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林业科学 ›› 2018, Vol. 54 ›› Issue (9): 104-113.doi: 10.11707/j.1001-7488.20180913

• 论文与研究报告 • 上一篇    下一篇

抗绕实蝇沙棘无性系的筛选及相关化学物质和酶活性分析

刘建凤1, 董礼龙2, 曹丹丹1, 罗红梅2, 魏建荣1   

  1. 1. 河北大学生命科学学院 保定 071002;
    2. 中国林业科学研究院沙漠林业实验中心 磴口 015200
  • 收稿日期:2017-11-17 修回日期:2018-05-05 出版日期:2018-09-25 发布日期:2018-09-10
  • 基金资助:
    国家自然科学基金项目(31370647);新疆林业厅项目(新林计字2015-610)。

Screening of Sea-Buckthorn (Hippophae rhamnoides) Clones Resisting Fruit Fly (Rhagoletis batava obseuriosa)(Diptera: Tephritidae) and Analysis of the Related Biochemicals and Enzyme Activities

Liu Jianfeng1, Dong Lilong2, Cao Dandan1, Luo Hongmei2, Wei Jianrong1   

  1. 1. College of Life Science, Hebei University Baoding 071002;
    2. Experimental Center of Desert Forestry, CAF Dengkou 015200
  • Received:2017-11-17 Revised:2018-05-05 Online:2018-09-25 Published:2018-09-10

摘要: [目的]筛选抗绕实蝇的沙棘无性系植株,分析不同抗、感植株间相关生化物质种类和含量的差异,为进一步研究抗性机制提供理论基础。[方法]首先通过林间调查和室内生测试验,筛选抗性植株,然后以抗、感沙棘无性系为材料,利用气相色谱-质谱联用仪(GC-MC)和高效液相色谱(HPLC)等,在成虫羽化前和产卵盛期检测抗、感无性系植株相关化学物质含量和酶活性。[结果]1)通过林间调查果实受害率及开展沙棘绕实蝇产卵选择性试验,筛选出抗沙棘绕实蝇的沙棘无性系。2)在产卵盛期,抗虫无性系果实和叶片中总酚含量、可溶性蛋白质和脯氨酸含量均显著高于敏感无性系,在果实中分别高出101.34%、50.21%和20.12%,叶片中分别高出121.22%、30.43%和40.68%。3)抗、感无性系果实的挥发性物质种类及含量在2种无性系植株间具有显著差异,其中,抗虫无性系的酯类物质种类及含量显著低于敏感无性系,而醇类物质含量明显高于敏感无性系。4)抗虫无性系果实和叶片中非挥发性物质山奈酚含量明显高于敏感无性系,分别高出29.40%和40.80%;果实中异鼠李素含量较敏感无性系高出43.20%,而在叶片中的含量较敏感无性系低41.34%。5)产卵盛期,抗虫无性系果实中过氧化物酶(POD)、几丁质酶(Chi)、苯丙氨酸解氨酶(PAL)活性较敏感无性系分别高出98.21%、25.03%和20.10%,在叶片中POD和PAL则分别高出33.40%和36.30%。[结论]抗、感无性系果实、叶片中化学物质及酶活性均存在较大差异,由此推测,沙棘体内抗虫相关化学物质和酶活性与其抗沙棘绕实蝇的能力密切相关。结果初步揭示沙棘抗虫无性系的生理生化特征,为进一步研究沙棘抗虫性机制及开展抗虫性育种提供依据。

关键词: 沙棘, 沙棘绕实蝇, 抗虫性, 生化物质, 酶活性

Abstract: [Objective] Sea-buckthorn (Hippophae rhamnoides) is an important economic and ecological plant in the arid and semi-arid region of China, and Rhagoletis batava obseuriosa (Diptera:Tephritidae) (RBO) is a serious quarantine insect pest that damaged the plant fruit. In order to find the RBO-resistant clones, a field investigation and indoor test were carried out. The difference of the biochemical substances between the resistant- and sensitive-plants were investigated to provide the theoretical basis for further study of physiological and biochemical resistance mechanisms of sea-buckthorn to RBO.[Method] First, the difference of RBO adult's oviposition biology and fruit damage were compared between RBO-resistant clones and RBO-sensitive clones both in the field and in laboratory. Then, two stages:before the adult emergence and at the peak time of adult oviposition, were chosen to detect biochemical substances and enzyme activities in the fruits and leaves of the resistant- and sensitive-clones using gas chromatography-mass spectrometry (GC-MC) and high performance liquid chromatography (HPLC).[Result] 1) Sea-buckthorn clones resisting RBO were obtained by screening the fruit damage in the field and comparing the RBO ovposition rates indoors. 2) At the peak time of adult oviposition, the total phenolics, soluble protein and proline content in RBO-resistant clone were higher than those in RBO-sensitive clones, with increasing of 101.34%, 50.21% and 20.12% in the fruits, and 121.22%, 30.43% and 40.68% in the leaves, respectively. 3) The volatile compounds were significantly different in the fruits of the two clones. The types and contents of the esters in the RBO-resistant clones were significantly lower than those in the RBO-sensitive clones, while the content of alcohols was significantly higher than that in the RBO-sensitive clones. 4) The content of non-volatile kaempferol in RBO-resistant clones was significantly higher than that in the RBO-sensitive clones, with increase of 29.40% and 40.80% in fruits and leaves, respectively. The content of isorhamnetin in fruits of RBO-resistant was 43.20% higher than that in RBO-sensitive clones, while the content in the leaves was 41.34% lower than that in the RBO-sensitive clones. 5) At the peak time of adult oviposition, activity of the peroxidase (POD), chitinase and phenylalanine ammonialyase (PAL) in the fruits were 98.21%, 25.03%, and 20.10% higher, respectively, and the POD and PAL activities in the leaves of RBO resistant clones were 33.40% and 36.30% higher, respectively than those in RBO-sensitive clones.[Conclusion] There were significant differences in the secondary metabolites and enzyme activities in the sea-buckthorn fruits and leaves between the RBO-resistant clones and the RBO-sensitive clones. The results preliminarily revealed the physiological and biochemical characteristics of the RBO-resistance, and provided the basis for further study on the mechanism of the RBO-resistance and the new RBO-resistant clone breeding.

Key words: sea-buckthorn, Rhagoletis batava obseuriosa, insect-resistant, biochemicals, enzyme activity

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