千层金MbEGS基因的克隆与功能分析

doi:10.16420/j.issn.0513-353x.2021-0562

园艺学报 ›› 2022, Vol. 49 ›› Issue (8): 1747-1760.doi: 10.16420/j.issn.0513-353x.2021-0562

千层金MbEGS基因的克隆与功能分析

邱子文¹^,³, 刘林敏¹, 林永盛¹, 林晓洁¹, 李永裕¹^,²^,^*(), 吴少华¹^,²^,^*(), 杨超¹^,²^,^*()

1.福建农林大学园艺学院,福州 350002
2.福建农林大学园艺植物天然产物研究所,福州 350002
3.广东省潮州市农业科技发展中心,广东潮州 521000

收稿日期:2021-11-07 修回日期:2022-03-24 出版日期:2022-08-25 发布日期:2022-09-05
通讯作者: 李永裕,吴少华,杨超 E-mail:lilin3182@163.com;348279953@qq.com;55753827@qq.com
基金资助:
国家自然科学基金项目(31902067);国家自然科学基金项目(31501694);福建省自然科学基金项目(2022J01138);福建省科技计划项目(2018N0003);福建农林大学科技创新专项基金项目(CXZX2017171)

Cloning and Functional Analysis of the MbEGS Gene from Melaleuca bracteata

QIU Ziwen¹^,³, LIU Linmin¹, LIN Yongsheng¹, LIN Xiaojie¹, LI Yongyu¹^,²^,^*(), WU Shaohua¹^,²^,^*(), YANG Chao¹^,²^,^*()

1. College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002,China
2. Institute of Natural Products of Horticultural Plants,Fujian Agriculture and Forestry University,Fuzhou 350002,China
3. Agricultural Science and Technology Development Center of Chaozhou in Guangdong Province,Chaozhou,Guangodng 521000,China

Received:2021-11-07 Revised:2022-03-24 Online:2022-08-25 Published:2022-09-05
Contact: LI Yongyu,WU Shaohua,YANG Chao E-mail:lilin3182@163.com;348279953@qq.com;55753827@qq.com

摘要/Abstract

摘要：

在前期构建的千层金（Melaleuca bracteata）转录组数据库中鉴定并克隆得到2个丁香酚合成酶基因MbEGS1和MbEGS2,基因编码区（coding sequences,CDs）序列长度分别为963 和972 bp,分别编码320和323个氨基酸。序列比对和系统进化树分析表明,MbEGS1、MbEGS2与仙女扇（Clarkia breweri）CbIGS1蛋白序列相似性最高,达到65.92%和63.89%。qRT-PCR分析表明,二者在千层金的花、叶片和茎中均有表达,其中在花中表达量最高,叶中次之,茎中最低,与不同组织中丁香酚含量的变化类似,基因表达量与丁香酚含量呈显著正相关;MbEGS1在千层金的根中有表达,且表达量高于茎段,但低于叶片,而MbEGS2在根中不表达。采用不同浓度MeJA处理千层金发现,MeJA能够诱导MbEGS1和MbEGS2表达和丁香酚合成,0.1 mmol · L^-1 MeJA诱导效果最显著,其转录水平与丁香酚含量呈正相关。亚细胞定位结果表明,MbEGS1主要定位于细胞质膜和细胞核中,MbEGS2主要定位于细胞质膜中。在森林草莓中异位表达MbEGS1和MbEGS2的结果表明,过表达MbEGS1和MbEGS2促进了丁香酚含量的增加。以上结果表明,二者均参与了丁香酚的生物合成。

关键词: 千层金, 丁香酚合成酶, 基因克隆, 表达分析, 功能鉴定

Abstract:

Melaleuca bracteata is rich in essential oil,the main component of which is methyl eugenol with the content is as high as 86.5%. Eugenol is the precursor for the synthesis of methyl eugenol,but the biosynthetic pathway of essential oil is not clear. Based on the transcriptome databases of Melaleuca bracteata,two eugenol synthase genes（MbEGS1 and MbEGS2）were identified and cloned from Melaleuca bracteata. The coding sequences（CDS）of MbEGS1 and MbEGS2 were 963 and 972 bp in length,encoding 320 and 323 amino acids,respectively. Multiple sequence alignment and phylogenetic tree analysis showed that the deduced proteins MbEGS1 and MbEGS2 had the highest similarity （65.92% and 63.89%）with CbIGS1. qRT-PCR analysis showed that MbEGS1 and MbEGS2 were expressed in flowers,leaves and stems of Melaleuca bracteata. The highest expression level was observed in flowers,followed by leaves and stems,which was similar to the eugenol content in different tissues of Melaleuca bracteata,showing a significant positive correlation between the gene expression and eugenol content. MbEGS1 was expressed in roots of Melaleuca bracteata,and the expression level was higher than that in stems and lower than that in leaves. However,MbEGS2 was not expressed in roots. When MeJA was used to treat Melaleuca bracteata,MeJA could induce the expression of MbEGS1 and MbEGS2 and eugenol synthesis,which showed a trend of first increase and then decrease. Treatment with MeJA at 0.1 mmol · L^-1 had the most significant effect on gene expression,and their transcription levels were positively correlated with the eugenol content. Subcellular localization showed that MbEGS1 was mainly located in plant cell membrane and nucleus,and MbEGS2 was mainly located in plant cell membrane. Ectopic expression of MbEGS1 and MbEGS2 showed that overexpression of MbEGS1 and MbEGS2 promoted the synthesis of eugenol in Fragaria vesca（Yellow Wonder）. The results show that both MbEGS1 and MbEGS2 genes are involved in eugenol biosynthesis.

