Functional Analysis of FmCCoAOMT Gene in Fraxinus mandshurica During Lignin Synthesis and Abiotic Stress

doi:10.7525/j.issn.1673-5102.2023.05.013

Abstract

Abstract:

To clarify the function of FmCCoAOMT gene， the expression pattern of FmCCoAOMT gene was analyzed by qRT-PCR using Fraxinus mandshurica as material in this study. The results showed that FmCCoAOMT gene was tissue specific. The expression level was the highest in the xylem of both female and male perennial plants， and was 9.4 times higher than that in the bark of new shoots in male plants， and was 13.1 times higher than that of leaf in female plants. The leaf disk method was used to obtain transgenic tobacco plants. Compared with the wild type， the lignin content of overexpressing FmCCoAOMT was increased by 41%， while the cellulose and hemicellulose contents were decreased by 32% and 52%， respectively. Tobacco overexpressing FmCCoAOMT after abiotic stress treatment compared to wild type， the activity of POD and SOD increased， the content of MDA decreased， and the content of reactive oxygen decreased after stress treatment. After NaCl treatment， POD activity was 83% higher than that of control group. After NaCl， ABA and mannitol treatments， SOD activity increased by 56.6%， 44.2% and 19.0%， and MDA content decreased by 77.2%， 11.7% and 47.6%， respectively. The results indicated that FmCCoAOMT gene might be involved in lignin synthesis of F. mandshurica， promoted abiotic stress tolerance.

Key words: Fraxinus mandshurica, Lignin, FmCCoAOMT, Abiotic stress

CLC Number:

Q786

Chaoran SHAN, Xiaohu CHEN, Yunfei DING, Wei ZHAO, Han LU, Shangzhu GAO, Fenghui QI, Yaguang ZHAN, Fansuo ZENG. Functional Analysis of FmCCoAOMT Gene in Fraxinus mandshurica During Lignin Synthesis and Abiotic Stress[J]. Bulletin of Botanical Research, 2023, 43(5): 768-778.

