Dataset Information

Comparative Transcriptomic and Proteomic Analysis to Deeply Investigate the Role of Hydrogen Cyanamide in Grape Bud Dormancy.

ABSTRACT: Hydrogen cyanamide (HC) is an agrochemical compound that is frequently used to break bud dormancy in grapevines grown under mild winter conditions globally. The present study was carried out to provide an in-depth understanding of the molecular mechanism associated with HC releasing bud dormancy in grapevines. For this purpose, RNA-seq based transcriptomic and tandem mass tag (TMT)-based proteomic information was acquired and critically analyzed. The combined results of transcriptomic and proteomic analysis were utilized to demonstrate differential expression pattern of genes at the translational and transcriptional levels. The outcome of the proteomic analysis revealed that a total of 7135 proteins (p-value ? 0.05; fold change ? 1.5) between the treatments (HC treated versus control) were identified, out of which 6224 were quantified. Among these differentially expressed proteins (DEPs), the majority of these proteins were related to heat shock, oxidoreductase activity, and energy metabolism. Metabolic, ribosomal, and hormonal signaling pathways were found to be significantly enriched at both the transcriptional and translational levels. It was illustrated that genes associated with metabolic and oxidoreductase activity were mainly involved in the regulation of bud dormancy at the transcriptomic and proteomic levels. The current work furnishes a new track to decipher the molecular mechanism of bud dormancy after HC treatment in grapes. Functional characterization of key genes and proteins will be informative in exactly pinpointing the crosstalk between transcription and translation in the release of bud dormancy after HC application.

SUBMITTER: Khalil-Ur-Rehman M

PROVIDER: S-EPMC6679053 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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Comparative Transcriptomic and Proteomic Analysis to Deeply Investigate the Role of Hydrogen Cyanamide in Grape Bud Dormancy.

Khalil-Ur-Rehman Muhammad M Wang Wu W Dong Yang Y Faheem Muhammad M Xu Yanshuai Y Gao Zhihong Z Guo Shen Zhen Z Tao Jianmin J

International journal of molecular sciences 20190718 14

Hydrogen cyanamide (HC) is an agrochemical compound that is frequently used to break bud dormancy in grapevines grown under mild winter conditions globally. The present study was carried out to provide an in-depth understanding of the molecular mechanism associated with HC releasing bud dormancy in grapevines. For this purpose, RNA-seq based transcriptomic and tandem mass tag (TMT)-based proteomic information was acquired and critically analyzed. The combined results of transcriptomic and proteo ...[more]

PMID: 31323865

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Project description:BackgroundBud dormancy is an important biological phenomenon of perennial plants that enables them to survive under harsh environmental circumstances. Grape (Vitis vinifera) is one of the most grown fruit crop worldwide; however, underlying mechanisms involved in grape bud dormancy are not yet clear. This work was aimed to explore the underlying molecular mechanism regulating bud dormancy in grape.ResultsWe have performed transcriptome and differential transcript expression analyses of "Shine Muscat" grape buds using the Illumina RNA-seq system. Comparisons of transcript expression levels among three stages of dormancy, paradormancy (PD) vs endodormancy (ED), summer buds (SB) vs ED and SB vs PD, resulted in the detection of 8949, 9780 and 3938 differentially expressed transcripts, respectively. Out of approximately 78 million high-quality generated reads, 6096 transcripts were differentially expressed (log2 ratio ≥ 1, FDR ≤ 0.001). Grape reference genome was used for alignment of sequence reads and to measure the expression level of transcripts. Furthermore, findings obtained were then compared using two different databases; Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), to annotate the transcript descriptions and to assign a pathway to each transcript. KEGG analysis revealed that secondary metabolites biosynthesis and plant hormone signaling was found most enriched out of the 127 total pathways. In the comparisons of the PD vs ED and SB vs ED stages of grape buds, the gibberellin (GA) and abscisic acid (ABA) pathways were found to be the most enriched. The ABA and GA pathways were further analyzed to observe the expression pattern of differentially expressed transcripts. Transcripts related to the PP2C family (ABA pathway) were found to be up-regulated in the PD vs ED comparison and down-regulated in the SB vs ED and SB vs PD comparisons. GID1 family transcripts (GA pathway) were up-regulated while DELLA family transcripts were down-regulated during the three dormancy stages. Differentially expressed transcripts (DEGs) related to redox activity were abundant in the GO biological process category. RT-qPCR assay results for 12 selected transcripts validated the data obtained by RNA-seq.ConclusionAt this stage, taking into account the results obtained so far, it is possible to put forward a hypothesis for the molecular mechanism underlying grape bud dormancy, which may pave the way for ultimate improvements in the grape industry.

