Published June 17, 2024 | Version v1

Characterisation of the Cinnamomum parthenoxylon (Jack) Meisn (Lauraceae) transcriptome using Illumina paired-end sequencing and EST-SSR markers development for population genetics

  • 1. Graduate University of Science and Technology (GUST), Vietnam Academy of Science and Technology, Hanoi, Vietnam|Join Vietnam–Russia Tropical Science and Technology Research Center, Hanoi, Vietnam
  • 2. Join Vietnam–Russia Tropical Science and Technology Research Center, Hanoi, Vietnam
  • 3. Jiangsu Vocational Institute of Architectural Technology, School of Architectural Decoration, Xuzhou 221100, Jiangsu, China
  • 4. Institute of Ecology and Biological Resource, Vietnam Academy of Science and Technology, Hanoi, Vietnam
  • 5. Institute of Technology, Hanoi University of Industry (HaUI), Hanoi, Vietnam
  • 6. Department of Horticulture, Faculty of Agriculture, Gomal University, Dera Ismail Khan, Pakistan

Description

Cinnamomum parthenoxylon is an endemic and endangered species with significant economic and ecological value in Vietnam. A better understanding of the genetic architecture of the species will be useful when planning management and conservation. We aimed to characterize the transcriptome of C. parthenoxylon, develop novel molecular markers, and assess the genetic variability of the species. First, transcriptome sequencing of five trees (C. parthenoxylon) based on root, leaf, and stem tissues was performed for functional annotation analysis and development of novel molecular markers. The transcriptomes of C. parthenoxylon were analyzed via an Illumina HiSeqTM 4000 sequencing system. A total of 27,363,199 bases were generated for C. parthenoxylon. De novo assembly indicated that a total of 160,435 unigenes were generated (average length = 548.954 bp). The 51,691 unigenes were compared against different databases, i.e. COG, GO, KEGG, KOG, Pfam, Swiss-Prot, and NR for functional annotation. Furthermore, a total of 12,849 EST-SSRs were identified. Of the 134 primer pairs, 54 were randomly selected for testing, with 15 successfully amplified across nine populations of C. parthenoxylon. We uncovered medium levels of genetic diversity (PIC = 0.52, Na = 3.29, Ne = 2.18, P = 94.07%, Ho = 0.56 and He = 0.47) within the studied populations. The molecular variance was 10% among populations and low genetic differentiation (Fst = 0.06) indicated low gene flow (Nm = 2.16). A reduction in the population size of C. parthenoxylon was detected using BOTTLENECK (VP population). The structure analysis suggested two optimal genetic clusters related to gene flow among the populations. Analysis of molecular variance (AMOVA) revealed higher genetic variation within populations (90%) than among populations (10%). The UPGMA approach and DAPC divided the nine populations into three main clusters. Our findings revealed a significant fraction of the transcriptome sequences and these newlydeveloped novel EST-SSR markers are a very efficient tool for germplasm evaluation, genetic diversity and molecular marker-assisted selection in C. parthenoxylon. This study provides comprehensive genetic resources for the breeding and conservation of different varieties of C. parthenoxylon.

Files

BDJ_article_123405.pdf

Files (1.1 MB)

Name Size
md5:352a83ba0c51335185ce5958a4a82b15
1.1 MB Preview Download

System files (234.3 kB)

