Roles of RNA m1A and m5C Methylation Modifications in the Fumonisin Biosynthesis of Fusarium verticillioides

doi:10.13560/j.cnki.biotech.bull.1985.2024-0213

Abstract

Abstract:

【Objective】Fusarium verticillioides is a serious plant pathogen fungus, which greatly reduces grain yield. It also produces class 2B carcinogen fumonisin, which threatens human and animal health. To explore the relationship between RNA methylation modification and fumonisin, the role of RNA methylated transferase in fumonisin synthesis was analyzed.【Method】 HPLC was applied to determine the fumonisin synthesis ability of F. verticillium strains from different regions. Six kinds of RNA methylation modification detection methods, including m⁶A, m^lA, m⁵C, Gm, m⁷G, and Um, were established by QuEChERS pretreatment combined with ultra-high performance liquid chromatograph-tandem mass spectrometry（UPLC-MS/MS）. Then the RNA methylation modifications of different toxin-producing fungi were detected, and the RNA methylation modification genes related to fumonisin synthesis were identified by bioinformatics and RT-qPCR.【Result】It was found that there were significant differences in fumonisin synthesis ability of F. verticillium strains in different regions, the detection method for RNA methylation modification was successfully constructed, the methylation of RNA m^lA and m⁵C was negatively correlated with fumonisin synthesis, and RT-qPCR showed that Fvalyref gene negatively regulated fumonisin biosynthesis.【Conclusion】The methylation modification of RNA m⁵C and its Reader gene Fvalyref negatively regulate fumonisin biosynthesis.

Key words: Fusarium verticillioides, RNA methylation modification, detection, Fumonisin

HOU Zhi-han, HAO Nan, LI Jia-qi, ZHAO Bin, LIU Ying-chao. Roles of RNA m¹A and m⁵C Methylation Modifications in the Fumonisin Biosynthesis of Fusarium verticillioides[J]. Biotechnology Bulletin, 2024, 40(9): 282-290.

