【作者】 秦栋；

【导师】 翟衡； 束怀瑞；

【作者基本信息】 山东农业大学 ， 果树学， 2009， 博士

【摘要】 本实验于2006年～2009年在山东泰安小津口苹果园和山东农业大学园艺实验站进行,以能避过晚霜危害,比金冠、富士、瑞林等晚萌发20天左右的酸王苹果(Malus domestica cv.‘Avrolles’)为试材,以瑞林(Malus domestica cv.‘Judeline’)作对照,探讨了酸王自然休眠解除至萌发过程中的生态学(光照和温度)和生理生化变化(内源激素、呼吸代谢、抗氰呼吸、活性氧代谢和酚类物质等),主要结果如下:1、利用光照培养箱清水扦插法和水分状态法界定酸王自然解除休眠的时间为1月底(泰安),瑞林为12月底,酸王比瑞林晚一个月左右,用0～7.2℃模型估算酸王解除自然休眠的需冷量为1200～1500 h,瑞林为600～850 h,酸王是瑞林的2倍左右。物候观察表明,酸王在泰安4月中下旬萌发,瑞林在3月中下旬萌发,以5℃作为苹果休眠解除积累有效积温的起始温度,估算酸王萌发所需有效积温(∑T,℃)为320～380℃,瑞林(∑T)为100～150℃,酸王是瑞林的3倍左右;以生长度小时(GDH℃)计算需热量,酸王萌发时(露绿)需热量为7800～9300 GDH℃,瑞林为4500～5500 GDH℃,酸王是瑞林的1.7倍左右,表明酸王萌发不但需要较高的需冷量,而且需要较高的热量。补充酸王的需冷量,能减少萌发的需热量,但并非对等关系,补充5d和10d需冷量仅能替代80%、60%的需热量,日照长短对萌发的影响不大。2.内源激素测定结果表明,随着自然休眠的解除,芽内生长促进型激素GA3、IAA和ZR逐步升高,生长抑制型激素ABA逐步下降。在整个萌发过程中,酸王芽内GA3的浓度显著高于同时期的瑞林,萌发前GA3及GA3/ABA分别是瑞林的1.55和2.08倍;酸王芽内ZR的浓度积累上升时间比瑞林长26d, ZR及ZR/ABA分别是瑞林的2.12倍和1.55倍,(GA3 + IAA + ZR)/ABA是瑞林的1.59倍,表明酸王萌发需要较高浓度的GA3、ZR等生长促进型激素;萌发时较高浓度的GA3和ZR使酸王的GA3/ABA、ZR/ABA、(GA3 + IAA + ZR)/ABA比值显著高于瑞林。3、酸王的总呼吸和EMP途径呼吸强度低于瑞林,最高点分别是瑞林的84.6%和84.7%;PPP途径增加的时间比瑞林长21d,且运行活性高于瑞林,上升幅度是瑞林的2.02倍;随着芽自然休眠解除和萌发的进行,作为呼吸底物的可溶性糖含量升高,淀粉含量下降,酸王芽内可溶性糖含量显著低于同时期的瑞林。4、加入电子传递途径的抑制剂KCN和SHAM,芽内呼吸减弱,表明芽内同时存在细胞色素途径和抗氰呼吸途径,且细胞色素途径呼吸及细胞色素途径呼吸对总呼吸的贡献率(ρ’Vcyt/Vt)始终高于抗氰呼吸,表明细胞色素途径仍是电子传递的主要途径。酸王芽内抗氰呼吸的速率、抗氰呼吸对总呼吸的贡献率(ρValt/ Vt)及抗氰呼吸的实际运行系数(ρ)在整个萌发过程中始终高于瑞林,表明酸王芽的萌发需要较高活性的抗氰支路传递电子并提供抵御外界低温的能量。5、在萌发的过程中,酸王芽内O2-的产生速率及H2O2的水平均低于瑞林,抗氧化酶活性及抗氧化物质含量也低于瑞林,表明酸王芽内活性氧代谢水平低于瑞林。萌发过程中O2-.主要来自于线粒体呼吸电子渗漏,POD主要在萌发前期清除H2O2,而CAT主要在萌发后期;抗氧化物质谷胱甘肽(GSH)和抗坏血酸(AsA)在萌发过程中逐步上升,表明在清除活性氧的过程中启动了抗坏血酸-谷胱甘肽循环。6、随着休眠的加深,芽内总酚含量及PAL酶活性上升;随着休眠解除总酚含量下降,PAL酶活性下降,PPO酶活性上升,表明总酚是芽萌发的抑制物。酸王芽内的总酚含量比瑞林高25%左右,在萌发过程中PAL和PPO酶活性高于瑞林,表明酸王的晚萌发与总酚的代谢有关。利用液相色谱检测到六种单酚:没食子酸,表儿茶素、槲皮素、根皮素、阿魏酸和绿原酸,其中槲皮素含量最高,其次是(-)表儿茶素,没食子酸含量最少,酸王芽内(-)表儿茶素含量是瑞林的1.31倍,根皮素含量是瑞林的76.8%。阿魏酸随着休眠的加深而升高,并随萌发而降低,是萌发的抑制物。更多还原

