玉米木质素合成途径基因ZmCCoAOMT1功能研究及转录组分析

doi:10.13304/j.nykjdb.2024.0186

摘要/Abstract

摘要：

青贮玉米是一种优良的饲料作物，是畜牧业的优质粗饲料来源。咖啡酰辅酶A-O-甲基转移酶（caffeoyl-CoA-O-methyltransferase， CCoAOMT）是木质素合成途径中的关键甲基转移酶。从玉米自交系B73的EMS突变体库获得了ZmCCoAOMT1基因的单碱基突变体，通过表型分析、生理生化分析、基因表达分析验证了ZmCCoAOMT1的功能。研究发现，ZmCCoAOMT1突变造成茎秆木质化程度降低、木质素含量显著减少、G型木质素（guaiacyl lignin）和S型木质素（syringa lignin）含量下降、体外干物质消化率提高。此外，突变体茎秆中ZmCCoAOMT1的表达量也显著低于野生型。转录组分析表明，ZmCCoAOMT1基因的突变可能调控苯丙烷代谢途径中相关基因的表达，进而引起木质素含量和单体组成发生变化。因此，ZmCCoAOMT1基因是培育高消化率优质青贮玉米品种的重要基因资源，研究结果为木质素合成代谢途径遗传机理研究提供了重要理论依据。

关键词: 青贮玉米, 木质素, 转录组分析, ZmCCoAOMT1

Abstract:

Silage corn with high-quality roughage source for animal husbandry is an excellent forage crop. Caffeoyl-CoA-O-methyltransferase （CCoAOMT）， a key methyltransferase， plays an important role in the lignin synthesis. In this study， the EMS single-base mutant of ZmCCoAOMT1 gene was obtained from EMS mutant library of corn inbred line B73， and the function of ZmCCoAOMT1 was demonstrated based on phenotypic analysis， physiological and biochemical analysis and gene expression analysis. It was found that the mutation of ZmCCoAOMT1 reduced the degree of lignification， significantly decreased the stem lignin content， and declined G （guaiacyl lignin） and S （syringa lignin） monomers contents， whereas increased in vitro digestibility of dry matter. The expression of ZmCCoAOMT1 in the stalks of the mutant was significantly reduced compared to the wild type. Transcriptome analysis analysis showed that the mutation of ZmCCoAOMT1 gene might change the expression of related genes involved in the phenylpropanoid metabolic pathway， leading to variation in lignin content and monomer composition. Therefore， ZmCCoAOMT1 gene would be valuable in corn breeding for high-digestibility and high-quality silage， and above results provided an important genetic basis for elucidating the mechanism of lignin anabolic pathway.

Key words: silage corn, lignin, transcriptome analysis, ZmCCoAOMT1

中图分类号:

S513

徐佳睿, 王逸茹, 赵绍赓, 李坤, 郑军. 玉米木质素合成途径基因ZmCCoAOMT1功能研究及转录组分析[J]. 中国农业科技导报, 2024, 26(5): 30-43.

Jiarui XU, Yiru WANG, Shaogeng ZHAO, Kun LI, Jun ZHENG. Functional Study and Transcriptome Analysis of Corn Gene ZmCCoAOMT1 Involved in Lignin Synthesis Pathway[J]. Journal of Agricultural Science and Technology, 2024, 26(5): 30-43.

