scholarly journals Full-length transcriptomic identification of R2R3-MYB family genes related to secondary cell wall development in Cunninghamia lanceolata (Chinese fir)

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Hebi Zhuang ◽  
Sun-Li Chong ◽  
Borah Priyanka ◽  
Xiao Han ◽  
Erpei Lin ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Cell Wall ◽  
Secondary Wall ◽  
Secondary Cell Wall ◽  
Evolutionary Relationship ◽  
Wood Formation ◽  
Chinese Fir ◽  
Full Length ◽  
Cell Wall Development ◽  
R2r3 Myb

Abstract Background R2R3-MYB is a class of transcription factor crucial in regulating secondary cell wall development during wood formation. The regulation of wood formation in gymnosperm has been understudied due to its large genome size. Using Single-Molecule Real-Time sequencing, we obtained full-length transcriptomic libraries from the developmental stem of Cunninghamia lanceolata, a perennial conifer known as Chinese fir. The R2R3-MYB of C. lanceolata (hereafter named as ClMYB) associated with secondary wall development were identified based on phylogenetic analysis, expression studies and functional study on transgenic line. Results The evolutionary relationship of 52 ClMYBs with those from Arabidopsis thaliana, Eucalyptus grandis, Populus trichocarpa, Oryza sativa, two gymnosperm species, Pinus taeda, and Picea glauca were established by neighbour-joining phylogenetic analysis. A large number of ClMYBs resided in the woody-expanded subgroups that predominated with the members from woody dicots. In contrast, the woody-preferential subgroup strictly carrying the members of woody dicots contained only one candidate. The results suggest that the woody-expanded subgroup emerges before the gymnosperm/angiosperm split, while most of the woody-preferential subgroups are likely lineage-specific to woody dicots. Nine candidates shared the same subgroups with the A. thaliana orthologs, with known function in regulating secondary wall development. Gene expression analysis inferred that ClMYB1/2/3/4/5/26/27/49/51 might participate in secondary wall development, among which ClMYB1/2/5/26/27/49 were significantly upregulated in the highly lignified compression wood region, reinforcing their regulatory role associated with secondary wall development. ClMYB1 was experimentally proven a transcriptional activator that localised in the nucleus. The overexpression of ClMYB1 in Nicotiana benthamiana resulted in an increased lignin deposition in the stems. The members of subgroup S4, ClMYB3/4/5 shared the ERF-associated amphiphilic repression motif with AtMYB4, which is known to repress the metabolism of phenylpropanoid derived compounds. They also carried a core motif specific to gymnosperm lineage, suggesting divergence of the regulatory process compared to the angiosperms. Conclusions This work will enrich the collection of full-length gymnosperm-specific R2R3-MYBs related to stem development and contribute to understanding their evolutionary relationship with angiosperm species.

scholarly journals Full-Length Transcriptomic Identification of R2R3-MYB Family Genes Related to Secondary Cell Wall Development in Cunninghamia Lanceolata (Chinese Fir)

2021 ◽  
Author(s):  
Hebi Zhuang ◽  
Sun-Li Chong ◽  
Priyanka Borah ◽  
Xiao Han ◽  
Erpei Lin ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Cell Wall ◽  
Secondary Wall ◽  
Secondary Cell Wall ◽  
Evolutionary Relationship ◽  
Wood Formation ◽  
Chinese Fir ◽  
Full Length ◽  
Cell Wall Development ◽  
R2r3 Myb

Abstract Background: R2R3-MYB is a class of transcription factor crucial in regulating secondary cell wall development during wood formation. The regulation of wood formation in gymnosperm has been understudied due to its large genome size. Using Single-Molecule Real-Time sequencing, we obtained full-length transcriptomic libraries from the developmental stem of Cunninghamia lanceolata, a perennial conifer known as Chinese fir. The R2R3-MYB of C. lanceolata (hereafter named as ClMYB) associated with secondary wall development were identified based on phylogenetic analysis, expression studies and functional study on transgenic line. Results: The evolutionary relationship of 52 ClMYBs with those from Arabidopsis thaliana, Eucalyptus grandis, Populus trichocarpa, Oryza sativa, two gymnosperm species, Pinus taeda, and Picea glauca were established by neighbour-joining phylogenetic analysis. A large number of ClMYBs resided in the woody-expanded subgroups that predominated with the members from woody dicots. In contrast, the woody-preferential subgroup strictly carrying the members of woody dicots contained only one candidate. The results suggested that the woody-expanded subgroup emerges before the gymnosperm/angiosperm split, while most of the woody-preferential subgroups are likely lineage-specific to woody dicots. Nine candidates shared the same subgroups with the A. thaliana orthologs, with known function in regulating secondary wall development. Gene expression analysis inferred that ClMYB1/2/3/4/5/26/27/49/51 might participate in secondary wall development, among which ClMYB1/2/5/26/27/49 were significantly upregulated in the highly lignified compression wood region, reinforcing their regulatory role associated with secondary wall development. ClMYB1 was experimentally proven a transcriptional activator that localised in the nucleus. The overexpression of ClMYB1 in Nicotiana benthamiana resulted in an increased lignin deposition in the stems. The members of subgroup S4, ClMYB3/4/5 shared the ERF-associated amphiphilic repression motif with AtMYB4, which is known to repress the metabolism of phenylpropanoid derived compounds. They also carried a core motif specific to gymnospermm lineage, suggesting divergence of the regulatory process compared to the angiosperms.Conclusions: This work will enrich the collection of full-length gymnosperm-specific R2R3-MYBs related to stem development and contribute to understanding their evolutionary relationship with angiosperm species.

