| The cDNA encoding full-length of CCoAOMT1, CCoAOMT2, C4H, 4CL genes were cloned from cDNA prepared from tissue culture seedings of Moso bamboo using RT-PCR and RACE methods. Its nucleotide sequences and the encoded amino acid sequences were analyzed. The important component to explicit the functions of gene is to carry out the study of gene expression with tissues,for instance, winter-shoot, whole spring-shoot, top-springshoot, mid-springshoot, base-shoot, leaf, leaf sheath, root, 1-year-old stem, 2-year-old stem and 3-year-old stem, using technology of fluorescence quantitative real-time PCR(QRT-PCR). To determine their expression properities and confirm their involvement in the lignin biosythesis in bamboo genes.(1)The whole open reading frame of CCoAOMT1 gene is 1045 bp encoding 262 amina acids. From the analysis of the sequence through the website of SMART and ExPASy against NCBI database, it is found that CCoAOMT1 is belong to p450 family and there are nine EGF-like domain signature 1, fifteen VWFC domain signature,one Integrins beta chain cysteine-rich domain signature, two Integrins beta chain cysteine-rich domain signature, two 2Fe-2S ferredoxins, iron-sulfur binding region signature, one 4Fe-2S ferredoxins, iron-sulfur binding region signature, one Anaphylatoxin domain signature, two Thiolases active site, four C-terminal cystine knot signatureone Insulin-like growth factor- binding protein (IGFBP) N-terminal domain signature.The amino acid sequence showed high similarity with the corresponding genes from monocots Bambusa oldhamii, oryza and zea mays at 99%, 88% and 88%,respectively. Expression analysis of CoAOMT1 by real time quantitative PCR showed the expression level in different tissues from high to low was in the following order: top-springshoot and mid-springshoot, base-shoot, leaf sheath, winter-shoot , 2-year-old stem, 1-year-old stem, root, whole spring-shoot, leaf, and the lowest expression level in the 3-year-old stem. (2)The whole open reading frame of CCoAOMT2 gene is 1131 bp encoding 262 amina acids. From the analysis of the sequence through the website of SMART and ExPASy, it is found that CCoAOMT2 is also belong to p450 family and there are nine EGF-like domain signature 1, fifteen VWFC domain signature, one Integrins beta chain cysteine-rich domain signature, four 2Fe-2S ferredoxins, iron-sulfur binding region signature, two 4Fe-2S ferredoxins, iron-sulfur binding region signature, three Anaphylatoxin domain signature, two Thiolases active site, three C-terminal cystine knot signature.The amino acid sequence showed high similarity with the corresponding genes from monocots oryza and zea mays at 88% and 87%,respectively. Expression analysis of CoAOMT2 by real time quantitative PCR showed the expression level in different tissues from high to low was in the following order: top-springshoot and mid-springshoot, winter-shoot, base-shoot, 2-year-old stem, 3-year-old stem, whole spring-shoot, and the lowest expression level in the leaf sheath, 1-year-old stem, root, leaf.(3)The whole open reading frame of C4H gene is 1006 bp encoding 502 amina acids. From the analysis of the sequence through the website of SMART and ExPASy, it is found that C4H is belong to p450 family and there are twelve EGF-like domain signature 1, two C-terminal cystine knot signature, twelve VWFC domain signature, one Integrins beta chain cysteine-rich domain signature, six 2Fe-2S ferredoxins, iron-sulfur binding region signature, one Thiolases active site, one 4Fe-2S ferredoxins, iron-sulfur binding region signature, one Mammalian defensins signature, three Anaphylatoxin domain signature.The amino acid sequence showed high similarity with the corresponding genes from monocots Sorghum Moench, oryza and zea mays at 88%, 88% and 87%,respectively. Expression analysis of C4H by real time quantitative PCR showed the expression level in different tissues from high to low was in the following order: winter-shoot, top-spring shoot, mid-spring shoot, leaf, 3-year-old stem, root, leaf sheath, 2-year-old stem, whole spring-shoot, 1-year-old stem, the base-shoot.(4) The whole open reading frame of 4CL gene is 1792 bp encoding 543 amina acids. From the analysis of the sequence through the website of SMART and ExPASy, it is found that 4CL is belong to AMP-binding family and there are ten EGF-like domain signature 1, two C-te rminal cystine knot signature, twenty VWFC domain signature, four Integrins beta chain cystei ne-rich domain signature, six 2Fe-2S ferredoxins, iron-sulfur binding region signature, two 4Fe-2S ferredoxins, iron-sulfur binding region signature, three Mammalian defensins signature, two Anaphylatoxin domain signature, two Insulin-like growth factor-binding protein (IGFBP) N-terminal domain signature, one Janus-faced atracotoxin (J-ACTX) family signature.The amino acid sequence showed high similarity with the corresponding genes from monocots Neosinocalamus affinis, oryza and zea mays at 100%, 97% and 83%,respectively. Expression analysis of 4CL by real time quantitative PCR showed the expression level in different tissues from high to low was in the following order: winter-shoot, mid-springshoot, top-springshoot, leaf, 2-year -old stem, 3-year-old stem, leaf sheath, root, 1-year-old stem, base-springshoot, the lowest expr ession level in the whole spring-shoot. This indicates that the cloned CCoAOMT1,CCoAOMT2,C4H,4CL involved in the cell wall deposition during the development of vascular tissues thus could be used in modulation of lignin biosynthesis in bamboo through genetic engineering.
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