图/表 12

表 1 研究所用的引物

Table 1 Primers used in this study

引物名称Primer name	正向序列Forward sequence （5’-3’）	反向序列Reverse sequence （5’-3’）
ZmCCoAOMT1-EMS-F1/R1	ATCAACCGCGAGAACTACGA	GCTGCGGGTCGTCTATTATG
GAPDH-F/R	AGGATATCAAGAAAGCTATTAAGGC	GTAGCCCCACTCGTTGTCG
ZmCCoAOMT1-Q-F1/R1	ACTTCGTGCTCGTCCTCAAC	GCTGCGGGTCGTCTATTATG
ZmCCoAOMT2-Q-F1/R1	GGCCACAAGATCGACTTCC	GTGTTGTCGTAGCCGATGAG
ZmCCoAOMT3-Q-F1/R1	CTTCGACTTCGCCTTCGTC	CGAGAACCTCCTGTCGATGT
ZmCCoAOMT4-Q-F1/R1	CGTCAGCGAAGAAGAGGTG	CTCTCGAGGTCAGCCTCAAC
Zm20961-F/R	GCCTAATCGGATCAGCACAAAC	TAACCCTCTCCATCACGCAAAA
Zm02901-F/R	GCCTTCGACAACGTCTACTACA	GCTCATCTTGGCGTTATTGGTC
Zm14603-F/R	CAGAGAATCCGTCTACCAGAGC	TGTACTGGAAGAAAGGGCAGTC
Zm52336-F/R	CGACCGTCTTCGAGAACAACTA	AGTTAACGGAGTGATGTCACCC
Zm32405-F/R	GTCATCGACGACATCAAGGAGG	TGAGAGAACAGCGAGATCCTTG
Zm40581-F/R	CCTGGACAACTCCTTCTACCAC	TTAGTATAGGCGGTGGTTGACG
Zm37359-F/R	GGAAGACTGGACTCCAAGAAGG	GGGTTGTACTTGGACGTCATCT
Zm32854-F/R	TTAGTGGAGGGCCTTACTACGA	GGCGAACTTGTGAATGATGGAG
Zm32406-F/R	TTCGCGAAGTCGATGGTGAG	GTTAAACCTTCGGCAGTTGAGC
Zm46184-F/R	CTCTCTGCTCAGGCTCTTCTTC	CTTGATGGTGTCGATGACCTCA
Zm04387-F/R	AATGACCAGCTGTGCTTGGATA	TGACGGCACAATACCCACTTTA
Zm07828-F/R	TCTGAACTCCTCATCCACAAGC	TTCCTCATCGGGATGGAGTAGT
Zm48354-F/R	TCGTCAAGATGCAGAAGGAACA	TGTTCCTTCTGCATCTTGACGA
Zm13212-F/R	GCCTTCGACAACGTCTACTACA	GCTCATCTTGGCGTTATTGGTC
Zm50572-F/R	TGAGAAAGGAAGTGGAGAAGGC	AACCATAGACCCAGCAGTCATG

图 1 ZmCCoAOMT1突变体的鉴定A：ZmCCoAOMT1突变位置。B：野生型和ccoaomt1突变体测序结果，红框为突变位点；C：氨基酸序列比对；D： ZmCCoAOMT1基因在野生型和ccoaomt1突变体中的表达量，**表示在P<0.01水平与野生型差异显著（t检验）

Fig. 1 Identification of ZmCCoAOMT1 mutantA： Mutation location of the ZmCCoAOMT1； B： Sequencing of WT and ccoaomt1 mutant， red box indicates mutation location； C： Amino acid sequence alignment； D： Expression levels of ZmCCoAOMT1 in WT and ccoaomt1 mutant， ** represents significant difference compared with WT at P<0.05 level （t-test）

图 2 ZmCCoAOMT家族系统发育分析

Fig. 2 Analysis of the evolutionary tree of the corn ZmCCoAOMT family

图 3 ZmCCoAOMT1对玉米株型的影响A：野生型和ccoaomt1突变体在开花后20 d的表型；B：野生型和ccoaomt1突变体开花后20 d的株高（PH）和穗位高（EH）比较

Fig. 3 Effect of ZmCCoAOMT1 on corn plant architectureA： Phenotype of WT and ccoaomt1 mutant at 20 d after flowering. B： Comparison of plant height （PH） and ear height （EH） at 20 d after flowering between wild-type and ccoaomt1 mutant

图 4 野生型和ccoaomt1突变体的木质素Wiesner染色A：野生型；B： ccoaomt1突变体

Fig. 4 Wiesner staining of lignin in ccoaomt1 mutant and WTA： WT； B： ccoaomt1 mutant

图5 ccoaomt1对木质素含量、组成及消化品质的影响A：成熟期野生型和ccoaomt1突变体地上部3～4茎节的木质素含量；B：成熟期野生型和ccoaomt1突变体地上部3~4茎节的酸性洗涤木质素（ADL）、酸性洗涤纤维（ADF）、中性洗涤纤维（NDF）、体外干物质消化率（IVDMD）差异； C：成熟期野生型和ccoaomt1突变体地上部3~4茎节的木质素单体（G型、S型、H型）差异。*和**分别表示与野生型相比在P<0.05和P<0.01水平差异显著（t检验）