Key words: Melaleuca bracteata, eugenol synthase, gene cloning, expression analysis, functional identification

中图分类号:

S687

邱子文, 刘林敏, 林永盛, 林晓洁, 李永裕, 吴少华, 杨超. 千层金MbEGS基因的克隆与功能分析[J]. 园艺学报, 2022, 49(8): 1747-1760.

QIU Ziwen, LIU Linmin, LIN Yongsheng, LIN Xiaojie, LI Yongyu, WU Shaohua, YANG Chao. Cloning and Functional Analysis of the MbEGS Gene from Melaleuca bracteata[J]. Acta Horticulturae Sinica, 2022, 49(8): 1747-1760.

图/表 9

表1 本研究中所用的特异性引物序列

Table 1 Sequence of specific primers used in this study

引物用途 Primer use	引物名称 Primer name	引物序列（5′-3′） Primer sequence
基因克隆 Gene cloning	MbEGS1-F1	ATGGCAGGAGAGGCCGAGAAAA
	MbEGS1-R1	TCATTCGAAAGCAGCCCTAGCTG
	MbEGS2-F1	ATGACGATCATAAGCAGTAGTTGCA
	MbEGS2-R1	TCATTCCAATACAGCGCTTGC
实时荧光定量引物 qRT-PCR primer	MbEGS1-QF	TTCGGGAGTGAGGAGGATAA
	MbEGS1-QR	TGGGGATGAAGGAGGTTGTT
	MbEGS2-QF	CCCCACCTTTGTCTTCGCTC
	MbEGS2-QR	AACCTCTTTATGTTGCCCGC
	GAPDH-F	GAAGGAATACAAGCCCGACC
	GAPDH-R	GACCCTCAACAATGCCAAAT
亚细胞定位和瞬时过表达 Subcellular localization and transient overexpression	MbEGS1-YF1	ATGGCAGGAGAGGCCGAG
	MbEGS1-YR1	TCATTCGAAAGCAGCCCTAGC
	MbEGS2-YF2	ATGACGATCATAAGCAGTAGTTGC
	MbEGS2-YR2	TCATTCCAATACAGCGCTTG
	MbEGS1-YF2	AGTGGTCTCTGTCCAGTCCTATGGCAGGAGAGGCCGAG
	MbEGS1-YR2	GGTCTCAGCAGACCACAAGTTCATTCGAAAGCAGCCCTAGC
	MbEGS2-YF2	AGTGGTCTCTGTCCAGTCCTATGACGATCATAAGCAGTAGTTGC
	MbEGS2-YR2	GGTCTCAGCAGACCACAAGTTCATTCCAATACAGCGCTTG

图1 千层金不同组织（A）和MeJA处理后叶片（B）中的丁香酚含量不同小写字母表示差异显著（P < 0.05）。下同。

Fig. 1 Eugenol content in different tissue（A）and the leaves（B）under different treatment of Melaleuca bracteata Different lowercase letters indicate significant differences（P < 0.05）. The same below.

图2 MbEGS1和MbEGS2的克隆

Fig. 2 MbEGS1 and MbEGS2 cloning

图3 MbEGS1（A）、MbEGS2（B）基因序列及其推导的氨基酸序列

Fig. 3 MbEGS1（A）and MbEGS2（B）and their deduced amino acid sequences

图4 不同物种基于MbEGS同源蛋白的进化树 Mb：千层金;Cb：仙女扇;Ob：罗勒;Ph：矮牵牛;Tp：北美乔柏;La：羽扇豆;Lc：刚玉亚麻;Lu：亚麻;Fi：连翘;At：拟南芥;Lp：宿根亚麻;Zm：玉米;Ps：豌豆;Ms：紫花苜蓿;Ca：鹰嘴豆;Gm：大豆;Nt：烟草;Ole：油橄榄;Bp：垂枝桦;Pop：毛果杨;Pyrc：西洋梨;Pt：火炬松;Gb：银杏。

