StCDF1 transcription factor: "The jack of all trades" and its nitrogen signalling regulation in potato [RNA-seq]
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ABSTRACT: Due to the sessile nature of plants, they are under increased risk of environmental threats,aggravated by climate-change induces abiotic stress. However, plants have adapted toachieve growth development and reproduction by modulating their molecular stressresponse. Transcription factors (TFs) encoded belonging to DNA-BINDING WITH ONEFINGER (DOF) as CYCLING DOF FACTOR (CDFs) have been implicated in diverseprocesses, including nitrogen response. In potato,StCDF1TF plays a central role as a clockoutputthat regulatestuberizationand we previously demonstrated that it is also involved indrought response together with its antisense lncRNAthroughstomata regulation.Using arecently developed high-throughput DNA-affinity purification sequencing technology(DAP-Seq) we were able to identify the molecular targets of StCDF1. We further combinedDAP-Seq & RNA-Seq data from overexpressed and knockdownStCDF1transgenic plants,to specify its downstream candidate targets. We found that in addition to its canonical roleas a repressor, StCDF1 can also act as an activator of target genes involved in a range ofbiological processes, including the regulation of nitrogen metabolism. Finally, wedemonstrated that plants overexpressingStCDF1are more sensitive to lownitrogenconditions compared to the knockoutStCDF1
Project description:Due to the sessile nature of plants, they are under increased risk of environmental threats,aggravated by climate-change induces abiotic stress. However, plants have adapted toachieve growth development and reproduction by modulating their molecular stressresponse. Transcription factors (TFs) encoded belonging to DNA-BINDING WITH ONEFINGER (DOF) as CYCLING DOF FACTOR (CDFs) have been implicated in diverseprocesses, including nitrogen response. In potato,StCDF1TF plays a central role as a clockoutputthat regulatestuberizationand we previously demonstrated that it is also involved indrought response together with its antisense lncRNAthroughstomata regulation.Using arecently developed high-throughput DNA-affinity purification sequencing technology(DAP-Seq) we were able to identify the molecular targets of StCDF1. We further combinedDAP-Seq & RNA-Seq data from overexpressed and knockdownStCDF1transgenic plants,to specify its downstream candidate targets. We found that in addition to its canonical roleas a repressor, StCDF1 can also act as an activator of target genes involved in a range ofbiological processes, including the regulation of nitrogen metabolism. Finally, wedemonstrated that plants overexpressingStCDF1are more sensitive to lownitrogenconditions compared to the knockoutStCDF1
Project description:Sets of seven 2-week old potato plants carrying the nematode resistance gene H1, grown from tuber ‘chits’ in sandy loam at a constant temperature of 20 ºC and a light cycle of 16 hour light/8 hour dark, were each inoculated in the roots evenly with 2000 juveniles of the virulent potato cyst nematode Globodera pallida, or with the avirulent G. rostochiensis pathotype Ro1, or with water. Plants were manually watered throughout the duration of the experiment. 5, 17 and 33 days after inoculation, the roots were carefully washed and root tissue samples were individually flash-frozen in liquid nitrogen and stored at –80 ºC. Total RNA isolations were performed using the Qiagen RNeasy kit. All samples were treated with DNase. The experiment was replicated twice. Keywords: Direct comparison
Project description:The purpose of this study is to survey gene expression of potato (solanum tuberosum gilroy) in a variety of tissues and life cycle timepoints. Potato plants were grown from true seeds, planted in either soil or MS media, and grown in growth chambers. Four biological replicates each of twenty different tissue samples were collected at the appropriate timepoints. The samples were immediately frozen at -80. The tissues were then ground into a powder in liquid nitrogen, and RNA was extracted using Qiagen's QiaShredder columns and Qiagen's Rneasy MinElute Kit. The RNA was amplified using Arcturus' RiboAmp RNA Amplification Kit. 1ug of amplified RNA was used in TIGR's indirect labelleling procedure. All hybridaztions were applied to TIGR's 10kV4 potato array. Keywords: Loop design
Project description:To screen genes related to the development of sweet potato tuberous roots, the high throughput sequencing of different stages of sweet potato tuberous roots was performed. The fibrous roots (FR; roots at 20 dap), developing tuberous roots (DR; roots at 60 dap) and mature tuberous roots (MR; roots at 120 dap) of Ipomoea batatas (L.) Taizhong 6 and MBP3 overexpressed lines were used for transcriptome analysis. Totally, we identified 5488 differentially expressed genes between different stage tuberous roots of Taizhong6 and 14312 differentially expressed genes between the tuberous roots of Taizhong6 and MBP3 overexpressed lines, by calculating the gene FPKM in each sample and conducting differential gene analysis. This study provides a foundation for the mechanism analysis of sweet potato tuberous root development.
