Abstract
Ginger (Zingiber officinale Rosc.), a well-known member of family Zingiberaceae, is bestowed with number of medicinal properties which is because of the secondary metabolites, essential oil and oleoresin, it contains in its rhizome. The drug yielding potential is known to depend on agro-climatic conditions prevailing at the place cultivation. Present study deals with comparative transcriptome analysis of two sample of elite ginger variety Suprabha collected from two different agro-climatic zones of Odisha. Transcriptome assembly for both the samples was done using next generation sequencing methodology. The raw data of size 10.8 and 11.8 GB obtained from analysis of two rhizomes S1Z4 and S2Z5 collected from Bhubaneswar and Koraput and are available in NCBI accession number SAMN03761169 and SAMN03761176 respectively. We identified 60,452 and 54,748 transcripts using trinity tool respectively from ginger rhizome of S1Z4 and S2Z5. The transcript length varied from 300 bp to 15,213 bp and 8988 bp and N50 value of 1415 bp and 1334 bp respectively for S1Z4 and S2Z5. To the best of our knowledge, this is the first comparative transcriptome analysis of elite ginger cultivars Suprabha from two different agro-climatic conditions of Odisha, India which will help to understand the effect of agro-climatic conditions on differential expression of secondary metabolites.
Keywords: Zingiber officinale, Suprabha, Agro-climatic condition, Illumina, Transcriptome
| Specification: | |
| Organism/cell line/tissue | Ginger (Zingiber officinale cv. Suprabha) rhizome |
| Sex | N/A |
| Sequencer or array type | Illumina Nextseq 500 |
| Data format | Raw data |
| Experimental factors | Transcriptome profiling of elite ginger cv. Suprabha from two different agro climatic zones of Odisha. |
| Experimental features | Fresh and healthy rhizomes of Zingiber officinal Rose. cv. Suprabha, grown in two different agro climatic zones of Odisha were harvested for RNA isolation, de novo transcriptome assembly and protein annotations. |
| Consent | N/A |
| Sample source location | S1Z4: Center of Biotechnology, Siksha ‘O’ Anusandhan University, Kalinga Nagar, Ghatikia, Bhubaneswar, Odisha. S2Z5: High Altitude Research Station of Orissa University of Agriculture & Technology, Pottangi-764039, Koraput (Dist), Orissa. |
1. Direct link to deposited data
http://www.ncbi.nlm.nih.gov/biosample/?term=SAMN03761176 for Ginger cultivar S1Z4.
http://www.ncbi.nlm.nih.gov/biosample/?term=SAMN03761169 for Ginger cultivar S2Z5.
2. Introduction
Ginger (Zingiber officinale, Rose.), a pan-tropical plant of South East Asian origin, belongs to family Zingiberaceae. Even though, the crop is able to grow in different climatic conditions, essential oil and oleoresin synthesized in its rhizomes are reported to vary with climate and soil type of the area of cultivation [1]. Agro-climatic conditions at different localities are known to vary greatly across a state like Odisha, Eastern India and thus classified into ten different zones. Agro-climatic conditions are known to influence the production of secondary metabolites in ginger rhizome when same cultivar is grown in two different locations [2]. Therefore, in the present study we conducted de novo transcriptome assembly for two ginger cv. Suprabha rhizome samples S1Z4 and S2Z5 collected respectively from two different locations of the state; (i) Bhubaneswar of agro-climatic zone 4 (Climate: Hot and humid and Soil type: Saline, lateritic alluvial, red, mixed Red and Black); (ii) Koraput belonging to agro-climatic zone 5 (Climate: Hot and moist sub humid and Soil type: Brown forest, lateritic alluvial, red, mixed Red and Black) using next generation sequencing.
3. Experimental design, materials and methods
3.1. Plant materials
Fresh, healthy rhizome of Zingiber officinale, Rose. (cv. Suprabha) sample S1Z4 and S2Z5 were harvested from the underground soil of the High Altitude Research Station, Koraput and medicinal plant garden of Center of Biotechnology, Siksha ‘O’ Anusandhan University, Bhubaneswar, Odisha. Rhizomes are rinsed thoroughly with sterile distilled water, immediately dipped into RNA stabilizer solution (Xcelris Genomics, India) and stored in liquid nitrogen until further experiments.
3.2. RNA isolation, library preparation and sequencing
RNA isolation and transcriptome library construction was performed according to the Illumina TruSeq RNA library protocol and sequencing was done using Illumina Nextseq 500 at Genotypic Technology's Genomics facility, Genotypic Technology (P) Limited Bangalore.
3.3. Transcriptome de novo assembly, annotation and classification
Raw data of size 10.8 GB and 11.8 GB approximately was obtained from both the ginger variety S1Z4 and S2Z5. De novo assembly of Illumina Nextseq 500 processed data was performed using trinityrnaseq [3] for k-mers = 25 has been selected for downstream analysis. Detailed statistics of transcriptome de novo assembly are presented in Table 1. The number of total generated transcripts (≥ 300 bp) was 60,452 and 54,748 with a median transcript length of 393 bp and 1164 bp and N50 value of 1415 and 1334 respectively for ginger cultivar S1Z4 and S2Z5. Transcripts were annotated using NCBI BLAST 2.2.29 [4] with the proteins viridiplantae taken from uniprot database. For annotation, we have considered transcripts having length ≥ 300 bp, followed by clustering these transcripts with 95% indent using CD-HIT [5] which resulted into COG's. Unannotated transcripts were considered for Pfam domain analysis. We obtained 54,322 and 48,483 proteins of which only 38,243 and 36,678 are annotated for sample S1Z4 and S2Z5 respectively. The first comparative transcriptome analysis of elite ginger cultivars S1Z4 and S2Z5 from two different agro-climatic conditions of Odisha, India will help to understand the effect of agro-climatic conditions on differential expression of secondary metabolites in addition to genetic marker development.
Table 1.
Summary of de novo assembled cv. Suprabha transcriptome.
| Features | S1Z4 | S2Z5 |
|---|---|---|
| Total trinity transcripts generated | 60,452 | 54,748 |
| Maximum transcript length (bp) | 15,213 | 8988 |
| Median transcript length (bp) | 393 | 1164 |
| Average transcript Length (bp) | 1009.9 ± 830.6 | 986.7 ± 716.5 |
| Total transcripts Length (bp) | 6,10,51,850 | 5,40,19,323 |
| Total transcripts ≥ 500 bp | 40,026 | 37,791 |
| Total transcripts > 1 Kb | 21,743 | 20,305 |
| Total transcripts > 5 Kb | 232 | 36 |
| N50 | 1415 | 1334 |
| GC (%) | 45.06 | 44.81 |
Note: S1Z4-Sample 1 of cv. Suprabha harvested from zone 4; S2Z5-Sample 2 of cv. Suprabha harvested from zone 5.
Acknowledgements
The encouragement and support by Siksha ‘O’ Anusandhan University, Bhubaneswar, to carry out the present work is highly acknowledged.
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