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Changes of Gene Expression in Ovule of Chinese Fir (Cunninghamia lanceolata) Induced by Colchicine |
PAN Yao-Yao, WU Bo-Hui, TONG Zai-Kang, LU Yong-Quan* |
The State Key Laboratory of Subtropical Silviculture/College of Forestry and Biotechnology, Zhejiang A&F University, Hangzhou 311300, China |
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Abstract Chinese fir (Cunninghamia lanceolata) has the characteristics of naked ovule, which provides favorable conditions for colchicine to enter the ovule quickly and improve induction efficiency. In order to study the induction effect of colchicine on ovule of Chinese fir, the gene expression changes were analyzed by transcriptome sequencing technique, and the expression characteristic of related genes were verified by qRT-PCR. The results showed that, for colchicine treatment group and control group, 6.39 and 6.96 Gb original data were obtained by transcriptome sequencing, respectively. A total of 60 790 unigenes were obtained after de novo assembly, and 1 430 differentially expressed genes were screened between the two groups, from which 4 spindle formation-related genes, 8 DNA replication-related genes, and 6 cell cycle-related genes were obtained and all showed significant down-regulated expression. qRT-PCR analysis showed that the gene expression trends of NEDD1 (neural precursor cell expressed, developmentally down-regulated gene 1), MCM3 (minichromosome maintenance complex component 3), MCM7, CDC6 (cell division cycle 6) and ATXR6 (trithorax-related protein 6) were consistent with those of transcriptome data. In this study, colchicine was directly acted on ovules and differentially expressed genes were screened, which provides basic information for further studying the molecular mechanism on polyploidy induction by colchicine.
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Received: 29 March 2019
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Corresponding Authors:
luyongquan@zafu.edu.cn
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1 姜蓓, 唐亮, 路兆庚, 等. 2017. 杉木大孢子叶球发育过程中形态结构的变化[J]. 植物科学学报, 35(4): 469-476. (Jiang P, Tang L, Lu Z G, et al.2017. Morphological and structural changes during female cone development in Cunninghamia lanceolata[J]. Plant Science Journal, 35(4): 469-476.) 2 金术超, 曹立成. 2015. 再议秋水仙素诱导变异的机制[J]. 生物学教学, 40(12): 64-64. (Jin S C, Cao L C.2015. Further discussion on the mechanism of inducing variation of colchicine[J]. Biology Teaching, 40(12): 64-64.) 3 康向阳, 张平冬, 高鹏, 等. 2004. 秋水仙碱诱导白杨三倍体新途径的发现[J]. 北京林业大学学报, 26(1): 1-4. (Kang X Y, Zhang P D, Gao P, et al.2004. Discovery of a new way of poplar triploids induced with colchicine after pollination[J]. Journal of Beijing Forestry University, 26(1): 1-4.) 4 李云, 朱之悌, 田砚亭, 等. 2001. 秋水仙碱处理白杨雌花芽培育三倍体植株的研究[J]. 林业科学, 37(5): 68-74. (Li Y, Zhu Z T, Tian Y T, et al.2001. Studies on obtaining triploids by colchicine treating female flower buds of white poplar[J]. Scientia Silvae Sinicae, 37(5): 68-74.) 5 徐忠东, 吴琴. 1999. 微管蛋白的研究进展[J]. 安徽教育学院学报(自然科学版), (2): 73-74. (Xu Z D, Wu Q.1999. Progress in the study of tubulin[J]. Journal of Anhui Institute of Education (Natural Science Edition), (2): 73-74. 6 Bell S P, Dutta A.2003. DNA replication in eukaryotic cells[J]. Annual Review of Biochemistry, 71(1): 333-374. 7 Bell S P, Stillman B.1992. ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex[J]. Nature (London), 357(6374): 128-134. 8 DeLuca, J G, Dong Y, Hergert P, et al.2005. hHec1 and Nuf2 are core components of the kinetochore outer plate essential for organizing microtubule attachment sites[J]. Molecular Biology of the Cell, 16(2): 519-531. 9 Diffley J F X, Cocker J H, Dowell S J, et al.1994. Two steps in the assembly of complexes at yeast replication origins in vivo[J]. Cell, 78(2): 303-316. 