Figures/Tables 9

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References 37

1	ESLICK E M， BEILBY M J， MOON A R.A study of the native cell wall structures of the marine alga Ventricaria ventricosa（Siphonocladales，Chlorophyceae） using atomic force microscopy［J］.Microscopy，2014，63（2）：131-140.
2	YE Z H， YORK W S， DARVILL A G.Important new players in secondary wall synthesis［J］.Trends in Plant Science，2006，11（4）：162-164.
3	魏建华，宋艳茹.木质素生物合成途径及调控的研究进展［J］.植物学报，2001，43（8）：771-779.
	WEI J H， SONG Y R.Recent advances in study of lignin biosynthesis and manipulation［J］.Bulletin of Botany，2001，43（8）：771-779.
4	YE Z H.Association of caffeoyl coenzyme A 3-O-methyltransferase expression with lignifying tissues in several dicot plants［J］.Plant Physiology，1997，115（4）：1341-1350.
5	WANG G F， BALINT-KURTI P J.Maize homologs of CCoAOMT and HCT，two key enzymes in lignin biosynthesis，form complexes with the NLR Rp1 protein to modulate the defense response［J］.Plant Physiology，2016，171（3）：2166-2177.
6	PAKUSCH A E， KNEUSEL R E， MATERN U.S-adenosyl-L-methionine：trans-caffeoyl-coenzyme A 3-O-methyltransferase from elicitor-treated parsley cell suspension cultures［J］.Archives of Biochemistry and Biophysics，1989，271（2）：488-494.
7	李雪平，高志民，彭镇华，等.绿竹咖啡酰辅酶A-O-甲基转移酶基因的克隆与分析［J］.分子植物育种，2008，6（3）：587-592.
	LI X P， GAO Z M， PENG Z H，et al.Cloning and characterization of CCoAOMT gene from bambusa oldhamii［J］.Molecular Plant Breeding，2008，6（3）：587-592.
8	吴晓宇，胡尚连，曹颖，等.慈竹CCoAOMT基因的克隆及生物信息学分析［J］.南京林业大学学报（自然科学版），2012，36（3）：17-22.
	WU X Y， HU L S， CAO Y，et al.Cloning of CCoAOMT gene in Neosinocalamus affinis and its bioinformatics analysis［J］.Journal of Nanjing Forestry University（Natural Sciences Edition），2012，36（3）：17-22.
9	姚连梅，胡晓晴，周菲，等.白桦反义CCoAOMT基因调控木质素生物合成［J］.植物研究，2019，39（1）：123-130.
	YAO L H， HU X Q， ZHOU F，et al.Antisense CCoAOMT gene regulates lignin biosynthesis in Betula platyphylla ［J］.Bulletin of Botanical Research，2019，39（1）：123-130.
10	RAES J， ROHDE A， CHRISTENSEN J H，et al.Genome-wide characterization of the lignification toolbox in Arabidopsis ［J］.Plant Physiology，2003，133（3）：1051-1071.
11	刘红梅，胡尚连，卢学琴，等.青杨CCoAOMT基因的克隆及其生物信息学分析［J］.湖北农业科学，2014，53（11）：2670-2674.
	LIU H M， HU S L， LU X Q， et al.Cloning and bioinformatics analysis of CCoAOMT gene in Populus cathayana Rehd［J］.Hubei Agricultural Sciences，2014，53（11）：2670-2674.
12	MARTZ F， MAURY S， PINCON G，et al.cDNA cloning，substrate specificity and expression study of tobacco caffeoyl-CoA 3-O-methyltransferase，a lignin biosynthetic enzyme［J］.Plant Molecular Biology，1998，36（3）：427-437.
13	LI X Y， CHEN W J， ZHAO Y，et al.Downregulation of caffeoyl-CoA O-methyltransferase（CCoAOMT） by RNA interference leads to reduced lignin production in maize straw［J］.Genetics and Molecular Biology，2013，36（4）：540-546.
14	FU Y Y， ZHU Y Y， YANG W，et al.Isolation and functional identification of a Botrytis cinerea-responsive caffeoyl-CoA O-methyltransferase gene from Lilium regale wilson［J］.Plant Physiology and Biochemistry，2020，157：379-389.
15	YANG Q， HE Y J， KABAHUMA M，et al.A gene encoding maize caffeoyl-CoA O-methyltransferase confers quantitative resistance to multiple pathogens［J］.Nature Genetics，2017，49（9）：1364-1372.
16	WANG Q X， JIN Q， MA Y Y，et al.Iron toxicity-induced regulation of key secondary metabolic processes associated with the quality and resistance of Panax ginseng and Panax quinquefolius ［J］.Ecotoxicology and Environmental Safety，2021，224：112648.
17	YAMAGUCHI M， VALLIYODAN B， ZHANG J，et al.Regulation of growth response to water stress in the soybean primary root.I.Proteomic analysis reveals region-specific regulation of phenylpropanoid metabolism and control of free iron in the elongation zone［J］.Plant，Cell & Environment，2010，33（2）：223-243.
18	GIORDANO D， PROVENZANO S， FERRANDINO A，et al.Characterization of a multifunctional caffeoyl-CoA O-methyltransferase activated in grape berries upon drought stress［J］.Plant Physiology and Biochemistry，2016，101：23-32.
