木豆类黄酮代谢通路关键基因家族的鉴定与表达分析

doi:10.3969/j.issn.1674-7968.2021.12.003

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Abstract Flavonoids are important secondary metabolites in plants, and they play an important role in plant growth and resistance to adversity and stress. Studies on the function and metabolic regulation of plant flavonoids (flavonoids, flavonols, etc.) have attracted more and more attention, especially in woody plants rich in flavonoid metabolites. The research on the synergistic interaction between key enzymes in flavonoid metabolic pathway will provide guidance for the better development and utilization of plant flavonoid functional components. Pigeon pea (Cajanus cajan) is a pioneer woody legume tree growing in karst soils with high acid aluminum content, which is rich in secondary metabolites such as flavonoids. In order to identify the key pathways of flavonoid biosynthesis, this study screened and identified 8 key enzyme gene families in pigeon pea, namely phenylalanine ammonia-lyase (PAL), cinnamic acid-4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonol synthase (FLS) and dihydroflavonol 4-reductase (DFR). Bioinformatics software such as MEGA 7.0, MEME, TBtools v1.089 and PlantCare were used to analyze the phylogeny, gene structure, protein structure and cis-acting elements of the promoter, as well as the regulation of gene expression in the metabolic pathway under stress conditions. Phylogenetic analysis showed that some members of the PAL, C4H, 4CL and CHI gene families could cluster together with the genes in soybean (Glycine max), while the CHS, FLS and DFR gene families only showed sequence similarity among family members. Protein structure analysis showed that each gene family contained a special conserved domain. Analysis of promoter cis-acting elements showed that the genes containing methyl jasmonate (MeJA) response elements accounted for more than 50% of the total genes in 6 gene families. Meanwhile, in view of the habitat characteristics of pigeon pea, RNA-seq analysis was conducted on the 8 gene families under aluminum stress and methyl jasmonate treatment in the study. The results showed that CcPAL2, CcC4H1, CcCHS5~7 and CcCHS12 with the higher relative expression levels were all significantly up-regulated under aluminum stress, while CcCHI4~5 and CcFLS7 all showed a significant down-regulation trend, indicating that chalcone in pigeon pea might accumulate under aluminum stress. It was speculated that chalcone might play a role in aluminum resistance. Under MeJA treatment, CcC4H1, CcCHS5, CcCHS7, and CcCHI4 were significantly up-regulated, while CcF3H was down-regulated 0.6 times that of the control. At the same time, CcFLS1, CcFLS5, and CcFLS6 in the downstream pathway were significantly up-regulated. The above results indicated that MeJA may also promote the accumulation of naringin and flavonol, it was speculated that the substances might play an important role in the MeJA signaling pathway. The expression changes of the above flavonoid biosynthetic enzyme genes at different stages were verified by qPCR and consistent with the transcriptome sequencing results.This study reveals the expression patterns of key enzymes in flavonoid biosynthesis under different stresses and provides reference for further exploration of the regulatory role of flavonoid substances such as chalcones and flavonols in stress.

Key words： Flavonoid metabolism pathway Gene family analysis Aluminum stress Methyl jasmonate treatment

Received: 22 March 2021

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Corresponding Authors: **yang.qing1020@163.com; yujie_fu@163.com

About author:: * These authors contributed equally to this work

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	DU Ting-Ting
	SONG Zhi-Hua
	DONG Bi-Ying
	CAO Hong-Yan
	LIU Teng-Yue
	YANG Wan-Long
	QI Meng
	CHEN Ting
	WANG Meng-Ying
	MENG Dong
	YANG Qing
	FU Yu-Jie

Cite this article:

DU Ting-Ting,SONG Zhi-Hua,DONG Bi-Ying, et al. Identification and Expression Analysis of Key Gene Families in Flavonoid Metabolism Pathway in Pigeon Pea (Cajanus cajan)[J]. 农业生物技术学报, 2021, 29(12): 2289-2303.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2021.12.003 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2021/V29/I12/2289

