Cloning, subcellular localization and expression analysis of telomere genes CcTBP1 and CcPOT1 in ancient Cinnamomum camphora

Liu Lipan; Yang Aihong; Liu Tengyun; Zhong Yongda; Zhou Hua; Yu Faxin

doi:10.12171/j.1000-1522.20210179

Journal of Beijing Forestry University > 2022 > 44(12): 1-11. > DOI: 10.12171/j.1000-1522.20210179

Liu Lipan, Yang Aihong, Liu Tengyun, Zhong Yongda, Zhou Hua, Yu Faxin. Cloning, subcellular localization and expression analysis of telomere genes CcTBP1 and CcPOT1 in ancient Cinnamomum camphora[J]. Journal of Beijing Forestry University, 2022, 44(12): 1-11. DOI: 10.12171/j.1000-1522.20210179

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Cloning, subcellular localization and expression analysis of telomere genes CcTBP1 and CcPOT1 in ancient Cinnamomum camphora

Key Laboratory of Horticultural Plant Genetic Improvement of Jiangxi Province, Institute of Biological Resources,Jiangxi Academy of Sciences, Nanchang 330096, Jiangxi, China

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Received Date: May 10, 2021
Revised Date: September 08, 2021
Accepted Date: September 18, 2022
Available Online: September 20, 2022
Published Date: December 24, 2023

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Abstract

Abstract

Objective Telomere genes play an important role in the process of plant tissue senescence and differentiation. In this study, the telomere genes CcTBP1 and CcPOT1 of ancient Cinnamomum camphora were cloned and analyzed by bioinformatics, subcellular localization and detection of their expression level in ancient C. camphora and its rejuvenating cuttings, to preliminarily reveal the function of telomere genes in the process of adventitious root formation of ancient C. camphora rejuvenating cuttings, and lay a foundation for exploring the telomere regulation mechanism of ancient C. camphora rejuvenating cuttings.
Method In this study, leaf RNA was extracted from 1200 years old ancient C. camphora. The telomere genes CcTBP1 and CcPOT1 were cloned by PCR and homologous cloning, as well the bioinformatics analysis was conducted to their sequences. The over-expression vector was constructed for subcellular localization analysis. The CcTBP1 and CcPOT1 expression level of different tissues and adventitious root formation process was analyzed by qRT-PCR in ancient C. camphora and its rejuvenation cutting seedlings.
Result (1) Two telomere genes were cloned, named CcTBP1 and CcPOT1, respectively. The CDS sequence of CcTBP1 gene was 2 163 bp and encoded 720 amino acids. The CDS sequence of CcPOT1 gene was 1 407 bp and encoded 468 amino acids. (2) The results of sequence similarity and phylogenetic tree analysis showed that the sequence similarity of CcTBP1 protein and Cinnamomum kanehirae RWR76720.1 was 98%, and their genetic relationship was the closest; the sequence similarity of CcPOT1 protein and Cinnamomum kanehirae RWR96964.1 was as high as 99%, and their genetic relationship was the closest. (3) Subcellular localization analysis showed that CcTBP1 protein was located in the nucleus, and CcPOT1 protein was located in the cytoplasm and nucleus. (4) The qRT-PCR expression analysis in root, stem and leaf showed that the expression level of CcTBP1 gene was highest in root of ancient C. camphora, the expression level of CcPOT1 gene was highest in root of rejuvenation cutting seedlings. Expression analysis showed CcTBP1 was down-regulated tendency during adventitious root formation process, and the expression level of ancient C. camphora was higher than rejuvenation cutting seedlings. And CcPOT1 gene was up-regulated tendency during adventitious root formation process, and the expression level of ancient C. camphora was lower than rejuvenation cutting seedlings.
Conclusion The telomere genes CcTBP1 and CcPOT1 play an important regulation function in the adventitous root formation process of ancient C. camphora rejuvenation.
- ancient Cinnamomum camphora,
- telomere gene,
- cloning,
- subcellular localization,
- expression analysis