Figures/Tables 8

References 41

[1]	Deepa N, Achar PN, Sreenivasa MY. Current perspectives of biocontrol agents for management of Fusarium verticillioides and its fumonisin in cereals-a review[J]. J Fungi, 2021, 7(9): 776.
[2]	Han SL, Wang MM, Ma ZY, et al. Fusarium diversity associated with diseased cereals in China, with an updated phylogenomic assessment of the genus[J]. Stud Mycol, 2023, 104: 87-148. doi: 10.3114/sim.2022.104.02 pmid: 37351543
[3]	Iqbal N, Czékus Z, Poór P, et al. Plant defence mechanisms against mycotoxin Fumonisin B1[J]. Chem Biol Interact, 2021, 343: 109494.
[4]	Carvajal-Moreno M. Mycotoxin challenges in maize production and possible control methods in the 21st century[J]. J Cereal Sci, 2022, 103: 103293.
[5]	Winter G, Pereg L. A review on the relation between soil and mycotoxins: effect of aflatoxin on field, food and finance[J]. Eur J Soil Sci, 2019, 70(4): 882-897. doi: 10.1111/ejss.12813
[6]	Desmond J. A modest proposal: a response to the marketing challenges presented by the crisis confronting humanity in respect to the requirement to feed nine billion by 2050[J]. J Mark Manag, 2013, 29(13/14): 1631-1643.
[7]	Dai YQ, Huang KL, Zhang BY, et al. Aflatoxin B1-induced epigenetic alterations: an overview[J]. Food Chem Toxicol, 2017, 109(Pt 1): 683-689. doi: S0278-6915(17)30342-3 pmid: 28645871
[8]	Teng PC, Liang YW, Yarmishyn AA, et al. RNA modifications and epigenetics in modulation of lung cancer and pulmonary diseases[J]. Int J Mol Sci, 2021, 22(19): 10592.
[9]	马士清, 彭金英, 伊成器. RNA修饰检测技术[J]. 生命科学, 2018, 30(4): 440-446.
	Ma SQ, Peng JY, Yi CQ. The detection methods of RNA modifications[J]. Chin Bull Life Sci, 2018, 30(4): 440-446.
[10]	Yu HW, Raza SHA, Zhang WZ, et al. Research progress of m⁶A regulation during animal growth and development[J]. Mol Cell Probes, 2022, 65: 101851.
[11]	Ghazi T, Nagiah S, Chuturgoon AA. Fusaric acid induces hepatic global m⁶A RNA methylation and differential expression of m⁶A regulatory genes in vivo - a pilot study[J]. Epigenetics, 2022, 17(6): 695-703.
[12]	Růžička K, Zhang M, Campilho A, et al. Identification of factors required for m⁶ A mRNA methylation in Arabidopsis reveals a role for the conserved E3 ubiquitin ligase HAKAI[J]. New Phytol, 2017, 215(1): 157-172. doi: 10.1111/nph.14586 pmid: 28503769
[13]	Zou MJ, Mu Y, Chai X, et al. The critical function of the plastid rRNA methyltransferase, CMAL, in ribosome biogenesis and plant development[J]. Nucleic Acids Res, 2020, 48(6): 3195-3210. doi: 10.1093/nar/gkaa129 pmid: 32095829
[14]	Zhang C, Zhang H, Zhu QQ, et al. Overexpression of global regulator LaeA increases secondary metabolite production in Monascus purpureus[J]. Appl Microbiol Biotechnol, 2020, 104(7): 3049-3060. doi: 10.1007/s00253-020-10379-4 pmid: 32043189
[15]	Wang B, Li XJ, Tabudravu J, et al. The chemical profile of activated secondary metabolites by overexpressing LaeA in Aspergillus niger[J]. Microbiol Res, 2021, 248: 126735.
[16]	Wang YS, Chen YC, Zhang J, et al. Overexpression of llm1 affects the synthesis of secondary metabolites of Aspergillus cristatus[J]. Microorganisms, 2022, 10(9): 1707.
[17]	Yadav PK, Rajasekharan R. The m⁶A methyltransferase Ime4 epitranscriptionally regulates triacylglycerol metabolism and vacuolar morphology in haploid yeast cells[J]. J Biol Chem, 2017, 292(33): 13727-13744.
[18]	Zhang YW, Wang YM, Fan JL, et al. Aspergillus fumigatus Elongator complex subunit 3 affects hyphal growth, adhesion and virulence through wobble uridine tRNA modification[J]. PLoS Pathog, 2022, 18(11): e1010976.
[19]	Ren ZY, Tang BZ, Xing JJ, et al. MTA1-mediated RNA m⁶ A modification regulates autophagy and is required for infection of the rice blast fungus[J]. New Phytol, 2022, 235(1): 247-262.
[20]	Huang DY, Cui LQ, Sajid A, et al. The epigenetic mechanisms in Fusarium mycotoxins induced toxicities[J]. Food Chem Toxicol, 2019, 123: 595-601. doi: S0278-6915(18)30798-1 pmid: 30599843
[21]	刘静. 拟轮枝镰孢ORPs家族主效蛋白鉴定及其功能解析[D]. 保定: 河北农业大学, 2023.
	Liu J. Identification and functional analysis of major protein from ORPs family of Fusarium verticillioides[D]. Baoding: Hebei Agricultural University, 2023.
