4-羟基-3-甲氧基肉桂酸乙酯在拟南芥中的作用位点研究

doi:10.7668/hbnxb.20190170

摘要/Abstract

摘要： 为了探究4-羟基-3-甲氧基肉桂酸乙酯的除草作用方式，在前期蛋白组学研究结果的基础之上，对4-羟基-3-甲氧基肉桂酸乙酯的潜在除草靶标基因进行筛选。利用实时定量PCR （RT-qPCR）技术，测定茎叶喷施质量浓度为1 000 mg/L的4-羟基-3-甲氧基肉桂酸乙酯0，15，30，45 min后，拟南芥野生型株系和6种T-DNA插入株系的拟南芥中AT2G43030和AT1G66240基因的相对表达量，并比较拟南芥野生型株系和6种T-DNA插入株系的受损程度，分析AT2G43030和AT1G66240基因在4-羟基-3-甲氧基肉桂酸乙酯除草过程中的作用。结果表明：相比野生型拟南芥株系而言， AT2G43030和AT1G66240基因的6种T-DNA插入纯合株系对4-羟基-3-甲氧基肉桂酸乙酯更为敏感，茎叶喷施4-羟基-3-甲氧基肉桂酸乙酯24 h后不同T-DNA株系的叶片损伤程度相较野生型株系受损更为严重，证明AT2G43030和AT1G66240基因与拟南芥对4-羟基-3-甲氧基肉桂酸乙酯的防御系统有关，同时也说明4-羟基-3-甲氧基肉桂酸乙酯的除草作用机制与植株的光合作用和细胞膜功能有关。

关键词: 4-羟基-3-甲氧基肉桂酸乙酯, 拟南芥, 除草剂, 作用位点, AT2G43030基因, AT1G66240基因

Abstract: In order to explore the herbicidal action of 4-hydroxy-3-methoxy cinnamic acid ethyl, the potential herbicidal target gene of ethyl 4-hydroxy-3-methoxy cinnamic acid ethyl was screened, based on previous results of proteomics studies. Real-time quantitative PCR(RT-qPCR) was used to determine,the relative quantitative expression levels of AT2G43030 and AT1G66240 genes of wild-type and six T-DNA insertion lines of A. thaliana, which were sprayed with 1 000 mg/L 4-hydroxy-3-methoxy cinnamic acid ethyl for 0, 15, 30 and 45 min, respectively. The damage degrees of wild-type and other six T-DNA insertion lines of A. thaliana were compared, and the roles of AT2G43030 and AT1G66240 genes in the herbicidal process of 4-hydroxy-3-methoxy cinnamic acid ethyl were analyzed. The results showed that the six T-DNA insertion homozygous lines with AT2G43030 and AT1G66240 genes were more sensitive to 4-hydroxy-3-methoxy cinnamic acid ethyl, and the degree of leaf damage in different T-DNA strains sprayed with 4-hydroxy-3-methoxy cinnamic acid ethyl for 24 h was more severe, compared with the wild-type A. thaliana, which proved that the AT2G43030 and AT1G66240 genes were related to the defense system of Arabidopsis to 4-hydroxy-3-methoxy cinnamic acid ethyl. This indicated that the mechanism of herbicidal action of 4-hydroxy-3-methoxy cinnamic acid ethyl was related to photosynthesis and cell membrane function of plants.

Key words: 4-hydroxy-3-methoxy cinnamic acid ethyl, Arabidopsis thaliana, Herbicide, Action site, AT2G43030 gene, AT1G66240 gene

中图分类号:

Q78
S682.03

王稳, 靳丽宇, 张利辉. 4-羟基-3-甲氧基肉桂酸乙酯在拟南芥中的作用位点研究[J]. 华北农学报, 2020, 35(1): 168-174. doi: 10.7668/hbnxb.20190170.

