【作者】 胡亚东；

【导师】 陈昆松；

【作者基本信息】 浙江大学 ， 植物学， 2004， 硕士

【摘要】 花青苷是存在于植物体内的一类天然色素，果实发育过程中的花青苷积累常常受到内、外环境的影响，其含量水平直接影响果实表观品质和商品性。本试验以“玉露”桃和“湖景蜜露”桃（Prunus persica L．）果实为试材，采用普通桃果袋（CK）、105^#果袋（日本引进）和108^#果袋（日本引进）三种不同纸质果袋套袋处理，研究其对桃果实花青苷形成的影响及其相关生理学基础。主要研究结果为： 1．针对桃果实的生物学特性，改进了花青苷提取、检测方法。研究确定了花青苷提取液为0.05％盐酸-甲醇，HPLC检测的梯度洗脱条件：流动相A：0.02 M醋酸铵缓冲液（pH 3.0），流动相B：甲醇：洗脱梯度为：0-5 min，流动相B从0-25％；5-15 min，使B从25-65％；从15-17 min，使B从65-100％，并维持2 min（从17到19 min）；从19-22 min使B从100％变为0，并维持3 min（即到第25 min结束一个循环）。经改进的方法操作简便易行，方法回收率高（94.37％），检测限（S／N=3）为0.2 μg／g，RSD（n=5）为2.94％，峰面积与浓度在0.0625-0.25 mg／mL范围内线性关系(r=0.997^**)。 2．桃果实中的花青苷以矢车菊-3-葡萄糖苷（Cyanidin-3-glu）为主，约占98％。随着果实逐渐发育成熟，果皮中PAL活性趋于上升，花青苷积累增加，同时叶绿素降解。果实开始着色（30／6）之前花青苷含量变化平稳，三个处理之间无显著差异；30／6之后，花青苷含量增加迅速，108^#处理果实的花青苷积累显著高于105^#和对照果；与对照相比，105^#处理对果实花青苷合成的促进效应主要表现在采收前3 d。 3．三种纸质果袋处理之间PAL活性存在显著差异，其中以108^#纸质果袋处理的PAL活性最高，105^#次之，CK最低。 4．无论在果皮中，还是在果肉中，30／6之前，三种纸质果袋处理的蔗糖增加较快，30／6之后转为缓慢上升，果糖和葡萄糖在果实发育过程的变化相对平缓；采收时（16／7）105^#、108^#果袋处理的果实中葡萄糖和果糖均高于CK。 5．果实中花青苷水平与蔗糖、果糖、葡萄糖和PAL活性均呈极显著正相关关系。在果皮中，花青苷积累与蔗糖、果糖、葡萄糖、PAL活性变化的相关系数分别为0.7928^**、0.8383^**、0.8078^**和0.8643^**；在果肉中，花青苷积累与蔗糖、果糖、葡萄糖变化的相关系数分别为0.7945^**、0.7924^**、0.8663^**。摘要 6.尽管108“果袋处理的桃果实具有较高的队L活性、花青昔积累，但由于采用了双色设计（顶部白色，底部橙色），导致果实着色不够均匀，因此在改善果实表观品质方面，105”果袋处理整果着色均一程度明显优于108”和CK果袋，综合效果以105“果袋处理最佳。 7.采前去袋有利于花青营积累，在采前10d内，套袋果实的去袋时间越早，队L活性越高，花青普积累越多。 8.三种不同纸质果袋透光率和光谱分析结果显示，108“果袋的总透光率和各不同波长范围透光率均显著大于105“果袋和CK果袋，同时也表现出相对较高的花青昔积累。从105”果袋和CK果袋的比较来看，在红光（610一70Onm）和远红光（700·80Onm）区域透光率与CK相比差异不明显;而在近紫外光（300一400nm）和蓝光（400一510 nm）区域内，105”果袋透光率低于CK果袋。推测较低的近紫外光和蓝光透光率可能更有利于桃果实着色均匀。更多还原

【Abstract】 Anthocyanins are a kind of natural pigments in plant, and play an important role in fruit coloration. Using "Yulu" and "Hujingmilu" (Ptiinus persica L.) peach fruits as materials in present experiment, we studied the role of peach fruit anthocyanin biosynthesis with different paper material bags, CK(the bag used in local), 105 and 108, analyzed related physiological changes, and postulated the regulation mechanism of anthocyanin. The obtained results are described as follows:1. According to the biological characteristics of peach fruit, we improved a method for anthocyanin extracting and detecting. An effective, feasible system for anthocyanin analysis was built, with extracted by 0.05% methanol-HCl solution for 24 hours at 4 and detected by the gradient elution of high-performance liquid chromatography (HPLC) with the mobile phase A: 0.02 M ammonium acetate buffer (pH 3.0) and the mobile phase B: methanol. The gradient elution was as following process: from 0 B to 25% B in 5 min, 25% B to 65% B in 10 min, 65% B to 100% B in 2 min, 100% B for 2 min (from 17-19 min), and then from 100% B to 0, and 0 B for 3 min (end an recycle in 25th min). Under 1 mL/min flow-rate condition, the limit determination in this system (S/N=3) was 0.2ug/g. There was a significant relationship between HPLC peak area and anthocyanin concentration from 0.0625 to 0.25 mg/mL was observed, with the correlative coefficient of 0.997 and RSD=2.94%.2. The results showed that the anthocyanin mainly was Cyanidin-3-glu in peach fruit peel accounting for nearly 98%. During the development of peach fruit, the activity of PAL tended to increase with the accumulation of anthocyanin and was accompanied with chlorophyll degradation. Prior to peach fruit coloration (30/6), the anthocyanin content changed steadily and there was no significant difference among three bags. However, after date of 30/6 theanthocyanin accumulated rapidly and markedly, the fruit with 108 bag whose anthocyanin content was obviously higher than that with 105# and CK. The benefit effect on anthocyanin synthesis with 105# bag was appeared 3 d before harvest when compared with CK.3. The significant difference in PAL activity of peach fruit existed among 3 various bags treatments. The PAL activity was highest in fruit with 108# bag, followed by 105# bag, and37lowest in CK.4. Either in peach fruit peel or flesh tissue, the sucrose content increased rapidly (before 30/6) and then accumulated steadily. As for fructose and glucose, whose content was no obvious change during fruit development. However, 105 and 108# bags could more effectively promote fructose and glucose accumulation than CK.5. The relationships between sugar content, PAL activity and anthocyanin accumulation were significant positive correlation. The correlative coefficients between anthocyanin accumulation and sucrose, fructose, glucose content and PAL activity were 0.7928**/0.7945** (in fruit peel/in fruit flesh), 0.8383**/0.7924**, 0.8078**/0.8663** and 0.8643**, respectively.6. For the fruit with 105* bag, which produced more even coloration than that with 108# bag and CK, thus showed better appearance quality. Although higher PAL activity, more anthocyanin accumulation were detected in peach fruit with 108# bagging, it tended to result in uneven fruit coloration due to the bag with two kinds of color (white and orange) design. So, 105* bag treatment was optimum for improving peach fruit coloration.7. The anthocyanin content in bagging peach fruit was obviously affected by the time for bag removing. Within 10 d before harvest, earlier removing resulted in high PAL activity and anthocyanin content.8. It was postulated that lower adjacent ultraviolet and blue light were more related to even coloration of the fruit. The detecting results of light penetration rate with three different bags showed that 108# bag allowed higher total light and different spectrum light penetration than 105* bag and CK. Moreover, the peach fruit with 108# bag had more anthocyanin更多还原