Skip to main content
SpringerLink
Log in

Cloning, expression analysis and molecular marker development of cinnamyl alcohol dehydrogenase gene in common wheat

  • Original Article
  • Published:
Protoplasma Aims and scope Submit manuscript

Abstract

In common wheat, stem strength is one of the key factors for lodging resistance, which is influenced by lignin content. Cinnamyl alcohol dehydrogenase (CAD) is a vital enzyme in the pathway of lignin biosynthesis. Cloning and marker development of the CAD gene could be helpful for lodging resistance breeding. In this study, the full-length genomic DNA sequence of CAD gene in wheat was cloned by using homologous strategy. A marker 5-f2r2 was developed based on CAD sequence and used to genotype 258 wheat lines. Four haplotype combinations of CAD genes were identified in 258 wheat lines. Correction analyses among the CAD gene expression, CAD activity, and stem strength indicated significant positive correlation between CAD gene expression and CAD activity, between wheat CAD activity and wheat stem strength. The haplotype combination B is significantly associated with the lower enzyme activity and weak stem strength, which was supported by the level of CAD gene expression. The CAD activity and stem strength of wheat could be distinguished to some extent using this pair of specific primer 5-f2r2 designed in this study, indicating that the sequence targeted site (STS) marker 5-f2r2 could be used in marker assistant selection (MAS) breeding.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25(17):3389–3402

    Article  CAS  Google Scholar 

  • Barakat A, Bagniewska-Zadworna A, Choi A, Plakkat U, DiLoreto DS, Yellanki P, Carlson JE (2009) The cinnamyl alcohol dehydrogenase gene family in Populus: phylogeny, organization, and expression. BMC Plant Biol 9:26

    Article  Google Scholar 

  • Baucher M, Halpin C, Petit-conil M, Boerjan W (2003) Lignin: genetic engineering and impact on pulping. Crit Rev Biochem Mol Biol 38(4):305–350

    Article  CAS  Google Scholar 

  • Berry PM, Spink J, Sterling M, Pickett AA (2003) Methods for rapidly measuring the lodging resistance of wheat cultivars. J Agron Crop Sci 189:390–401

    Article  Google Scholar 

  • Chen XG (2011) Relationship between stem characteristics and lodging and its regulation study in wheat. Dissertation, Shandong Agricultural University

  • Chen F, Srinivasa Reddy MS, Temple S, Jackson L, Shadle G, Dixon RA (2006) Multi-site genetic modulation of monolignol biosynthesis suggests new routes for formation of syringyl lignin and wall-bound ferulic acid in alfalfa (Medicago sativa L.). Plant J 48(1):113–124

    Article  CAS  Google Scholar 

  • Chen W, VanOpdorp N, Fitzl D, Tewari J, Friedemann P, Greene T, Thompson S, Kumpatla S, Zheng P (2012) Transposon insertion in a cinnamyl alcohol dehydrogenase gene is responsible for a brown midrib1 mutation in maize. Plant Mol Biol 80(3):289–297

    Article  CAS  Google Scholar 

  • Fan YY, Yang GT, Chen J, Jiang F, Muslim Q, Chen YJ, Hu YG (2017) The physical chemical characters and lodging resistance of rice stem with silicon potassium collocation application. Acta Botanica Boreali-Occidentalia Sinica 37:751–757

    Google Scholar 

  • Foulkes MJ, Slafer GA, Davies WJ, Berry PM, Sylvester-Bradley R, Martre P, Calderini DF, Griffiths S, Reynolds MP (2011) Raising yield potential of wheat. III. Optimizing partitioning to grain while maintaining lodging resistance. J Exp Bot 2:469–486

    Article  Google Scholar 

  • Fu CX, Xiao XR, Xi YJ, Ge YX, Chen F, Joseph B, Richard AD, Wang ZY (2011) Down-regulation of cinnamyl alcohol dehydrogenase (CAD) leads to improved saccharification efficiency in switchgrass. Bioenerg Res 4(3):153–164

    Article  Google Scholar 

  • Fu LP, Xiao YG, Yan J, Liu JD, Wen WE, Zhang Y, Xia XC, He ZH (2019) Characterization of TaCOMT genes associated with stem lignin content in common wheat and development of a gene-specific marker. J Integr Agric 18(05):939–947

    Article  CAS  Google Scholar 

  • Goffner D, Joffroy I, Grima-Pettenati J, Halpin C, Knight ME, Schuch W, Boudet AM (1992) Purification and characterization of isoforms of cinnamyl alcohol dehydrogenase from Eucalyptus xylem. Planta 188(1):48–53

    Article  CAS  Google Scholar 

  • Guo D, Chen F, Inoue K, Blount JW, Dixon RA (2001) Downregulation of caffeic acid 3-O-methyltransferase and caffeoyl CoA 3-O-methyltransferase in transgenic alfalfa. impacts on lignin structure and implications for the biosynthesis of G and S lignin. Plant Cell 13(1):73–88

    Article  CAS  Google Scholar 

  • Humphreys JM, Chapple C (2002) Rewriting the lignin roadmap. Current Opinion in Plant Biology 5(3):224–229

    Article  CAS  Google Scholar 

  • Jin Y, Zhang C, Liu W, Qi H, Chen H, Cao S (2014) The cinnamyl alcohol dehydrogenase gene family in melon (Cucumis melo L.): bioinformatic analysis and expression patterns. PLoS One 9(7):e101730

    Article  Google Scholar 

  • Jones L, Ennos AR, Turner SR (2001) Cloning and characterization of irregular xylem4 (irx4): a severely lignin-deficient mutant of Arabidopsis. Plant J 26:205–216

    Article  CAS  Google Scholar 

  • Kang HW, Cho YG, Yoon UH, Eun MY (1998) A rapid DNA extraction method for RFLP and PCR analysis from a single dry seed. Plant Mol Biol Rep 16:90

    Article  Google Scholar 

  • Khobra R, Sareen S, Meena BK, Kumar A, Tiwari V, Singh GP (2019) Exploring the traits for lodging tolerance in wheat genotypes: a review. Physiol Mol Biol Plants 25:589–600

    Article  CAS  Google Scholar 

  • Kim SJ, Kim MR, Bedgar DL, Moinuddin SG, Cardenas CL, Davin LB, Kang C, Lewis NG (2004) Functional reclassification of the putative cinnamyl alcohol dehydrogenase multigene family in Arabidopsis. Proc Natl Acad Sci U S A 101:1455–1460

    Article  CAS  Google Scholar 

  • Li X, Weng JK, Chapple C (2008) Improvement of biomass through lignin modification. Plant J 54(4):569–581

    Article  CAS  Google Scholar 

  • Lynch D, Lidgett A, Mcinnes R, Huxley H, Jones E, Mahoney N, Spangenberg G (2002) Isolation and characterisation of three cinnamyl alcohol dehydrogenase homologue cDNAs from perennial ryegrass (Loliumperenne L.). J Plant Physiol 159:653–660

    Article  CAS  Google Scholar 

  • Ma QH (2009) The expression of caffeic acid 3-O-methyltransferase in two wheat genotypes differing in lodging resistance. J Exp Bot 60(9):2763–2771

    Article  CAS  Google Scholar 

  • Ma QH (2010) Functional analysis of a cinnamyl alcohol dehydrogenase involved in lignin biosynthesis in wheat. J Exp Bot 61(10):2735–2744

    Article  CAS  Google Scholar 

  • Nguyen TN, Son S, Jordan MC, Levin DB, Ayele BT (2016) Lignin biosynthesis in wheat (Triticum aestivum L.): its response to waterlogging and association with hormonal levels. BMC Plant Biol 16:28

    Article  Google Scholar 

  • Pandey B, Pandey VP, Dwivedi UN (2011) Cloning, expression, functional validation and modeling of cinnamyl alcohol dehydrogenase isolated from xylem of Leucaena leucocephala. Protein Expr Purif 79(2):197–203

    Article  CAS  Google Scholar 

  • Ralph J, Lundquist K, Brunow G, Lu FC, Kim H, Schatz PF, Marita JM, Hatfield RD, Ralph SA, Christensen JH, Boerjan W (2004) Lignins: Natural polymers from oxidative coupling of 4-hydroxyphenyl- propanoids. Phytochem Rev 3:29–60

    Article  CAS  Google Scholar 

  • Rohde A, Morreel K, Ralph J, Goeminne G, Hostyn V, De Rycke R, Kushnir S, Van Doorsselaere J, Joseleau JP, Vuylsteke M, Van Driessche G, Van Beeumen J, Messens E, Boerjan W (2004) Molecular phenotyping of the pal1 and pal2 mutants of Arabidopsis thaliana reveals far-reaching consequences on phenylpropanoid, amino acid, and carbohydrate metabolism. Plant Cell 16(10):2749–2771

    Article  CAS  Google Scholar 

  • Saathoff AJ, Hargrove MS, Haas EJ, Tobias CM, Twigg P, Sattler S, Sarath G (2012) Switchgrass PviCAD1: understanding residues important for substrate preferences and activity. Appl Biochem Biotechnol 168(5):1086–1100

    Article  CAS  Google Scholar 

  • Saballos A, Ejeta G, Sanchez E, Kang C, Vermerris W (2009) A genomewide analysis of the cinnamyl alcohol dehydrogenase family in sorghum [Sorghum bicolor (L.) Moench] identifies SbCAD2 as the brown midrib6 gene. Genetics 181(2):783–795