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References

[1]	Shakirov E V, Salzberg S L, Alam M, et al. Analysis of Carica papaya telomeres and telomere-associated proteins: insights into the evolution of telomere maintenance in Brassicales[J]. Tropical Plant Biology, 2008, 1(3−4): 202−215.
[2]	McKnight T D, Riha K, Shippen D E. Telomeres, telomerase, and stability of the plant genome[J]. Plant Molecular Biology, 2002, 48(4): 331−337. doi: 10.1023/A:1014091032750
[3]	Shay J W, Wright W E. Telomeres and telomerase: three decades of progress[J]. Nature Reviews Genetics, 2019, 20(5): 299−309. doi: 10.1038/s41576-019-0099-1
[4]	Rossi M, Gorospe M. Noncoding RNAs controlling telomere homeostasis in senescence and aging[J]. Trends in Molecular Medicine, 2020, 26(4): 422−433. doi: 10.1016/j.molmed.2020.01.010
[5]	Hwang M G, Chung I K, Kang B G, et al. Sequence-specific binding property of Arabidopsis thaliana telomeric DNA binding protein 1 (AtTBP1)[J]. FEBS Letters, 2001, 503(1): 35−40. doi: 10.1016/S0014-5793(01)02685-0
[6]	Choi J Y, Abdulkina L R, Yin J, et al. Natural variation in plant telomere length is associated with flowering time[J]. The Plant Cell, 2021, 33(4): 1118−1134. doi: 10.1093/plcell/koab022
[7]	Fajkus J, Sykorova E, Leitch A R. Telomeres in evolution and evolution of telomeres[J]. Chromosome Research, 2005, 13(5): 469−479. doi: 10.1007/s10577-005-0997-2
[8]	Procházková S P, Schořová Š, Fajkus J. Telomere- and telomerase-associated proteins and their functions in the plant cell[J]. Frontiers in Plant Science, 2016, 7: 851.
[9]	de Lange T. Shelterin: the protein complex that shapes and safeguards human telomeres[J]. Genes & Development, 2005, 19(18): 2100−2110.
[10]	Yu E Y, Kim S E, Kim J H, et al. Sequence-specific DNA recognition by the Myb-like domain of plant telomeric protein RTBP1[J]. The Journal of Biological Chemistry, 2000, 275(31): 24208−24214. doi: 10.1074/jbc.M003250200
[11]	Hong J P, Byun M Y, Koo D H, et al. Suppression of rice telomere binding protein 1 results in severe and gradual developmental defects accompanied by genome instability in rice[J]. The Plant Cell, 2007, 19(6): 1770−1781. doi: 10.1105/tpc.107.051953
[12]	Shakirov E V, McKnight T D, Shippen D E. POT1-independent single-strand telomeric DNA binding activities in Brassicaceae[J]. The Plant Journal, 2009, 58(6): 1004−1015. doi: 10.1111/j.1365-313X.2009.03837.x
[13]	Baumann P, Cech T R. Pot1, the putative telomere end-binding protein in fission yeast and humans[J]. Science, 2001, 292: 1171−1175. doi: 10.1126/science.1060036
[14]	Tani A, Murata M. Alternative splicing of Pot1 (protection of telomere)-like genes in Arabidopsis thaliana[J]. Genes & Genetic Systems, 2005, 80(1): 41−48.
[15]	Rossignol P, Collier S, Bush M, et al. Arabidopsis POT1A interacts with TERT-V(I8), an N-terminal splicing variant of telomerase[J]. Journal of Cell Science, 2007, 120(20): 3678−3687. doi: 10.1242/jcs.004119
[16]	赵育新. 侧柏古树组织培养再生及其端粒相关基因克隆研究[D]. 北京: 中国林业科学研究院, 2015. Zhao Y X. Researches on tissue culture of old Platycladus orientalis and cloning relevant genes of telomere[D]. Beijing: Chinese Academy of Forestry, 2015.