Fig. 4 Evolutionary tree based on MbEGS homologous protein of different species Mb：Melaleuca bracteata;Cb：Clarkia breweri;Ob：Ocimum basilicum;Ph：Petunia × hybrida;Tp：Thuja plicata;La：Lupinus albus;Lc：Linum corymbulosum;Lu：Linum usitatissimum;Fi：Forsythia × intermedia;At：Arabidopsis thaliana;Lp：Linum perenne;Zm：Zea mays;Ps：Pisum sativum;Ms：Medicago sativa;Ca：Cicer arietinum;Gm：Glycine max;Nt：Nicotiana tabacum;Ole：Olea europaea;Bp：Betula pendula;Pop：Populus trichocarpa;Pyrc：Pyrus communis;Pt：Pinus taeda;Gb：Ginkgo biloba.

图5 MbEGS在不同组织和不同浓度MeJA处理下的转录表达模式分析

Fig. 5 Analysis of transcriptional expression patterns of MbEGS in different tissues and MeJA treatments

图6 MbEGS1和MbEGS2在烟草叶片细胞中的亚细胞定位

Fig. 6 Subcellular localization of MbEGS1 and MbEGS2 in leaf cells of Nicotiana benthamiana

图7 草莓转基因果实中MbEGS1和MbEGS2的相对表达量

Fig. 7 Relative expression levels of MbEGS1 and MbEGS2 in transgenic strawberry fruit

表2 森林草莓转化果实丁香酚含量

Table 2 Content of eugenol in strawberry transgenic fruits

表达载体 Expression vector	丁香酚含量/（μg · g^-1） Eugenol content
pCam1304-MCS35S（对照Control）	0.235 ± 0.004 a
pCam1304-MCS35S-MbEGS1	0.263 ± 0.009 b
pCam1304-MCS35S-MbEGS2	0.327 ± 0.019 c

