Functional characterization of fifteen hundred transcripts from Ziarat juniper (Juniperus excelsa M.Bieb)

Humaira Abdul Wahid, Muhammad Younas Khan Barozai, Muhammad Din

Abstract


Background: Ziarat juniper (Juniperus excelsa M.Bieb) is an evergreen and dominant species of Balochistan juniper forests. This forest is providing many benefits to regional ecosystems and surrounding populations. No functional genomics study is reported for this important juniper plant. This research is aimed to characterize the Ziarat juniper functional genome based on the analyses of 1500 transcripts.  

Methods: Total RNA from shoot of Juniperus excelsa was extracted and subjected for transcriptome sequencing using Illumina HiSeq 2000 with the service from Macrogen, Inc., South Korea. The Illumina sequenced data was subjected to bioinformatics analysis. Quality assessment and data filtration was performed for the removal of low-quality reads, ambiguous reads and adaptor sequences. The high-quality clean reads data was deposited in the Sequence Read Archive (SRA) at NCBI, and used for downstream processes. Fifteen hundred transcripts were randomly chosen and used for functional characterization.

Results: As a result of homology search 80.3% transcripts showed significant similarities and were placed  in significant similarities category, 19.3% transcripts showed low similarities and assigned to the ‘‘unclassified’’ category while 0.4% transcripts are defined as no hits. The functional characterization results showed that most (18%) of the transcripts are involved in metabolism, followed by 11.7% in transcription and 11.5% as structural protein. 8.8% transcripts are engaged in stress response, whereas the transcripts involved in growth and development constituted 6.7%. Transcripts involved in signal transduction represented 5.6%, while 3.5% facilitating transport and 34.1% are involved in hypothetical functions.

Conclusion: The functional annotation data produced in this study will be very useful for future functional genome analysis of Juniperus excelsa. 


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Gulacti T, Ramazan E, Osman C, Candan J, Cennet C, Chai HB, Pezzuto JM. Diterpenes from the berries of Juniperus excels. Phytochemistry, (1999); 50(7): 1195-119.

Farjon, A. World checklist and bibliography of conifers. Royal Botanic Gardens, Kew chemistry of natural compounds, (1998); 44(1): 2008.

Rafi, M. (1965). Vegetation types of Balochistan Province. Pakistan Govt, Printing Press, Lahore, Pakistan.

Weli AM, Jamila RK, AL-Hinai, Jawaher MA, Al-Mjrafi, Jawaher RS, Mohammad AH, Saeed S, Aktar MS. Effect of different polarities leaves crude extracts of Omani juniperus excels on antioxidant, antimicrobial and cytotoxic activities and their biochemical screening. Asian Pacific Journal of Reproduction, (2014); 3(3): 218-223.

Khan M, Khan AU, Rehman NU, Zafar MA, Hazrat A, Gilani AH. Cardiovascular effects of Juniperus excelas, are mediated through multiple pathways. Clinical and Experimental Hypertension, (2012); 34: 209-216.

Sela F, Karapandzova M, Stefkov G, Cvetkovikj I, Kulevanova S. Chemical composition and antimicrobial activity of essential oils of Juniperus excelsa Bieb. (Cupressaceae) grown in R. Macedonia. Pharmacognosy Research, (2015); 7(1): 74-80.

Unlu M, Vardar-Unlu G, Vural N, Domnez E, Cakmak O. Composition and antimicrobial activity of Juniperus excelsa essential oil. Chemistry of Natural Compounds, (2008); 44: 129-31.

Patel RK, Jain M. NGS QC toolkit: a toolkit for quality control of next generation sequencing data. PLoS One, (2012); 7 (2).

Morozova O, Marra MA. Applications of next-generation sequencing technologies in functional genomics. Genomics, (2008); 92(5): 255-64.

Wahid HA, Barozai MYK, Din M. Optimization of total RNA extraction protocol from Ziarat Juniper (Juniperus excelsa M.Bieb). Pure and Applied Biology, (2015); 4(2):275-279.

Bohnerta HJ, Ayoubid P, Borcherta C, Bressanc RA, Burnapd R, Cushmane JC, et al. A genomics approach towards salt stress tolerance. Plant Physiology and Biochemistry, (2001); 39: 295-311.

Fu XH, Huang YL, Deng SL, Zhou RC, Yang GL, Ni XW, Li WJ, Shi SH. Construction of a SSH library of Aegiceras corniculatum under salt stress and expression analysis of four transcripts. Plant Science, (2005); 169: 147–154.