Project description:BACKGROUND:Grape buds exhibit non-uniform, or delayed, break in early spring in subtropical regions because the accumulation of chilling is insufficient. Hydrogen cyanamide (H2CN2, HC) can partially replace chilling to effectively promote bud sprouting and is used widely in warm winter areas. However, the exact underlying mechanism of grape bud release from endodormancy induced by HC remains elusive. RESULTS:In this study, the transcriptome of grape winter buds under in vitro conditions following HC and water treatment (control) was analyzed using RNA-seq technology. A total of 6772 differentially expressed genes (DEGs) were identified. Furthermore, the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that starch and sucrose metabolism and plant hormone signaling transduction were most enriched out of the 50 total pathways. HC treatment induced the upregulated expression of sucrose synthase (SUS), sucrose phosphate synthase (SPS), α-amylase (AM), and β-amylase (BM) and downregulated expression of sucrose invertase (INV), hexokinase (HK), fructokinase (FK), soluble starch synthase (SS), and granule-bound starch synthase (GBSS). Hence, the starch concentration in the HC-treated group was significantly lower than that in control, whereas soluble sugar content in the HC-treated group increased quickly and was higher than that in control between 0 and 8 d. The concentration of indoleacetic acid (IAA) and zeatin (ZT) increased, whereas that of abscisic acid (ABA) and gibberellin (GA) decreased in HC treated group, which coincided with the expression level of genes involved in above hormone signals. The content of hydrogen peroxide (H2O2) and enzyme activity of superoxide dismutase (SOD) and peroxidase (POD) were increased in grape buds with HC treatment, whereas catalase (CAT) activity was decreased. HC treatment increased the expression of POD, SOD, primary amine oxidase (PAO), polyamine oxidase (PAOX), and glutathione peroxidase (GSH-Px). CONCLUSION:Based on these results, it is possible to propose a mechanistic model that underlies the regulation of endodormancy release in grapevine buds by exogenous HC application.

Project description:Bud endodormancy induction response of two genotypes (‘Seyval’ a hybrid white wine grape and V. riparia, PI588259 a native north american species) was compared under long (15h) and short (13h) photoperiod. Three separate replicates (5 plants/replicate) were treated to generate paradormant (LD) and same aged endodormancy-induced (SD) buds for transcriptomic, proteomic and metabolomic analysis. Potted, spur-pruned two to six-year-old vines were removed from cold storage (Seyval 3-19-07; V. riparia 3/26/07) and grown under a LD (15 h) at 25/20 + 3C day/night temperatures (D/N). When vines reached 12-15 nodes (3-25-07) they were randomized into LD or SD treatments with 25/20 + 3C D/N in climate controlled greenhouses with automated photoperiod system (VRE Greenhouse Systems). Three replications (5 vines/replication) were harvested between 5/07-6/07 and then again in 5/08-6/08 for a total of six replications. All treatments are repeated at the same time every year and harvested at the same time of day each year to minimize biological noise. At 1, 3, 7, 14, 21, 28 and 42 days of LD and SD treatment, buds were harvested from nodes 3 to 12 of each separate replicate, immediately frozen in liquid nitrogen, and placed at -80C for future RNA, protein and metabolite extraction. These time points encompass early reversible phases as well as key time points during transition to irreversible endodormancy development. After photoperiod treatments and bud harvests, all pruned vines were returned to LD and monitored for bud endodormancy. The endodormant vines were identified after 28 days and moved to cold storage. The nondormant vines were allowed to grow again and induced into dormancy at a later date. Acknowledgement:This study was funded by NSF Grant DBI0604755 and funds from the South Dakota Agriculture Experiment Station. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Anne Fennell. The equivalent experiment is VV10 at PLEXdb.] 84 Samples of two genotypes compared under long and short photoperiod, 3 replicates each. Seyval (SV, Seyve Villard 5-276) : LD, 15h : 1 days (3-replications) Seyval (SV, Seyve Villard 5-276) : LD, 15h : 3 days (3-replications) Seyval (SV, Seyve Villard 5-276) : LD, 15h : 7 days (3-replications) Seyval (SV, Seyve Villard 5-276) : LD, 15h : 14 days (3-replications) Seyval (SV, Seyve Villard 5-276) : LD, 15h : 21 days (3-replications) Seyval (SV, Seyve Villard 5-276) : LD, 15h : 28 days (3-replications) Seyval (SV, Seyve Villard 5-276) : LD, 15h : 42 days (3-replications) Seyval (SV, Seyve Villard 5-276) : SD, 13h : 1 days (3-replications) Seyval (SV, Seyve Villard 5-276) : SD, 13h : 3 days (3-replications) Seyval (SV, Seyve Villard 5-276) : SD, 13h : 7 days (3-replications) Seyval (SV, Seyve Villard 5-276) : SD, 13h : 14 days (3-replications) Seyval (SV, Seyve Villard 5-276) : SD, 13h : 21 days (3-replications) Seyval (SV, Seyve Villard 5-276) : SD, 13h : 28 days (3-replications) Seyval (SV, Seyve Villard 5-276) : SD, 13h : 42 days (3-replications) V. riparia (VR, PI588259) : LD, 15h : 1 days (3-replications) V. riparia (VR, PI588259) : LD, 15h : 3 days (3-replications) V. riparia (VR, PI588259) : LD, 15h : 7 days (3-replications) V. riparia (VR, PI588259) : LD, 15h : 14 days (3-replications) V. riparia (VR, PI588259) : LD, 15h : 21 days (3-replications) V. riparia (VR, PI588259) : LD, 15h : 28 days (3-replications) V. riparia (VR, PI588259) : LD, 15h : 42 days (3-replications) V. riparia (VR, PI588259) : SD, 13h : 1 days (3-replications) V. riparia (VR, PI588259) : SD, 13h : 3 days (3-replications) V. riparia (VR, PI588259) : SD, 13h : 7 days (3-replications) V. riparia (VR, PI588259) : SD, 13h : 14 days (3-replications) V. riparia (VR, PI588259) : SD, 13h : 21 days (3-replications) V. riparia (VR, PI588259) : SD, 13h : 28 days (3-replications) V. riparia (VR, PI588259) : SD, 13h : 42 days (3-replications)