参考文献 55

[1]	Akashi T, Koshimizu S, Aoki T, Ayabe S. 2006. Identification of cDNAs encoding pterocarpan reductase involved in isoflavan phytoalexin biosynthesis in Lotus japonicus by EST mining. FEBS Letters, 580 (24):5666-5670. doi: 10.1016/j.febslet.2006.09.016 URL
[2]	An Yang. 2016. Isolation and characterization of eugenol synthase genes in Prunus mume‘Sanlunyudie’[M. D. Dissertation]. Beijing: Beijing Forestry University. (in Chinese)
	安阳. 2016. 梅花丁子香酚合酶基因的克隆及功能分析[硕士论文]. 北京: 北京林业大学.
[3]	Anand A, Jayaramaiah R H, Beedkar S D, Singh P A, Joshi R S, Mulani F A, Dholakia B B, Punekar S A, Gade W N, Thulasiram H V, Giri A P. 2016. Comparative functional characterization of eugenol synthase from four different Ocimum species:implications on eugenol accumulation. BBA- roteins and Proteomics, 1864 (11):1539-1547.
[4]	Ando T, Nomura M, Tsukahara J, Watanabe H, Kokubun H, Tsukamoto T, Hashimoto G, Marchesi E, Kitching I J. 2001. Reproductive isolation in a native population of Petunia sensu Jussieu(Solanaceae). Annals of Botany, 88 (3):403-413. doi: 10.1006/anbo.2001.1485 URL
[5]	Aragüez I, Osorio S, Hoffmann T, Rambla J L, Medina-Escobar N, Granell A, Botella M Á, Schwab W, Valpuesta V. 2013. Eugenol production in achenes and receptacles of strawberry fruits is catalyzed by synthases exhibiting distinct kinetics. Plant physiology(Bethesda), 163 (2):946-958.
[6]	Asbahani A E, Miladi K, Badri W, Sala M, Addi E H A, Casabianca H, Mousadik A E, Hartmann D, Jilale A, Renaud F N R, Elaissari A. 2015. Essential oils:from extraction to encapsulation. International Journal of Pharmaceutics, 483 (1-2):220-243. doi: 10.1016/j.ijpharm.2014.12.069 pmid: 25683145
[7]	Bari R, Jones J D G. 2009. Role of plant hormones in plant defence responses. Plant Molecular Biology, 69 (4):473-488. doi: 10.1007/s11103-008-9435-0 URL
[8]	Carson C F, Hammer K A, Riley T V. 2006. Melaleuca alternifolia(tea tree)oil:a review of antimicrobial and other medicinal properties. Clinical Microbiology Reviews, 19 (1):50-62. pmid: 16418522
[9]	Cho H, Son S Y, Rhee H S, Yoon S H, Lee-Parsons C W T, Park J M. 2008. Synergistic effects of sequential treatment with methyl jasmonate,salicylic acid and yeast extract on benzophenanthridine alkaloid accumulation and protein expression in Eschscholtzia californica suspension cultures. Journal of Biotechnology, 135 (1):117-122. doi: 10.1016/j.jbiotec.2008.02.020 URL
[10]	Claus W, Irene S, Bettina H, Gerd H, Claudia K, Helmut M, Jana N, Ivo F, Otto M. 2006. The wound response in tomato-role of jasmonic acid. Journal of Plant Physiology, 163 (3):297-306. doi: 10.1016/j.jplph.2005.10.014 URL
[11]	Dryden M S, Dailly S, Crouch M. 2004. A randomized,controlled trial of tea tree topical preparations versus a standard topical regimen for the clearance of MRSA colonization. Journal of Hospital Infection, 56 (4):283-286. pmid: 15066738
[12]	Farag M A, Paré P W. 2002. C 6 -Green leaf volatiles trigger local and systemic VOC emissions in tomato. Phytochemistry, 61 (5):545-554. doi: 10.1016/S0031-9422(02)00240-6 URL
[13]	Fernandes M J G, Pereira R B, Pereira D M, Fortes A G, Castanheira E M S, Gonçalves M S T. 2020. New eugenol derivatives with enhanced insecticidal activity. International journal of molecular sciences, 21 (23):9257. doi: 10.3390/ijms21239257 URL
[14]	Gang D R, Kasahara H, Xia Z Q, Mijnsbrugge K V, Bauw G, Boerjan W, Montagu M V, Davin L B, Lewis N G. 1999. Evolution of plant defense mechanisms:relationships of phenylcoumaran benzylic ether reductases to pinoresinol-lariciresinol and isoflavone reductases. Journal of Biological Chemistry, 274 (11):7516-7527. doi: 10.1074/jbc.274.11.7516 pmid: 10066819
[15]	Hart P H, Brand C, Carson C F, Riley T V, Prager R H, Finlay-Jones J J. 2000. Terpinen-4-ol,the main component of the essential oil of Melaleuca alternifolia(tea tree oil),suppresses inflammatory mediator production by activated human monocytes. Inflammation Research, 49 (11):619-626. pmid: 11131302