Name Size
md5:126b0e4258b47fe331a2bd0c690c9e62
234.3 kB Download

Linked records

Additional details

References

  • Abeysinghe PD, Samarajeewa NG, Li G, Wijesinghe KG (2014) Preliminary investigation for the identification of Sri Lankan Cinnamomum species using randomly amplified polymorphic DNA (RAPD) and sequence related amplified polymorphic (SRAP) markers. Journal of the National Science Foundation of Sri Lanka 42: 175‑182.
  • Apweiler R, Bairoch A, Wu C, Barker W, Boeckmann B, Ferro S, Gasteiger E, Huang H, Lopez R, Magrane M, Martin M, Natale D, O'donovan C, Donovan NR, L.Yeh L (2004) UniProt: the Universal Protein knowledgebase. Nucleic Acids Research 32 (Suppl. 1). https://doi.org/10.1093/nar/gkh131
  • Ashburner M, Ball CA, Blake JA, Botstein D, Cherry JM (2000) Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nature Genetics 25: 25‑29. https://doi.org/10.1038/75556
  • Bolger AM, Marc L, Bjoern U (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 15: 2114‑2120. https://doi.org/10.1093/bioinformatics/btu170
  • Botstein D, White RL, Skolnick M, Davis RW (1980) Construction of a genetic linkage map in man using restriction fragment length polymorphisms. American Journal of Human Genetics 32: 314.
  • Brutting C, Wesche K, Meyer S (2012) Genetic diversity of six arable plants in relation to their Red List status. Biodiversity and Conservation 21: 745‑761. https://doi.org/10.1007/s10531-011-0212-z
  • Chen J, Li R, Xia Y, Bai G, Guo P, Wang Z, Zhang H, Siddique KH (2017) Development of EST-SSR markers in flowering Chinese cabbage (Brassica campestris L ssp. chinensis var. utilis Tsen et Lee) based on de novo transcriptomic assemblies. PLOS One 12 (9). https://doi.org/10.1371/journal.pone.0184736
  • Chen LY, Cao YN, Yuan N, Nakamura K, Wang GM, Qiu YX (2015) 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. Molecular Breeding 35: 1‑15.
  • Chybicki IJ, Burczyk J (2009) Simultaneous estimation of null alleles and inbreeding coefficients. Journal of Heredity 100: 106‑113. https://doi.org/10.1093/jhered/esn088
  • Cui B, Vu DD, Vu DG, Bui TT, Rahman SU, Pham MP, Nguyen MT, Nguyen VS, Shah SN, Tran VH (2022) Genetic diversity and population structure of Cinnamomum balansae Lecomte inferred by microsatellites. Open Life Sciences 17: 323‑332. https://doi.org/10.1515/biol-2022-0037
  • Deng YY, Li JQ, Wu SF, Zhu YP, Chen YW, He FC (2006) Integrated nr database in protein annotation system and its localization. Computer Engineering 32: 71‑72.
  • Earl DA, Vonholdt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conservation Genetics Resources 4: 359‑361. https://doi.org/10.1007/s12686-011-9548-7
  • Evangelistella C, Valentini A, Ludovisi R, Firrincieli A, Fabbrini F, Scalabrin S, Cattonaro F, Morgante M, Mugnozza GS, Keurentjes JJ (2017) De novo assembly, functional annotation, and analysis of the giant reed (Arundo donax L.) leaf transcriptome provide tools for the development of a biofuel feedstock. Biotechnology for Biofuels 10: 138. https://doi.org/10.1186/s13068-017-0828-7
  • Evanno GS, Regnaut SJ, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: A simulation study. Molecular Ecology 14: 2611‑2620. https://doi.org/10.1111/j.1365-294X.2005.02553.x
  • Excoffier L, Laval G, Schneider S (2005) Arlequin (version 3.0): an integrated software package for population genetics data analysis. Evolutionary bioinformatics 1: 47‑50.
  • Finkeldey R, Hattemer HH (2007) Tropical forest genetics. Springer https://doi.org/10.1007/978-3-540-37398-8
  • Finn RD (2005) Pfam: the protein families database. American Cancer Society URL: http://pfam.xfam.org/
  • Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nature Biotechnology 29: 644‑652. https://doi.org/10.1038/nbt.1883
  • Gu C, Xu S, Wang Z, Liu L, Zhang Y, Deng Y, Huang S (2018) De novo sequencing, assembly, and analysis of Iris lactea var. chinensis roots' transcriptome in response to salt stress. Plant Physiology and Biochemistry 125: 1‑12. https://doi.org/10.1016/j.plaphy.2018.01.019
  • Hai TN, Nghi DH, Phuong LD (2016) Study on the sylvicuture characters of Cinnamomum balansae Lecomte at Ben En National Park. Journal of Forestry Science and Technology 6: 176‑185.
  • Hamrick J, Godt M (1996) Conservation genetics of endemic plant species. Chapman & Hall https://doi.org/10.1007/978-1-4757-2504-9
  • Hedrick PW (2005) A standardized genetic differentiation measure. Evolution 59 (8): 1633‑1638. https://doi.org/10.1554/05-076.1
  • 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. Molecular Genetics & Genomics 291: 849‑862. https://doi.org/10.1007/s00438-015-1147-y
  • Jakobsson M, Rosenberg NA (2007) CLUMPP: a cluster matching and permutation program for dealing with label switching and multimodality in analysis of population structure. Bioinformatics 23: 1801‑1806. https://doi.org/10.1093/bioinformatics/btm233
  • Jia XP, Deng YP, Sun XB, Liang LJ, Su JL (2016) De novo assembly of the transcriptome of Neottopteris nidus using Illumina paired-end sequencing and development of EST-SSR markers. Molecular Breeding 36: 94. https://doi.org/10.1007/s11032-016-0519-2
  • Jombart T, Devillard S, Balloux F (2010) Discriminant analysis of principal components: a new method for the analysis of genetically structured populations. BMC Genetics 11: 94. https://doi.org/10.1186/1471-2156-11-94
  • Kalinowski ST, Taper ML, Marshall TC (2007) Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Molecular Ecology 16: 1099‑1106. https://doi.org/10.1111/j.1365-294X.2007.03089.x
  • Kameyama Y, Furumichi J, Li JX, Tseng YH (2012) Development of microsatellite markers for Cinnamomum camphora (Lauraceae). American Journal of Botany 99: 1‑3.
  • Kameyama Y, Furumichi J, Li J, Tseng YH (2017) Natural genetic differentiation and human-mediated gene flow: the spatiotemporal tendency observed in a long-lived Cinnamomum camphora (Lauraceae) tree. Tree Genetics & Genomes 13: 38. https://doi.org/10.1007/s11295-017-1119-y
  • Kanehisa M, Goto S, Kawashima S, Okuno Y, Hattori M (2004) The KEGG resource for deciphering the genome. Nucleic Acids Research 32 (Suppl 1): 277‑280. https://doi.org/10.1093/nar/gkh063
  • Kaur S, Pembleton LW, Cogan NO, Savin KW, Leonforte T, Paull J, Materne M, Forster JW (2012) Transcriptome sequencing of field pea and faba bean for discovery and validation of SSR genetic markers. BMC Genomics 13: 104. https://doi.org/10.1186/1471-2164-13-104
  • Koonin EV, Fedorova ND, Jackson JD, Jacobs AR (2004) A comprehensive evolutionary classification of proteins encoded in complete eukaryotic genomes. Genome Biology 5: 7. https://doi.org/10.1186/gb-2004-5-3-p7
  • Liu J, Möller M, Provan J, Gao L, Poudel RC, Li D (2013) Geological and ecological factors drive cryptic speciation of yews in a biodiversity hotspot. New Phytologist 199 (4): 1093‑1108. https://doi.org/10.1111/nph.12336
  • Li Z, Zhong Y, Yu F, Xu M (2018) Novel SSR marker development and genetic diversity analysis of Cinnamomum camphora based on transcriptome sequencing. Plant Genetic Resources 16 (6): 568‑571. https://doi.org/10.1017/S147926211800014X