【Abstract】 This experiment was carried out in‘xiaojinkou’apple orchard of Tai’an and Horticultural experiment station of Shandong Agricultural University from 2006 to 2009. Avrolles (Malus domestica), which germinates 20d later than some other apple varieties (such as Golden Delicious, Fuji, Judeline(Malus domestica), etc.) and can avoid late frost damage, was used to research the ecological features (light and temperature) and physiological and biochemical changes (endogenous hormones, respiratory metabolism and phenolic substances, etc.) from natural dormancy releasing to germination. Taking Judeline for control, the main results were as follows:1. Methods of water cultivation in the artificial lighting incubator and water state determination were used to test the dormancy state of Avrolles and Judeline. The date of bud dormancy releasing of Avrolles was at the end of January, while Judeline at the end of December, one-month difference between the two varieties. Using 0～7.2℃model to estimate the chilling requirement of natural dormancy releasing, Avrolles’chilling requirement (1200～1500 h) was, about two times higher than that of Judelin (600～850 h). The phenological observation showed that Avrolles geminated at mid-and-late April, while Judeline at mid-and-late March. 5°C was usually treated as the initial temperature to calculate the efficiently accumulative temperature of germination. The sum of efficiently accumulative temperature (∑T) was 320～380°C and 100～150℃for Avrolles and Judeline, respectively. When growth degree hours (GDH℃) was used to calculate the heat requirement of germination, Avrolles was 7800～9300 GDH℃at the green opened in buds, which was 1.7 times higher than that in Judeline. These showed that Avorlles required not only higher chilling requirement, but also higher heat requirement. The heat requirement of germination in Avrolles reduced when the chilling requirement was added, but the relation was not reciprocity, adding more 5d and 10d chilling requirement, the heat requirement only replaced 80% and 60%, respectively. The length of sunshine had little effect on germination.2. The content of endogenous hormone in buds showed that growth-promoting hormones, such as GA3, IAA and ZR, increased while growth-inhibiting hormones, such as ABA, declined, with the natural dormancy released. During the procedure of germination, GA3 concentration of Avrolles was significantly higher than that of Judeline at the same period. In addition, GA3 concentration and the ratio of GA3/ABA were 1.55 and 2.08 times higher than that of Judeline before germination, respectively. The time for ZR concentration increasing in Avrolles was 26d longer than in Judeline. The value of ZR and ZR/ABA were 2.12 and 1.55 times higher than that of Judeline, respectively, while the ratio of GA3 + IAA + ZR)/ABA was 1.59 times as large as that of Judeline. That indicated that a relatively higher concentration of growth-promoting hormones (GA3, ZR etc. ) was required in the germination process of Avrolles, which led to the high ratio of GA3/ABA, ZR/ABA and (GA3 + IAA + ZR)/ABA, compared with Judeline3. The total and EMP respiration intensity of Avrolles were less than that of Judeline, only 84.6% and 84.7% at the highest point, respectively. PPP accumulation in Avrolles was 21 days longer than that in Judeline. Furthermore, the running activity was higher and the increased amplitude was 2.02 times than that in Judeline. With the bud dormancy releasing and germination proceeding, the content of soluble sugar which as the substrate of respiration, increased, while starch content declined for transforming into soluble sugars. Soluble sugar content of Avrolles was significantly lower than that of Judeline at the same period.4. The respiration rate of buds declined with adding inhibitor of electron transfer (KCN and SHAM), which implied the existence of both cytochorome pathway and cyanide-resistant pathway in the buds. In contribution rate to total respiration (ρ’Vcyt/Vt), the cytochorome pathway was always higher than cyanide-resistant pathway, which indicated that cytochorome pathway was the main electron transfer pathway. During the whole germination process, the respiration rate of cyanide-resistant pathway,ρValt/Vt and the coefficient of running(ρ)in buds of Avrolles were always higher than that of Judeline, which indicated that the germination of Avrolles required high activity of electron transfer in cyanide-resistant pathway to supply energy to resist the low external temperature.5. Compared with Judeline, Avrolles had the low O2-. production and H2O2 level, and the activity of antioxidase and content of substance resisting oxidation in the process of germination, which indicated that the metabolism rate of active oxygen in the buds of Avrolles was lower than that of Judeline. O2-. mainly comes from the electron leak of mitochondrion respiration. POD played an important role in eliminating H2O2 at the early stage of germination, while CAT at the late stage of germination. Content of GSH and AsA increased in the process of germination which indicated that the AsA-GSH Cycle had been started in the process of the elimination of active oxygen. 6. With the dormancy deepening, content of total phenolics and activity of PAL rised. As the dormancy released, content of total phenolics and activity of PAL declined while activity of PPO increased, which indicated that total phenolics was a germination inhibitor. Compared with Judeine, Avrolles had the high the content of total phenolics, the activity of PAL and PPO, of which the total phinolics content was 25% higher. That indicated that the late-germination of Avrolles was related to metabolism of total phenolics. By HPLC, the content of six monophenols, i.e. gallic acid, epicatechin, meletin, phloretin, ferulic acid and caffetannic acid, were measured. The content of meletin followed by epicatechin was the highest, while the content of gallic acid was the lowest. The content of epicatechin in the buds of Avrolles was 1.31 times higher than that of Judeline, the content of phloretin was 76.8% lower than that of Judeline. The content of ferulic acid increased as the dormancy strengthened while it declined in the process of germination, which indicated that ferulic acid was an inhibitor of germination.更多还原