[16]	Hoffmann T, Kalinowski G, Schwab W. 2006. RNAi-induced silencing of gene expression in strawberry fruit(Fragaria × ananassa)by agroinfiltration:a rapid assay for gene function analysis. The Plant Journal:for Cell and Molecular Biology, 48 (5):818-826. doi: 10.1111/j.1365-313X.2006.02913.x URL
[17]	Hoffmann T, Kurtzer R, Skowranek K, Kießling P, Fridman E, Pichersky E, Schwab W. 2011. Metabolic engineering in strawberry fruit uncovers a dormant biosynthetic pathway. Metabolic Engineering, 13 (5):527-531. doi: 10.1016/j.ymben.2011.06.002 pmid: 21689779
[18]	Jiao Long, Bian Lei, Luo Zongxiu, Li Zhaoqun, Xin Zhaojun, Xiu Chunli, Cai Xiaoming, Chen Zongmao. 2020. Comparison of tea plant volatiles exogenously induced by jasmonates or salicylates elicitors. Acta Horticulturae Sinica, 47 (5):927-938. (in Chinese)
	焦龙, 边磊, 罗宗秀, 李兆群, 辛肇军, 修春丽, 蔡晓明, 陈宗懋. 2020. 茉莉酸、水杨酸类激发子外源诱导的茶树挥发物比较. 园艺学报, 47 (5):927-938.
[19]	Juan A A J, Kaae S C, Kaae S S, Henrik N, Ole W. 2017. DeepLoc:prediction of protein subcellular localization using deep learning. Bioinformatics (Oxford,England), 33 (24):4049. doi: 10.1093/bioinformatics/btx548 URL
[20]	Koeduka T. 2014. The phenylpropene synthase pathway and its applications in the engineering of volatile phenylpropanoids in plants. Plant Biotechnology, 31 (5):401-407. doi: 10.5511/plantbiotechnology.14.0801a URL
[21]	Koeduka T, Fridman E, Gang D R, Vassão D G, Jackson B L, Kish C M, Orlova I, Spassova S M, Lewis N G, Noel J P, Baiga T J, Dudareva N, Pichersky E. 2006. Eugenol and isoeugenol,characteristic aromatic constituents of spices,are biosynthesized via reduction of a coniferyl alcohol ester. Proceedings of the National Academy of Sciences of the United States of America, 103 (26):10128-10133.
[22]	Koeduka T, Louie G V, Orlova I, Kish C M, Ibdah M, Wilkerson C G, Bowman M E, Baiga T J, Noel J P, Dudareva N, Pichersky E. 2008. The multiple phenylpropene synthases in both Clarkia breweri and Petunia hybrida represent two distinct protein lineages. The Plant Journal, 54 (3):362-374. doi: 10.1111/j.1365-313X.2008.03412.x pmid: 18208524
[23]	Koeduka T, Orlova I, Baiga T J, Noel J P, Dudareva N, Pichersky E. 2009. The lack of floral synthesis and emission of isoeugenol in Petunia axillaris subsp. parodii is due to a mutation in the isoeugenol synthase gene. The Plant Journal, 58 (6):961-969. doi: 10.1111/j.1365-313X.2009.03834.x pmid: 19222805
[24]	Li Haixin. 2015. Effects of methyl jasmonate and light intensity on the denisity of glandular trichomes and glandular trichome secretions of Dendranthema indicum var. aromaticum[M. D. Dissertation]. Harbin:Northeast Forestry University. (in Chinese)
	李海芯. 2015. 茉莉酸甲酯及光照强度处理对神农香菊(Dendranthema indicum var. aromaticum)毛状体及腺毛分泌物的影响[硕士论文]. 哈尔滨: 东北林业大学.
[25]	Li Y Y, Ye Z M, Wang W T. 2018. Composition analysis of essential oil from Melaleuca bracteata leaves using ultrasound-assisted extraction and its antioxidative and antimicrobial activities. BioResources, 13 (4):8488-8504.
[26]	Niha D, Sarangapani S, Amarr R V, Nadimuthu K, Deepa P, Jingjing J, NamHai C, Sarojam R. 2020. Characterization of a sweet basil acyltransferase involved in eugenol biosynthesis. Journal of Experimental Botany, 71 (12):3638-3652. doi: 10.1093/jxb/eraa142 pmid: 32198522
[27]	Pasay C, Mounsey K, Stevenson G, Davis R, Arlian L, Morgan M, Vyszenski-Moher D, Andrews K, McCarthy J. 2010. Acaricidal activity of eugenol based compounds against Scabies mites. PLoS ONE, 5 (8):e12079.
[28]	Porto C D, Decorti D. 2009. Ultrasound-assisted extraction coupled with under vacuum distillation of flavour compounds from spearmint (carvone-rich)plants:Comparison with conventional hydrodistillation. Ultrasonics-Sonochemistry, 16 (6):795-799. doi: 10.1016/j.ultsonch.2009.03.010 URL
