Cloning and Expression of Premnaspirodiene Oxygenase Gene from <i>Dendrobium officinale</i>

LIN Jiangbo; ZOU Hui; WANG Weiying; DAI Yimin

doi:10.19303/j.issn.1008-0384.2021.06.003

Volume 36 Issue 6

Jun. 2021

Turn off MathJax

Article Contents

Article Navigation > Fujian Journal of Agricultural Sciences > 2021 > 36(6): 636-641

LIN J B, ZOU H, WANG W Y, et al. Cloning and Expression of Premnaspirodiene Oxygenase Gene from Dendrobium officinale [J]. Fujian Journal of Agricultural Sciences，2021，36（6）：636−641 doi: 10.19303/j.issn.1008-0384.2021.06.003

Citation:

LIN J B, ZOU H, WANG W Y, et al. Cloning and Expression of Premnaspirodiene Oxygenase Gene from Dendrobium officinale [J]. Fujian Journal of Agricultural Sciences，2021，36（6）：636−641 doi: 10.19303/j.issn.1008-0384.2021.06.003

Citation:

PDF( 807 KB)

Cloning and Expression of Premnaspirodiene Oxygenase Gene from Dendrobium officinale

doi: 10.19303/j.issn.1008-0384.2021.06.003

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

Received Date: 2021-03-31
Rev Recd Date: 2021-06-03

Available Online: 2021-07-13

Publish Date: 2021-06-28

Abstract

Abstract

Objective To decipher the gene function, phytoalexin biosynthesis, and disease resistance mechanism of Dendrobium officinale (Do), the premnaspirodiene oxygenase gene (PO) was cloned and its expressions in the plant in various organs at different growth stages studied. Method RACE-PCR and RT-PCR were employed to clone the full-length cDNA and ORF of DoPO. ProtParam was used for the physiochemical determination, BLAST P on the amino acid homology, and MEGA 6.0 for the phylogenetic tree construction. The expressions of DoPO in different organs at flowering stage as well as in leaves at growth stages were examined by qPCR. Result The DoPO encoded 498 amino acids, 1 704 bp in length with an 1 497 bp ORF. The unstable protein had a molecular weight of 56.914 kDa belonging to the P450 superfamily. It was phylogenetically clustered in the same branch with Phalaenopsis equestris (XP 020571355) and Apostasia shenzhenica (PKA52400) and most closely related to P. equestris. The relative DoPO expression in the leaves was significantly higher than that in the stems, roots, or flowers at flowering stage (P＜0.01); and during a year, that in October significantly higher than that in either August or December (P＜0.01) and that in December significantly higher than in August (P＜0.05). Conclusion The full-length cDNA of DoPO was successfully cloned. Its relative expression in leaf peaked in October and was highest in leaves among the organs at flowering stage.
- Dendrobium officinale,
- premnaspirodiene oxygenase,
- gene cloning,
- expression analysis

FullText(HTML)

References(28)