The R2R3 MYB transcription factor MYB189 negatively regulates secondary cell wall biosynthesis in Populus

2019 ◽  
Vol 39 (7) ◽  
pp. 1187-1200 ◽  
Author(s):  
Bo Jiao ◽  
Xin Zhao ◽  
Wanxiang Lu ◽  
Li Guo ◽  
Keming Luo
Keyword(s):  
Cell Wall ◽  
Transcription Factor ◽  
Secondary Wall ◽  
Secondary Cell Wall ◽  
Populus Trichocarpa ◽  
Wood Formation ◽  
Myb Transcription Factor ◽  
Polymerase Chain ◽  
Xylem Fibers ◽  
R2r3 Myb

Abstract Secondary cell wall (SCW) biosynthesis during wood formation in trees is controlled by a multilevel regulatory network that coordinates the expression of substantial genes. However, few transcription factors involved in the negative regulation of secondary wall biosynthesis have been characterized in tree species. In this study, we isolated an R2R3 MYB transcription factor MYB189 from Populus trichocarpa, which is expressed predominantly in secondary vascular tissues, especially in the xylem. A novel repression motif was identified in the C-terminal region of MYB189, which indicates this factor was a transcriptional repressor. Overexpression (OE) of MYB189 in Arabidopsis and poplar resulted in a significant reduction in the contents of lignin, cellulose and hemicelluloses. Vascular development in stems of MYB189 OE lines was markedly inhibited, leading to a dramatic decrease in SCW thickness of xylem fibers. Gene expression analyses showed that most of the structural genes involved in the biosynthesis of lignin, cellulose and xylans were significantly downregulated in MYB189-overexpressing poplars compared with the wild-type control. Chromatin immunoprecipitation-quantitative real-time polymerase chain reaction and transient expression assays revealed that MYB189 could directly bind to the promoters of secondary wall biosynthetic genes to repress their expression. Together, these data suggest that MYB189 acts as a repressor to regulate SCW biosynthesis in poplar.

Lignification of spruce tracheid secondary cell walls related to longitudinal hardness and modulus of elasticity using nano-indentation

2002 ◽  
Vol 80 (10) ◽  
pp. 1029-1033 ◽  
Author(s):  
W Gindl ◽  
H S Gupta ◽  
C Grünwald
Keyword(s):  
Cell Wall ◽  
Modulus Of Elasticity ◽  
Cell Walls ◽  
Secondary Cell Wall ◽  
Lignin Content ◽  
Wood Formation ◽  
Nano Indentation ◽  
Secondary Cell Walls ◽  
Cell Wall Development ◽  
The One

The lignin content and the mechanical properties of lignifying and fully lignified spruce tracheid secondary cell walls were determined using UV microscopy and nano-indentation, respectively. The average lignin content of developing tracheids was 0.10 g·g–1, as compared with 0.21 g·g–1 in mature tracheids. The modulus of elasticity of developing cells was on average 22% lower than the one measured in mature, fully lignified cells. For the longitudinal hardness, a larger difference of 26% was observed. As lignifying cells in the cambial zone are undergoing cell wall development, spaces in the cellulose–hemicellulose structure are filled with lignin and the density of the cell wall is believed to increase. It is therefore suggested that the observed difference in modulus of elasticity between developing and fully lignified cell walls is due to the filling of spaces with lignin and an increase of the packing density of the cell wall during lignification. Although remarkably less stiff than the composite polysaccharide structure in the secondary cell wall, lignin may be considered equally hard. Therefore, the observed increase in lignin content may contribute directly to the measured increase of hardness.Key words: secondary cell wall, hardness, lignin, modulus of elasticity, wood formation.

scholarly journals MYB Transcription Factors and Its Regulation in Secondary Cell Wall Formation and Lignin Biosynthesis during Xylem Development

2021 ◽  
Vol 22 (7) ◽  
pp. 3560
Author(s):  
Ruixue Xiao ◽  
Chong Zhang ◽  
Xiaorui Guo ◽  
Hui Li ◽  
Hai Lu
Keyword(s):  
Cell Wall ◽  
Transcription Factors ◽  
Secondary Wall ◽  
Secondary Cell Wall ◽  
Lignin Biosynthesis ◽  
Cell Wall Formation ◽  
Long Distance ◽  
Secondary Cell Wall Formation ◽  
Myb Transcription Factors ◽  
Wall Formation