10 Dunphy W G.1994. The decision to enter mitosis[J]. Trends in Cell Biology, 4(6): 202-207. 11 Fumiyuki S, Yasunari T, Hirak Kumar B, et al.2006. Asf1 is required for viability and chromatin assembly during DNA replication in vertebrate cells[J]. Journal of Biological Chemistry, 281(19): 13817-13827. 12 Grabherr M G, Haas B J, Moan Y, et al.2011. Full-length transcriptome assembly from RNA-Seq data without a reference genome[J]. Nature Biotechnology, 29(7): 644-652. 13 Graves D E, Velea L M.2000. Intercalative binding of small molecules to nucleic acids[J]. Current Organic Chemistry, 4(9): 915-929. 14 Groth A, Corpet A, Cook A J L, et al.2007. Regulation of replication fork progression through histone supply and demand[J]. Science, 318(5858): 1928-1931. 15 Jasencakova Z, Scharf A N D, Ask K, et al.2010. Replication stress interferes with histone recycling and predeposition marking of new histones[J]. Molecular Cell, 37(5): 736-743. 16 Ma W, Baumann C, Viveiros M M.2010. NEDD1 is crucial for meiotic spindle stability and accurate chromosome segregation in mammalian oocytes[J]. Developmental Biology, 339(2): 439-450. 17 Manning J A, Shalini S, Risk J M, et al.2010. A direct interaction with NEDD1 regulates γ-tubulin recruitment to the centrosome[J]. PLOS ONE, 5(3): e9618. 18 Mccleland M L, Kallio M J, Barrett-wilt G A, et al.2004. The vertebrate Ndc80 complex contains Spc24 and Spc25 homologs, which are required to establish and maintain kinetochore-microtubule attachment[J]. Current Biology, 14(2): 131-137. 19 Nebel B R.1937. Mechanism of polyploidy through colchicine[J]. Nature, 140(3556): 1101-1101. 20 Polo S E, Almouzni G.2006. Chromatin assembly: A basic recipe with various flavours[J]. Current Opinion in Genetics & Development, 16(2): 104-111. 21 Rajnish B, Wei Q, Hongtao Y.2004. Identification of two novel components of the human NDC80 kinetochore complex[J]. Journal of Biological Chemistry, 279(13): 13076-13085. 22 Raynaud C, Sozzani R, Glab N, et al.2010. Two cell-cycle regulated SET-domain proteins interact with proliferating cell nuclear antigen (PCNA) in Arabidopsis[J]. Plant Journal, 47(3): 395-407. 23 Riou-Khamlichi C, Huntley R, Jacqmard A, et al.1999. Cytokinin activation of Arabidopsis cell division through a D-type cyclin[J]. Science, 283(5407): 1541-1544. 24 Schulz, L L. Tyler J K.2006.The histone chaperone ASF1 localizes to active DNA replication forks to mediate efficient DNA replication[J]. The FASEB Journal, 20(3):488-90. 25 Sun S C, Lee S E, Xu Y N, et al.2010. Perturbation of Spc25 expression affects meiotic spindle organization, chromosome alignment and spindle assembly checkpoint in mouse oocytes[J]. Cell Cycle, 9(22): 4552-4559. 26 Shin J, Jeong G, Park J Y, et al.2018. MUN (MERISTEM UNSTRUCTURED), encoding a SPC24 homolog of NDC80 kinetochore complex, affects development through cell division in Arabidopsis thaliana[J]. Plant Journal for Cell & Molecular Biology, 93(6): 977-991. 27 Tsurimoto T.1998. PCNA, a multifunctional ring on DNA[J]. Biochimica et Biophysica Acta, 1443(1-2): 23-39. 28 Tyler J K, Adams C R, Chen S R, et al.1999. The RCAF complex mediates chromatin assembly during DNA replication and repair[J]. Nature, 402(6761): 555-560. 29 Watanabe N, Broome M, Hunter T.1995. Regulation of the human WEE1Hu CDK tyrosine 15-kinase during the cell cycle[J]. EMBO Journal, 14(9): 1878-1891. 30 Xu X, Rochette P J, Feyissa E A, et al.2014. MCM10 mediates RECQ4 association with MCM2-7 helicase complex during DNA replication[J]. EMBO Journal, 28(19): 3005-3014. 31 Zeng C J T, Y-R Julie L, Bo L.2009. The WD40 repeat protein NEDD1 functions in microtubule organization during cell division in Arabidopsis thaliana[J]. Plant Cell, 21(4): 1129-1140. |
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