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01.08.2014 | Original Paper | Ausgabe 4/2014

Cellulose 4/2014

Eight distinct cellulose synthase catalytic subunit genes from Betula luminifera are associated with primary and secondary cell wall biosynthesis

Zeitschrift:
Cellulose > Ausgabe 4/2014
Autoren:
Huahong Huang, Cheng Jiang, Zaikang Tong, Longjun Cheng, Muyuan Zhu, Erpei Lin
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1007/​s10570-014-0261-z) contains supplementary material, which is available to authorized users.
Huahong Huang, Jiang Cheng and Zaikang Tong have contributed equally to this work. Erpei Lin is the corresponding author.

Abstract

Cellulose, a significant proportion of the plant cell wall, is the main determining factor of the wood qualities of a timber tree. Cellulose synthase (CESA), an essential catalyst for cellulose biosynthesis, is encoded by the CesA gene family. We isolated eight full-length CesA cDNAs from Betula luminifera on the basis of transcriptome sequencing. The predicted proteins comprised 985–1,103 amino acids, sharing 85–92 % identity with the corresponding PtiCESAs from Populus trichocarpa. Each protein contained typical motifs of plant CESA proteins. Phylogenetic analysis showed that BlCESA1, BlCESA3 and BlCESA4 belonged to three clades linked to secondary cell wall synthesis, and BlCESA2, BlCESA5, BlCESA6, BlCESA7 and BlCESA8 belonged to three clades associated with primary cell wall synthesis. Quantitative reverse transcription PCR and in situ mRNA hybridization indicated that transcripts of BlCesA1, BlCesA3 and BlCesA4 were most abundant in the xylem of the lignified stem and had similar expression patterns when treated with hormones and sucrose. Thus, the proteins encoded by these three genes might be subunits of the secondary wall CesA complex. However, the expression patterns of BlCesA1, BlCesA3 and BlCesA4 were different in response to mechanical stress. These three genes showed much higher expression levels in the tension wood than in the control, and they showed different expression patterns in the opposite wood, suggesting that these genes play different roles during secondary wall formation. BlCesA2, BlCesA5, BlCesA6 and BlCesA7 were predominantly expressed in the leaf and phloem. BlCesA8, which is most similar to BlCesA6, was strongly expressed in the leaf, phloem and cambium. Interestingly, these two genes showed distinct expression patterns when treated with hormones, sucrose and bending. Thus, in addition to biosynthesis of the primary cell wall, BlCesA8 might have other functions.

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Zusatzmaterial
Tension experiment and sampling of xylem in B. luminifera. A. Tension experiment of B. luminifera. Bent tree with harvesting zone (open box). Tension wood on upper side; opposite wood on lower side. B. Xylem of harvesting zone was divided into three equal parts. The upper was the TW sample; the lower was the OW sample. Scale bars: (A) 5.0 cm, (B) 2.0 cm. (TIFF 5868 kb)
10570_2014_261_MOESM1_ESM.tif
Amino acid alignment and motif analysis of the eight predicted BlCESA protein sequences. Identical conserved residues at a particular site are shown with a black background. A gray background highlights non-identical conserved residues with similar properties. TM, transmembrane region. (EPS 1782 kb)
10570_2014_261_MOESM2_ESM.eps
Sectioning of the up-right current-year branches from B. luminifera. A, B, C and D represent handmade transverse sections of the first, third, fifth and eighth internodes, respectively. Lignified walls stain greenish blue or bright blue, and most primary (non-lignified) walls stain pinkish-purple. co, cortex; ph, phloem; cz, cambial zone; xy, xylem; pi, pith. Scale bars: 40.0 μm. (TIFF 8017 kb)
10570_2014_261_MOESM3_ESM.tif
Relative expression levels of three positive genes in two tissues after treatment with different hormones. (A) Effect of IAA treatments on BlPIN1 expression; (B) effect of GA3 treatments on BlGA20ox1 expression; (C) effect of BR treatments on BlHDZ1 expression. Expression levels in phloem and xylem after 1/2 MS treatment were used individually as a reference and were set at 1. The relative expression level was log2 transformed; > 0 means upregulated, = 0 means unregulated, and < 0 means downregulated. *Gene expression is significantly regulated (p ≤ 0.01, |log2-fold change| ≥ 1). Error bars represent the standard deviation of four biological replicates. (TIFF 5680 kb)
10570_2014_261_MOESM4_ESM.tif
Supplementary material 5 (DOCX 18 kb)
10570_2014_261_MOESM5_ESM.docx
Supplementary material 6 (DOCX 16 kb)
10570_2014_261_MOESM6_ESM.docx
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