Cloning and Expression of Mannose-1-phosphate Guanyltransferase Gene in <i>Anoectochilus roxburghii</i>

LI Heping; LIN Jiangbo; HUANG Huiming; ZOU Hui; DAI Yimin

doi:10.19303/j.issn.1008-0384.2023.05.004

Volume 38 Issue 5

May 2023

Turn off MathJax

Article Contents

Article Navigation > Fujian Journal of Agricultural Sciences > 2023 > 38(5): 537-544

LI H P, LIN J B, HUANG H M, et al. Cloning and Expression of Mannose-1-phosphate Guanyltransferase Gene in Anoectochilus roxburghii [J]. Fujian Journal of Agricultural Sciences，2023，38（5）：537−544 doi: 10.19303/j.issn.1008-0384.2023.05.004

Citation:

LI H P, LIN J B, HUANG H M, et al. Cloning and Expression of Mannose-1-phosphate Guanyltransferase Gene in Anoectochilus roxburghii [J]. Fujian Journal of Agricultural Sciences，2023，38（5）：537−544 doi: 10.19303/j.issn.1008-0384.2023.05.004

Citation:

PDF( 1757 KB)

Cloning and Expression of Mannose-1-phosphate Guanyltransferase Gene in Anoectochilus roxburghii

doi: 10.19303/j.issn.1008-0384.2023.05.004

Institute of Subtropical Agriculture, Fujian Academy of Agricultural Sciences, Zhangzhou, Fujian　363005, China

Received Date: 2022-12-22
Rev Recd Date: 2023-04-06

Available Online: 2023-05-24

Publish Date: 2023-05-28

Abstract

Abstract

Objective Cloning and expression of mannose-1-phosphate guanyltransferase (GMP) gene that regulates the polysaccharide synthesis in Anoectochilus roxburghii were studied. Method ArGMP cDNA and genome sequences were cloned, and bioinformatics analyzed using online software. Gene expression pattern was then determined by qRT-PCR. Result The ORF of ArGMP was 1 086 bp encoded 361 amino acids with a length of 1 760 bp that contained 3 introns. It has an accession number of OQ030271 in GenBank. A stable hydrophilic protein free of a transmembrane structure, it was closely related to those of Dendrobium officinale, Apostasia shenzhenica, and Phalaenopsis equestris. Expressed differently in different tissues, the highest level was found in the flowers. The expression varied under different temperatures that peaked at 25 ℃, decreased significantly after 3 h at 35 ℃, and was severely inhibited beyond that. However, it was not affected by the stress exerted by NaCl in different concentrations. Conclusion The cDNA and sequence of GMP gene from A. roburghii were successfully cloned to display a tissue-specific expression pattern that could be significantly affected by temperature but not salt stress.
- Anoectochilus roxburghii,
- mannose-1-phosphate guanyltransferase,
- polysaccharide,
- gene expression

FullText(HTML)

References(26)

