Project description:Orchid Dendrobium Sonia (Sonia) has wide demand in floral market throughout the world due to its vibrant coloured flowers, flowering recurrence and dense inflorescence. Effects of different monochromatic light qualities (red, far-red, blue, white) on different developmental and growth responses such as seed gemination, shoot and root growth, chlorophyll and carotenoid accumulation were studied in Sonia. Asymbiotic seed germination was the highest under blue or white light (80%, p<0.001) and least under far-red (55%, p<0.001) suggesting that it could either be controlled by an exceptional novel function of Cryptochrome 1 or the blue wavelengths perceived by PHYA or PHYB in Sonia. All analyses were done in 6-months-old plants till 1 year of age. Shoot length increased significantly in white or red light (3.7-fold, 3.5-fold respectively) while was least under far-red (decreased by 1.6-fold) indicating the major regulatory role of PHYB for shoot growth. Root growth in red in Sonia was more similar to hypocotyl adventitious root (HAR) formation genetically considering PHYB transcript expression. We suggest that the root growth (HAR) under red in Sonia is partly attributed to PHYB, negatively regulated with HY5 transcription factor and positively associated with auxin biosynthesis and accumulation gene BABY BOOM (BBM2) and efflux carriers such as LIKE AUXIN RESISTANT 2 and 3 (LAX2, LAX3). Highest chlorophyll content under far-red and blue might be a hyper-response of SAR under FR light in Sonia. Taxonomic tree analysis finds Sonia closer to Phalenopsis and Dendrobium catenatum Lindl. The three phytochromes and one cryptochrome sequences which were similar to PHYA, B, C and CRY1 respectively. CRY1 was retrieved with one more isoform CRY1_X2, with all these sequences more similar to those of Oryza.sativa. Further study can clarify the indicating reason of a probable gene loss which is evident from the absence of any sequence similar to CRY2 in root RNA isolates of Dendrobium Sonia.

Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of gene expression profiles of cucumber under short-term chilling stress. The goals of this study are to transcriptome analysis of cucumber leaves under chilling stress. Methods: mRNA profiles of seedlings exposed to an air temperature of 6°C in the absence of light at 0, 2, 6, and 12 h were generated by deep sequencing, in triplicate, using Illumina Hiseq platform. The reference genome and gene model annotation files were downloaded from the genome website （http://cucurbitgenomics.org/）. An index of the reference genome was built using Bowtie v.2.2.3 and paired-end clean reads were aligned to the reference genome using TopHat v.2.0.12. qRT–PCR validation was performed using SYBR Green assays. Results: A total of 55.7 million clean reads was generated. Based on the threshold values of absolute value of log2 ratio ≥ 1 and FDR ≤ 0.05, a total of 2113 DEGs was identified at three time points (2, 6, and 12 h). A total of 30 genes was detected at all time points. The number of DEGs increased with time. In total, 100 TFs from 22 families in three subsets were detected. And 19 kinase families were identified in three subsets. The DEGs identified by RNA sequencing were confirmed by qRT-PCR analysis, indicating that the data were reliable. These findings provide information that can be useful for investigating the molecular mechanisms underlying the response to chilling stress in cucumber and other plants. Conclusions: The results presented here reveal changes in the transcriptome profile of cucumber in response to chilling stress. Exposure to a low temperature induced genes involved in hormone regulation, lipid metabolism, and photosynthesis, including NAC, WRKY, AP2/ERF, ERD, MYB as well as zinc finger TFs and protein kinases such as receptor-like protein kinase, MAPK, and CDK. Most TFs were upregulated whereas CDKs were downregulated. These findings provide information that can be useful for investigating the molecular mechanisms underlying the response to chilling stress in cucumber and other plants.

Project description:Background. Ovary culture has been a useful way to generate double haploid (DH) plant in cucumber (Cucumis sativus L.). However, the rate of embryo induction and the ability for induced embryo to grow into normal embryo are quite low. Moreover, the s mechanism of cucumber embryogenesis remains ambiguous. In this study, the molecular basis for cucumber embryogenesis was explored to set up basis for a more efficient ovary culture method. Differentially expressed genes during embryogenesis process, including the early stages of embryo formation, embryo maturation and shoot formation, were investigated using transcriptomic sequencing. Methods. Based on the cytological observation of cucumber ovary culture, the ovary culture can be divided into three stages:early embryo development, embryo maturation (from pre-embryos to cotyledon embryos) and the shoot formation stage. six key time points were selected for transcriptome sequencing and analysis. Results. We firstly conducted cytological observations which suggest that cell enlargement is the symbol for gametophytes to switch to sporophyte development pathway during early embryogenesis stage. In this stage, RNA-seq revealed 3468 up-regulated genes, including hormone signal transduction genes, hormone response genes and stress-induced genes. The reported embryogenesis-related genes BBM, HSP90 and AGL were also actively expressed during this stage. The total 480 genes that function in protein complex binding, microtubule binding, tetrapyrrole binding, tubulin binding and other microtubule activities were continuously up-regulated during the embryo maturation stage, indicating that the cytoskeleton structure was continuously being built and maintained by the action of microtubule-binding proteins and enzyme modification during embryo development. In shoot formation stage, 1383 genes were up-regulated, which were mainly enriched in phenylpropanoid biosynthesis, plant hormone signal transduction, phenylalanine metabolism, and starch and sucrose metabolism. The shoot formation stage might be regulated by 6 transcription factors that contained a B3 domain, 9 genes in the AP2/ERF family and 2 genes encoded WUS homologous domain proteins. Conclusions. These findings offer a valuable framework for explaining the transcriptional regulatory mechanism underlying embryogenesis during cucumber ovary culture.

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Cucumber

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