摘要
本文以高丛蓝莓品种“蓝金”、“伯克利”、“北陆”和“日出”等为实验材料,系统研究了高浓度CO2冲击、不同O2和CO2浓度箱式气调对采后蓝莓果实的生理生化代谢、果实品质和贮藏特性等影响,提出了适合于蓝莓果实采后生理特性的高CO2冲击处理时间和气调贮藏条件；研究了外源乙烯和外源水杨酸两种具有信号作用的生理调控物质诱导采后蓝莓果实自身抗性的产生,筛选出适宜的处理方法和浓度。分析了蓝莓在不同贮藏和处理条件下果实中的多酚氧化酶(PPO)、过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)和脂氧合酶(LOX)等酶活性变化,以及蓝莓果实外观颜色与感官品质的相关性,揭示了蓝莓果实衰老机理、腐烂发生及其控制；同时,还分析了不同蓝莓品种的抗氧化物质含量及其抗氧化活性,以及引起蓝莓采后主要病害种类与防治方法。为形成采后蓝莓果实商业化处理、贮藏保鲜及流通技术的集成奠定了理论基础。
本文的主要研究内容包括：不同北高丛蓝莓品种抗氧化物质及其抗氧化活性研究；纳他霉素对葡萄孢霉(Botrytis cinerea)引起的蓝莓灰霉病防治效果的研究；高CO2冲击处理对采后蓝莓生理代谢及品质的影响；箱式气调贮藏对采后蓝莓生理生化变化的影响：外源乙烯处理对采后蓝莓感官性状和呼吸代谢的影响；外源水杨酸处理对采后蓝莓果实苯丙烷代谢系统的影响。试验结果表明：
1.不同北高丛蓝莓品种抗氧化物质及抗氧化活性存在较大差异：以蓝金、日出、塞拉、柳叶4个北高丛蓝莓品种为试验材料,对其总花色苷、可溶性多糖、维生素C以及还原型谷胱甘肽等4种抗氧化物质的含量进行了测定,并测试了代表抗氧化活性的DPPH自由基清除能力和超氧化物歧化酶(SOD)活性,同时分析比较了4种抗氧化物质与DPPH自由基清除能力之间的相关性关系。研究结果表明：总花色苷含量、维生素C含量以及还原型谷胱甘肽含量与DPPH自由基清除能力间存在显著正相关关系(P<0.05)。4个品种中柳叶的总花色苷含量、维生素C含量、还原型谷胱甘肽含量以及DPPH自由基清除能力均显著高于其他3个品种(P<0.05),说明柳叶品种的抗氧化性最强。
2.蓝莓采后主要病害及其防治。葡萄孢霉(Botrytis cinerea)引起的蓝莓灰霉病是蓝莓采后的主要病害,低浓度的纳他霉素处理(200-600mg/L)对蓝莓灰霉病有很好的防治效果。纳他霉素能诱导蓝莓果实中与抗病性相关的PPO、POD、PAL活性增加,减少花色苷的损失,有效地抵御病原菌的入侵。相同浓度(600mg/L)的三种果蔬保鲜剂：扑海因、纳他霉素、甲基托布津对于病害的防治效果为：扑海因>纳他霉素>甲基托布津。但其中扑海因和甲基托布津是化学防腐剂,易使病菌产生抗药性,并具有一定毒性,而纳他霉素作为一种天然食品防腐剂能安全、高效地控制蓝莓采后灰霉病的发生。
3.蓝莓果实对高浓度CO2冲击处理有很强的忍耐力。蓝莓果实可忍耐96h以下的高CO2冲击处理(体积分数为99.9%C02),48h和96h的高C02冲击处理降低了果肉pH值和果实腐烂率,保持了果实硬度,诱导POD活性上升并使LOX活性维持在较低水平,从而减慢了蓝莓果实呼吸代谢速率,保持了果实品质。而144h的高CO2冲击处理对果实造成不可逆伤害,导致无氧呼吸和异味的产生以及腐烂的加剧。研究结果认为高C02冲击处理作为一种高效、安全的采后技术,可有效抑制蓝莓果实的生理生化代谢,控制采后果实腐烂的发生。
4.蓝莓果实适合于较高C02浓度的箱式气调贮藏：与空气对照相比,5%02+10%CO2；5%O2+20%C02和5%02+30%C023种CO2浓度的箱式气调贮藏条件均可启动酶促防御系统,使POD、过氧化氢酶(CAT)和SOD的活性得到提高,同时也减慢了还原型谷胱甘肽(GSH)含量下降的速率,增加了Vc含量。3种浓度的CO2处理还可有效抑制呼吸速率和腐烂的发生,提高了果实木质素含量和硬度,其中5%02+30%C02更有利于延缓衰老进程,贮藏期可延长至95d。高C02冲击处理和高C02箱式气调的研究结果表明,蓝莓对高浓度CO2有很强的忍耐力,保鲜效果显著。
5.适当浓度的外源乙烯处理能增加蓝莓果实的抗病性：与空气对照相比,0.1ml/L、1.0ml/L和10ml/L的乙烯密闭处理48h均可提高蓝莓果实PPO、POD、PAL等与采后蓝莓果实抗病相关酶的活性,但10ml/L的乙烯处理增加了内源乙烯的合成,加速了呼吸代谢速率,果实衰老并引发腐烂发生,而0.1ml/L和1.0ml/L的乙烯处理提高了蓝莓果实的抗性和贮藏效果。去除蜡质前后蓝莓果实的外观亮度差值和感官品质间存在着极显著相关性关系(P<0.01)。
6.外源水杨酸处理促进了采后蓝莓果实苯丙烷代谢的进程：0.5mmol/L和1.0mmol/L的水杨酸处理均可促进蓝莓果实苯丙烷代谢的进程,从而使苯丙烷代谢的终产物—木质素含量增加,进而增加了果实硬度并可有效地抵御微生物的入侵,降低果实腐烂,延长保质期。
The highbush cultivars of blueberry(Vaccinium corymbosum L.), such as'Bluegold', 'Northland','Berkeley' and 'Sunrise' were studied their postharvest physio-biochemical metabolism, quality and storability during the storage periods, in order to determine the suitable high CO2 shock treatment, the sound concentrations of O2 and CO2 in controlled atmosphere and the storage time for the blueberry fruits. Moreover, the physiological regulation substances, including ethylene and salicylic acid, were used as signal substances properly to induce the generation of resistance. In this experiment, polyphenol