References

[1]	中国科学院中国植物志编辑委员会. 中国植物志-第六卷, 第一分册-蕨类植物门[M]. 北京: 科学出版社, 1999: 204-227.
[2]	YANG Z G, ZHANG X H, YANG L W, et al. Protective effect of Anoectochilus roxburghii polysaccharide against CCl₄-induced oxidative liver damage in mice [J]. International Journal of Biological Macromolecules, 2017, 96: 442−450. doi: 10.1016/j.ijbiomac.2016.12.039
[3]	马玉芳, 郑小香, 郑乃珍, 等. 金线莲多糖协同ConA对小鼠脾淋巴细胞分泌Th1、Th2型细胞因子及其mRNA表达量的影响 [J]. 中国食品学报, 2018, 18(4):72−78. doi: 10.16429/j.1009-7848.2018.04.011 MA Y F, ZHENG X X, ZHENG N Z, et al. Regulatory effects of Anoectochilus roxburghii polysaccharide(ARP) on ConA-induced cytokines secretion and mRNA expression by murine splenocytes [J]. Journal of Chinese Institute of Food Science and Technology, 2018, 18(4): 72−78.(in Chinese) doi: 10.16429/j.1009-7848.2018.04.011
[4]	GUO Y L, YE Q, YANG S L, et al. Therapeutic effects of polysaccharides from Anoectochilus roxburghii on type II collagen-induced arthritis in rats [J]. International Journal of Biological Macromolecules, 2019, 122: 882−892. doi: 10.1016/j.ijbiomac.2018.11.015
[5]	YU X L, LIN S E, ZHANG J Q, et al. Purification of polysaccharide from artificially cultivated Anoectochilus roxburghii (wall. ) Lindl. by high-speed counter current chromatography and its antitumor activity [J]. Journal of Separation Science, 2017, 40(22): 4338−4346. doi: 10.1002/jssc.201700340
[6]	TANG T T, DUAN X Y, KE Y, et al. Antidiabetic activities of polysaccharides from Anoectochilus roxburghii and Anoectochilus formosanus in STZ-induced diabetic mice [J]. International Journal of Biological Macromolecules, 2018, 112: 882−888. doi: 10.1016/j.ijbiomac.2018.02.042
[7]	王常青, 严成其, 王勇, 等. 台湾金线莲多糖的分离纯化及其体外抑瘤活性研究 [J]. 中国生化药物杂志, 2008, 28(2):93−96. WANG C Q, YAN C Q, WANG Y, et al. Isolaiton, purification and in vitro anti-tumor activities of Anoectochilus formosanus polysaccharide [J]. Chinese Journal of Biochemical Pharmaceutics, 2008, 28(2): 93−96.(in Chinese)
[8]	林守二, 黄丽英, 俞晓玲. 柱前衍生化HPLC法分析人工栽培金线莲中多糖的单糖组成 [J]. 福建医科大学学报, 2016, 50(3):148−154. doi: 10.3969/j.issn.1672-4194.2016.03.002 LIN S E, HUANG L Y, YU X L. Analyze monosaccharide composition of polysaccharide in artificial cultivated Anoectochilus roxburghii by pre-column derivatization HPLC [J]. Journal of Fujian Medical University, 2016, 50(3): 148−154.(in Chinese) doi: 10.3969/j.issn.1672-4194.2016.03.002
[9]	NARASAKI C T, MERTENS K, SAMUEL J E. Characterization of the GDP-D-mannose biosynthesis pathway in Coxiella burnetii: The initial steps for GDP-β-D-virenose biosynthesis [J]. PLoS One, 2011, 6(10): e25514. doi: 10.1371/journal.pone.0025514
[10]	PENG L, LI J, LIU Y, et al. Effects of mixed carbon sources on galactose and mannose content of exopolysaccharides and related enzyme activities in Ganoderma lucidum [J]. RSC Advances, 2016, 6(45): 39284−39291. doi: 10.1039/C6RA04798J
[11]	赵丽娜, 徐军伟. 过表达GDP-D-甘露糖焦磷酸化酶(GMP)基因提高灵芝多糖的生产 [J]. 菌物学报, 2021, 40(5):1099−1109. ZHAO L N, XU J W. Overexpression of GDP-D-mannose pyrophosphorylase (GMP) gene improves the production of polysaccharides in Ganoderma lingzhi [J]. Mycosystema, 2021, 40(5): 1099−1109.(in Chinese)
[12]	ZHOU J S, BAI Y, DAI R J, et al. Improved polysaccharide production by homologous co-overexpression of phosphoglucomutase and UDP glucose pyrophosphorylase genes in the mushroom Coprinopsis cinerea [J]. Journal of Agricultural and Food Chemistry, 2018, 66(18): 4702−4709. doi: 10.1021/acs.jafc.8b01343
[13]	HOWLETT R, READ N, VARGHESE A, et al. Streptomyces coelicolor strains lacking polyprenol phosphate mannose synthase and protein O-mannosyl transferase are hyper-susceptible to multiple antibiotics [J]. Microbiology, 2018, 164(3): 369−382. doi: 10.1099/mic.0.000605
