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De novo assembly of the seed transcriptome and search for potential EST-SSR markers for an endangered, economically important tree species: Elaeagnus mollis Diels

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Abstract

Seeds of Elaeagnus mollis (Elaeagnaceae) produce an edible oil and contain more vitamin E (Ve) than major oil-seed crops. Despite its economic value, there is no information on its genome sequence. Here, we used the Illumina platform to determine the seed transcriptome of E. mollis to identify the genes related to Ve biosynthesis and potential simple sequence repeat (SSR) markers. In total, 100,999 unigenes were obtained with an average length of 605 bp and N50 of 985 bp. Of these unigenes, 52,256 (51.7%) were annotated in at least one public database (NT, NR, PFAM, SwissProt, KOG, KO, and GO) in searches using blastn/x. The unigene annotation identified 15 unigenes encoding six enzymes (GGR, HPPD, HPT/VTE2, MPBQ-MT/VTE3, TC/VTE1, and γ-TMT/VTE4) putatively involved in Ve biosynthesis. In addition, 16,810 SSRs distributed in 14,057 unigenes were mined. Of these, 2820, 583, and 3423 SSRs were located in the 5′–UTR, coding sequence (CDS), and 3′–UTR regions, respectively, while the remaining 9984 SSRs had undetermined physical locations. The largest group of repeat motifs comprised mononucleotide repeats (70.76%), followed by dinucleotide (15.59%) and trinucleotide (12.10%) repeats. AG/CT (8.69%) and AAG/CTT (4.15%) were the main dinucleotide and trinucleotide repeats, respectively. Furthermore, 9597 SSR-specific primer pairs were designed. Among 100 primer pairs selected randomly to determine their usefulness, 53 proved to be efficient. To the our best of knowledge, this work is the first study of the E. mollis transcriptome and constitutes valuable genomics data for future genetic engineering studies to alter the amount of Ve. The identified potential EST-SSR markers can be used for population genetics studies and assisted-breeding of E. mollis.

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References

  • Cao X, Zhang G (2017) Application of the hyper-accurate mapping algorithm FANSe for next-generation sequencing in non-model organisms. Sci Sin Vitae 47:702–707

    Article  Google Scholar 

  • Chase MW, Christenhusz MJM, Fay MF, Byng JW, Judd WS, Soltis DE, Mabberley DJ, Sennikov AN, Soltis PS, Stevens PF (2016) An update of the Angiosperm Phylogeny Group classification for the orders and families of flowering plants: APG IV. Bot J Linn Soc 181:1–20

    Article  Google Scholar 

  • Chen SY, Li HJ, Liu GS (2006) Progress of vitamin E metabolic engineering in plants. Transgenic Res 15:655–665

    Article  CAS  Google Scholar 

  • Chen HL, Wang LX, Wang SH, Liu CJ, Blair MW, Cheng XZ (2015a) Transcriptome sequencing of mung bean (Vigna radiate L.) genes and the identification of EST-SSR markers. PLoS ONE 10:e0120273

    Article  Google Scholar 

  • Chen LY, Cao YN, Yuan N, Nakamura K, Wang GM, Qiu YX (2015b) Characterization of transcriptome and development of novel EST-SSR makers based on next-generation sequencing technology in Neolitsea sericea (Lauraceae) endemic to East Asian land-bridge islands. Mol Breed 35:187

    Article  CAS  Google Scholar 

  • Dorp KV, Hölzl G, Plohmann C, Eisenhut M, Abraham M, Weber APM, Hanson AD, Dörmann P (2015) Remobilization of phytol from chlorophyll degradation is essential for tocopherol synthesis and growth of Arabidopsis. Plant Cell 27:2846–2859

    Google Scholar 

  • Du X, Hussain N, Li ZL, Chen XY, Hua SJ, Zhang DQ, Jiang LX (2015) Effect of gibberellin on the biosynthesis of tocopherols in oilseed rape (Brassica napus L.) and Arabidopsis. J Agr Food Chem 63:360–369

    Article  CAS  Google Scholar 

  • Gailing O, Bodénès C, Finkeldey R, Kremer A, Plomion C (2013) Genetic mapping of EST-derived simple sequence repeats (EST-SSRs) to identify QTL for leaf morphological characters in a Quercus robur full-sib family. Tree Genet Genomes 9:1361–1367