WANG Wen, JIN Liyu, ZHANG Lihui. The Role of Arabidopsis thaliana AT2G43030 and AT1G66240 Genes in the Herbicidal Process of 4-hydroxy-3-methoxy Cinnamic Acid Ethyl[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(1): 168-174. doi: 10.7668/hbnxb.20190170.

http://www.hbnxb.net/CN/Y2020/V35/I1/168

参考文献

[1] Mouncey N J, Otani H, Udwary D, Yoshikuni Y. New voyages to explore the natural product galaxy[J]. Journal of Industrial Microbiology & Biotechnology, 2019, 46(3-4):273-279.doi:10.1007/s10295-018-02122-w.
[2] Zhou J, Liu K, Xin F X, Ma J F, Xu N, Zhang W M, Fang Y, Jiang M, Dong W L. Recent insights into the microbial catabolism of aryloxyphenoxy-propionate herbicides:microbial resources, metabolic pathways and catabolic enzymes[J]. World Journal of Microbiology and Biotechnology, 2018, 34(8):117.doi:10.1007/s11274-018-2503-y.
[3] 陈世国, 强胜. 生物除草剂研究与开发的现状及未来的发展趋势[J]. 中国生物防治学报, 2015, 31(5):770-779.doi:10.16409/j.cnki.2095-039x.2015.05.017.
Chen S G, Qiang S. The status and future directions of bioherbicide study and development[J]. Chinese Journal of Biological Control, 2015, 31(5):770-779.
[4] Lee D L, Knudsen C G, Michaely W J, Chin H L, Nguyen N H, Carter C G, Cromartie T H, Lake B H, Shribbs J M, Fraser T. The structure activity relationships of the triketone class of HPPD herbicides[J]. Pest Management Science, 1998, 54(4):377-384.doi:10.1002/(SICI)1096-9063(199812)54:4<377∷AID-PS827>3.0.
CO;2-0.
[5] Duke S O. Why have no new herbicide modes of action appeared in recent years?[J]. Pest Management Science, 2012, 68(4):505-512.doi:10.1002/ps.2333.
[6] Redlick C, Syrovy L D, Duddu H S N, Benaragama D, Johnson E N,Willenborg C J,Shirtiffe S J. Developing an integrated weed management system for herbicide-resistant weeds using lentil(Lens culinaris) as a model crop[J]. Weed Science, 2017, 65(6):778-786.doi:10.1017/wsc.2017.47.
[7] Fleet B, Malone J, Preston C, Gill G. Target-site point mutation conferring resistance to trifluralin in rigid ryegrass(Lolium rigidum)[J]. Weed Science, 2018, 66(2):246-253.doi:10.1017/wsc.2017.67.
[8] Fu Y B, Mu Y, Lei H, Wang P,Li X, Leng Q, Han L, Qu X D,Wang Z Y,Huang X S. Design, synthesis and evaluation of novel tacrine-ferulic acid hybrids as multifunctional drug candidates against alzheimer's disease[J]. Molecules, 2016, 21(10):1338-1348.doi:10.3390/molecules21101338.
[9] Totani N, Tateishi S, Takimoto T, Shinohara R,Sasaki H. Ferulic acid esters and weight-loss promoting effects in rats[J]. Journal of Oleo Science, 2012, 61(6):331-336.doi:10.5650/jos.61.331.
[10] 张明月, 刘策, 杨娟, 乔欣, 张利辉, 董金皋. 4-羟基-3-甲氧基肉桂酸乙酯的生物活性及其除草作用机制[J]. 植物保护学报, 2018, 45(3):543-551.doi:10.13802/j.cnki.zwbhxb.2018.2016210.
Zhang M Y, Liu C, Yang J, Qiao X, Zhang L H, Dong J G.The biological activity of 4-hydroxy-3-methoxycinnamate acid ethyl ester and its herbicidal mechanism[J]. Journal of Plant Protection, 2018, 45(3):543-551.
