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儿茶素是茶树重要的次生代谢产物,也是茶叶的重要功能物质,具有多种药理活性。因此正确评价茶叶及其制品中儿茶素含量,了解儿茶素生物合成及其调控手段具有重要的意义。利用香草醛盐酸比色法、HPLC等方法,研究蔗糖诱导子处理对茶儿茶素生物合成的影响,并通过qRT-PCR技术分析蔗糖处理对茶树儿茶素生物合成过程中基因表达的影响。结果显示,无论是茶树愈伤组织还是茶树幼苗经蔗糖处理后其儿茶素含量及组分都有所增加,但是高浓度的蔗糖会抑制茶树愈伤组织的生长,特别是在液体培养的早期添加蔗糖。添加适当浓度的蔗糖能增加茶树愈伤组织中非酯型儿茶素C和EC的含量;蔗糖处理后茶树幼苗中非酯型儿茶素EGC和EC含量明显上升,而酯型儿茶素EGCG相对含量有所下降。qRT-PCR分析显示蔗糖诱导了茶树幼苗中C4H、CHS、CHI、F3H、LAR、ANR、DFR、F3’H、F3’5’H基因表达,其中变化最明显的是F3H,其次是ANR、CHS。这进一步证实了F3H、ANR为茶树儿茶素合成的关键基因。
Catechin is an important secondary metabolite of tea, tea is also an important functional material, with a variety of pharmacological activities. Therefore, a correct evaluation of tea and its products in catechins content, understanding of catechin biosynthesis and its regulatory means of great significance. The effects of sucrose elicitor treatment on the biosynthesis of tea catechins were studied by vanillin-HCl colorimetric and HPLC methods. The effects of sucrose treatment on the gene expression during the biosynthesis of tea catechins were analyzed by qRT-PCR. The results showed that the content of catechins and their components increased after both the callus of tea tree and the seedlings of tea tree were treated with sucrose, but the high concentration of sucrose inhibited the growth of tea callus, especially in the early stage of liquid culture Add sucrose. Adding proper concentration of sucrose increased the content of non-esterified catechins C and EC in the callus of tea plant; the content of non-esterified catechins EGC and EC in tea seedlings increased significantly after sucrose treatment, while the content of ester-type catechin EGCG Relative content decreased. qRT-PCR analysis showed that sucrose induced the expression of C4H, CHS, CHI, F3H, LAR, ANR, DFR, F3’H and F3’5’H in tea seedlings. F3H was the most obvious change, followed by ANR, . This further confirmed that F3H, ANR is a key gene in the synthesis of tea catechins.