19	CHUN H J， BAEK D， CHO H M，et al.Lignin biosynthesis genes play critical roles in the adaptation of Arabidopsis plants to high-salt stress［J］.Plant Signaling & Behavior，2019，14（8）：1625697.
20	CHUN H J， LIM L H， CHEONG M S，et al.Arabidopsis CCoAOMT1 plays a role in drought stress response via ROS- and ABA-dependent manners［J］.Plants，2021，10（5）：831.
21	赵兴堂，夏德安，曾凡锁，等.水曲柳生长性状种源与地点互作及优良种源选择［J］.林业科学，2015，51（3）：140-147.
	ZHAO X T， XIA D A， ZENG F S，et al.Provenances by sites interaction of growth traits and provenance selection of Fraxinus mandshurica ［J］.Scientia Silvae Sinicae，2015，51（3）：140-147.
22	梁楠松.小黑杨bHLH转录因子基因的克隆及遗传转化［D］.哈尔滨：东北林业大学，2014.
	LIANG N S.Cloning and gentic transformation of bHLH transcription factor gene in Populus xiaohei T.S.hwang et liang［D］.Harbin：Northeast Forestry University，2014.
23	纪欣童，于磊，詹亚光.水曲柳BZR1基因克隆及其表达模式分析［J］.植物研究，2021，41（5）：744-752.
	JI X T， YU L， ZHAN Y G.Cloning and expression analysis of BRASSINAZOLE RESISTANTANT1（BZR1） gene from Fraxinus mandshurica ［J］.Bulletin of Botanical Research，2001，41（5）：744-752.
24	齐凤慧，陈思齐，付德山，等.水曲柳FmSOC1基因烟草中遗传转化［J］.西北林学院学报，2017，32（5）：98-103.
	QI F H， CHEN S Q， FU D S，et al.Genetic transformation of SOC1 gene from Ash to Tobacco［J］.Journal of Northwest Forestry University，2017，32（5）：98-103.
25	尹增芳，樊汝汶.植物细胞壁的研究进展［J］.植物研究，1999，41（4）：407-414.
	YIN Z F， FAN R W.The study progress on the cell wall［J］.Bulletin of Botanical Research，1999，41（4）：407-414.
26	常瑞娜，汪杏芬，陈鸿鹏.五节芒CCoAOMT和4CL的克隆和表达分析［J］.华北农学报，2012，27（4）：29-35.
	CHANG R N， WANG X F， CHEN H P.Cloning and expression analysis of CCoAOMT and 4CL genes from Miscanthus floridulus ［J］.Acta Agriculturae Boreali-Sinica，2012，27（4）：29-35.
27	章霄云，郭安平，贺立卡，等.木质素生物合成及其基因调控的研究进展［J］.分子植物育种，2006，4（3）：431-437.
	ZHANG X Y， GUO A P， HE L K，et al.Advances in study of lignin biosynthesis and its genetic manipulation［J］.Molecular Plant Breeding，2006，4（3）：431-437.
28	WAGNER A， TOBIMATSU Y， PHILLIPS L，et al.CCoAOMT suppression modifies lignin composition in Pinus radiata ［J］.The Plant Journal，2011，67（1）：119-129.
29	ZHAO D Q， LUAN Y T， SHI W B，et al.A Paeonia ostii caffeoyl-CoA O-methyltransferase confers drought stress tolerance by promoting lignin synthesis and ROS scavenging［J］.Plant Science，2021，303：110765.
30	PANG S L， ONG S S， LEE H H，et al.Isolation and characterization of CCoAOMT in interspecific hybrid of Acacia auriculiformis×Acacia mangium：a key gene in lignin biosynthesis［J］.Genetics and Molecular Research，2014，13（3）：7217-7238.
31	黄春琼，刘国道，郭安平.反义CCoAOMT基因调控烟草木质素的生物合成［J］.安徽农业科学，2008，36（19）：8026-8027，8043.
	HUANG C Q， LIU G D， GUO A P.Lignin biosynthesis regulated by antisense CCoAOMT gene in Tobacco［J］.Journal of Anhui Agricultural Sciences，2008，36（19）：8026-8027，8043.
32	LE GALL H， PHILIPPE F， DOMON J M，et al.Cell wall metabolism in response to abiotic stress［J］.Plants，2015，4（1）：112-166.
33	吕笑言，王宇光，金英.甜菜BvM14-CCoAOMT基因的克隆、表达及生物信息学分析［J］.黑龙江大学自然科学学报，2018，35（3）：317-323.
	LÜ X Y， WANG Y G， JIN Y.Cloning，Expression and bioinformatics analyses of BvM 14 -CCoAOMT gene［J］.Journal of Natural Science of Heilongjiang University，2018，35（3）：317-323.
34	GHOSH R， CHOI B， JEONG M J，et al.Comparative transcriptional analysis of caffeoyl-coenzyme A 3-O-methyltransferase from Hibiscus cannabinus L.，during developmental stages in various tissues and stress regulation［J］.Plant Omics，2012，5（2）：184-193.
35	LIU S J， HUANG Y H， HE C J，et al.Cloning，bioinformatics and transcriptional analysis of caffeoyl-coenzyme A 3-O-methyltransferase in switchgrass under abiotic stress［J］.Journal of Integrative Agriculture，2016，15（3）：636-649.
36	LIU C J， MIAO Y C， ZHANG K W.Sequestration and transport of lignin monomeric precursors［J］.Molecules，2011，16（1）：710-727.
37	SONG J L， WANG Z Y， WANG Y H，et al.Overexpression of Pennisetum purpureum CCoAOMT contributes to lignin deposition and drought tolerance by promoting the Accumulation of flavonoids in Transgenic Tobacco［J］.Frontiers in Plant Science，2022，13：884456.

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