Dataset Information

Comparative RNA-seq based transcriptomic analysis of bud dormancy in grape.

ABSTRACT: Bud 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.We 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.At 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.

SUBMITTER: Khalil-Ur-Rehman M

PROVIDER: S-EPMC5244717 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature

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Comparative RNA-seq based transcriptomic analysis of bud dormancy in grape.

Khalil-Ur-Rehman Muhammad M Sun Long L Li Chun-Xia CX Faheem Muhammad M Wang Wu W Tao Jian-Min JM

BMC plant biology 20170119 1

<h4>Background</h4>Bud 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.<h4>Results</h4>We have performed transcriptome and differential transcript expression ana ...[more]

PMID: 28103799

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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:American ginseng (Panax quinquefolius L.) is an important herb that is cultivated in China, North American, and South Korea. It is propagated from seed, but the seed has deep dormancy characteristics described as morphophysiological dormancy. Two-stage temperature stratification, a warm (15-20°C) and cold (2°C) stratification period of 6 months, has been used successfully for seed dormancy release. However, little is known about the molecular mechanisms of seed dormancy release in the stratification process. In this study, seed development after pollination and seed development in the dormancy release process were investigated in American ginseng. The transcriptome during seed dormancy release was analyzed using RNA-Seq technology and 78,207 unigenes (mean length 531 bp) were generated. Based on similarity searches of public databases, 54,292 of the unigenes (69.4%) were functionally annotated. Further, three digital gene expression (DGE) libraries were sequenced and differences in gene expression at three stages during seed cold stratification were examined. The greatest number of differentially expressed genes occurred in the 90DCS versus 180DCS libraries, while the lowest number of differentially expressed genes occurred in the 135DCS verus 180DCS libraries. GO enrichment analysis revealed that 59, 29, and 39 GO terms were significantly enriched in the biological process, molecular function, and cell component GO categories, respectively. There were 25,190 genes with KEGG pathway annotation in the three DGE libraries and their enrichment pathways were compared. The gene expressions of 30 selected unigenes were validated using quantitative PCR. This study is the first to provide the transcriptome sequences for seed dormancy release in American ginseng, and demonstrates the successful use of DGE profiling data for analyzing transcriptomic variation during dormancy release. These data provide a basis for future researches of seed dormancy in morphophysiological dormancy seeds in non-model plants.

Dataset Information

Comparative RNA-seq based transcriptomic analysis of bud dormancy in grape.

Publications

Comparative RNA-seq based transcriptomic analysis of bud dormancy in grape.

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