Fig. 5 Effects of ccoaomt1 on lignin content， composition and digestive qualityA： Differences of lignin content in the shoots of three or four stalk nodes between wild type and ccoaomt1 mutant at corn maturity stage； B： Differences of acid detergent lignin（ADL）， acid detergent fiber （ ADF ）， neutral detergent fiber （NDF ） and in vitro digestibility （IVDMD ） in 3~4 shoots of corn between wild type and ccoaomt1 mutants； C： Difference of lignin monomer （G-type， S-type， H-type） in the shoots of wild-type and ccoaomt1 mutants at maturity stage. * and ** represent significant differences compared with WT at P<0.05 and P<0.01 levels （t-test）， respectively

图 6 野生型和突变体 ZmCCoAOMT基因不同时期表达量A： V9期；B吐丝期。*和**分别表示与野生型相比在P<0.05和P<0.01水平差异显著（t检验）

Fig. 6 Expression levels of ZmCCoAOMT genes at different times in wild type and mutantA： V9 stage； B： Silking stage. * and ** represent significant differences compared with WT at P<0.05 and P<0.01 levels （t-test）， respectively

图 7 差异表达基因的qRT-PCR验证A： RNA-seq数据的qRT-PCR验证结果；B： RNA-Seq和qRT-PCR结果相关性分析

Fig. 7 qRT-PCR verification of differentially expressed genesA： qRT-PCR verification of RNA-seq data，； B： Correlation analysis of RNA-Seq and qRT-PCR data

图 8 不同时期野生型和突变体的差异表达基因分析A：差异表达基因数量；B：差异表达基因韦恩图；4个组分别是Ⅰ（V9-野生型）、Ⅱ（V9-ccoaomt1突变体）、Ⅲ（吐丝期-野生型）、Ⅳ（吐丝期-ccoaomt1突变体）。

Fig. 8 Analysis of differentially expressed genes in wild type and mutant at different stagesA： Number of differentially expressed genes； B： Venn of differentially expressed genes； the 4 groups were Ⅰ （V9-WT）， Ⅱ （V9-ccoaomt1 mutant）， Ⅲ （silking-WT） and Ⅳ （silking-ccoaomt1 mutant）

图9 V9时期和吐丝期野生型和突变体差异表达基因的GO富集分析A：V9期；B：吐丝期

Fig. 9 GO enrichment analysis of differentially expressed genes in wild type and mutant at V9 and silking stagesA： V9 stage； B：Silking stage

图 10 V9时期和吐丝期野生型和突变体差异表达基因的KEGG富集分析A：V9时期；B：吐丝期

Fig. 10 KEGG analysis of differentially expressed genes of wild-type and mutant at V9 and silking stagesA： V9 stage； B：Silking stage

表 3 木质素合成相关的差异表达基因

Table 3 Differentially expressed genes associated with lignin synthesis

基因ID Gene ID	差异倍数 log2FC	假定值 Postulate value	调节 Regulate	蛋白 Protein
Zm00001d002898	-1.311 629 353	0.016 167	下调 Down	过氧化物酶12 Peroxidase 12
Zm00001d004443	-2.003 251 187	0.195 030	下调 Down	肉桂醇脱氢酶6 CAD 6
Zm00001d005279	-1.150 411 362	0.419 242	下调Down	过氧化物酶51 Peroxidase 51
Zm00001d007161	-1.111 930 889	0.335 002	下调 Down	过氧化物酶42 Peroxidase 42
Zm00001d008173	-2.638 001 029	0.009 849	下调 Down	过氧化物酶2 Peroxidase 2
Zm00001d008898	1.233 030 247	0.002 509	上调 Up	过氧化物酶4 Peroxidase 4
Zm00001d009373	1.494 820 928	0.001 897	上调 Up	过氧化物酶72 Peroxidase 72
Zm00001d014606	-4.768 298 025	0.055 246	下调 Down	过氧化物酶45 Peroxidase 45
Zm00001d018620	1.666 833 782	0.019 950	上调 Up	过氧化物酶39 Peroxidase 39
Zm00001d019534	-1.111 649 783	0.018 069	下调 Down	丝氨酸羧肽酶 SC-PLs
Zm00001d022451	-0.261 406 941	0.454 268	下调 Down	胱硫醚β裂解酶29 CBL 29
Zm00001d020401	-2.7 444 206	0.111 547	下调 Down	肉桂醇脱氢酶8 CAD8
Zm00001d024119	-1.690 949 412	0.473 358	下调Down	过氧化物酶40 Peroxidase 40
Zm00001d026268	-1.179 850 994	0.531 814	下调 Down	莽草酸羟基肉桂酰转移酶 HCT
Zm00001d026357	-4.078 721 983	0.010 213	下调 Down	过氧化物酶1 Peroxidase 1
Zm00001d028347	-1.309 639 992	0.274 804	下调 Down	过氧化物酶53 Peroxidase 53
Zm00001d030220	-2.225 592 941	0.021 711	下调 Down	咖啡酰莽草酸酯酶 CSE
Zm00001d032405	-1.604 153 903	0.019 927	下调 Down	过氧化物酶56 Peroxidase 56
Zm00001d032467	-1.372 207 308	0.152 413	下调 Down	细胞P450氧化酶 Cytochrome P450 84A1
Zm00001d032854	-3.70 874 872	0.00 016	下调 Down	过氧化物酶59 Peroxidase 59
Zm00001d035055	-2.536 037 556	0.013 541	下调 Down	过氧化物酶47 Peroxidase 47
Zm00001d037359	-1.152 148 355	0.715 147	下调 Down	过氧化物酶11 Peroxidase 11
Zm00001d045101	-1.762 321 261	1.41E-07	下调 Down	肉桂酰辅酶A还原酶1 CCR1
Zm00001d047066	-2.134 983 298	0.034 705	下调 Down	过氧化物酶57 Peroxidase 57
Zm00001d053554	4.15 134 339	0.0 965 01	上调 Up	过氧化物酶70 Peroxidase 70