    Article  Google Scholar 

  • Georgiadou EC, Ntourou T, Goulas V, Manganaris GA, Kalaitzis P, Fotopoulos V (2015) Temporal analysis reveals a key role for VTE5 in vitamin E biosynthesis in olive fruit during on-tree development. Front Plant Sci 6:871

    Article  Google Scholar 

  • Götz S, Garcíagómez JM, Terol J, Williams TD, Nagaraj SH, Nueda MJ, Robles M, Talón M, Dopazo J, Conesa A (2008) High-throughput functional annotation and data mining with the Blast2GO suite. Nucleic Acids Res 36:3420

    Article  Google Scholar 

  • Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng QD, Chen ZH, Mauceli E, Hacohen N, Gnirke A, Rhind N, Palma F, Birren BW, Chad Nusbaum, Lindblad-Toh K, Friedman N, Regev A (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotechnol 29:644–652

    Article  CAS  Google Scholar 

  • Harish MC, Dachinamoorthy P, Balamurugan S, Murugan SB, Sathishkumar R (2013) Enhancement of α-tocopherol content through transgenic and cell suspension culture systems in tobacco. Acta Physiol Plant 35:1121–1130

    Article  CAS  Google Scholar 

  • Hu Z, Zhang T, Gao XX, Wang Y, Zhang Q, Zhou HJ, Zhao GF, Wang ML, Woeste KE, Zhao P (2016) De novo assembly and characterization of the leaf, bud, and fruit transcriptome from the vulnerable tree Juglans mandshurica for the development of 20 new microsatellite markers using Illumina sequencing. Mol Genet Genom 291:849–862

    Article  CAS  Google Scholar 

  • Iseli C, Jongeneel CV, Bucher P (1999) ESTScan: a program for detecting, evaluating, and reconstructing potential coding regions in EST sequences. Proc Int Conf Intell Syst Mol Biol 99:138–148

    Google Scholar 

  • Kan LN, Wang L, Ding QZ, Wu YW, Ouyang J (2017) Flash extraction and physicochemical characterization of oil from Elaeagnus mollis Diels seeds. J Oleo Sci 66:345–352

    Article  CAS  Google Scholar 

  • Kong SL, Abdullah SNA, Chai LH, Amiruddin MD (2016) Molecular cloning, gene expression profiling and in silico sequence analysis of vitamin E biosynthetic genes from the oil palm. Plant Gene 5:100–108

    Article  CAS  Google Scholar 

  • Li DJ, Zhi D, Bi Q, Liu XH, Men ZH (2012) De novo assembly and characterization of bark transcriptome using Illumina sequencing and development of EST-SSR markers in rubber tree (Hevea brasiliensis Muell. Arg.). BMC Genom 13:192

    Article  CAS  Google Scholar 

  • Liang SH, Yang RN, Dong CW, Yang QP (2015) Physicochemical properties and fatty acid profiles of Elaeagnus mollis Diels nut oils. J Oleo Sci 64:1267–1272

    Article  CAS  Google Scholar 

  • Liu FL, Hu ZM, Liu WH, Li JJ, Wang WJ, Liang ZR, Sun XT (2016) Distribution, function and evolution characterization of microsatellite in Sargassum thunbergii (Fucales, Phaeophyta) transcriptome and their application in marker development. Sci Rep 6:18947

    Article  CAS  Google Scholar 

  • Liu CC, Ying D, Guan XL, Fu Q, Zhang Z, Hu ZH, Zheng J, Lu YZ, Li W (2017a) De novo transcriptomic analysis and development of EST-SSRs for Sorbus pohuashanensis (Hance) Hedl. PLoS ONE 12:e0179219

    Article  Google Scholar 

  • Liu HC, Cao GL, Han YP, Jiang ZF, Zhao HW, Li WB (2017b) Identification of the QTL underlying the vitamin E content of soybean seeds. Plant Breed 136:147–154

    Article  CAS  Google Scholar 

  • Mao XZ, Tao C, Olyarchuk JG, Wei LP (2005) Automated genome annotation and pathway identification using the KEGG Orthology (KO) as a controlled vocabulary. Bioinformatics 21(19):3787–3793

    Article  CAS  Google Scholar 

  • Mène-Saffrané L, Pellaud S (2017) Current strategies for vitamin E biofortification of crops. Curr Opin Biotech 44:189–197