[11] 张利辉, 张洋, 司贺龙,董金皋,张金林. 瓜果腐霉毒素除草活性物质的初步分离[J]. 河北农业大学学报, 2008,31(5):71-74.doi:10.3969/j.issn.1000-1573.2008.05.016.
Zhang L H, Zhang Y, Si H L, Dong J G, Zhang J L. Primary isolation of herbicidal substances in crude toxin produced by Pythium aphanider-matum[J]. Journal of Agricultural University of Hebei, 2008,31(5):71-74.
[12] 杨鹏,杨娟,王伟,张明月,张利辉,董金皋.阿魏酸衍生物的合成及其除草活性测定[J]. 植物保护学报, 2015,42(6):1050-1056.doi:10.13802/j.cnki.zwbhxb.2015.06.029.
Yang P, Yang J, Wang W, Zhang M Y, Zhang L H,Dong J G. Synthesis and herbicidal activity of the derivatives of ferulis acid[J]. Journal of Plant Protection, 2015,42(6):1050-1056.
[13] Zhang L H, Zhang J L, Liu Y C,Cao Z Y,Han J M,Yang J,Dong J G. Isolation and structural speculation of herbicide-active compounds from the metabolites of Pythium aphanidermatum[J]. Journal of Integrative Agriculture, 2013, 12(6):1026-1032.doi:10.1016/S2095-3119(13)60480-3.
[14] Zhang M Y, Liu C, Yang J, Yang P, Zhang L H, Dong J G. Analysis of the herbicidal mechanism of 4-hydroxy-3-methoxy cinnamic acid ethyl ester using iTRAQ and Real-time PCR[J]. Journal of Proteomics, 2017, 159:47-53.doi:10.1016/j.jprot.2017.02.014.
[15] Schmid M, Davison T S, Henz S R, Pape U J, Demar M, Vingron M, Schölkopf B, Weigel D, Lohmann J U. A gene expression map of Arabidopsis thaliana development[J]. Nature Genetics, 2005, 37(5):501-506.doi:10.1038/ng1543.
[16] Rutschow H, Ytterberg A J, Friso G F, Nilsson R, Wijk K J V. Quantitative proteomics of a chloroplast SRP54 sorting mutant and its genetic interactions with CLPC1 in Arabidopsis[J]. Plant physiology, 2008, 148(1):156-175.doi:10.1104/pp.108.124545.
[17] Li M, Wang Q Q, Song X F, Guo J J, Wu J R, Wu R N. iTRAQ-based proteomic analysis of responses of Lactobacillus plantarum fs5-5 to salt tolerance[J]. Annals of Microbiology, 2019, 69(4):377-394.doi:10.1007/s13213-018-1425-0.
[18] Pan L, Zhang J, Wang J Z, Qin Y, Bai L Y, Dong L Y. iTRAQ-based quantitative proteomic analysis reveals proteomic changes in three fenoxaprop-P-ethyl-resistant Beckmannia syzigachne biotypes with differing ACCase mutations[J]. Journal of Proteomics, 2017, 160:47-54.doi:10.1016/j.jprot.2017.03.018.
[19] Yan Y, Liu Q K, Zang X, Bat-Erdene U, Nguyen C, Gan J H, Zhou J H, Jacobsen S E, Tang Y. Resistance-gene-directed discovery of a natural-product herbicide with a new mode of action[J]. Nature, 2018, 559:415-418.doi:10.1038/s41586-018-0319-4.
[20] Wallace M D, Waraich N F, Debowski A W, Corrol M G, Maxwell A, Mylne J S, Stubbs K A. Developing ciprofloxacin analogues against plant DNA gyrase:a novel herbicide mode of action[J]. Chemical communications, 2018, 54(15):1869-1872.doi:10.1039/C7CC09518J.
[21] Wright A A, Sasidharan R, Koski L, Rodriguez-carres M, Peterson D G, Nandula V K, Ray J D, Bond J A, Shaw D R. Transcriptomic changes in Echinochloa colona in response to treatment with the herbicide imazamox[J]. Planta, 2018, 247(2):369-379.doi:10.1007/s00425-017-2784-7.

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