oxidase (PPO), peroxidase (POD), lipoxygenase (LOX) and phenylalnine ammonialyase (PAL) were analyzed during different storage conditions, and the relativity between fruit surface color changes and sensory evaluation were studied, and senescence mechanism, occurrence and control of decay were revealed. Meanwhile, the content of biological active substances and antioxidant capacity in various cultivars of blueberry fruits were examined, as well as the main diseases of postharvest blueberry and its control methods were also evaluated in storage. The results will be beneficial to provide integrative technologies for commercial storage of blueberry fruits.
This experiments included following contents:(1) Study on the biological active substances and antioxidant capacity in various cultivars of northern highbush blueberry fruits; (2) Effects of Natamycin treatments on the control of blueberry gray mold that caused Botrytis cinerea; (3) Effects of high CO2 shock treatments on physiological metabolism and quality of postharvest blueberry fruits; (4) Effects of plastic box modified atmosphere storage on the physiological and biochemical changes of postharvest blueberry fruits; (5) Effects of exogenous ethylene treatments on sensory properties and metabolism of postharvest blueberry fruits; (6) Effects of exogenous salicylic acid treatment on phenylproanoid metabolic system of postharvest blueberry fruits. The results showed as following:
1. There were certain differences in biological active substances and antioxidant capacity between various cultivars of northerm highbush blueberry fruits:Four northern highbush blueberry cultivars, including Bluegold, Sunrise, Sierra and Willow, were used as experimental materials. The content of antioxidant substances such as total anthocyanin. soluble polysaccharide, vitamin C and reduced glutathione were analyzed in the four northern highbush blueberry cultivars. Moreover, DPPH free radical scavenging ability and superoxide dismutase (SOD) activity which behalf of antioxidant ability were investigated. The correlative relationship between DPPH free radical scavenging ability and four kinds of antioxidant substances were researched. The results indicated that DPPH free radical scavenging ability was significantly related with total anthocyanin content, vitamin C content and reduced glutathione content (P<0.05). The total anthocyanin content, vitamin C content, reduced glutathione content and DPPH free radical scavenging ability in Willow cultivar were significantly higher than that of other three cultivars (P<0.05), it is for this reason that Willow cultivar has the highest antioxidant ability.