[17]	Zhou Y, Yan W. Conservation and applications of camphor tree (Cinnamomum camphora) in China: ethnobotany and genetic resources[J]. Genetic Resources and Crop Evolution, 2016, 63(6): 1049−1061. doi: 10.1007/s10722-015-0300-0
[18]	汪松, 解焱. 中国物种红色目录[M]. 北京: 高等教育出版社, 2004. Wang S, Xie Y. China species red list[M]. Beijing: Higher Education Press, 2004.
[19]	裴东, 谷瑞升. 树木复幼的研究概述[J]. 植物学通报, 2005, 22(6): 753−760. Pei D, Gu R S. A review on the rejuvenation of mature trees[J]. Chinese Bulletin of Botany, 2005, 22(6): 753−760.
[20]	Liu S, Sun Z, Xu M. Identification and characterization of long non-coding RNAs involved in the formation and development of poplar adventitious roots[J]. Industrial Crops and Products, 2018, 118: 334−346. doi: 10.1016/j.indcrop.2018.03.071
[21]	秦爱丽, 简尊吉, 马凡强, 等. 母树年龄、生长调节剂、容器与基质对崖柏嫩枝扦插的影响[J]. 林业科学, 2018, 54(7): 40−50. doi: 10.11707/j.1001-7488.20180705 Qin A L, Jian Z J, Ma F Q, et al. Effects of the mother tree age, growth regulator, containers and substrates on softwood cutting propagation of Thuja sutchuenensis[J]. Scientia Silvae Sinicae, 2018, 54(7): 40−50. doi: 10.11707/j.1001-7488.20180705
[22]	Chang Y, Song X, Zhang Q, et al. Genome-wide identification of WOX gene family and expression analysis during rejuvenational rhizogenesis in walnut (Juglans regia L.)[J]. Forests, 2020, 11(1): 16.
[23]	孙晓梅, 韩华, 王笑山, 等. 杂种落叶松连续繁殖与插穗生根关系的生理研究[J]. 林业科学研究, 2012, 25(4): 486−491. doi: 10.3969/j.issn.1001-1498.2012.04.012 Sun X M, Han H, Wang X S, et al. Physiological research on the relationship between repeated cutting and rooting of Larix[J]. Forest Research, 2012, 25(4): 486−491. doi: 10.3969/j.issn.1001-1498.2012.04.012
[24]	Mather K A, Jorm A F, Parslow R A, et al. Is telomere length a biomarker of aging? a review[J]. The Journals of Gerontology: Series A, 2011, 66A(2): 202−213.
[25]	Díaz-Sala C. Molecular dissection of the regenerative capacity of forest tree species: special focus on conifers[J]. Frontiers in Plant Science, 2019, 9:1943.
[26]	Díaz-Sala C. A perspective on adventitious root formation in tree species[J]. Plants, 2020, 9(12): 1789. doi: 10.3390/plants9121789
[27]	Li S W. Molecular bases for the regulation of adventitious root generation in plants[J]. Frontiers in Plant Science, 2021, 12: 614072. doi: 10.3389/fpls.2021.614072
[28]	Wei M, Liu Q, Wang Z, et al. PuHox52-mediated hierarchical multilayered gene regulatory network promotes adventitious root formation in Populus ussuriensis[J]. New Phytologist, 2020, 228(4): 1369−1385. doi: 10.1111/nph.16778
[29]	Gailis A, Samsone I, Šēnhofa S, et al. Silver birch (Betula pendula Roth.) culture initiation in vitro and genotype determined differences in micropropagation[J]. New Forests, 2021: 1−16.
[30]	倪妍妍, 常二梅, 刘建锋, 等. 不同树龄侧柏接穗光合生理的比较研究[J]. 西北林学院学报, 2017, 32(1): 19−24. doi: 10.3969/j.issn.1001-7461.2017.01.03 Ni Y Y, Chang E M, Liu J F, et al. Comparison on photosynthetic physiology in various age scions of Platycladus orientalis[J]. Journal of Northwest Forestry University, 2017, 32(1): 19−24. doi: 10.3969/j.issn.1001-7461.2017.01.03
[31]	Sun H, Wu S, Wu L. White oak (Quercus fabri Hance) regenerated stump sprouts show few senescence symptoms during 40 years of growth in a natural forest[J/OL]. Forest Ecosystems, 2021, 8(1): 11[2021−03−11]. https://doi.org/10.1186/s40663-021-00292-1.