[29]	Qiu Ziwen, Lin Yongsheng, Xiang Xuwen, Wu Shaohua, Li Yongyu, Yang Chao. 2020. Effects of medium and rooting agents on cuttings of Melaleuca bracteata. Journal of Forest and Environment, 40 (6):648-653. (in Chinese)
	邱子文, 林永盛, 向旭文, 吴少华, 李永裕, 杨超. 2020. 基质和生根剂处理对千层金扦插繁育的影响. 森林与环境学报, 40 (6):648-653.
[30]	Ramos T C P M, Santos E P S., Ventura M, Pina J C, Cavalheiro A A, Fiorucci A R, Silva M S. 2021. Eugenol and TBHQ antioxidant actions in commercial biodiesel obtained by soybean oil and animal fat. Fuel, 286 (1):119374.
[31]	Robert C S, Michel A. H, David G C. 2006. Regulation of volatile benzenoid biosynthesis in petunia flowers. Trends in Plant Science, 11 (1):20-25. doi: 10.1016/j.tplants.2005.09.009 URL
[32]	Shukla R P, Prasad V G. 1985. Population fluctuations of the oriental fruit fly,Dacus dorsalis Hendel in relation to hosts and abiotic factors. International Journal of Pest Management, 31 (4):273-275.
[33]	Sparkes I A, Runions J, Kearns A, Hawes C. 2006. Rapid,transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nature Protocols:Recipes for Researchers, 1 (4):2019-2025.
[34]	Sudhir K, Glen S, Koichiro T. 2016. MEGA7:molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33 (7):1870-1874. doi: 10.1093/molbev/msw054 pmid: 27004904
[35]	Sun Xiaoling, Cai Xiaoming, Ma Chunlei, Wang Guochang, Chen Zongmao. 2011. Kinetics of polyphenol oxidase activity in tea leaves induced by exogenous application of meja and mechanical damage. Acta Botanica Boreali-Occidentalia Sinica, 31 (9):1805-1810. (in Chinese)
	孙晓玲, 蔡晓明, 马春雷, 王国昌, 陈宗懋. 2011. 茉莉酸甲酯和机械损伤对茶树叶片多酚氧化酶时序表达的影响. 西北植物学报, 31 (9):1805-1810.
[36]	Tongpil M, Hiroyuki K, Diana L B, Buhyun Y, Paulraj K L, David R G, Steven C H, HaJeung P, Jacqueline L H, Laurence B D, Norman G L, ChulHee K. 2003. Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases. The Journal of biological chemistry, 278 (50):50714-50723. doi: 10.1074/jbc.M308493200 URL
[37]	Wang Haiping, Yan Huijun, Zhang Hao, Jian Hongying, Wang Qigang, Qiu Xianqin, Li Shubin, Zhou Ningning, Tang Kaixue. 2012. Cloning and expression analysis of eugenol synthase gene RcEGS1 in Rosa chinensis‘Pallida’. Acta Horticulturae Sinica, 39 (7):1387-1394. (in Chinese)
	王海萍, 晏慧君, 张颢, 蹇洪英, 王其刚, 邱显钦, 李淑斌, 周宁宁, 唐开学. 2012. 月季(Rosa chinensis)丁香酚合成酶基因RcEGS1的克隆及其表达分析. 园艺学报, 39 (7):1387-1394.
[38]	Wang Lichun, Ren Qin, Xu Zhichun, Li Hailin, Li Zhenyu. 2008. Effects of methyl jasmonate on Pinus massoniana needle terpenes volatiles and the growth of Dendrolimus punctatus. Journal of Beijing Forestry University, 30 (1):79-84. (in Chinese)
	王立春, 任琴, 许志春, 李海林, 李镇宇. 2008. 茉莉酸甲酯对马尾松松针萜烯类挥发物及马尾松毛虫生长发育的影响. 北京林业大学学报, 30 (1):79-84.
[39]	Wang W T, Huang X Q, Yang H X, Niu X Q, Li D X, Yang C, Li L, Zou L T, Qiu Z W, Wu S H, Li Y Y. 2019. Antibacterial activity and anti-quorum sensing mediated phenotype in response to essential oil from Melaleuca bracteata leaves. International Journal of Molecular Sciences, 20 (22):5696. doi: 10.3390/ijms20225696 URL
[40]	Wang X Q, He X Z, Lin J Q, Shao H, Chang Z Z, Dixon R A. 2006. Crystal structure of isoflavone reductase from alfalfa(Medicago sativa L.). Journal of Molecular Biology, 358 (5):1341-1352. doi: 10.1016/j.jmb.2006.03.022 URL
[41]	Wilkins M R, Gasteiger E, Bairoch A, Sanchez J C, Williams K L, Appel R D, Hochstrasser D F. 1999. Protein identification and analysis tools in the ExPASy server. Methods in Molecular Biology(Clifton,N. J.), 112:531-552.
[42]	Wongwicha W, Tanaka H, Shoyama Y, Putalun W. 2014. Methyl jasmonate elicitation enhances glycyrrhizin production in Glycyrrhiza inflata hairy roots cultures. Zeitschrift für Naturforschung C, 66 (7-8):423-428. doi: 10.1515/znc-2011-7-815 URL
[43]	Yahyaa M, Berim A, Nawade B, Ibdah M, Dudareva N, Ibdah M. 2019. Biosynthesis of methyleugenol and methylisoeugenol in Daucus carota leaves:characterization of eugenol/isoeugenol synthase and O-methyltransferase. Phytochemistry, 159:179-189. doi: 10.1016/j.phytochem.2018.12.020 URL