表 3 木质素合成相关的差异表达基因

Table 3 Differentially expressed genes associated with lignin synthesis

基因ID Gene ID	差异倍数 log2FC	假定值 Postulate value	调节 Regulate	蛋白 Protein
Zm00001d002898	-1.311 629 353	0.016 167	下调 Down	过氧化物酶12 Peroxidase 12
Zm00001d004443	-2.003 251 187	0.195 030	下调 Down	肉桂醇脱氢酶6 CAD 6
Zm00001d005279	-1.150 411 362	0.419 242	下调Down	过氧化物酶51 Peroxidase 51
Zm00001d007161	-1.111 930 889	0.335 002	下调 Down	过氧化物酶42 Peroxidase 42
Zm00001d008173	-2.638 001 029	0.009 849	下调 Down	过氧化物酶2 Peroxidase 2
Zm00001d008898	1.233 030 247	0.002 509	上调 Up	过氧化物酶4 Peroxidase 4
Zm00001d009373	1.494 820 928	0.001 897	上调 Up	过氧化物酶72 Peroxidase 72
Zm00001d014606	-4.768 298 025	0.055 246	下调 Down	过氧化物酶45 Peroxidase 45
Zm00001d018620	1.666 833 782	0.019 950	上调 Up	过氧化物酶39 Peroxidase 39
Zm00001d019534	-1.111 649 783	0.018 069	下调 Down	丝氨酸羧肽酶 SC-PLs
Zm00001d022451	-0.261 406 941	0.454 268	下调 Down	胱硫醚β裂解酶29 CBL 29
Zm00001d020401	-2.7 444 206	0.111 547	下调 Down	肉桂醇脱氢酶8 CAD8
Zm00001d024119	-1.690 949 412	0.473 358	下调Down	过氧化物酶40 Peroxidase 40
Zm00001d026268	-1.179 850 994	0.531 814	下调 Down	莽草酸羟基肉桂酰转移酶 HCT
Zm00001d026357	-4.078 721 983	0.010 213	下调 Down	过氧化物酶1 Peroxidase 1
Zm00001d028347	-1.309 639 992	0.274 804	下调 Down	过氧化物酶53 Peroxidase 53
Zm00001d030220	-2.225 592 941	0.021 711	下调 Down	咖啡酰莽草酸酯酶 CSE
Zm00001d032405	-1.604 153 903	0.019 927	下调 Down	过氧化物酶56 Peroxidase 56
Zm00001d032467	-1.372 207 308	0.152 413	下调 Down	细胞P450氧化酶 Cytochrome P450 84A1
Zm00001d032854	-3.70 874 872	0.00 016	下调 Down	过氧化物酶59 Peroxidase 59
Zm00001d035055	-2.536 037 556	0.013 541	下调 Down	过氧化物酶47 Peroxidase 47
Zm00001d037359	-1.152 148 355	0.715 147	下调 Down	过氧化物酶11 Peroxidase 11
Zm00001d045101	-1.762 321 261	1.41E-07	下调 Down	肉桂酰辅酶A还原酶1 CCR1
Zm00001d047066	-2.134 983 298	0.034 705	下调 Down	过氧化物酶57 Peroxidase 57
Zm00001d053554	4.15 134 339	0.0 965 01	上调 Up	过氧化物酶70 Peroxidase 70

