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核桃(Juglans regia Linnaeus)又叫胡桃,是我国重要的经济林树种。其坚果具有较高的营养价值,有良好的保健作用。我国核桃的生产与出口位居世界第一,根据FAO资料统计,在2020年,我国的核桃种植面积为48.71万公顷,核桃产量高达110万吨[1]。其中核桃的种植区主要分布在新疆、云南、四川、陕西等地,而新疆拥有独特的地理环境造就了核桃优良的品种,因此核桃产业的发展被当地高度重视。苹果蠹蛾(Cydia pomonella Linnaeus)是世界性检疫害虫,主要为害苹果(Malus pumila Mill)、桃(Prunus persica Linnaeus)、海棠(Malus spectabilis Borkh)、核桃等仁果类和坚果类果树[2]。在中国新疆和田县,于2018年首次发现苹果蠹蛾为害核桃,并造成20%~39%的产量损失,近几年来苹果蠹蛾在核桃产区不断扩散。目前该虫的防治主要依靠化学防治,这与绿色防控和生产绿色食品的原则相悖。因此有效利用植物的抗虫性对害虫的防治具有着重要意义,也符合当前绿色环保的防控策略。
植物与昆虫相互作用是生物多样性的驱动力,在漫长的协同进化过程中,植物已形成相应的防御措施,以确保植物自身的正常生长[3-4]。最直接的方式是植物通过产生多种代谢物直接作用植食性昆虫或威慑植食性昆虫来保护自身,如植物代谢物质胡桃醌可以毒杀舞毒蛾(Lymantria dispar Linnaeus)和甘蓝夜蛾(Mamestra brassicae Linnaeus)幼虫,进而提高植物对植食性昆虫的抗性[5]。另外当植食性昆虫为害时,植物可以改变自身的营养物质来抑制植食性昆虫的生长发育[6-7]。已有研究表明营养物质是影响昆虫生长发育与繁殖的重要因素,尤其是植物体内的氮和糖[8-9]。植物被害虫为害后,为了应对害虫的胁迫,植物的相关防御酶被启动,酶活性的变化则反应寄主植物的抗虫反应[10]。已有研究表明过氧化氢酶(catalase,CAT)、过氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)等在植食性昆虫胁迫时诱导酶活性的改变,并生成相关的防御体系[11-13]。在田间观察发现被苹果蠹蛾为害后,核桃果皮蛀孔处组织褐化,部分细胞死亡,通过室内饲喂发现与取食核仁相比取食桃外果皮的苹果蠹蛾生长发育已受到严重影响。此外,本课题组在野外调查发现核桃不同品种被蛀率之间存在着较大的差异。因此,通过研究苹果蠹蛾蛀食核桃果皮营养物质、代谢物质及酶活性的动态变化,探明核桃果皮对苹果蠹蛾取食胁迫的防御反应,为抗苹果蠹蛾核桃品种的选育提供更科学的理论依据。
核桃果皮对苹果蠹蛾为害的防御响应
Defensive Response of Walnut Husk to Cydia Pomonella
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摘要:
目的 研究苹果蠹蛾为害后诱导核桃果皮产生的防御应答响应。 方法 采用紫外分光光度法与酶标仪微量法分析核桃果皮营养物质、次生代谢物质的含量,防御酶的活性以及苹果蠹蛾与核桃的互作关系。 结果 核桃果皮被苹果蠹蛾蛀食后可溶性糖与可溶性蛋白的含量随着时间推移逐渐降低,在为害24 h时可溶性蛋白含量与对照相比差异显著(P<0.05);在为害48 h时可溶性糖与对照相比差异显著(P<0.05)。次生代谢物质胡桃醌、单宁的含量随时间的推移呈先上升后下降的趋势,其中胡桃醌在为害48 h时含量达到最高,为976.68 µg·g−1,是对照的1.44倍;单宁含量在24 h时达到高峰,为4.11 mg·g−1,是对照的1.33倍;类黄酮的含量呈逐渐上升趋势,在24 h时与对照相比差异显著(P<0.05)。在为害12 h时,CAT活性与对照相比差异显著(P<0.05),在72 h时CAT活性达峰值,为55.97 U·min−1·g−1,是对照的1.45倍;POD活性呈先下降后上升的趋势,在72 h时达最高值,是对照的1.62倍;SOD活性呈先上升后下降的趋势,在24 h时达到峰值,为623.69 U·g−1,是对照的1.98倍。 结论 核桃果皮主要通过调节体内的营养物质、次生代谢物质以及防御酶活性的变化,对苹果蠹蛾的为害产生应激反应,进而发挥防御作用。 Abstract:Objective To Investigate the defensive response to walnut husk after Cydia pomonella infestation. Method UV Spectrophotometry and Microplate Reader were used to analyze nutrient substances, secondary metabolites content, defense enzyme activity and interaction between Cydia pomonella and walnut Result The content of soluble sugar and soluble protein decreased gradually over time after the walnut husk was eaten by C. pomonella. The soluble protein content at 24 h of damage was significantly different from the control (P<0.05); There was a significant difference between the soluble sugar and the control at 48 hours of damage (P<0.05). The contents of secondary metabolites juglone and tannin increased first and then decreased. Among them, the content of juglone quinone reached the highest at 48 h, which was 976.68 µg·g−1 FW, and was 1.44 times that of the control; The tannin content reached a peak at 24 h at 4.11 mg·g−1, which was 1.33 times that of the control; The content of flavonoids showed a gradual upward trend, and the difference was significant compared with the control at 24 h (P<0.05). At 12 h of damage, there was a significant difference in CAT activity compared with the control (P<0.05). At 72 h, the CAT activity peaked at 55.97 U·min−1·g−1, which was 1.45 times that of the control; The POD activity showed a trend of first decreasing and then increasing. At 72 h, the POD activity reached the highest value, which was 1.62 times that of the control; The SOD activity increased firts and then decreased. At 24 h, the SOD activity reached a peak value of 623.69 U·g−1, which was 1.98 times that of the control. Conclusion The walnut husk mainly produces a stress response to the damage of C. pomonella by regulating the changes of nutrients, secondary metabolites and defense enzyme activities in the body, and then plays a defensive role. -
Key words:
- Cydia pomonella
- / walnut husk
- / nutrients
- / metabolites
- / defense enzyme activity
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