[22]	Tian F, Lee SY, Woo SY, et al. Effect of plant-based compounds on the antifungal and antiaflatoxigenic efficiency of strobilurins against Aspergillus flavus[J]. J Hazard Mater, 2021, 415: 125663.
[23]	Bryła M, Roszko M, Szymczyk K, et al. Fumonisins in plant-origin food and fodder—a review[J]. Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2013, 30(9): 1626-1640.
[24]	Yang X, Gao J, Liu Q, et al. Co-occurrence of mycotoxins in maize and maize-derived food in China and estimation of dietary intake[J]. Food Addit Contam Part B Surveill, 2019, 12(2): 124-134.
[25]	王燕, 董燕婕, 岳晖, 等. 山东省玉米真菌毒素污染状况调查及分析[J]. 粮油食品科技, 2016, 24(3): 69-73.
	Wang Y, Dong YJ, Yue H, et al. Investigatin and analysis on mycotoxins contamination of maize in Shandong Province[J]. Sci Technol Cereals Oils Foods, 2016, 24(3): 69-73.
[26]	Hu L, Liu HW, Yang J, et al. Free and hidden fumonisins in raw maize and maize-based products from China[J]. Food Addit Contam Part B Surveill, 2019, 12(2): 90-96.
[27]	Covarelli L, Stifano S, Beccari G, et al. Characterization of Fusarium verticillioides strains isolated from maize in Italy: fumonisin production, pathogenicity and genetic variability[J]. Food Microbiol, 2012, 31(1): 17-24. doi: 10.1016/j.fm.2012.02.002 pmid: 22475938
[28]	Li LL, He ZQ, Shi Y, et al. Role of epigenetics in mycotoxin toxicity: a review[J]. Environ Toxicol Pharmacol, 2023, 100: 104154.
[29]	Yang C, Wu DD, Lin H, et al. Role of RNA modifications, especially m⁶A, in aflatoxin biosynthesis of Aspergillus flavus[J]. J Agric Food Chem, 2024, 72(1): 726-741.
[30]	Liang LK, Wang XY, Lan HE, et al. Comprehensive analysis of aflatoxin B₁ biosynthesis in Aspergillus flavus via transcriptome-wide m⁶A methylome response to cycloleucine[J]. J Hazard Mater, 2024, 461: 132677.
[31]	Shafik AM, Zhou HQ, Lim J, et al. Dysregulated mitochondrial and cytosolic tRNA m¹A methylation in Alzheimer's disease[J]. Hum Mol Genet, 2022, 31(10): 1673-1680.
[32]	Boo SH, Ha H, Kim YK. m¹A and m⁶A modifications function cooperatively to facilitate rapid mRNA degradation[J]. Cell Rep, 2022, 40(10): 111317.
[33]	Qi ZY, Zhang C, Jian H, et al. N¹-Methyladenosine modification of mRNA regulates neuronal gene expression and oxygen glucose deprivation/reoxygenation induction[J]. Cell Death Discov, 2023, 9(1): 159.
[34]	Trixl L, Lusser A. The dynamic RNA modification 5-methylcytosine and its emerging role as an epitranscriptomic mark[J]. Wiley Interdiscip Rev RNA, 2019, 10(1): e1510-e1527.
[35]	Zhao Y, Xing C, Peng HL. ALYREF(Aly/REF export factor): a potential biomarker for predicting cancer occurrence and therapeutic efficacy[J]. Life Sci, 2024, 338: 122372.
[36]	Klec C, Knutsen E, Schwarzenbacher D, et al. ALYREF, a novel factor involved in breast carcinogenesis, acts through transcriptional and post-transcriptional mechanisms selectively regulating the short NEAT1 isoform[J]. Cell Mol Life Sci, 2022, 79(7): 391. doi: 10.1007/s00018-022-04402-2 pmid: 35776213
[37]	Wang N, Chen RX, Deng MH, et al. m⁵C-dependent cross-regulation between nuclear reader ALYREF and writer NSUN2 promotes urothelial bladder cancer malignancy through facilitating RABL6/TK1 mRNAs splicing and stabilization[J]. Cell Death Dis, 2023, 14(2): 139.
[38]	Berson A, Goodman LD, Sartoris AN, et al. Drosophila Ref1/ALYREF regulates transcription and toxicity associated with ALS/FTD disease etiologies[J]. Acta Neuropathol Commun, 2019, 7(1): 65.
[39]	Kow RL, Black AH, Saxton AD, et al. Loss of aly/ALYREF suppresses toxicity in both tau and TDP-43 models of neurodegeneration[J]. GeroScience, 2022, 44(2): 747-761. doi: 10.1007/s11357-022-00526-2 pmid: 35122183
[40]	Sultana S, Bao WX, Shimizu M, et al. Frequency of three mutations in the fumonisin biosynthetic gene cluster ofFusarium fujikuroi that are predicted to block fumonisin production[J]. World Mycotoxin J, 2021, 14(1): 49-60.
[41]	Janevska S, Ferling I, Jojić K, et al. Self-protection against the sphingolipid biosynthesis inhibitor fumonisin B₁ is conferred by a FUM cluster-encoded ceramide synthase[J]. mBio, 2020, 11(3): e00455-20.