References

[1]	JEANDET P, CLÉMENT C, COUROT E, et al. Modulation of phytoalexin biosynthesis in engineered plants for disease resistance [J]. International Journal of Molecular Sciences, 2013, 14(7): 14136−14170. doi: 10.3390/ijms140714136
[2]	陈东菊, 安敏敏, 李丽兰, 等. 植保素及其在增强作物抗性中的作用 [J]. 分子植物育种, 2017, 15(2):774−780. CHEN D J, AN M M, LI L L, et al. The mechanism of phytoalexin enhancing crop disease resistance [J]. Molecular Plant Breeding, 2017, 15(2): 774−780.(in Chinese)
[3]	吴劲松. 植物对病原微生物的“化学防御”: 植保素的生物合成及其分子调控机制 [J]. 应用生态学报, 2020, 31(7):2161−2167. WU J S. The “chemical defense” of plants against pathogenic microbes: Phytoalexins biosynthesis and molecular regulations [J]. Chinese Journal of Applied Ecology, 2020, 31(7): 2161−2167.(in Chinese)
[4]	TOYOMASU T. Recent advances regarding diterpene cyclase genes in higher plants and fungi [J]. Bioscience, Biotechnology, and Biochemistry, 2008, 72(5): 1168−1175. doi: 10.1271/bbb.80044
[5]	POLONI A, SCHIRAWSKI J. Red card for pathogens: Phytoalexins in Sorghum and maize [J]. Molecules (Basel, Switzerland), 2014, 19(7): 9114−9133. doi: 10.3390/molecules19079114
[6]	CHEN K N, ERH M H, SU N W, et al. Soyfoods and soybean products: From traditional use to modern applications [J]. Applied Microbiology and Biotechnology, 2012, 96(1): 9−22. doi: 10.1007/s00253-012-4330-7
[7]	李端, 周立刚, 王蓟花, 等. 茄科植保素的研究进展 [J]. 天然产物研究与开发, 2004, 16(1):84−87, 79. doi: 10.3969/j.issn.1001-6880.2004.01.024 LI D, ZHOU L G, WANG J H, et al. Progress on phytoalexin research of solanaceous plants [J]. Natural Product Research and Development, 2004, 16(1): 84−87, 79.(in Chinese) doi: 10.3969/j.issn.1001-6880.2004.01.024
[8]	DESENDER S, ANDRIVON D, VAL F. Activation of defence reactions in Solanaceae: Where is the specificity? [J]. Cellular Microbiology, 2007, 9(1): 21−30. doi: 10.1111/j.1462-5822.2006.00831.x
[9]	BACK K, CHAPPELL J. Cloning and bacterial expression of a sesquiterpene cyclase from Hyoscyamus muticus and its molecular comparison to related terpene cyclases [J]. The Journal of Biological Chemistry, 1995, 270(13): 7375−7381. doi: 10.1074/jbc.270.13.7375
[10]	TAKAHASHI S, YEO Y S, ZHAO Y X, et al. Functional characterization of premnaspirodiene oxygenase, a cytochrome P450 catalyzing regio- and stereo-specific hydroxylations of diverse sesquiterpene substrates [J]. Journal of Biological Chemistry, 2007, 282(43): 31744−31754. doi: 10.1074/jbc.M703378200
[11]	MAHESH KUMAR P, MURUGAN K, KOVENDAN K, et al. Mosquito larvicidal and pupicidal efficacy of Solanum xanthocarpum (Family: Solanaceae) leaf extract and bacterial insecticide, Bacillus thuringiensis, against Culex quinquefasciatus Say (Diptera: Culicidae) [J]. Parasitology Research, 2012, 110(6): 2541−2550. doi: 10.1007/s00436-011-2797-2
[12]	PARDHI P, JAIN A P, GANESHPURKAR A, et al. Anti-microbial, Anti-oxidant and anthelmintic activity of crude extract of Solanum xanthocarpum [J]. Pharmacognosy Journal, 2010, 2(11): 400−404. doi: 10.1016/S0975-3575(10)80022-7
[13]	SATYAL P, MAHARJAN S, SETZER W N. Volatile constituents from the leaves, fruits (berries), stems and roots of Solanum xanthocarpum from Nepal [J]. Natural Product Communications, 2015, 10(2): 1934578X1501000.
[14]	DESJARDINS A E, MCCORMICK S P, PLAISTED R L, et al. Association between solavetivone production and resistance to Globodera rostochiensis in potato [J]. Journal of Agricultural and Food Chemistry, 1997, 45(6): 2322−2326. doi: 10.1021/jf960830p