    Article  Google Scholar 

  • Wang YL, Qin YY, Du Z, Yan GQ (2012) Genetic diversity and differentiation of the endangered tree Elaeagnus mollis Diels (Elaeagnus L.) as revealed by simple sequence repeat (SSR) markers. Biochem Syst Ecol 40:25–33

    Article  Google Scholar 

  • Wang L, Yang Y, Zhao Y, Yang S, Udikeri S, Liu T (2016a) De Novo Characterization of the root transcriptome and development of EST-SSR markers in Paris polyphylla Smith var. yunnanensis, an endangered medical plant. J Agric Sci Technol 18:437–452

    Google Scholar 

  • Wang PL, Yang LP, Zhang EH, Qin ZH, Wang HY, Liao YY, Wang XY, Gao L (2016b) Characterization and development of EST-SSR markers from a cold-stressed transcriptome of Centipedegrass by Illumina paired-end sequencing. Plant Mol Biol Rep 35:1–9

    CAS  Google Scholar 

  • Wang WC, Chen SY, Zhang XZ (2017a) Characterization of the complete chloroplast genome of Elaeagnus mollis, a rare and endangered oil plant. Conserv Gene Resour 9:439–442

    Article  Google Scholar 

  • Wang XD, Guo K, Guo JL, Juan LI, Lu JC (2017b) Chemical constituents from leaves of Elaeagnus mollis. Chin Tradit Herbal Drugs 48:236–240

    Google Scholar 

  • Xing W, Liao JY, Cai MY, Xia QF, Liu Y, Zeng W, Jin XL (2017) De novo assembly of transcriptome from Rhododendron latoucheae Franch using Illumina sequencing and development of new EST-SSR markers for genetic diversity analysis in Rhododendron. Tree Genet Genom 13:53

    Article  Google Scholar 

  • Yan GQ, Tian LH, Yang LY (2004) Studies on the browning in callus induction of Elaeagnus mollis. Acta Bot Boreal-Occident Sin 24:1384–1389

    CAS  Google Scholar 

  • Yan LP, Liu CL, Wu DJ, Li L, Shu J, Sun C, Xia Y, Zhao LJ (2016) De novo transcriptome analysis of Fraxinus velutina using Illumina platform and development of EST-SSR markers. Biol Plant 61:210–218

    Article  Google Scholar 

  • Yang W, Cahoon RE, Hunter SC, Zhang C, Han J, Borgschulte T, Cahoon EB (2011) Vitamin E biosynthesis: functional characterization of the monocot homogentisate geranylgeranyl transferase. Plant J Cell Mol Biol 65:206–217

    Article  CAS  Google Scholar 

  • Ye ZY, Wang ZS, Li YX, Rao GD, Zhang JG (2016) Development and characterization of microsatellite markers in the Elaeagnus mollis Diels. Acta Bot Boreal-Occident Sin 36:274–279

    Google Scholar 

  • Zhang JM, Zhang F (2015) Population structure and genetic variation of the endangered species Elaeagnus mollis Diels (Elaeagnaceae). Genet Mol Res 14:5950–5957

    Article  CAS  Google Scholar 

  • Zhang LW, Li YR, Tao AF, Fang PP, Qi JM (2015) Development and characterization of 1906 EST-SSR narkers from unigenes in jute (Corchorus spp.). PLoS ONE 10:e0140861

    Article  Google Scholar 

  • Zhou Q, Luo D, Ma LC, Xie WG, Wang Y, Wang YR, Liu ZP (2016) Development and cross-species transferability of EST-SSR markers in Siberian wildrye (Elymus sibiricus L.) using Illumina sequencing. Sci Rep 6:20549

    Article  CAS  Google Scholar 

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Correspondence to Yulin Liu.

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Project funding: The work was supported by the National Natural Science Foundation of China (31600549) and Doctor Faculty Inaugurating Project of Northwest A&F University (2452015296).

The online version is available at http://www.springerlink.com

Corresponding editor: Tao Xu.

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Liu, Y., Li, S., Wang, Y. et al. De novo assembly of the seed transcriptome and search for potential EST-SSR markers for an endangered, economically important tree species: Elaeagnus mollis Diels. J. For. Res. 31, 759–767 (2020). https://doi.org/10.1007/s11676-019-00917-w

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