[14]	李鹏佳, 焦茂娟, 王启军, 等. 魔芋GDP-甘露糖焦磷酸化酶基因及其启动子的克隆与分析 [J]. 西南大学学报(自然科学版), 2019, 41(3):14−22. doi: 10.13718/j.cnki.xdzk.2019.03.003 LI P J, JIAO M J, WANG Q J, et al. Cloning and characterization of GDP-mannose pyrophosphorylase gene in konjac and its promoter [J]. Journal of Southwest University (Natural Science Edition), 2019, 41(3): 14−22.(in Chinese) doi: 10.13718/j.cnki.xdzk.2019.03.003
[15]	郑云柯, 胡翔宇, 宋希强, 等. 石斛属植物基因组DNA提取方法的对比 [J]. 热带生物学报, 2015, 6(2):168−172. doi: 10.3969/j.issn.1674-7054.2015.02.011 ZHENG Y K, HU X Y, SONG X Q, et al. Optimized extraction method for genomic DNA from Dendrobium Species (Orchidaceae) [J]. Journal of Tropical Biology, 2015, 6(2): 168−172.(in Chinese) doi: 10.3969/j.issn.1674-7054.2015.02.011
[16]	林江波, 王伟英, 邹晖, 等. 金线莲3个持家基因表达稳定性分析 [J]. 福建农业学报, 2018, 33(11):1125−1129. doi: 10.19303/j.issn.1008-0384.2018.11.001 LIN J B, WANG W Y, ZOU H, et al. Expression stabilities of three housekeeping genes of Anoectochilus roxburghii [J]. Fujian Journal of Agricultural Sciences, 2018, 33(11): 1125−1129.(in Chinese) doi: 10.19303/j.issn.1008-0384.2018.11.001
[17]	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^−△△Ct Method. Methods,2001, 25(4), 402-408.
[18]	CRONJE C, GEORGE G M, FERNIE A R, et al. Manipulation of l-ascorbic acid biosynthesis pathways in Solanum lycopersicum: Elevated GDP-mannose pyrophosphorylase activity enhances l-ascorbate levels in red fruit [J]. Planta, 2012, 235(3): 553−564. doi: 10.1007/s00425-011-1525-6
[19]	申艳红, 陈晓静, 卢秉国, 等. 番木瓜GDP-D-甘露糖焦磷酸化酶基因的克隆及分析 [J]. 福建农林大学学报(自然科学版), 2011, 40(2):172−177. doi: 10.13323/j.cnki.j.fafu(nat.sci.).2011.02.017 SHEN Y H, CHEN X J, LU B G, et al. Cloning and analysis of GDP-D-mannose pyrophosphorylase gene from papaya [J]. Journal of Fujian Agriculture and Forestry University (Natural Science Edition), 2011, 40(2): 172−177.(in Chinese) doi: 10.13323/j.cnki.j.fafu(nat.sci.).2011.02.017
[20]	范林洪, 李锐, 童永鳌, 等. 金发草GDP-D-甘露糖焦磷酸化酶基因的克隆及功能分析 [J]. 四川大学学报(自然科学版), 2015, 52(3):682−688. FAN L H, LI R, TONG Y A, et al. Cloning and analysis of GDP-D-mannose pyrophosphorylase gene from Pogonatherum paniceum [J]. Journal of Sichuan University (Natural Science Edition), 2015, 52(3): 682−688.(in Chinese)
[21]	WANG C K, PENG D Y, ZHU J H, et al. Transcriptome analysis of Polygonatum cyrtonema Hua: Identification of genes involved in polysaccharide biosynthesis [J]. Plant Methods, 2019, 15(1): 1−14. doi: 10.1186/s13007-018-0385-5
[22]	SAWAKE S, TAJIMA N, MORTIMER J C, et al. KONJAC1 and 2 are key factors for GDP-mannose generation and affect l-ascorbic acid and glucomannan biosynthesis in Arabidopsis [J]. The Plant Cell, 2015, 27(12): 3397−3409. doi: 10.1105/tpc.15.00379
[23]	肖瑶, 周天山, 李佼, 等. 茶树GDP-D-甘露糖焦磷酸化酶基因cDNA全长的克隆与表达分析 [J]. 茶叶科学, 2015, 35(1):55−63. doi: 10.3969/j.issn.1000-369X.2015.01.010 XIAO Y, ZHOU T S, LI J, et al. Cloning and expression analysis of GDP-D-mannose pyrophosphorylase c DNA in Camellia sinensis [J]. Journal of Tea Science, 2015, 35(1): 55−63.(in Chinese) doi: 10.3969/j.issn.1000-369X.2015.01.010
[24]	李超汉, 李青竹, 史庆华, 等. 超表达番茄GDP-D-甘露糖焦磷酸化酶基因马铃薯对温度胁迫的响应 [J]. 中国农业科学, 2011, 44(23):4952−4961. doi: 10.3864/j.issn.0578-1752.2011.23.024 LI C H, LI Q Z, SHI Q H, et al. Effect of tomato GMPase overexpression on tolerance of potato plants to temperature stress [J]. Scientia Agricultura Sinica, 2011, 44(23): 4952−4961.(in Chinese) doi: 10.3864/j.issn.0578-1752.2011.23.024
[25]	程智慧, 关志华, 薛琳, 等. 叶面喷施NaCl对加工番茄果实耐挤压性和品质的影响 [J]. 西北农林科技大学学报(自然科学版), 2007, 35(2):97−100, 105. doi: 10.13207/j.cnki.jnwafu.2007.02.021 CHENG Z H, GUAN Z H, XUE L, et al. Effects of foliage application of sodium chloride on fruit compression tolerance and quality in processing tomato [J]. Journal of Northwest A & F University (Natural Science Edition), 2007, 35(2): 97−100, 105.(in Chinese) doi: 10.13207/j.cnki.jnwafu.2007.02.021
[26]	鲁少尉, 齐飞, 李天来. NaCl胁迫对番茄叶片光合特性及蔗糖代谢的影响 [J]. 北方园艺, 2012(9):14−18. LU S W, QI F, LI T L. Effect of NaCl stress on photosynthetic characteristics and sucrose metabolizing in tomato leaf [J]. Northern Horticulture, 2012(9): 14−18.(in Chinese)