[1] 付林, 孙炳夏, 翟佳黛, 等. 2019. 5'位羟基异戊烯基查尔酮类天然产物的合成及其抗菌活性研究[J]. 有机化学, 40(01): 201-222.
(Fu L, Sun B X, Zhai J D, et al.2019. 5'-hydroxyisopentenyl chalcone natural products synthesis and antibacterial activity[J]. Chinese Journal of Organic Chemistry, 40(01): 201-222.)
[2] 何贝轩, 薛英茹, 涂燕华, 等. 2018. CtCHS4响应茉莉酸甲酯诱导促进了红花醌式查尔酮类化合物的积累[J]. 药学学报, 53(04): 636-645.
(He B X, Xue Y R, Tu Y H, et al.2018. CtCHS4 promoted the accumulation of safflower quinoid chalcones in response to methyl jasmonate induction[J]. Acta Pharmaceutica Sinica, 53(04): 636-645.)
[3] 李程勋, 李爱萍, 徐晓俞, 等. 2018. 浅谈木豆的抗逆机制及在福建的应用前景[J]. 作物杂志, 2018(04): 28-31.
(Li C X, Li A P, Xu X Y, et al. 2018. Discussion on the mechanism of stress resistance of pigeonpea and application prospect in Fujian Province[J]. Crops, (04): 28-31.)
[4] 马鑫彤, 朱枚桦, 王南, 等. 2021. 红花类黄酮生物合成通路 2ODD基因家族鉴定与表达分析.
(Ma X T, Zhu M H, Wang N, et al.2021. Identification and expression analysis of 2ODD gene family involved in flavonoid biosynthesis pathway in safflower.) DOI: 10.16438/j.0513-4870.2021-0633
[5] 王婵婵. 2010. 大豆(Glycine max)苯丙氨酸解氨酶基因的克隆和生物信息学分析[D]. 硕士学位论文, 东北农业大学, 导师: 马凤鸣, pp. 24-29.
(Wang C C.2010. Gene cloning and bioinformatics analysis of phenylalanine ammonia-lyase in soybean (Glycine max L.)[D]. Thesis for M.S., Northeast Agricultural University, Suppervisor: Ma F M, pp. 24-29.)
[6] 赵继发, 沙伟, 马天意. 2021. 类黄酮物质在植物逆境胁迫中的研究进展[J]. 高师理科学刊, 41(07): 53-59.
(Zhao J F, Sha W, Ma T Y, et al.2021. Research progress of flavonoids in plants stresses[J]. Journal of Science of Teachers' College and University, 41(07): 53-59.)
[7] Allamin I A, Halmi M I E, Yasid N A, et al.2020. Rhizodegradation of petroleum oily sludge-contaminated soil using Cajanus cajan increases the diversity of soil microbial community[J]. Scientific Reports, 10(1): 1-11.
[8] Barcelo J, Poschenrieder C.2002. Fast root growth responses, root exudates, and internal detoxification as clues to the mechanisms of aluminium toxicity and resistance: A review[J]. Environmental and Experimental Botany, 48(1): 75-92.
[9] Brunner I, Sperisen C.2013. Aluminum exclusion and aluminum tolerance in woody plants[J]. Frontiers in Plant Science, 4: 172.
[10] Chen W, Zhang M, Zhang G, et al.2019. Differential regulation of anthocyanin synthesis in apple peel under different sunlight intensities[J]. International Journal of Molecular Sciences, 20(23): 6060.
[11] Gu H, Wang Y, Xie H, et al.2020. Drought stress triggers proteomic changes involving lignin, flavonoids and fatty acids in tea plants[J]. Scientific Reports, 10(1): 1-11.
[12] Han Y, Ding T, Su B, et al.2016. Genome-wide identification, characterization and expression analysis of the chalcone synthase family in maize[J]. International Journal of Molecular Sciences, 17(2): 161.
[13] Han Y, Huang K, Liu Y, et al.2017. Functional analysis of two flavanone-3-hydroxylase genes from Camellia sinensis: A critical role in flavonoid accumulation[J]. Genes, 8(11): 300.
[14] Ito M, Ichinose Y, Kato H, et al.1997. Molecular evolution and functional relevance of the chalcone synthase genes of pea[J]. Molecular and General Genetics, 255(1): 28-37.
[15] Jenkins G I, Long J C, Wade H K, et al.2001. UV and blue light signalling: Pathways regulating chalcone synthase gene expression in Arabidopsis[J]. New Phytologist, 151(1): 121-131.
[16] Khoury C K, Castañeda-Alvarez N P, Achicanoy H A, et al.2015. Crop wild relatives of pigeonpea (Cajanus cajan (L.) Millsp.): Distributions, ex situ conservation status, and potential genetic resources for abiotic stress tolerance[J]. Biological Conservation, 184: 259-270.
[17] Krueger R J.2007. Flavonoids: Chemistry, biochemistry and applications[J]. Economic Botany, 61(1): 101-101.
[18] Lepiniec L, Debeaujon I, Routaboul J M, et al.2006. Genetics and biochemistry of seed flavonoids[J]. Annual Review of Plant Biology, 57: 405-430.
[19] Li W, Tan L, Zou Y, et al.2020. The effects of ultraviolet A/B treatments on anthocyanin accumulation and gene expression in dark-purple tea cultivar 'ziyan'(Camellia sinensis)[J]. Molecules, 25(2): 354.
[20] Lin L M, Guo H Y, Song X, et al.2021. Adaptive evolution of chalcone isomerase superfamily in fagaceae[J]. Biochemical Genetics, 59(2): 491-505.