[44]	Yan Huijun, Zhang Hao, Jian Hongying, Wang Qigang, Qiu Xianqin, Zhang Ting, Tang Kaixue. 2012. Cloning and expression analysis of eugenol synthase gene(RhEGS1)in cut rose(Rosa hybrida). Scientia Agricultura Sinica, 45 (3):590-597. (in Chinese)
	晏慧君, 张颢, 蹇洪英, 王其刚, 邱显钦, 张婷, 唐开学. 2012. 月季丁香酚合成酶基因RhEGS1的克隆及表达分析. 中国农业科学, 45 (3):590-597.
[45]	Yan P, Zeng Y J, Shen W T, Tuo D C, Li X Y, Zhou P. 2020. Nimble cloning:a simple,versatile,and efficient system for standardized molecular cloning. Frontiers in Bioengineering and Biotechnology, 7:460. doi: 10.3389/fbioe.2019.00460 URL
[46]	Yang Chao, Zhang Shujie, Liu Wenchong, Wang Wenting, Wang Zhiheng, Wu Shaohua, Li Yongyu. 2019. Study on the changes of contents and compositions of essential oil in Melaleuca bracteata. Natural Product Research and Development, 31 (3):489-495. (in Chinese)
	杨超, 张恕杰, 刘文嵩, 王文婷, 王志恒, 吴少华, 李永裕. 2019. 千层金精油含量和成分变化规律研究. 天然产物研究与开发, 31 (3):489-495.
[47]	Yang Yi, Zhang Cuiping, Liu Xing, Wei Yue, Wu Nengbiao. 2018. Effects of methyl jasmonate on metabolism of topical alkaloids and expression of relate genes in Atropa belladonna. China Journal of Chinese Materia Medica, 43 (20):4044-4049. (in Chinese) doi: 10.19540/j.cnki.cjcmm.20180726.018 pmid: 30486528
	杨怡, 张翠平, 刘兴, 韦悦, 吴能表. 2018. 茉莉酸甲酯对颠茄托品烷类生物碱代谢及相关基因表达的影响. 中国中药杂志, 43 (20):4044-4049. pmid: 30486528
[48]	Yano S, Suzuki Y, Yuzurihara M, Kase Y, Takeda S, Watanabe S, Aburada M, Miyamoto K. 2006. Antinociceptive effect of methyleugenol on formalin-induced hyperalgesia in mice. European Journal of Pharmacology, 553 (1):99-103. doi: 10.1016/j.ejphar.2006.09.020 URL
[49]	Zhang Fengjuan, Jin Youju. 2007. Comparison of volatiles from Anoplophora glabripenni(Motsch.)and methyl jasmonate(MeJA)-applied Acer mono Maxim to identify wound signal transduction pathways. Acta Ecologica Sinica, 27 (7):2990-2996. (in Chinese)
	张风娟, 金幼菊. 2007. 茉莉酸甲酯喷施和光肩星天牛Anoplophora glabripennis(Motschulsky)咬食后五角枫释放的挥发物. 生态学报, 27 (7):2990-2996.
[50]	Zhang P, Zhang E L, Xiao M, Chen C, Xu W J. 2013. Enhanced chemical and biological activities of a newly biosynthesized eugenol glycoconjugate,eugenol α-D-glucopyranoside. Applied Microbiology and Biotechnology, 97 (3):1043-1050. doi: 10.1007/s00253-012-4351-2 pmid: 22923067
[51]	Zhang Runji, Huang Xiying, Xiang Quanju, Zeng Xianfu, Li Xinzhu, Zhang Xiaoping, Gu Yunfu. 2020. Effects of exogenous methyl jasmonate induction on polysaccharide metabolism and key enzyme gene expression in Lentinula edodes. Mycosystema, 39 (12):2338-2345. (in Chinese)
	张润吉, 黄熙瀛, 向泉桔, 曾先富, 李昕竺, 张小平, 辜运富. 2020. 外源茉莉酸甲酯诱导对香菇多糖代谢及关键酶基因表达的影响. 菌物学报, 39 (12):2338-2345.
[52]	Zhang T X, Huo T T, Ding A Q, Hao R J, Wang J, Cheng T R, Bao F, Zhang Q X. 2019. Genome-wide identification,characterization,expression and enzyme activity analysis of coniferyl alcohol acetyltransferase genes involved in eugenol biosynthesis in Prunus mume. PLoS ONE, 14 (10):e223974.
[53]	Zhang Xiaoxia, Hong Bo, Jing Liangliang, Cao Shuang, Jia Yanxia. 2020. Insect resistance of tomato induced by exogenous methyl jasmonate to Bemisia tabaci. Chinese Journal of Ecology, 39 (11):3651-3657. (in Chinese)
	张晓霞, 洪波, 景亮亮, 曹爽, 贾彦霞. 2020. 外源茉莉酸甲酯诱导番茄对烟粉虱的抗虫性. 生态学杂志, 39 (11):3651-3657.
[54]	Zhang Yanfeng, Xie Yingping, Xue Jiaoliang, Wang Xu. 2009. Attraction to the Ladybeetle by the volatiles of persimmon trees induced with methyl jasmonate and japanese wax scale attacking. Scientia Silvae Sinicae, 45 (1):90-96. (in Chinese)
	张艳峰, 谢映平, 薛皎亮, 王旭. 2009. 茉莉酸甲酯和日本龟蜡蚧诱导柿树挥发物对红点唇瓢虫的吸引. 林业科学, 45 (1):90-96.
[55]	Zhu Jinrong, Huang Baokang, Wu Jinzhong, Qin Luping. 2008. Effect of methyl jasmonate(MJ)on the secondary metabolites of Mentha haplocalyx. Lishizhen Medicine and Materia Medica Research, 19 (8):2011-2012. (in Chinese)
	朱金荣, 黄宝康, 吴锦忠, 秦路平. 2008. 茉莉酸甲酯胁迫处理对薄荷次生代谢产物的影响. 时珍国医国药, 19 (8):2011-2012.

