茶树发根在LG0培养基和添加1βmg/L IBA的1/2MS培养基上生长迅速,但只在一个培养于LG0培养基的发根克隆中检测到高含量的茶氨酸,含量达到23.12βmg/g鲜重,高于未转化根的18.77βmg/g鲜重。除茶氨酸外,该发根克隆中也检测到高含量的天门冬氨酸、谷氨酸、天门冬酰胺、谷氨酰胺和精氨酸。发根克隆培养于添加1βmg/L IBA的1/2MS培养基,茶氨酸的含量则显著降低,说明培养基组成也是影响发根中茶氨酸的含量的因素之一。茶树发根中也含有少量的儿茶素类单体,主要为儿茶素和表儿茶素。
Hairy roots induced by Agrobacterium rhizogenes wild strain 15834 were propagated on LG0 medium and half MS medium containing 1βmg/L IBA to promote rapid growth. Although rapid growth was obtained in half MS media containing 1βmg/L IBA, high content of theanine (23.12βmg/g fresh weight) was only detected in one hairy root clone cultured on LG0 medium and higher than the non-transformed root (18.77βmg/g fresh weight). High content of aspartic acid, glutamic acid, glutamine, histidine and arginine were also detected in this hairy root clones. Medium constitute also affected the contents of amino acid in hairy root clone, the content of theanine were reduced in two hairy root clones when they were cultivated on half MS with 1βmg/L IBA. Low content of simple catechins, catechin (C) and epicatechin (EC), were also detected in hairy roots.
[1] Mukai T, Horie H, Goto T.Differences in free amino acids and total nitrogen contents among various prices of green tea[J]. Tea Research Journal. 1992(76):45~50. (in Japanese with English abstract).
[2] Luneja L R, Chu D C, Okubo T, et al. L-theanine-a unique amino acid of green tea and its relaxation effect in humans[J]. Trends in Food Science & Technology, 1999(10): 199~204.
[3] 郭丽芸, 刘毅, 贾晓娟, 等. 酶法拆分DL-茶氨酸及其分离纯化[J]. 茶叶科学, 2006, 26(1): 31~36.
[4] Orihara Y, Huruya T.Production of theanine and otherγ- glutamyl derivatives by Camellia sinensis cultured cells[J]. Plant Cell Reports, 1990(9): 65~68.
[5] Matsuura T, Kakuda T, Kinoshita T, et al. Theanine formation by tea suspension cells[J]. Biosci Biotechnol Biochem, 1992(56): 1175~1181.
[6] Takihara T, Matsuura T, Sakane I, et al. Effects of plant growth regulators and carbon sources on theanine formation in callus culture of tea(Camellia sinensis)[J]. Biosci Biotechnol Biochem. 1994(58): 1519.
[7] Kawagishi H and Sugiyama K. Facile and large-scale synthesis of L-theanine[J]. Biosci Biotechnol Biochem, 1992, 56: 689.
[8] 钟俊辉, 陶文沂. 愈伤组织培养及茶氨酸的积累[J]. 无锡轻工大学学报, 1997, 16(3): 1~7.
[9] 袁弟顺, 林丽明, 孙威江, 等. 不同品种茶树愈伤组织的培养与茶氨酸的积累[J], 福建农林大学学报(自然科学版), 2004, 33(2): 178~181.
[10] 陈瑛, 陶文沂. 几种激素对茶愈伤组织合成茶氨酸的影响[J]. 无锡轻工大学学报, 1998, 17(1): 74~77.
[11] Giri A and Narasu M L. Transgenic hairy roots: recent trends and applications[J]. Biotechnology Advances, 2000(18): 1~22.
[12] Konishi S and Takahashi E. The metabolism and regulation of theanine and related compounds in tea plant[J]. Journal of the Science of Soil and Manure, Japan, 1969(40):479~484.(in Japanese)
[13] Zehra M, Banerjee S, Mathur A K, et al. Induction of hairy roots in tea(Camellia sinensis L.) using Agrobacterium rihizogenes[J]. Current Science, 1996(70): 84~86.
[14] 奚彪, 刘祖生, 梁月荣, 等. 发根农杆菌介导的茶树遗传转化[J]. 茶叶科学, 1997(17)(增刊): 155~156.
[15] 高秀清, 成浩, 牛爱军. 茶树毛状根的诱导[J]. 特产研究, 2004, 26(2): 30~32.
[16] 张广辉, 梁月荣, 陆建良. 发根农杆菌介导的茶树发根高频诱导与遗传转化[J]. 茶叶科学, 2006, 26(1): 1~10.
[17] Torrgrosa L, Bouquet A.Agrobacterium rhizogenes and A. tumefaciens co-transformation to obtain grapevine hairy roots producing the coat protein of grapevine chrome mosaic nepovirus[J]. Plant Cell, Tissue Organ culture, 1997(49): 53~62.
[18] Peris-Vicente J, Gimeno-Adelantado J V, Domenech-Carbo M T, et al. Characterization of proteinaceous glues in old paintings by separation of the o-phtalaldehyde derivatives of their amino acids by liquid chromatography with fluorescence detection[J]. Talanta, 2006(68): 1648~1654.
[19] George E M, Yuerong L, Jianliang L.Expression of basic genes involved in tea polyphenol synthesis in relation to accumulation of catechins and total tea polyphenols[J]. Journal of Science Food Agriculture, 2006(86): 459~464.
[20] 王关林, 方宏筠. 植物基因工程(第二版)[M]. 北京:科学出版社, 2002.
[21] Matsuura T, Kakuda T, Kinoshita T, et al.Production of theanine by callus culture of tea [A]. In: Proceedings of the International Symposium on Tea Science, Shizuoka. Japan, 1991: 432~435.
浙公网安备 33019902000101号 