菌株编号 Strain number	菌株来源 Strain source	FB₁含量 Content of FB₁/（mg·g^-1）
87	14-SD-94-6	145.20
64	HK12-5	116.90
114	14-17-15	228.20
126	14-SD-91-13	116.80
81	HN 22-2	112.30
104	25-4	89.55
50	LZ-2-81	103.57
80	NMG-1-2	92.44
84	124-3	64.73
86	HN 24-2	99.39
63	S-9-7	101.82
6	M-I1-46	59.29
38	M-10-48	ND
1	LZ-2-77甘	2.18
49	P-HB-T3-4	2.19
55	NMG-3-3	ND
109	M-3-20	ND
36	26-4	ND
18	14-SD-22-20	ND
37 22	14-SD-91-9 M-4-78	ND ND
40 139	ZL-1-6 14-SD-91-9	ND 1.81
33	P-C1-32-1	ND

菌株编号 Strain number	菌株来源 Strain source	FB₁含量 Content of FB₁/（mg·g^-1）
87	14-SD-94-6	145.20
64	HK12-5	116.90
114	14-17-15	228.20
126	14-SD-91-13	116.80
81	HN 22-2	112.30
104	25-4	89.55
50	LZ-2-81	103.57
80	NMG-1-2	92.44
84	124-3	64.73
86	HN 24-2	99.39
63	S-9-7	101.82
6	M-I1-46	59.29
38	M-10-48	ND
1	LZ-2-77甘	2.18
49	P-HB-T3-4	2.19
55	NMG-3-3	ND
109	M-3-20	ND
36	26-4	ND
18	14-SD-22-20	ND
37 22	14-SD-91-9 M-4-78	ND ND
40 139	ZL-1-6 14-SD-91-9	ND 1.81
33	P-C1-32-1	ND

核苷Nucleotide	母离子Precursorion（m/z）	子离子Production（m/z）	保留时间Residence time/ ms	去簇电压DP/V	碰撞电压CE /V
Um	259.3	113.1	0.94	17	14
Gm	298.2	152.1	1.10	8	16
m¹A	282.1	150.1	0.72	36	30
m⁶A	282.2	150.2	2.40	39	20
m⁵C	258.3	126.1	0.68	24	18
m⁷G	298.1	166.1	0.76	20	18

核苷Nucleotide	母离子Precursorion（m/z）	子离子Production（m/z）	保留时间Residence time/ ms	去簇电压DP/V	碰撞电压CE /V
Um	259.3	113.1	0.94	17	14
Gm	298.2	152.1	1.10	8	16
m¹A	282.1	150.1	0.72	36	30
m⁶A	282.2	150.2	2.40	39	20
m⁵C	258.3	126.1	0.68	24	18
m⁷G	298.1	166.1	0.76	20	18

核苷 Nucleotide	检出限LOD /（μg·kg^-1）	基质效应 Matrix effect/%	线性方程 Linearity equation	决定系数R² Determination coefficient	平均回收率/相对标准偏差 Average recovery（%）/ RSD
核苷 Nucleotide	检出限LOD /（μg·kg^-1）	基质效应 Matrix effect/%	线性方程 Linearity equation	决定系数R² Determination coefficient	1 μg/kg	20 μg/kg	500 μg/kg
Um	0.002	5.41	y =4E+06x - 2964.3	0.9979	91.9/4.7	84.4/4.5	91.9/3
Gm	0.001	2.27	y =2E+06x - 5329.46	0.9999	97.6/6.6	86.2/7.8	109.7/4.9
m¹A	0.0009	2.16	y = 7E+07x - 161526	0.9990	72.5/4.9	75.1/1.9	74.4/7.6
m⁶A	0.0006	2.13	y = 7E+07x - 30294	0.9981	88.5/7.0	78.7/5.9	116.5/1.4
m⁵C	0.0003	2.51	y = 3E+06x - 1896.4	0.9993	70.3/6.5	71.0/4.7	74.1/7.2
m⁷G	0.0006	1.95	y = 6E+07x - 141064	0.9995	74.0/8.9	72.1/7.7	70.9/9.5

Roles of RNA m¹A and m⁵C Methylation Modifications in the Fumonisin Biosynthesis of Fusarium verticillioides