千层金MbEGS基因的克隆与功能分析

Cloning and Functional Analysis of the MbEGS Gene from Melaleuca bracteata

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 55

相关文章 15

编辑推荐

Metrics

本文评价

访问总数:　当日访问总数:　当前在线人数:

[1]	赵雪艳, 王琪, 王莉, 王方圆, 王庆, 李艳. 基于比较转录组的延胡索组织差异性表达分析[J]. 园艺学报, 2023, 50(1): 177-187.
[2]	王沙, 张心慧, 赵玉洁, 李变变, 招雪晴, 沈雨, 董建梅, 苑兆和. 石榴花青苷合成相关基因PgMYB111的克隆与功能分析[J]. 园艺学报, 2022, 49(9): 1883-1894.
[3]	高彦龙, 吴玉霞, 张仲兴, 王双成, 张瑞, 张德, 王延秀. 苹果ELO家族基因鉴定及其在低温胁迫下的表达分析[J]. 园艺学报, 2022, 49(8): 1621-1636.
[4]	郑林, 王帅, 刘语诺, 杜美霞, 彭爱红, 何永睿, 陈善春, 邹修平. 柑橘响应黄龙病菌侵染的NAC基因的克隆及表达分析[J]. 园艺学报, 2022, 49(7): 1441-1457.
[5]	马维峰, 李艳梅, 马宗桓, 陈佰鸿, 毛娟. 苹果POD家族基因的鉴定与MdPOD15的功能分析[J]. 园艺学报, 2022, 49(6): 1181-1199.
[6]	张凯, 麻明英, 王萍, 李益, 金燕, 盛玲, 邓子牛, 马先锋. 柑橘HSP20家族基因鉴定及其响应溃疡病菌侵染表达分析[J]. 园艺学报, 2022, 49(6): 1213-1232.
[7]	刘瑶瑶, 吴严严, 石岩, 毛天宇, 包满珠, 张俊卫, 张杰. 垂枝与直枝梅花PmTAC1启动子序列差异与垂枝性状的关系初探[J]. 园艺学报, 2022, 49(6): 1327-1338.
[8]	梁晨, 孙如意, 向锐, 孙艺萌, 师校欣, 杜国强, 王莉. 葡萄生长调控因子GRF家族基因的鉴定及表达分析[J]. 园艺学报, 2022, 49(5): 995-1007.
[9]	肖学宸, 刘梦雨, 蒋梦琦, 陈燕, 薛晓东, 周承哲, 吴兴健, 吴君楠, 郭寅生, 叶开温, 赖钟雄, 林玉玲. 龙眼褪黑素合成途径SNAT、ASMT和COMT家族基因鉴定及表达分析[J]. 园艺学报, 2022, 49(5): 1031-1046.
[10]	高玮林, 张力曼, 薛超玲, 张垚, 刘孟军, 赵锦. 枣E类MADS基因在花和果中的表达及其蛋白互作研究[J]. 园艺学报, 2022, 49(4): 739-748.
[11]	刘梦雨, 蒋梦琦, 陈燕, 张舒婷, 薛晓东, 肖学宸, 赖钟雄, 林玉玲. 龙眼GDSL酯酶/脂肪酶基因的全基因组鉴定及表达分析[J]. 园艺学报, 2022, 49(3): 597-612.
[12]	姜翠翠, 方智振, 周丹蓉, 林炎娟, 叶新福. ‘芙蓉李’糖转运蛋白家族基因鉴定及表达分析[J]. 园艺学报, 2022, 49(2): 252-264.
[13]	宋蒙飞, 查高辉, 陈劲枫, 娄群峰. 黄瓜株型性状分子基础研究进展[J]. 园艺学报, 2022, 49(12): 2683-2702.
[14]	王智宇, 常贝贝, 刘琦, 程晓帆, 杜晓云, 于晓丽, 宋来庆, 赵玲玲. 苹果溶质转运蛋白基因MdSLC35F2-like表达与花青苷积累的研究[J]. 园艺学报, 2022, 49(11): 2293-2303.
[15]	黄仁维, 任迎虹, 祁伟亮, 曾睿, 刘欣宇, 邓彬艳. 桑树MaERF105-Like的克隆及其在干旱胁迫下的表达分析[J]. 园艺学报, 2022, 49(11): 2439-2448.

千层金MbEGS基因的克隆与功能分析

Cloning and Functional Analysis of the MbEGS Gene from Melaleuca bracteata

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 55

相关文章 15

编辑推荐

Metrics

本文评价

访问总数: 当日访问总数: 当前在线人数:

访问总数:　当日访问总数:　当前在线人数: