Basic HTML Version
张强等
, 2011,
玳玳花咖啡酸甲基转移酶基因的电子克隆及序列分析
,
分子植物育种
Vol.9 No.14 (doi: 10.5376/mpb.cn.2011.09.0014)
1106
TGGGTTCAACCAGTTC
-
3'
和
COMTW3
：
5'
-
TCCA
GTAGAACAAAAGACTCAAGC
-
3'
，以玳玳花花瓣
的
cDNA
为模板，进行
RT-PCR
验证。获得的
PCR
产
物用
AXYGEN
公司的
DNA
凝胶回收试剂盒回收，克
隆至
pGEM-T easy
载体，转化大肠杆菌，菌落
PCR
筛选阳性克隆送上海博尚生物公司测序。
3.5
序列分析
用
DNAStar
软件
EditSeq
程序对玳玳花
CaCOMT
进行氨基酸组成、分子质量、等电点分析，并通过
NCBI
的
CDD (Conserved Domain Database)
数据库
(
网
址为
http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.
cgi)
预测
CaCOMT
蛋白的保守结构域；利用
DNAM-
AN6.0
和
MEGA4.0
软件对
CaCOMT
进行序列分析并
与数据库中植物
OMTs
进行同源序列比对、聚类分
析，
NJ
法构建系统进化树。
作者贡献
张强是本研究的实验设计和实验研究的执行人，并和
张晓伟完成数据分析，论文初稿的写作；李艳敏和王慧娟及
王利民参与实验设计，试验结果分析；孟月娥是项目的构思
者及负责人，指导实验设计，数据分析，论文写作与修改。
全体作者都阅读并同意最终的文本。
致谢
本研究由河南省基础与前沿技术研究计划项目
(082300430100)
资助。
参考文献
Dixon R.A., Chen F., Guo D., and Parvathi K., 2001, The
biosynthesis of monolignols: a ‘‘metabolic grid’’, or
independent pathways to guaiacyl and syringyl units,
Phytochemistry, 57: 1069-1084
Guo D.J., Chen F., Inoue K., Blount J.W., and Dixon R.A., 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
Humphreys J.M. and Chapple C., 2002, Rewriting the lignin
roadmap, Current Opinion in Plant Biology, 5: 224-229
Ibrahim R.K., Bruneau A., and Bantignies B., 1998, Plant O-met-
hyltransferases: molecular analysis, common signature and
classification, Plant Mol. Biol., 36: 1-10
Joshi C.P., and Chiang V.L., 1998, Conserved sequence motifs in
plant S-adenosyl-L-methionine-dependent methyltransferases,
Plant Mol. Biol., 37: 663-674
Li L., Popko J.L., Umezawa T., and Chiang V.L., 2000,
5-Hydroxyconiferyl aldehyde modulates enzymatic
methylation for syringyl monolignol formation, a new view
of monolignol biosynthesis in angiosperms, J. Biol. Chem.,
275: 6537-6545
Osakabe K., Tsao C.C., Li L., Popko J.L., Umezawa T., and
Carraway D.T., 1999, Coniferyl aldehyde 5-hydroxylation
and methylation direct syringyl lignin biosynthesis in
angiosperms, Proc. Natl. Acad. Sci. USA, 96: 8955-8960
Pak F.E., Gropper S., Dai W.D., Havkin-Frenkel D., and
Belanger F.C., 2004, Characterization of a multifunctional
methyltransferase from the orchid Vanilla planifolia, Plant
Cell Rep., 22: 959-966
Parvathi K., Chen F., Guo D., Blount J.W., and Dixon R.A.,
2001, Substrate preferences of O-methyltransferases in
alfalfa suggest new pathways for 3-O-methylation of
monolignols, Plant J., 25: 193-202
Schroder G., Wehinger E. and Schroder J., 2002, Predicting the
substrates of cloned plant O-methyltransferases,
Phytochemistry, 59: 1-8
Wang J., and Pichersky E., 1999, Identification of specific residues
involved in substrate discrimination in two plant O-methy-
ltransferases, Arch. Biochem. Biophys., 368: 172-180
Wang J., and Pichersky E., 1998, Characterization of S-adenosyl-
L-methionine: (iso) eugenol O-methyltransferase involved
in floral scent production in Clarkia breweri, Arch. Biochem.,
Biophys., 349: 153-160
Wang J., Dudareva N., Bhakta S., Raguso R.A., and Pichersky E.,
1997, Floral scent production in Clarkia breweri (Onagraceae)
II. Localization and developmental modulation of the enzyme
S-adenosyl-l-methionine: (iso) eugenol O-methyltransferase
and phenylpropanoid emission, Plant Physiol., 114:213-221
Wu S., Watanabe N., Mita S., Ueda Y., Shibuya M., and
Ebizuka Y., 2003, Two O-methyltransferases isolated from
flower petals of Rosa chinensis var. spontanea involved in
scent biosynthesis, J. Biosci. Bioeng., 96: 119-128
Zubieta C., Kota P., Ferrer J-L., Dixon R.A., and Noel J.P., 2002,
Structural basis for the modulation of lignin monomer
methylation by caffeic acid/5-hydroxyferulic acid 3/5-O-me-
thyltransferase, Plant Cell, 14: 1265-1277