Liu X, Huang J, Zhang Y, Lu Y. Identification of differentially expressed sequences in bud differentiation of oriental hybrid lily cultivar ‘Sorbonne’ via suppression subtractive hybridization. African Journal of Biotechnology, (2012); 11(66):12990-12997.

Xu H, He X, Wang K, Chen L, Li K. Identification of early nitrate stress response genes in Spinach roots by suppression subtractive hybridization. Plant Molecular Biology Reporter, (2012); 30: 633-642.

Roohie RK, Umesha S. Identification of genes associated with black rot resistance in cabbage through suppression subtractive hybridization. Springer 3 Biotech, (2015); 5:1089-1100.

Fei J, Wang Y, Jiang Z, Cheng H, Zhang J. Identification of cold tolerance genes from leaves of mangrove plant Kandelia obovata by suppression subtractive hybridization. Ecotoxicology, (2015); 24:1686-1696.

Tian j, Belanger F, Huang B. Identification of heat stress-responsive genes in heat-adapted thermal Agrostis scabra by suppression subtractive hybridization. Journal of Plant Physiology, (2009); 166(6): 588-601.

Lu W, Tang X, Huo Y, Xu R, Qi S, Huang J, Zheng C, Wu CA. Identification and characterization of fructose 1,6-bisphosphate aldolase genes in Arabidopsis reveal a gene family with diverse responses to abiotic stresses. Gene, (2012)15; 503(1): 65-74.

Costa MA, Collins RE, Anterola AM, Cochrane FC, Davin LB, Lewis NG. An in silico assessment of gene function and organization of the phenylpropanoid pathway metabolic networks in Arabidopsis thaliana and limitations thereof, Phytochemistry, (2003); 64:1097-1112.

Kawasaki T, Koita H, Nakatsubo T, Hasegawa K, Wakabayashi K, Takahashi H, Umemura K, Umezawa T, Shimamoto K. Cinnamoyl-CoA reductase, a key enzyme in lignin biosynthesis, is an effector of small GTPase Rac in defense signaling in rice. Proceeding of National Academy of Science, (2006); 103: 230-235.

Li L, Cheng X, Lu S, Nakatsubo T, Umezawa T, Chiang VL. Clarification of cinnamoyl co-enzyme A reductase catalysis in monolignol biosynthesis of Aspen. Plant Cell Physiology, (2005); 46:1073-1082.

Ma QH. Characterization of a cinnamoyl-CoA reductase that is associated with stem development in wheat. Journal of Experimental Botany, (2007); 58: 2011-2021.

Andersen JR, Zein I, Wenzel G, Darnhofer B, Eder J, Ouzunova M, Lubberstedt T. Characterization of phenylpropanoid pathway genes within European maize (Zea mays L.) inbreds. BMC Plant Biology, (2008); 8: 2.

Dian WM, Jiang HW, Chen QS, Liu FY, Wu P. Cloning and characterization of the granule-bound starch synthase II gene in rice: gene expression is regulated by the nitrogen level, sugar and circadian rhythm. Planta, (2003); 218: 261-268.

Jiang HW, Dian WM, Liu FY, Wu P. Molecular cloning and expression analysis of three genes encoding starch synthase II in rice. Planta, (2004); 218: 1062-1070.

Barozai MYK, Wahid HA. In-silico identification and characterization of cumulative abiotic stress responding genes in potato (Solanum tuberosum L.). Pakistan Journal of Botany, (2012) 44: 57-69.

Cheuk A, Houde M. Genome wide identification of C1‑2i zinc finger proteins and their response to abiotic stress in hexaploid wheat. Molecular Genet Genomics, (2016); 291:873-890.

Hichri I, Barrieu F, Bogs J, Kappel C, Delrot S, Lauvergeat V.Recent advances in the transcriptional regulation of the flavonoid biosynthetic pathway. Journal of Experimental Botany, (2011); 62(8): 2465-2483.

Dubos C, Stracke R, Grotewold E, Weisshaar B, Martin C, Lepiniec L. MYB transcription factors in Arabidopsis. Trends in Plant Sciences, (2010); 15(10): 573-581.

Du H, Yang SS, Feng BR, Liu L, Tang YX, et al. Genome-wide analysis of the MYB transcription factor superfamily in soybean. BMC Plant Biology, (2012); 12: 106.

