采用mRNA差别显示的方法对茶树冷驯化过程中基因差异表达进行了初步研究,从58条差异片段得到稳定扩增的差异片段23条,用RT-PCR的方法鉴定其假阳性,得到5个阳性片段,其中3个片段在冷驯化过程中表达量增加,另外2个片段的表达量降低。经过BLAST比对分析,其中一个片段序列与高粱反式玉米素-O-葡糖基转移酶同源性较高;一个与黄瓜叶绿体合成相关蛋白基因同源性较高;一个为茶树核酮糖-1,5-双磷酸羧化酶/加氧酶小亚基基因片段;一个与细胞色素b亚基有较高同源性;还有两个片段序列比对没有同源序列,可能为新基因。
Differential gene expression during cold acclimation was studied by mRNA differential display reverse transcription-PCR (DDRT-PCR) in tea plant (Camellia sinensis). Twenty-three differentially expressed cDNA fragments were steadily re-amplified. Five fragments were confirmed by RT-PCR analysis. Among them, three were up-regulated while other two were down-regulated during the cold acclimation process. Sequencing of three differential fragments revealed homology to putative cis-zeatin-O-glucosyl-transferase of Sorghum （Sorghum bicolor）, chloroplast gene for chloroplast product of cucumber(Cucumis sativus),ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcS) mRNA of tea plant(Camellia sinensis), respectively, while the other two showed no homologous to known genes.
[1] 王新超, 杨亚军. 茶树抗性育种研究现状[J]. 茶叶科学, 2003, 23(2): 94~98.
[2] 杨亚军, 郑雷英, 王新超. 冷驯化和ABA对茶树抗寒力及其体内脯氨酸含量的影响[J]. 茶叶科学, 2004, 24(3): 177~182.
[3] 杨亚军, 郑雷英, 王新超. 低温对茶树叶片膜脂脂肪酸和蛋白质的影响[J]. 亚热带植物科学, 2005, 34(1): 6~10.
[4] Thomashow M F.Plant cold acclimation: freezing tolerance genes and regulatory mechanisms[J]. Annu Rev Plant Physiol Plant Mol Biol, 1999, 50: 571~599.
[5] Kalberer S, Wisniewski R, Arora R.Deacclimation and reacclimation of cold-hardy plants: Current understanding and emerging concepts[J]. Plant Science, 2006, 171: 3~16.
[6] Liang P, Pardee A B.Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction[J]. Science. 1992, 257:967~971.
[7] 童建华, 刘华英, 鲁旭东. mRNA差异显示技术及其在植物研究中的应用[J]. 农业与技术, 2004, 24(4): 109~111.
[8] 李卫, 孙中海, 章文才,等. 柑橘抗寒育种早期鉴定的一种指标[J]. 植物学报, 1998, 40(9): 827~830.
[9] Weiser C J.Cold resistance and injury in woody plant[J]. Science, 1970, 169: 1269~1273.
[10] Guy C L, Niemif K J, Brambl R.Altered gene expression during cold acclimation of spinach[J]. Proc Nati Acad Sci USA, 1985, 82(11): 3673~3677.
[11] Ruth C M, Machteld C M, Jeffrey E H, et al. A maize cytokinin gene encoding an O-glucosyltransferase specific to cis-zeatin[J]. Proc Nati Acad Sci USA, 2001, 98: 5922~5926.
[12] 师晨娟, 刘勇, 荆涛. 植物激素抗逆性研究进展[J]. 世界林业研究, 2006, 19(5): 21~26.
[13] 杨广容, 倪金卫. 冬秋季茶树抗寒性对光合呼吸作用的影响[J]. 福建茶叶, 2002, 4: 14~15.
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