  • Martins EM, Lamont RW, Martinelli G, Lira-Medeiros CF, Quinet A, Shapcott A (2015) Genetic diversity and population genetic structure in three threatened Ocotea species (Lauraceae) from Brazil's Atlantic Rainforest and implications for their conservation. Conservation Genetics 16: 1‑14. https://doi.org/10.1007/s10592-014-0635-7
  • MOST, VAST (2007) Vietnam red data book. Part II. Plants. Science and Technique Publishing House289‑290. [In Vietnamese].
  • Muriira NG, Xu W, Muchugi A, Xu JC, Liu AZ (2015) De novo sequencing and assembly analysis of transcriptome in the Sodom apple (Calotropis gigantea). BMC Genomics 16 https://doi.org/10.1186/s12864-015-1908-3
  • Nguyen KD (2010) Flora of Vietnam - Lauraceae. Science and Technics Publishing House 20: 22‑234.
  • Nguyen TM, Vu DD, Dang HP, Bui XT, Nguyen DM (2021) Population genetic structure and demographic history of the dipterocarp species Anisoptera costata Korth revealed by microsatellite analysis. Planta 253: 66. https://doi.org/10.1007/s00425-021-03584-6
  • Peakall R, Smouse PE (2010) GENALEX 6: genetic analysis in Excel. Population genetic software for teaching and research. Molecular Ecology Notes 6: 288‑295. https://doi.org/10.1111/j.1471-8286.2005.01155.x
  • Peng LP, Cai CF, Zhong Y, Xu XX, Xian HL, Cheng FY, Mao JF (2017) Genetic analyses reveal independent domestication origins of the emerging oil crop Paeonia ostii, a tree peony with a long-term cultivation history. Scientific Reports 7: 5340. https://doi.org/10.1038/s41598-017-04744-z
  • Pertea G, Huang XQ, Liang F, Antonescu V, Sultana R, Karamycheva S, Lee YD, White J, Cheung F, Parvizi B (2003) TIGR Gene Indices clustering tools (TGICL): a software system for fast clustering of large EST datasets. Bioinformatics 19: 651‑652. https://doi.org/10.1093/bioinformatics/btg034
  • Piry S, Luikart G, Cornuet J (1999) Computer note. BOTTLENECK: a computer program for detecting recent reductions in the effective size using allele frequency data. Journal of Heredity 90 (4): 502‑503. https://doi.org/10.1093/jhered/90.4.502
  • Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155 (2): 945‑959. https://doi.org/10.1093/genetics/155.2.945
  • Reed DH (2002) Population fitness is correlated with genetic diversity. Conservation Biology 17: 230‑237. https://doi.org/10.1046/j.1523-1739.2003.01236.x
  • Rosenberg NA (2004) Distruct: a program for the graphical display of population structure. Molecular Ecology Notes 4: 137‑138. https://doi.org/10.1046/j.1471-8286.2003.00566.x
  • Sahu J, Sarmah R, Dehury B, Sarma K, Sen P (2012) Mining for SSRs and FDMs from expressed sequence tags of Camellia sinensis. Bioinformation 8: 260‑266. https://doi.org/10.6026/97320630008260
  • Sandigawad AM, Patil CG (2013) Genetic diversity in Cinnamomum zeylanicum Blume. (Lauraceae) using random amplified polymorphic DNA (RAPD) markers. African Journal of Biotechnology 10: 3682‑3688.
  • Sebbenn AM, Licona JC, Mostacedo B, Degen B (2017) Gene flow in an overexploited population of Swietenia macrophylla King (Meliaceae) in the Bolivian Amazon. Silvae Genetica 61: 212‑220. https://doi.org/10.1515/sg-2012-0027
  • Slatkin M (1985) Gene flow in natural populations. Annual Review of Ecology & Systematics 16: 393‑430. https://doi.org/10.1146/annurev.es.16.110185.002141
  • Tai VA, Thin NN (2014) Survey and inventory of threatened plant species in Ha Giang Province, Vietnam. Academia Journal of Biology 36: 323‑329.
  • Takezaki N, Nei M, Tamura K (2010) POPTREE2: Software for constructing population trees from allele frequency data and computing other population statistics with Windows interface. Molecular Biology and Evolution 27 (4): 747‑752. https://doi.org/10.1093/molbev/msp312
  • Tatusov RL, Galperin MY, Natale DA, Koonin EV (2000) The COG database: a tool for genome-scale analysis of protein functions and evolution. Nucleic Acids Research 28: 33‑36. https://doi.org/10.1093/nar/28.1.33
  • Tsuneki S, Mori K, Kaneko S, Isagi Y, Murakami N, Kato H (2009) Identification and characterization of eight microsatellite loci in Machilus pseudokobu (Lauraceae), an endemic species of the Bonin Islands. Conservation Genetics 10: 2009‑2011. https://doi.org/10.1007/s10592-009-9881-5
  • Van Oosterhout C, Hutchinson WF, Wills DP, Shipley P (2004) MICRO‐CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Molecular Ecology Notes 4 (3): 535‑538. https://doi.org/10.1111/j.1471-8286.2004.00684.x
  • Vu DD, Bui TT, Nguyen MT, Vu DG, Nguyen MD, Bui VT, Huang X, Zhang Y (2017) Genetic diversity in two threatened species in Vietnam: Taxus chinensis and Taxus wallichiana. Journal of Forestry Research 28: 265‑272. https://doi.org/10.1007/s11676-016-0323-1
  • Wang Y, Liu K, Bi D, Zhou S, Shao J (2017) Characterization of the transcriptome and EST-SSR development in Boea clarkeana, a desiccation-tolerant plant endemic to China. PEER J 5: 3422. https://doi.org/10.7717/peerj.3422
  • Wang ZY, Fang BP, Chen JY, Zhang XJ, Luo ZX, Huang LF, Chen XL, Li YJ (2010) De novo assembly and characterization of root transcriptome using Illumina paired-end sequencing and development of cSSR markers in sweetpotato (Ipomoea batatas). BMC Genomics 11: 726. https://doi.org/10.1186/1471-2164-11-726
  • Weir BS, Cockerham CC (1984) Estimating F-statistics for the analysis of population structure. Evolution1358‑1370. https://doi.org/10.2307/2408641
  • Yan K, Wei Q, Feng R, Zhou W, Chen F (2017) Transcriptome analysis of Cinnamomum longepaniculatum by high-throughput sequencing. Electronic Journal of Biotechnology 28: 58‑66. https://doi.org/10.1016/j.ejbt.2017.05.006
  • Yan L, Fan P, Wambulwa M, Qi H, Chen Y, Wu Z, Milne R, Khan R, Luo Y, Gao L, Shen S, Rashid I, Khan SM, Maity D, Li D, Liu J (2024) Human‐associated genetic landscape of walnuts in the Himalaya: implications for conservation and utilization. Diversity and Distributions 30 (4). https://doi.org/10.1111/ddi.13809
  • Yan XQ, Zhang X, Lu M, He Y, An HM (2015) De novo sequencing analysis of the Rosa roxburghii fruit transcriptome reveals putative ascorbate biosynthetic genes and EST-SSR markers. Gene 561: 54‑62. https://doi.org/10.1016/j.gene.2015.02.054
  • Yue H, Wang L, Liu H, Yue WJ, Du XH, Song WN, Nie XJ (2016) De novo assembly and characterization of the transcriptome of broomcorn millet (Panicum miliaceum L.) for gene discovery and marker development. Frontiers in Plant Science 7: 1083.
  • Zhang HB, Xia EH, Huang H, Jiang JJ, Liu BY, Gao LZ (2015) De novo transcriptome assembly of the wild relative of tea tree (Camellia taliensis) and comparative analysis with tea transcriptome identified putative genes associated with tea quality and stress response. BMC Genomics 16: 298. https://doi.org/10.1186/s12864-015-1494-4
  • Zhong YD, Yang AH, Li ZT, Zhang H, Liu LP, Wu ZX, Li YQ, Liu TY, Xu M, Yu FX (2019) Genetic diversity and population genetic structure of Cinnamomum camphora in South China revealed by EST-SSR markers. Forests 10: 1019. https://doi.org/10.3390/f10111019
  • Zhu SS, Ding YQ, Yap ZY, Qiu YX (2016) De novo assembly and characterization of the floral transcriptome of an economically important tree species, Lindera glauca (Lauraceae), including the development of EST-SSR markers for population genetics. Molecular Biology Reports 43: 1243‑1250. https://doi.org/10.1007/s11033-016-4056-1