β-glucosidase and β-primeverosidase are considered the most important genes related to made tea aroma. An absolute quantification method for gene expression in tea plant using real-time PCR analysis was established and, the expression of β-glucosidase and β-primeverosidase genes in different leaves of Longjing 43 young shoot were determined using this method. The results indicate that the fourth leaf of the ‘five and a bud’ shoot has the highest copy number of β-glucosidase mRNA, which is 2.86E+08 copies/μl, the 4th leaf>the 3rd leaf>the 5th leaf>the 2nd leaf>one and a bud. And the highest β-primeverosidase expression is in the one and a bud, corresponding to 4.31E+06 copies/μl, one and a bud>the 2nd leaf>the 3rd>the 4th leaf> the 5th leaf. The expression intensity and pattern of these genes, which are closely related to tea aroma, are different in the young shoot. The results illuminate that the new developed method could be effectively used to detect gene expression quantitatively in tea plant.
ZHAO Li-ping
,
CHEN Liang
,
WANG Xin-chao
,
YAO Ming-zhe
. Quantitative Detection of β-glucosidase, β-primeverosidase Gene Expressions in Different Leaves of Tea Plantby Real-Time PCR Analysis[J]. Journal of Tea Science, 2006
, 26(1)
: 11
-16
.
DOI: 10.13305/j.cnki.jts.2006.01.002
[1] Takeo T.Production of linalool and geraniol by hydrolytic breakdown of bound forms in disrupted tea shoots[J]. Phytochemistry, l981, 120(9): 2l45~2147.
[2] Guo W, Yamauchi K, Watanabe N, et.al. A primeverosidase as a main glycosidase concerned with the alcoholic aroma formation in tea leaves[J]. Biosci Biotechnol Biochem 1995 59: 962~964.
[3] 夏涛, 童启庆, 董尚胜, 等. 红茶萎凋发酵中β-葡萄糖苷酶的活性变化[J]. 茶叶科学, l996, l6(1): 63~66.
[4] 骆耀平, 董尚胜, 童启庆, 等. 7个茶树品种新梢生育过程中β-葡萄糖苷酶活性变化[J]. 茶叶科学, l997, l7(增刊): l04~l07.
[5] 江昌俊, 李叶云, 王朝霞. 茶树鲜叶中β-葡萄糖苷酶提取条件的研究[J]. 南京农业大学学报, 2000, 23(2): 93~96.
[6] Mizutani M, Nakanishi H, Ema J, et al. Cloning of β-primeverosidase from tea leaves, a key enzyme in tea aroma formation[J]. Plant Physiology, 2002, 130: 2164~2176.
[7] Chen L, Zhao LP, Gao QK.Generation and analysis of expressed sequence tags from the tender shoot cDNA library of tea plant (Camellia sinensis)[J]. Plant Sci, 2005, 168(2): 359~363.
[8] Livak K J, Flood S J, Marmaro J, et al. Oligonucleotides with fluorescent dyes at opposite ends provide aquenched probe system useful for detecting PCR product and nucleic acid hybridization[J]. PCR Methods Appl, 1995, 4(6), 357~362.
[9] Gibson U E, Heid C A, Williams P M.A novel method for real time quantitative RT-PCR[J]. Genome Res, 1996, 6(10): 995~1001.
[10] Heid C A, Stevens J, Livak K J, et al. Real time quantitative PCR[J]. Genome Res, 1996, 6(10): 986~994.
[11] Wong Y W, Sia G M, Too H P.Quantification of mouse glial cell-line derived neurotrophic factor family receptor alpha 2 alternatively spliced isoforms by real time detection PCR using SYBR Green I[J]. Neuroscience Letters, 2002, 320: 141~145.
[12] 苏建中, 董强刚, 黄进肃, 等. 肺癌组织中白介素-8基因表达的实时定量PCR研究[J]. 肿瘤, 2004, 24(1): 3~5.
[13] Boxus M, Letellier C, Kerkhofs P.Real Time RT-PCR for the detection and quantitation of bovine respiratory syncytial virus[J]. Journal of Virological Methods. 2005, 125: 125~130.
[14] http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=nucleotide&cmd=search&term=AB088027
[15] Becker K, Pan D, Whitley CB.Real-time quantitative polymerase chain reaction to assess gene transfer[J]. Hum. Gene Ther. 1999, 10(15): 2559~2566.
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