参考文献 33

1	李海燕,魏建民,安小虎,等.青贮玉米的发展现状及栽培技术[J].畜牧与饲料科学, 2011, 32(6): 27,43.
2	戴忠民,高凤菊,王友平,等.青贮玉米的育种及发展趋势[J].玉米科学,2004, 12(4): 9-11.
	DAI Z M, GAO F J, WANG Y P, et al.. Silage maize breeding and its development trend [J]. J. Maize Sci., 2004, 12(4): 9-11.
3	王东军.青贮玉米育种目标与育种方法分析[J].新农业,2020, 1(15): 14-15.
4	ZHONG R, MORRISON W H, HIMMELSBACH D S, et al.. Essential role ofcaffeoyl coenzyme A-O-methyltransferase in lignin biosynthesis in woody poplar plants [J]. Plant Physiol., 2000, 124(2): 563-578.
5	薛永常,李金花,卢孟柱,等.木质素单体生物合成途径[J].林业科学, 2003, 39(6): 146-153.
	XUE Y C, LI J H, LU M Z, et al.. The lignin subunits biosynthesis pathway and its rewriting [J]. Sci. Silvae Sin., 2003, 39(6): 146-153.
6	KUMAR M, CAMPBELL L, TURNER S. Secondary cell walls: biosynthesis and manipulation [J]. J. Exp. Bot., 2016, 67(2): 515-531.
7	SCHMITT D, PAKUSCH A E, MATERN U. Molecular cloning induction and taxonomic distribution of caffeoyl-CoA 3-O-methyltransferase, an enzyme involved in disease resistance [J]. J. Biol. Chem., 1991, 266(26): 17416-17420.
8	YE Z H, KNEUSEL R E, MATERN U, et al.. An alternative methylation pathway in lignin biosynthesis in Zinnia [J]. Plant Cell, 1994, 6(10): 1427-1439.
9	YE Z H. Association of caffeoyl coenzyme A 3-O-methyltransferase expression with lignifying tissues in several dicot plans [J]. Plant Physiol., 1997, 115(4): 1341-1350.
10	王华美,于延冲,付春祥,等.木质素合成关键酶咖啡酰辅酶 A 氧甲基转移酶的研究进展[J].基因组学与应用生物学, 2014, 33(2): 458-466.
	WANG H M, YU Y C, FU C X, et al.. Progress of a key Enzyme-Caffeoyl-CoA 3-O-methyltransferase in lignin biosynthesis [J]. Genom. Appl. Biol., 2014,33(2): 458-466.
11	CHEN C, MEYERMANS H, BURGGRAEVE B, et al.. Cell-specific and conditional expression of caffeoyl-coenzyme A-3-O-methyltransferase in poplar [J]. Plant Physiol., 2000, 123(3): 853-868.
12	RAES J, ROHDE A, CHRISTENSEN J H, et al.. Genome-wide characterization of the lignification toolbox in Arabidopsis [J]. Plant Physiol., 2003, 133(3): 1051-1071.
13	DO C T, POLLET B, THÉVENIN J, et al.. Both caffeoyl Coenzyme A 3-O-methyltransferase 1 and caffeic acid O-methyltransferase 1 are involved in redundant functions for lignin, flavonoids and sinapoyl malate biosynthesis in Arabidopsis [J]. Planta, 2007, 226(5): 1117-1129.
14	PINÇON G, MAURY S, HOFFMANN L, et al.. Repression of O-methyltransferase genes in transgenic tobacco affects lignin synthesis and plant growth [J]. Phytochemistry, 2001, 57(7): 1167-1176.
15	GUO D, CEN F, INOUE K, et al.. Downregulation of cafeic acid 3-O-methyltransferase and cafeoyl CoA 3-O-methyltransferase in transgenic alfalfa: impacts on lignin structure and implications for the biosynthesis of G and S lignin [J]. Plant Cell, 2001, 13(1): 73-88.
16	KWON H, CHO D J, LEE H, et al.. CCOAOMT1, a candidate cargo secreted via VAMP721/722 secretory vesicles in Arabidopsis [J]. Biochem. Biophys. Res. Commun., 2020, 524(4): 977-982.
17	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/OL]. Plants, 2021, 10(5): 831 [2024-04-02]. .
