毛竹PeIRX10基因在木聚糖合成中的功能

doi:10.11707/j.1001-7488.20161110

Abstract

Abstract: [Objective] PeIRX10 gene was cloned and functionally analyzed for further understanding of the molecular mechanism of xylan synthesis in Phyllostachys edulis.[Method] According to the sequence information from the bamboo genome database,primers were designed. A homologous gene from P.edulis playing critical role in xylan synthesis in Arabidopsis thaliana was cloned and named as PeIRX10. The nucleotide sequence and encoded amino acid sequence of PeIRX10 were analyzed using bioinformatics methods. The expression pattern of PeIRX10 in different organs (root, stem, leaf, shoot and flower) was detected by qRT-PCR. The binary vector of PeIRX10 overexpression was constructed using Gateway technology and then transformed into the A.thaliana irx10l(-/-) irx10(+/-) mutant plants by floral dip. Transgenic plants with double mutant background were selected by BASTA and PCR identification. Finally, the conserved function of PeIRX10 in thexylan synthesis was done by phenotype observation, stem section, sugar composition analysis and xylan immunolocalization. [Result] An ORF sequence of PeIRX10(PH01004923G0080,http://www.bamboogdb.org/)from P. edulis was cloned with a length of 1 251 bp,encoding 416 amino acids, with a predicted molecular weiqht of about 46 821 Da and pI of about 6.26. The protein sequence encoded by PeIRX10 exhibited relatively high similarity to those of other plants (>80%), indicating that it belongs to GT47 family. qRT-PCR showed that PeIRX10 was highly expressed in shoots and culms of P. edulis. After overexpression of PeIRX10, complementation of irx10l(-/-)irx10(-/-) double mutant can restore the phenotype with evidence of normal plant height, stem diameter, leaf numbers and sizes. Moreover, the analysis of monosaccharides indicated that the content of xylose and other monosaccharides, such as arabinose, fucose, galactose and glucose in the complementation plants was not significantly different to those of the wild type. Immunostaining of cross sections detected by LM10 labeling revealed strong signals in xylem cells in the irx10l(-/-)irx10(-/-) double mutant when overexpressedby PeIRX10. [Conclusion] Overexpression of PeIRX10 in the irx10l(-/-)irx10(-/-) double mutant rescued the plant growth phenotype and restored the secondary wall thickness, which suggested that the PeIRX10 gene have a function similar to the IRX10 geneof A. thaliana and play an important role in the synthesis of xylan.

Key words: Phyllostachys edulis, xylan, secondary wall synthesis, PeIRX10

CLC Number:

Wang Jie, Li Jun, Tong Tingting, Shao Xiangjun, Song Lili, Wu Aimin. Functions of PeIRX10 Gene from Phyllostachys edulis in Xylan Synthesis[J]. Scientia Silvae Sinicae, 2016, 52(11): 79-87.

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Functions of PeIRX10 Gene from Phyllostachys edulis in Xylan Synthesis

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