The secondary wall is the main part of wood and is composed of cellulose, xylan, lignin, and small amounts of structural proteins and enzymes. Lignin molecules can interact directly or indirectly with cellulose, xylan and other polysaccharide molecules in the cell wall, increasing the mechanical strength and hydrophobicity of plant cells and tissues and facilitating the long-distance transportation of water in plants. MYBs (v-myb avian myeloblastosis viral oncogene homolog) belong to one of the largest superfamilies of transcription factors, the members of which regulate secondary cell-wall formation by promoting/inhibiting the biosynthesis of lignin, cellulose, and xylan. Among them, MYB46 and MYB83, which comprise the second layer of the main switch of secondary cell-wall biosynthesis, coordinate upstream and downstream secondary wall synthesis-related transcription factors. In addition, MYB transcription factors other than MYB46/83, as well as noncoding RNAs, hormones, and other factors, interact with one another to regulate the biosynthesis of the secondary wall. Here, we discuss the biosynthesis of secondary wall, classification and functions of MYB transcription factors and their regulation of lignin polymerization and secondary cell-wall formation during wood formation.

scholarly journals Functional Characterisation of the Poplar Atypical Aspartic Protease Gene PtAP66 in Wood Secondary Cell Wall Deposition

Forests ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 1002
Author(s):  
Shenquan Cao ◽  
Cong Wang ◽  
Huanhuan Ji ◽  
Mengjie Guo ◽  
Jiyao Cheng ◽  
...  
Keyword(s):  
Cell Wall ◽  
Secondary Cell Wall ◽  
Fluorescent Protein ◽  
Secondary Xylem ◽  
Populus Trichocarpa ◽  
Wood Formation ◽  
Protease Gene ◽  
Cellulose Content ◽  
Transcription Analysis ◽  
Functional Characterisation

Secondary cell wall (SCW) deposition is an important process during wood formation. Although aspartic proteases (APs) have been reported to have regulatory roles in herbaceous plants, the involvement of atypical APs in SCW deposition in trees has not been reported. In this study, we characterised the Populus trichocarpa atypical AP gene PtAP66, which is involved in wood SCW deposition. Transcriptome data from the AspWood resource showed that in the secondary xylem of P. trichocarpa, PtAP66 transcripts increased from the vascular cambium to the xylem cell expansion region and maintained high levels in the SCW formation region. Fluorescent signals from transgenic Arabidopsis plant roots and transiently transformed P. trichocarpa leaf protoplasts strongly suggested that the PtAP66-fused fluorescent protein (PtAP66-GFP or PtAP66-YFP) localised in the plasma membrane. Compared with the wild-type plants, the Cas9/gRNA-induced PtAP66 mutants exhibited reduced SCW thickness of secondary xylem fibres, as suggested by the scanning electron microscopy (SEM) data. In addition, wood composition assays revealed that the cellulose content in the mutants decreased by 4.90–5.57%. Transcription analysis further showed that a loss of PtAP66 downregulated the expression of several SCW synthesis-related genes, including cellulose and hemicellulose synthesis enzyme-encoding genes. Altogether, these findings indicate that atypical PtAP66 plays an important role in SCW deposition during wood formation.

scholarly journals PmMYB4, a Transcriptional Activator from Pinus massoniana, Regulates Secondary Cell Wall Formation and Lignin Biosynthesis

Forests ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1618
Author(s):  
Sheng Yao ◽  
Peizhen Chen ◽  
Ye Yu ◽  
Mengyang Zhang ◽  
Dengbao Wang ◽  
...  
Keyword(s):  
Cell Wall ◽  
Secondary Cell Wall ◽  
Paper Industry ◽  
Lignin Biosynthesis ◽  
Pinus Massoniana ◽  
Wood Formation ◽  
Genetically Engineered ◽  
Cellulose Synthesis ◽  
Masson Pine ◽  
Reading Frame

Wood formation originates in the biosynthesis of lignin and further leads to secondary cell wall (SCW) biosynthesis in woody plants. Masson pine (Pinus massoniana Lamb) is an economically important industrial timber tree, and its wood yield affects the stable development of the paper industry. However, the regulatory mechanisms of SCW formation in Masson pine are still unclear. In this study, we characterized PmMYB4, which is a Pinus massoniana MYB gene involved in SCW biosynthesis. The open reading frame (ORF) of PmMYB4 was 1473 bp, which encoded a 490 aa protein and contained two distinctive R2 and R3 MYB domains. It was shown to be a transcription factor, with the highest expression in semi-lignified stems. We overexpressed PmMYB4 in tobacco. The results indicated that PmMYB4 overexpression increased lignin deposition, SCW thickness, and the expression of genes involved in SCW formation. Further analysis indicated that PmMYB4 bound to AC-box motifs and might directly activate the promoters of genes (PmPAL and PmCCoAOMT) involved in SCW biosynthesis. In addition, PmMYB4-OE(over expression) transgenic lines had higher lignin and cellulose contents and gene expression than control plants, indicating that PmMYB4 regulates SCW mainly by targeting lignin biosynthetic genes. In summary, this study illustrated the MYB-induced SCW mechanism in Masson pine and will facilitate enhanced lignin and cellulose synthesis in genetically engineered trees.

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