[32]	D’Amico-Willman K M, Anderson E S, Gradziel T M, et al. Relative telomere length and telomerase reverse transcriptase (TERT) expression are associated with age in almond (Prunus dulcis [Mill.] D. A. Webb)[J]. Plants, 2021, 10(2): 189. doi: 10.3390/plants10020189
[33]	Aronen T, Virta S, Varis S. Telomere length in Norway spruce during somatic embryogenesis and cryopreservation[J]. Plants, 2021, 10(2): 416. doi: 10.3390/plants10020416
[34]	Byun M Y, Cui L H, Lee H, et al. Telomere association of Oryza sativa telomere repeat-binding factor like 1 and its roles in telomere maintenance and development in rice, Oryza sativa L.[J]. BMB Reports, 2018, 51(11): 578−583. doi: 10.5483/BMBRep.2018.51.11.122
[35]	Zentgraf U, Hinderhofer K, Kolb D. Specific association of a small protein with the telomeric DNA-protein complex during the onset of leaf senescence in Arabidopsis thaliana[J]. Plant Molecular Biology, 2000, 42(3): 429−438. doi: 10.1023/A:1006324008600
[36]	Theobald D L, Mitton-Fry R M, Wuttke D S. Nucleic acid recognition by OB-fold proteins[J]. Annual Review of Biophysics and Biomolecular Structure, 2003, 32: 115−133. doi: 10.1146/annurev.biophys.32.110601.142506
[37]	Theobald D L, Wuttke D S. Prediction of multiple tandem OB-fold domains in telomere end-binding proteins Pot1 and Cdc13[J]. Structure, 2004, 12(10): 1877−1879. doi: 10.1016/j.str.2004.07.015
[38]	Luo M, Teng X, Wang B, et al. Protection of telomeres 1 (POT1) of Pinus tabuliformis bound the telomere ssDNA[J]. Tree Physiology, 2019, 40(1): 119−127.
[39]	Shakirov E V, Song X, Joseph J A, et al. POT1 proteins in green algae and land plants: DNA-binding properties and evidence of co-evolution with telomeric DNA[J]. Nucleic Acids Research, 2009, 37(22): 7455−7467. doi: 10.1093/nar/gkp785
[40]	Kobayashi C R, Castillo-González C, Survotseva Y, et al. Recent emergence and extinction of the protection of telomeres 1c gene in Arabidopsis thaliana[J]. Plant Cell Reports, 2019, 38(9): 1081−1097. doi: 10.1007/s00299-019-02427-9
[41]	Shakirov E V, Perroud P F, Nelson A D, et al. Protection of telomeres 1 is required for telomere integrity in the moss Physcomitrella patens[J]. The Plant Cell, 2010, 22(6): 1838−1848. doi: 10.1105/tpc.110.075846
[42]	Baumann P, Podell E, Cech T R. Human Pot1 (protection of telomeres) protein: cytolocalization, gene structure, and alternative splicing[J]. Molecular and Cellular Biology, 2002, 22(22): 8079−8087. doi: 10.1128/MCB.22.22.8079-8087.2002
[43]	Wei C, Price C M. Cell cycle localization, dimerization, and binding domain architecture of the telomere protein cPot1[J]. Molecular and Cellular Biology, 2004, 24(5): 2091−2102. doi: 10.1128/MCB.24.5.2091-2102.2004
[44]	Chen L Y, Liu D, Songyang Z. Telomere maintenance through spatial control of telomeric proteins[J]. Molecular and Cellular Biology, 2007, 27(16): 5898−5909. doi: 10.1128/MCB.00603-07

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Cloning, subcellular localization and expression analysis of telomere genes CcTBP1 and CcPOT1 in ancient Cinnamomum camphora

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Cloning, subcellular localization and expression analysis of telomere genes CcTBP1 and CcPOT1 in ancient Cinnamomum camphora

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