously. The three selected genes are as follows: OsADF1 (LOC_Os02g44470), OsADF6 (LOC_Os04g46910), and OsADF9 (LOC_Os07g30090). These three ADFs are strongly transcriptional expressed in the rice mature anthers (stages 13) and bi-/tricelluler pollen. The triple mutant of these OsADFs does not produce self-fertilizing seeds due to the short length of the pollen tube (male-sterile). This data is about mature anther transcriptome data about the triple mutant of OsADFs (ADFmT). We sampled mature anther for the analysis.
Project description:We generated single loss-of-function knockout mutants targeting a specific rice gene, Ruptured Pollen Tube (RUPO, GTrD2; LOC_Os06g03610), a member of the Catharanthus roseus RLK1-like (CrRLK1L) subfamily. RUPO is strongly expressed in mature anthers and pollen, and its knockout results in reduced pollen ROS levels and impaired pollen tube elongation, leading to germination defects. Consequently, RUPO mutants fail to produce self-fertilized seeds. This dataset comprises mature anther transcriptome data from RUPO single mutants, with samples collected from mature anthers for analysis.
Project description:We created a double loss-of-function/knockout mutant targeting three rice genes simultaneously. The two selected genes are as follows: OsABCG16 (LOC_Os06g51460), OsABCG28 (LOC_Os11g22350). These two ABCGs are strongly transcriptional expressed in the rice mature anthers (stages 13) and bi-/tricelluler pollen. The double mutant of these OsABCGs does not produce self-fertilizing seeds due to the short length of the pollen tube inside the pistil (male-sterile). This data is about mature anther transcriptome data about the double mutant of OsABCGs. We sampled mature anther for the analysis.
Project description:Pollen tube growth is essential for successful fertilization and stable crop yields. We constructed loss-of-function/knock-out mutants that simultaneously target two rice genes using the CRISPR/Cas9 mutagenesis system. The selected OsRALF17 and OsRALF19 genes are strongly expressed in rice bicellular/tricellular pollen and have essential functions in the pollen tube growth. For the corresponding transcriptomic analysis, we sampled mature pollen anthers from a control group and an OsRALF17/19 knock-out mutant.
Project description:Rice anthers at anthesis stage from the wild type and osrac6-1 mutant anther (Dongjin cultivar) We collected the sample from our field and immediately froze the samples with liquid nitrogen.
Project description:Protein lysine acetylation (KAC) is a dynamic and reversible post-translational modification, playing important biological roles in many organisms.Here, we reported results from a proteomic investigation to detect KAC status of the developing rice anthers near the time of meiosis (RAM), providing strong biochemical evidence for roles of many KAC-affected proteins during rice anther development and meiosis. We identified a total of 1,354 KAC sites in 676 proteins.
Project description:Soil salinity is a major production constrain for agricultural crops, especially in Oryza sativa (rice). Analyzing physiological effect and molecular mechanism under salt stress is key for developing stress-tolerant plants. Roots system has a major role in coping with the osmotic change impacted by salinity and few salt-stress-related transcriptome studies in rice have been previously reported. However, transcriptome data sets using rice roots grown in soil condition are more relevant for further applications, but have not yet been available. The present work analyzed rice root and shoot physiological characteristics in response to salt stress using 250 mM NaCl for different timepoints. Subsequently, we identified that 5 day treatment is critical timepoint for stress response in the specific experimental design. We then generated RNA-Seq-based transcriptome data set with rice roots treated with 250 mM NaCl for 5 days along with untreated controls in soil condition using rice japonica cultivar Chilbo. We identified 447 upregulated genes under salt stress with more than fourfold changes (p value < 0.05, FDR < 0.05) and used qRT-PCR for six genes to confirm their salt-dependent induction patterns. GO-enrichment analysis indicated that carbohydrate and amino-acid metabolic process are significantly affected by the salt stress. MapMan overview analysis indicated that secondary metabolite-related genes are induced under salt stress. Metabolites profiling analysis confirmed that phenolics and flavonoids accumulate in root under salt stress. We further constructed a functional network consisting of regulatory genes based on predicted protein–protein interactions, suggesting useful regulatory molecular network for future applications.
Project description:In this study, in order to identify miRNA targets, a degradome library derived from anthers of the WT and GMS (Genetic Male Sterility) mutant representing three stages of development was constructed and sequenced, resulting in the generation of 24.6 million raw reads. After removal of low quality sequences and adapter sequences, 24.4 million clean reads were obtained and 98% were 20 or 21 nt in length as expected in that normally length distribution peak of degradome fragment is between 20 and 21 nt [Addo-Quaye C, Eshoo TW, Bartel DP, Axtell MJ: Endogenous siRNA and miRNA targets identified by sequencing of the Arabidopsis degradome. Curr Biol 2008, 18:758-762]. Identification of miRNA targets in the WT and GMS muant anthers. Anthers of the WT and GMS mutant representing three stages of development [the meiosis stage (WT: Mar-F-1; mutant: Mar-S-1) and tetrad stage (WT: Mar-F-2; mutant: Mar-S-2), together with the uninucleate microspore stage (WT: Mar-F-3; mutant: Mar-S-3) from the GMS M-bM-^@M-^XDong AM-bM-^@M-^Y mutant and its fertile wild type] were collected during early mornings.
Project description:Illumina sequencing was employed to examine the expression profiles of rice anther miRNAs from the a non-pollen male sterile line Wuxiang S (WXS), one of photo-thermo sensitive genical male sterile (PTGMS) line rice, during in the fertility transition stage. A total of 493 known miRNAs and 273 novel miRNAs were identified during rice anther development. Based on the number of sequencing reads, a total of 26 miRNAs were discovered to be significant difference expression between WXS(S, Sterility) and WXS(F, Fertility), and the results were partially validated by qRT-PCR. Among these, 11 miRNAs were decreased and 15 miRNAs were increased in WXS(S) compared with WXS(F). The expression patterns for targets of osa-miR156a-j, osa-miR3879, osa-miR159c/d/e, osa-miR171a/c/e/i, osa-miR398b, osa-miR164d, osa-miR528 and osa-miR408 were selectively examined, and the results showed that there was a negative correlation on the expression patterns between miRNAs and their targets. These targets have previously been reported to be related with pollen development and male sterility, suggesting that miRNAs might act as regulators of rice anthers. Furthermore, miRNA editing events were observed. The U-to-C and U-to-A editing phenomenon was validated by molecular cloning and sequencing. Examine small RNA profiles change of four tissues of the rice non-pollen male sterile line Wuxiang S under two different environments.