    Article  CAS  Google Scholar 

  • Sattler SE, Saathoff AJ, Haas EJ, Palmer NA, Funnell-Harris DL, Sarath G, Pedersen JF (2009) A nonsense mutation in a cinnamyl alcohol dehydrogenase gene is responsible for the Sorghum brown midrib6 phenotype. Plant Physiol 150(2):584–595

    Article  CAS  Google Scholar 

  • Shewry PR (2009) Wheat. J Exp Bot 60:1537–1553

    Article  CAS  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729

    Article  CAS  Google Scholar 

  • Tobias CM, Chow EK (2005) Structure of the cinnamyl-alcohol dehydrogenase gene family in rice and promoter activity of a member associated with lignification. Planta 220(5):678–688

    Article  CAS  Google Scholar 

  • Vanholme R, Demedts B, Morreel K, Ralph J, Boerjan W (2010) Lignin biosynthesis and structure. Plant Physiol 153(3):895–905

    Article  CAS  Google Scholar 

  • Vanholme R, Storme V, Vanholme B, Sundin L, Christensen JH, Goeminne G, Halpin C, Rohde A, Morreel K, Boerian W (2012) A systems biology view of responses to lignin biosynthesis perturbations in Arabidopsis. Plant Cell 24(9):3506–3529

    Article  CAS  Google Scholar 

  • Wang C, Hu D, Liu XB, She HZ, Ruan RW, Yang H, Yi ZL, Wu DQ (2015) Effects of uniconazole on the lignin metabolism and lodging resistance of culm in common buckwheat (Fagopyrum esculentum M.). Field Crops Res 180:46–53

    Article  Google Scholar 

  • Wang M, Zhu X, Wang K, Lu C, Luo M, Shan T, Zhang Z (2018) A wheat caffeic acid 3-O-methyltransferase TaCOMT-3D positively contributes to both resistance to sharp eyespot disease and stem mechanical strength. Sci Rep 8(1):6543

    Article  Google Scholar 

  • Zhang K, Qian Q, Huang Z, Wang Y, Li M, Hong L, Zeng D, Gu M, Chu C, Cheng Z (2006) GOLD HULL AND INTERNODE2 encodes a primarily multifunctional cinnamyl-alcohol dehydrogenase in rice. Plant Physiol 140(3):972–983

    Article  CAS  Google Scholar 

  • Zhong R, Morrison WH III, Negrel J, Ye ZH (1998) Dual methylation pathways in lignin biosynthesis. Plant Cell 10(12):2033–2046

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by grants from the National Key Research and Development Programme of China (2016YFD0101802, 2017YFD0100804), NSFC 31601298, the Agriculture Research System (CARS-03), the University Synergy Innovation Program of Anhui Province (GXXT-2019-033), and Jiangsu Collaborative Innovation Center for Modern Crop Production (JCIC-MCP).

Author information

Author notes

  1. Can Chen and Jingming Chang contributed equally to this work.

Authors and Affiliations

  1. School of Agronomy, Anhui Agricultural University, Hefei, 230036, China

    Can Chen, Jingming Chang, Sheng Wang, Jie Lu, Yi Liu, Hongqi Si & Chuanxi Ma

  2. Key Laboratory of Wheat Biology and Genetic Improvement on Southern Yellow & Huai River Valley, Ministry of Agriculture and Rural Affairs, Hefei, 230036, China

    Can Chen, Jingming Chang, Sheng Wang, Jie Lu, Yi Liu, Hongqi Si & Chuanxi Ma

  3. Biology Department, Saint Mary’s University, Halifax, NS, B3H 3C3, Canada

    Genlou Sun

  4. National United Engineering Laboratory for Crop Stress Resistance Breeding, Hefei, 230036, China

    Chuanxi Ma

  5. Anhui Key Laboratory of Crop Biology, Hefei, 230036, China

    Chuanxi Ma

Authors
  1. Can Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jingming Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sheng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jie Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yi Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hongqi Si
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Genlou Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Chuanxi Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Hongqi Si, Chuanxi Ma, and Can Chen conceived and designed research. Can Chen, Jingming Chang, and Yi Liu conducted experiments. Jingming Chang, Sheng Wang, and Jie Lu contributed new reagents or analytical tools. Can Chen, Jingming Chang, Sheng Wang, and Genlou Sun analyzed data and wrote the manuscript. All authors read and approved the manuscript.

Corresponding authors

Correspondence to Hongqi Si, Genlou Sun or Chuanxi Ma.

Ethics declarations

Competing interests

The authors declare that they have no conflict of interest.

Additional information

Handling Editor: Handling Editor: Peter Nick

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Supplementary Fig S1.

Phylogenetic analysis of CAD family. (PPTX 72 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, C., Chang, J., Wang, S. et al. Cloning, expression analysis and molecular marker development of cinnamyl alcohol dehydrogenase gene in common wheat. Protoplasma 258, 881–889 (2021). https://doi.org/10.1007/s00709-021-01607-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00709-021-01607-3

Keywords

Search

Navigation