18	XIA Y, LIU J, WANG Y, et al.. Ectopic expression of Vicia sativa Caffeoyl-CoA-O -methyltransferase (VsCCoAOMT) increases the uptake and tolerance of cadmium in Arabidopsis [J]. Environ. Exp. Bot., 2018, 145(1): 47-53.
19	FORNALÉ S, RENCORET J, GARCÍA C L, et al.. Changes in cell wall polymers and degradability in maize mutants lacking 3’‍- and 5’‍-O-methyltransferases involved in lignin biosynthesis [J]. Plant Cell Physiol., 2017, 58(2): 240-255.
20	BRENNER E A, ZEIN I, CHEN Y, et al.. Polymorphisms in O-methyltransferase genes are associated with stover cell wall digestibility in European maize (Zea mays L.) [J/OL]. BMC Plant Biol., 2010, 10:27 [2024-04-02]. .
21	YANG Q, HE Y J, KABAHUMA M, et al.. A gene encoding maize caffeoyl-CoA O-methyltransferase confers quantitative resistance to multiple pathogens [J]. Nat. Genet., 2017, 49(9): 1364-1372.
22	刘玄,王梅,李玉龙,等.玉米主要农艺性状相关性分析[J].安徽农学通报, 2021, 27(24): 39-41, 79.
23	GEIGER H, FUGGERERA H. Über den chemismus der wiesner-reaktion auf lignin on the chemistry of the wiesner reaction on lignin [J]. Zeitschrift Fur Naturforschung B, 1979, 34: 1471-1472.
24	马飞前.玉米茎秆纤维性状QTL定位[D].北京:中国农业科学院, 2014.
	MA F Q. Mapping of quantitative trait loci (QTL) for stalk fiber traits in maize [D]. Beijing:Chinese Academy of Agricultural Sciences, 2014.
25	HARMAN-WARE A E, FOSTER C, HAPPS R M, et al.. A thioacidolysis method tailored for higher-throughput quantitative analysis of lignin monomers [J]. Biotechnol. J., 2016, 11(10): 1268-1273.
26	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C(T)) method [J]. Methods, 2001, 25(4): 402-408.
27	CHALMEL F, LARDENOIS A, THOMPSON J D, et al.. GOAnno:GO annotation based on multiple alignment [J]. Bioinformatics, 2005, 21(9): 2095-2096.
28	KANEHISA M, GOTO S. KEGG: kyoto encyclopedia of genes and genomes [J]. Nucleic. Acids. Res., 2000, 28(1): 27-30.
29	YU G C, WANG L G, HAN Y Y, et al.. ClusterProfiler: an R package comparing biological themes among gene clusters [J]. OMICS, 2012,16(5): 284-287.
30	ZAKZESKI J, BRUIJNINCX P C A, JONGERIUS A L, et al.. The catalytic valorization of lignin for the production of renewable chemicals [J]. Chem. Rev., 2010, 110(6): 3552-3599.
31	龙国辉,武鹏雨,付嘉智,等.过氧化物酶调控木质素合成研究进展[J].现代农业科技, 2021(23): 47-49, 54.
	LONG G H, WU P Y, FU J Z, et al.. Research progress on regulation of peroxidase on lignin synthesis [J]. Modern Agric. Sci. Technol., 2021(23): 47-49.
32	DIXON R A, BARROS J. Lignin biosynthesis: old roads revisited and new roads explored [J/OL]. Open Biol., 2019,9(12):190215 [2024-04-02]..
33	VANHOLME R, CESARINO I, RATAJ K, et al.. Caffeoyl shikimate esterase (CSE) is an enzyme in the lignin biosynthetic pathway in Arabidopsis [J]. Science, 2013, 341(6150): 1103-1106.

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[8]	张庆芳,于宗莲. 一株高效木质素降解细菌的筛选及产酶条件的优化[J]. , 2014, 16(2): 143-148.
[9]	陈翟微刘长江. 利用白腐真菌提高秸秆利用率[J]. , 2001, 3(3): 53-56.