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 8

References 41

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	WANG Chao-min, HE Mei-dan, WANG Wen-zhi, YUAN Qian-hua, ZHANG Shu-zhen, SHEN Lin-bo. Establishment and Application of Real-time PCR for Sugarcane Striate Virus [J]. Biotechnology Bulletin, 2024, 40(6): 126-133.
[2]	SANG Sen-hua. Detection of NK Cell Cytotoxicity: Real-time Dynamic Imaging-Based Analysis [J]. Biotechnology Bulletin, 2024, 40(4): 77-84.
[3]	YANG Wei-jie, YANG Zhou-lin, ZHU Hao-dong, WEI Yu, LIU Jun, LIU Xun. Study on the Properties and Functions of LchAD Protein, a Key Module of Lichenysin Synthase [J]. Biotechnology Bulletin, 2024, 40(3): 322-332.
[4]	YANG Wen-li, ZHU Li-li, CHEN Jian, CHEN Yan-xin, YAO Juan, JIANG Da-gang. Research Progress in the Reference Materials of Crop Pathogens in China [J]. Biotechnology Bulletin, 2024, 40(2): 31-37.
[5]	LIU Xing-yu, LI Jie, ZHU Long-jiao, LI Xiang-yang, XU Wen-tao. Aptamer of Pseudomonas aeruginosa: Acquiring and Application [J]. Biotechnology Bulletin, 2024, 40(1): 186-193.
[6]	ZHANG Xue-ping, LU Yu-qing, ZHANG Yue-qian, LI Xiao-juan. Advances in Plant Extracellular Vesicles and Analysis Techniques [J]. Biotechnology Bulletin, 2023, 39(5): 32-43.
[7]	ZHOU Xi-wen, CHENG Ke, ZHU Hong-liang. Research Progress in the Approaches to in vivo RNA Secondary Structure Profiling in Plants [J]. Biotechnology Bulletin, 2023, 39(2): 51-62.
[8]	GUO Wen-bo, LU Yang, SUI Li, ZHAO Yu, ZOU Xiao-wei, ZHANG Zheng-kun, LI Qi-yun. Preparation and Application of Polyclonal Antibodies Against Beauveria bassiana Mycovirus BbPmV-4 Coat Protein [J]. Biotechnology Bulletin, 2023, 39(10): 58-67.
[9]	LI Hui-jie, DONG Lian-hua, CHEN Gui-fang, LIU Si-yuan, YANG Jia-yi, YANG Jing-ya. Establishment of Droplet Digital PCR Assay for Quantitative Detection of Pseudomonas cocovenenans in Foods [J]. Biotechnology Bulletin, 2023, 39(1): 127-136.
[10]	CHEN Xiao-lin, LIU Yang-er, XU Wen-tao, GUO Ming-zhang, LIU Hui-lin. Application of Synthetic Biology Based Whole-cell Biosensor Technology in the Rapid Detection of Food Safety [J]. Biotechnology Bulletin, 2023, 39(1): 137-149.
[11]	HU Hai-yang, YING Wan-qin, HE Jun, LV Zhi-xian, XIE Xiao-ping, DENG Zhong-liang. Establishment and Application of ERA Real-time Fluorescence Method for Rapid Detection of Mycoplasma pneumoniae [J]. Biotechnology Bulletin, 2022, 38(9): 264-270.
[12]	GAO Wei-xin, HUANG Huo-qing, ZHAO Jing, ZHANG Xin, YANG Ning, YANG Hao-meng. Construction and Activity Verification of Ribonucleoprotein Complex for Gene Editing [J]. Biotechnology Bulletin, 2022, 38(8): 60-68.
[13]	LAN Xin-yue, LIU Ning-ning, ZHU Long-jiao, CHEN Xu, CHU Hua-shuo, LI Xiang-yang, DUAN Nuo, XU Wen-tao. Tetracycline Bivalent Aptamer Non-enzyme Label-free Sensor [J]. Biotechnology Bulletin, 2022, 38(3): 276-284.
[14]	LUO Xue-cong, AN Meng-nan, WU Yuan-hua, XIA Zi-hao. Applications of Recombinase Polymerase Amplification in Plant Virus Detection [J]. Biotechnology Bulletin, 2022, 38(2): 269-280.
[15]	KONG De-zhen, NIE Ying-bin, XU Hong-jun, CUI Feng-juan, MU Pei-yuan, TIAN Xiao-ming. Effects of Blend Seeding on the Yield，Purity and Yield Advantage of F₁ in Three-line Hybrid Wheat [J]. Biotechnology Bulletin, 2022, 38(10): 132-139.