Project description:BackgroundHydrogen cyanamide (HC) and pruning (P) have frequently been used to break dormancy in grapevine floral buds. However, the exact underlying mechanism remains elusive. This study aimed to address the early mode of action of these treatments on accumulation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) and expression of related genes in the dormancy breaking buds of grapevine in the summer.ResultsThe budbreak rates induced by pruning (P), hydrogen cyanamide (HC), pruning plus hydrogen cyanamide (PHC) and water (control) after 8 days were 33, 53, 95, and 0 %, respectively. Clearly, HC was more effective in stimulating grapevine budbreak and P further enhanced its potency. In situ staining of longitudinal bud sections after 12 h of treatments detected high levels of ROS and nitric oxide (NO) accumulated in the buds treated with PHC, compared with HC or P alone. The amounts of ROS and NO accumulated were highly correlated with the rates of budbreak among these treatments, highlighting the importance of a rapid, transient accumulation of sublethal levels of ROS and RNS in dormancy breaking. Microarray analysis revealed specific alterations in gene expression in dormancy breaking buds induced by P, HC and PHC after 24 h of treatment. Relative to control, PHC altered the expression of the largest number of genes, while P affected the expression of the least number of genes. PHC also exerted a greater intensity in transcriptional activation of these genes. Gene ontology (GO) analysis suggests that alteration in expression of ROS related genes is the major factor responsible for budbreak. qRT-PCR analysis revealed the transient expression dynamics of 12 specific genes related to ROS generation and scavenge during the 48 h treatment with PHC.ConclusionOur results suggest that rapid accumulation of ROS and NO at early stage is important for dormancy release in grapevine in the summer, and the identification of the commonly expressed specific genes among the treatments allowed the construction of the signal transduction pathway related to ROS/RNS metabolism during dormancy release. The rapid accumulation of a sublethal level of ROS/RNS subsequently induces cell wall loosening and expansion for bud sprouting.

Dataset Information

Comparative Transcriptomic and Proteomic Analysis to Deeply Investigate the Role of Hydrogen Cyanamide in Grape Bud Dormancy.

Publications

Comparative Transcriptomic and Proteomic Analysis to Deeply Investigate the Role of Hydrogen Cyanamide in Grape Bud Dormancy.

Similar Datasets

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