Soler M, Leal E, Camargo O, Carocha V, Cassan-Wang H, Clemente HS, Savelli B, Charles A,Jorge A, Paiva P, Myburg AA, Grima-Pettenati J. The Eucalyptus grandis R2R3-MYB transcription factor family: evidence for woody growth-related evolution and function. New Phytologist, (2015); 206(4): 1364-1377.

Li X, Xue C, Li J, Qiao X, Li L, Yu L, Huang Y. Genome-Wide Identification, Evolution and Functional Divergence of MYB Transcription Factors in Chinese White Pear (Pyrus bretschneideri) Plant and Cell Physiology, (2016); 57(4): 824-847.

Rea PA. Plant ATP-Binding Cassette Transporters. Plant Biology, (2007); 58: 347-375.

Chen L, Ortiz-Lopez A, Jung A, Bush DR. ANT1, an Aromatic and Neutral Amino Acid Transporter in Arabidopsis. American Society of Plant Physiologists, Plant Physiology, (2001); 125 (4): 1813-1820.

Hollender C, Liu Z. Histone Deacetylase Genes in Arabidopsis Development. Journal of Integrative Plant Biology, (2008); 50 (7): 875-885.

Wu K, Tian L, Malik K, Brown D, Miki B. Functional analysis of HD2 histone deacetylase homologues in Arabidopsis thaliana. Plant Journal, (2000); 22: 19-27.

Dangl M, Brosch G, Haas H, Loidl P, Lusser A. Comparative analysis of HD2 type histone deacetylases in higher plants. Planta, (2001); 213: 280-285.

Keller B, Sauer N, Lamb CJ. Glycine-rich cell wall proteins in bean: gene structure and association of the protein with the vascular system. The EMBO Journal, (1988); 7(12): 3625-3633.

Lei M, Wu R. A novel glycine-rich cell wall protein gene in rice. Plant Molecular Biology, (1991); 16 (2): 187-198.

Fu Y, Yang Z. Rop GTPase: a master switch of cell polarity development in plants. Trends Plant Science, (2001); 6: 545-547.

Schultheiss H, Dechert C, Kogel K, Huckelhoven R. Functional analysis of barley RAC/ROP G-protein family members in susceptibility to the powdery mildew fungus. The Plant Journal, (2003); 36(5): 589-601.

Falchi R, Cipriani G, Marrazzo T, Nonis A, Vizzotto G, Ruperti B. Identification and differential expression dynamics of peach small GTPases encoding genes during fruit development and ripening. Journal of Experimental Botany, (2010); 61(10): 2829-2842.

Shiu SH, Bleecker AB. Receptor-like kinase from Arabidopsis form a monophyletic gene family related to animal receptor kinases. Proceeding of National Academy of Science, (2001); 98:10763-10768.

Sun XL, Wang GL. Genome-wide identification, characterization and phylogenetic analysis of the rice LRR-kinases. PLoS ONE, (2011); 6(3):160-179.

Zan YJ, Ji Y, Zhang Y, Yang SH, Song YJ, Wang JH. Genome-wide identification, characterization and expression analysis of populus leucine-rich repeat receptor-like protein kinase genes. BMC Genomics, (2013); 14:318.

Zhou F, Guo Y, Qiu L. Genome-wide identification and evolutionary analysis of leucine-rich repeat receptor-like protein kinase genes in soybean. BMC Plant Biology, (2016) 16:58.

Santhanagopalan I, Basha E, Ballard KN , Bopp NE, Vierling E. Model Chaperones: Small Heat Shock Proteins from Plants. The series Heat Shock Proteins, (2015); 8: 119-153.

Barozai MYK, Husnain T. Identification of biotic and abiotic stress up-regulated ESTs in Gossypium arboreum. Molecular Biology Reports, (2012) 39 (2): 1011-1018.

Wang R, Zhang Y, Kieffer M, Yu H, Kepinski S, Estelle M. HSP90 regulates temperature-dependent seedling growth in Arabidopsis by stabilizing the auxin co-receptor F-box protein TIR1. Natural communication, (2016); DOI: 10.1038/ncomms10269.

Zhuo C, Wang T, Lu S, Zhao Y, Li X, Guo Z. A cold responsive galactinol synthase gene from Medicago falcata is induced by myo-inositol and confers multiple tolerances to abiotic stresses. Physiologia Plantarum, (2013); 149(1):67-78.


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