[15]	YAO M K, DÉSILETS H, CHARLES M T, et al. Effect of mycorrhization on the accumulation of rishitin and solavetivone in potato plantlets challenged with Rhizoctonia solani [J]. Mycorrhiza, 2003, 13(6): 333−336. doi: 10.1007/s00572-003-0267-0
[16]	KAWAUCHI M, ARIMA T, SHIROTA O, et al. Production of sesquiterpene-type phytoalexins by hairy roots of Hyoscyamus albus co-treated with cupper culfate and methyl jasmonate [J]. Chemical and Pharmaceutical Bulletin, 2010, 58(7): 934−938. doi: 10.1248/cpb.58.934
[17]	KAWAUCHI M, DE ARIMA T H, KUROYANAGI M. Molecular cloning and transcriptional analysis of WRKY and solavetivone biosynthetic genes in the hairy roots of Hyoscyamus albus [J]. Plant Gene, 2016, 5: 78−86. doi: 10.1016/j.plgene.2016.01.001
[18]	廖俊杰, 李文静, 李进进, 等. 铁皮石斛有效成分与功效研究进展 [J]. 广东轻工职业技术学院学报, 2015, 14(3):16−19, 26. doi: 10.3969/j.issn.1672-1950.2015.03.004 LIAO J J, LI W J, LI J J, et al. Advances in the of constituents and pharmacological effects of Dendrobium candidum wall. ex. Lind [J]. Journal of Guangdong Industry Technical College, 2015, 14(3): 16−19, 26.(in Chinese) doi: 10.3969/j.issn.1672-1950.2015.03.004
[19]	施仁潮, 竹剑平, 李明焱. 铁皮石斛抗肿瘤作用的研究进展 [J]. 中国药学杂志, 2013, 48(19):1641−1644. SHI R C, ZHU J P, LI M Y. Research progress of anti-tumor effect of Dendrobium officinale kimura et migo [J]. Chinese Pharmaceutical Journal, 2013, 48(19): 1641−1644.(in Chinese)
[20]	吕圭源, 颜美秋, 陈素红. 铁皮石斛功效相关药理作用研究进展 [J]. 中国中药杂志, 2013, 38(4):489−493. LYU G Y, YAN M Q, CHEN S H. Review of pharmacological activities of Dendrobium officinale based on traditional functions [J]. China Journal of Chinese Materia Medica, 2013, 38(4): 489−493.(in Chinese)
[21]	吴蓓丽, 吴月国, 赵铮蓉, 等. 铁皮石斛免疫调节作用及相关活性成分多糖的研究进展 [J]. 中草药, 2019, 50(21):5373−5379. doi: 10.7501/j.issn.0253-2670.2019.21.034 WU B L, WU Y G, ZHAO Z R, et al. Advances in immunoregulatory effects and bioactive compounds of polysaccharides from Dendrobium officinale [J]. Chinese Traditional and Herbal Drugs, 2019, 50(21): 5373−5379.(in Chinese) doi: 10.7501/j.issn.0253-2670.2019.21.034
[22]	TAMURA K, STECHER G, PETERSON D, et al. MEGA6: molecular evolutionary genetics analysis version 6.0 [J]. Molecular Biology and Evolution, 2013, 30(12): 2725−2729. doi: 10.1093/molbev/mst197
[23]	林江波, 邹晖, 王伟英, 等. 铁皮石斛DoLIS基因克隆与茉莉酸甲酯诱导表达分析 [J]. 福建农业学报, 2020, 35(10):1071−1077. LIN J B, ZOU H, WANG W Y, et al. Cloning and expressions of LIS in Dendrobium officinale [J]. Fujian Journal of Agricultural Sciences, 2020, 35(10): 1071−1077.(in Chinese)
[24]	宋展, 高鑫, 吴冕, 等. 细胞色素P450酶的结构、功能与应用研究进展 [J]. 微生物学通报, 2020, 47(7):2245−2254. SONG Z, GAO X, WU M, et al. Structure, function, and application of cytochrome P450 enzymes [J]. Microbiology China, 2020, 47(7): 2245−2254.(in Chinese)
[25]	CHAPPLE C. Molecular-genetic analysis of plant cytochrome p450-dependent monooxygenases [J]. Annual Review of Plant Physiology and Plant Molecular Biology, 1998, 49: 311−343. doi: 10.1146/annurev.arplant.49.1.311
[26]	PEDRAS M S C, YAYA E E, GLAWISCHNIG E. The phytoalexins from cultivated and wild crucifers: Chemistry and biology [J]. Natural Product Reports, 2011, 28(8): 1381. doi: 10.1039/c1np00020a
[27]	AHUJA I, KISSEN R, BONES A M. Phytoalexins in defense against pathogens [J]. Trends in Plant Science, 2012, 17(2): 73−90. doi: 10.1016/j.tplants.2011.11.002
[28]	李珂, 马良, 杜鹏飞, 等. 玉米萜类植保素代谢关键基因对小斑病侵染的防御响应分析 [J]. 西北植物学报, 2015, 35(9):1776−1780. doi: 10.7606/j.issn.1000-4025.2015.09.1776 LI K, MA L, DU P F, et al. Gene expression of maize terpenoid phytoalexin metabolism in response to southern leaf blight [J]. Acta Botanica Boreali-Occidentalia Sinica, 2015, 35(9): 1776−1780.(in Chinese) doi: 10.7606/j.issn.1000-4025.2015.09.1776