[21] Liu M, Li X, Liu Y, et al.2013. Regulation of flavanone 3-hydroxylase gene involved in the flavonoid biosynthesis pathway in response to UV-B radiation and drought stress in the desert plant, Reaumuria soongorica[J]. Plant Physiology and Biochemistry, 73: 161-167.
[22] Nabavi S M, Šamec D, Tomczyk M, et al.2020. Flavonoid biosynthetic pathways in plants: Versatile targets for metabolic engineering[J]. Biotechnology Advances, 38: 107316.
[23] Parvin K, Hasanuzzaman M, Bhuyan M H M, et al.2019. Quercetin mediated salt tolerance in tomato through the enhancement of plant antioxidant defense and glyoxalase systems[J]. Plants, 8(8): 247.
[24] Peer W A, Murphy A S.2007. Flavonoids and auxin transport: Modulators or regulators?[J]. Trends in Plant Science, 12(12): 556-563.
[25] Petrussa E, Braidot E, Zancani M, et al.2013. Plant flavonoids-biosynthesis, transport and involvement in stress responses[J]. International Journal of Molecular Sciences, 14(7): 14950-14973.
[26] Salunkhe D K, Chavan J K, Kadam S S, et al.1986. Pigeonpea as an important food source[J]. Critical Reviews in Food Science and Nutrition, 23(2): 103-145.
[27] Singh P, Arif Y, Bajguz A, et al.2021.The role of quercetin in plants[J]. Plant Physiology and Biochemistry, 166: 10-19.
[28] Singh U, Jain K C, Jambunathan R, et al.1984. Nutritional quality of vegetable pigeonpeas (Cajanus cajan (L.) Mill sp.): Mineral and trace elements[J]. Journal of Food Science, 49(2): 645-646.
[29] Song X, Diao J, Ji J, et al.2016. Molecular cloning and identification of a flavanone 3-hydroxylase gene from Lycium chinense, and its overexpression enhances drought stress in tobacco[J]. Plant Physiology and Biochemistry, 98: 89-100.
[30] Song Z, Dong B, Yang Q, et al.2020. Screening of CBL genes in pigeon pea with focus on the functional analysis of CBL4 in abiotic stress tolerance and flavonoid biosynthesis[J]. Environmental and Experimental Botany, 177: 104102.
[31] Thompson E P, Wilkins C, Demidchik V, et al.2010. An Arabidopsis flavonoid transporter is required for anther dehiscence and pollen development[J]. Journal of Experimental Botany, 61(2): 439-451.
[32] Tu Y H, Liu F, Guo D D, et al.2016. Molecular characterization of flavanone 3-hydroxylase gene and flavonoid accumulation in two chemotyped safflower lines in response to methyl jasmonate stimulation[J]. BMC Plant Biology, 16(1): 1-12.
[33] Vadivel A K A, Krysiak K, Tian G, et al.2018. Genome-wide identification and localization of chalcone synthase family in soybean (Glycine max [L.] Merr)[J]. BMC Plant Biology, 18(1): 325.
[34] Vanhaelewyn L, Viczián A, Prinsen E, et al.2019. Differential UVR8 signal across the stem controls UV-B induced inflorescence phototropism[J]. The Plant Cell, 31(9): 2070-2088.
[35] Wang M, Zhang Y, Zhu C, et al.2021. EkFLS overexpression promotes flavonoid accumulation and abiotic stress tolerance in plant[J]. Physiologia Plantarum, 172(4): 1966-1982.
[36] Wen W, Alseekh S, Fernie A R.2020. Conservation and diversification of flavonoid metabolism in the plant kingdom[J]. Current Opinion in Plant Biology, 55: 100-108.
[37] Williams C A, Grayer R J.2004. Anthocyanins and other flavonoids[J]. Natural Product Reports, 21(4): 539-573.
[38] Winkel-Shirley B.2001. Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology[J]. Plant Physiology, 126(2): 485-493.
[39] Wolf L, Rizzini L, Stracke R, et al.2010.The molecular and physiological responses of Physcomitrella patens to ultraviolet-B radiation[J]. Plant Physiology, 153(3): 1123-1134.
[40] Xu G, Guo C, Shan H, et al.2012. Divergence of duplicate genes in exon-intron structure[J]. Proceedings of the National Academy of the USA, 109(4): 1187-1192.
[41] Yonekura-Sakakibara K, Higashi Y, Nakabayashi R.2019. The origin and evolution of plant flavonoid metabolism[J]. Frontiers in Plant Science, 10: 943.
[42] Zhou X W, Fan Z Q, Chen Y, et al.2013. Functional analyses of a flavonol synthase-like gene from Camellia nitidissima reveal its roles in flavonoid metabolism during floral pigmentation[J]. Journal of Biosciences, 38(3): 593-604.