Project description:Potato plants of the cultivar 'Atlantic', which is IHN-susceptible, were grown in the greenhouse under a 16-hour photoperiod and 22 C day/18 C night temperatures for 46 days, after which they were transferred to growth chambers with a 14-hour photoperiod and normal (20 C day/18 C night) temperatures. At 71 DAP (days after planting), half of the plants were subjected to high (28 C day/20 C night) temperatures for the remainder of the study. Tubers from both normal and high temperature regimes were harvested at two-week intervals, beginning at 76 DAP and ending at 118 DAP. For each RNA sample, equal amounts of peeled tuber tissue (sampled from the center of the tuber using a cork borer) was pooled from three randomly chosen plants. RNA was extracted using a hot-phenol and high salt (2.4 M) CTAB-based extraction buffer. Keywords: Loop design
Project description:Nitrogen (N) can be absorbed by plants, thereby affects plant physiological activity, interferes gene expression, alters metabolite content and influences plant growth. However, the molecular mechanism underlying the potato tuberization response to nitrogen remains unclear. The plants were cultivated in the pots using N-deficient, N-Routine and N-sufficient conditions. Physiological response analysis, transcriptomics and metabolomics were performed on potato stolon exposed to Nitrogen stress. Transcriptomics analysis revealed that 2756 differentially expressed genes (DEGs) responded to nitrogen stress. By using metabolomics analysis, a total of 600 d differentially accumulated metabolites (DAMs) were identified. Further correlation analysis of major DEGs and DAMs showed that 9 key DEGs were involved in alpha-linolenic acid metabolism, 16 key DEGs in starch and sucrose metabolism, 7 key DEGs in nitrogen metabolism, and 16 key DEGs in ABC transporters. Nitrogen deficiency significantly up-regulated the contents of sucrose, GDP-glucose and L-glutamic acid, and promoted the growth of stolon by up-regulating the expression of AMY, SBE, SS, SPS, AGPS and NR-related genes. However, High nitrogen is the opposite. In addition, high nitrogen treatment up-regulated EG, SUS and GDH related genes, accumulated a large number of 9 (S) -HpOTr E, 13 (S) -HpOTr E and L-Glutamine, ultimately affected the balance between plant growth and defense. In general, our study revealed the co-expressed genes and potential pathways related to potato tuber formation under different nitrogen conditions. These comprehensive analysis data provide a better understanding of improving potato tuber traits at the molecular and metabolic levels.
Project description:In the present study molecular interactions between potato plants, Colorado potato beetle (CPB) larvae and Potato virus YNTN (PVYNTN) were investigated by analyzing gene expression in potato leaves. Grant ID: J4-4165 Slovenian Research Agency ARRS Growth and defense trade-offs in multitrophic interaction between potato and its two major pests Grant ID: P4-0165 Slovenian Research Agency ARRS Biotechnology and Plant Systems Biology
Project description:Potato seedlings were subjected to cold, heat and salt stress. Expression profiles were captured at three different time-points, 3h, 9h and 27h from two different tissues, roots and leaves. The experiment was preformed independently three times. Commercially available true potato seeds (Variety Gilroy) were germinated on rafts floating on hydroponic medium in Magenta boxes. Plants were grown for 5 weeks prior to stress application under long day conditions (16h light and 8h dark) at 25C with gentle agitation. To initiate stress the medium was replaced with fresh medium pre-chilled to 4C (cold stress), pre-heated to 35C (heat stress) or supplemented with 100mM NaCl (salt stress). Cold and heat stress were maintained for the duration of the experiment by placing the Magenta boxes on ice or in a water-bath at 35C. For every individual sample two boxes of plants were used pooling a total of 6 plants per sample. For each time-point a single control sample was used by changing the media in a similar way as for the stress induction. A total of six boxes were combined for the pooled reference samples. Plants were harvested at the appropriate time and snap-frozen in liquid nitrogen. Roots and aerial tissue was separated prior to freezing. The tissue was stored at -80C freezer until isolation. Total RNA was isolated using RNeasy isolation kit. The intactness of the RNA was verified on gel and the concentration was adjusted to 3ug/ul by ethanol precipitation and re-suspension. Series_weblink: http://www.tigr.org/tdb/potato Keywords = potato, Abiotic stress Keywords: ordered
Project description:In the present study molecular interactions between potato plants, Colorado potato beetle (CPB) larvae and Potato virus YNTN (PVYNTN) were investigated by analyzing gene expression in potato leaves. mRNA samples of secondary PVYNTN-infected (CPB_PVY) and healthy potato plants (CPB_H) cultivar Igor and of RNAi coi1-silenced (CPB_coi1) and non-transformed (CPB_NT) potato plants cultivar Desiree collected 24 h post CPB infestation and respective control non-infested samples (CONT_PVY, CONT_H, CONT_coi1, CONT_NT).
Project description:Potato virus YNTN (PVYNTN), causing potato tuber ring necrosis disease, dramatically lowers the quantity and the quality of the potato yield all over the world. The cultivar Igor is one of the most susceptible cultivars, developing severe disease symptoms on plants as well as on tubers. Finding genes differentially expressed in the early response to infection, when the host response is more defense- than infection- related, could improve our understanding of the potato - PVYNTN interaction. Differential gene expression in early response of potato cv. Igor plants to PVYNTN infection was studied using potato TIGR cDNA-microarrays. Expression was compared between mock inoculated and virus infected plants 12 hours after inoculation, in four biological replicates. Keywords: direct comparison