2. The main diseases of postharvest blueberry fruits and its controll:Green mold (Botrytis cinerea) is the main pathogenic microorganism. Low concentrations of Natamycin treatment (200-600mg/L) had a good control effect of Botrytis cinerea. Natamycin induced the increase of PPO, POD, PAL that realted to the disease resistance, reduced the loss of anthocyanins and resisted the invasion of pathogens. The same concentration (600mg/L) of three fruit and vegetable preservatives, including Natamycin, Iprodione and Mildothane, played different roles in controlling the disease of blueberry fruits. Although the control efficiency of Natamycin ranged between Iprodione and Mildothane, as natural food preservation, Natamycin had a good perspective in inhibiting the disease of blueberry.
3. Blueberry fruits had good torlerance to high CO2 shock treatment:High CO2 shock treatment (99.9% CO2, v/v) that less than 96 h had no injury on blueberry fruits. The pH value in pulp and fruit rot rate were significantly decreased, fruit firmness was well maintained, the storage period was effectively extended to 50 days, POD activity was increased and the LOX activity was decreased after the high CO2 shock treatments for 48 h and 96 h. The results indicated that the treatments slowed down the rate of metabolism and maintained the fruit quality of blueberry fruits. In contrast,144 h high CO2 shock treatment caused irreversible damage, resulting in anaerobic respiration, off-flavor and fruit rot. As a shock-impact method, high CO2 shock treatment may be used before the storage of blueberry fruits to inhibit the physio-biochemical metabolism, maintain the fruits quality and control the happiness of fruit decay.
4. Plastic box modified atmosphere storage CA with high-CO2 concentrations was beneficial for blueberry fruits:Compared with CK (air condition), three kinds of plastic box modified atmosphere with high CO2, including 5%O2+10%CO2; 5%O2+20% CO2和5%O2+30%CO2, stimulated the defense system. Therefore, not only the activities of POD. CAT, SOD, PAL and PPO were increased, but also Vc and reduced glutathione (GSH) contents were kept in a higher lever. Moreover, the plastic box modified atmosphere storage with high CO2 effectively slowed down the respiration rate, inhibited the occurrence of decay and increased the lignin content and fruit firmness, especially 5%O2+30%CO2 was more conducive to delaying the senescence process and effectively extending the storage period to 95 days. It is suggested that blueberry fruits have a strong endurance to high CO2 and the plastic box modified atmosphere storage with high CO2 is perfect at blueberry fruits storage.
5. The proper concentration of exogenous ethylene treatments induced the disease resistance of blueberry fruits:Compared with control (air condition), three kinds of ethylene treatments increased the activities of PPO, POD and PAL. Nevertheless, 10ml/L ethylene treatment increased the biosynthesis of endogenous ethylene and accelerated the metabolism rate and senescence process, while 0.1 and 1.0ml/L ethylene treatments promoted the disease resistance ability and storability of blueberry fruits. The lightness discrepancy between before and after removal of wax was significantly related with the result of sensory evaluation (P<0.01).
6. Exogenous salicylic acid treatments accelerated the process of phenylproanoid metabolism of postharvest blueberry fruits:0.5mmol/L and 1.0mmol/L salicylic acid treatment promoted the process of phenylpropanoid metabolism. Therefore, the end product of phenylpropanoid metabolism, lignin content increased and it increased the fruit firmness
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