【作者】 程红；

【导师】 严善春；

【作者基本信息】 东北林业大学 ， 森林保护学， 2013， 博士

【摘要】 丁香属植物(Syringa spp.)为园林绿化的优良树种,其很多种类又兼具良好的药用功能,长期以来人们一直认为其少虫害、耐瘠薄。但笔者的调查却发现,危害丁香属植物的昆虫至少有11种,且致使其树势衰弱、甚至死亡的有2-3种。研究这些害虫的危害规律,抗虫机理,是保护丁香属植物资源和进一步挖掘丁香属植物利用潜能的必要前提。为此,笔者细致调查了从丁香展叶至其落叶过程中的害虫种类,并从化学生态学的角度分析了丁香对柳蛎盾蚧的抗性机理,深入探讨了丁香单宁、酚酸以及各种防御蛋白在丁香抗柳蛎盾蚧方面的作用,为更好地保护和利用丁香资源及抗虫品种的选育提供基础资料,也为柳蛎盾蚧的防治提供理论依据。调查发现,在哈尔滨地区至少有11种昆虫危害丁香属植物,除柳蝙蛾和饰棍蓟马外,其余9种均为首次报道。其中：蛀干类昆虫柳蝙蛾(Phassus excrescens Butler)危害紫丁香(Syringa oblata Lindl.)、红丁香(Syringa villosa Vahl.)、西南丁香(Syringa emodi Wall. ex G. Don)、小叶丁香(Syringa microphylla Diels)、洋丁香(Syringa vulgaris Linn.)、关东丁香(Syringa velutina Kom.)、大花重瓣洋丁香(Syringa vulgaris ’dahua’)、暴马丁香(Syringa reticulata (Blume) Hara var. mandshurica (Maxim.) Hara.)、朝鲜丁香(Syringa dilatata Nakai),寄主范围较广；食叶类昆虫丁香天蛾(Psilogramma increta (Walker,[1865]))为黑龙江省新纪录种,危害紫丁香、红丁香、西南丁香、小叶丁香、关东丁香、紫叶重瓣洋丁香(Syringa vulgaris’zihong’);刺吸性昆虫黑龙江粒粉蚧(Coccurra ussuriensis (Borchs))只在暴马丁香上被发现；康氏粉蚧(Pseudococcus comstocki (Kuwana,1902))危害暴马丁香及小叶丁香；柳蛎盾蚧(Lepidosaphes salicina Borchsenius)对紫丁香、红丁香、西南丁香、白丁香(Syringa oblata var. alba Hort. ex Rehd.)、朝鲜丁香、洋丁香及其变种危害较重。此外,还有危害紫丁香叶部的饰棍蓟马(Dendrothrips ornatus (Jablonowsky,1894))、梨剑纹夜蛾(Acronicta rumicis Linnaeus)及1种毒蛾科幼虫,危害暴马丁香叶部的2种蓟马及1种蚜虫。根据柳蛎盾蚧在丁香上的虫口密度,应用聚类分析法将13种(品种)丁香分为高抗类(什锦丁香等7种)、中抗类(西南丁香等3种)、易感类(紫丁香等2种)、高感类(红丁香)。分别在柳蛎盾蚧的未危害期(5月末)、危害盛期(6月末)、危害弱期(7月术)、危害末期(8月末)采集其叶片,用香兰素盐酸法、高效液相色谱法测定其内的单宁、酚酸含量,分析了丁香叶中单宁、总酚酸含量、酚酸种类等与柳蛎盾蚧危害的关系。结果表明：高抗类中不感虫丁香叶中的酚酸种类齐全,除小叶丁香外其总酚酸含量均显著高于其余种(品种)(p<0.05)。而高抗类中的感虫丁香,未危害期,朝鲜丁香单宁含量显著高于其余感虫丁香(p<0.05)；在危害盛期,除朝鲜丁香单宁增幅不显著外,其余种单宁、总酚酸含量均显著高于中抗、易感和高感类丁香,其增幅为81.75%-888.43%。中抗类丁香,在危害盛期其总酚酸含量增幅显著高于易感和高感类丁香,其单宁含量显著高于易感类丁香。易感和高感类丁香在未危害期其单宁和总酚酸含量相对较低,危害盛期其单宁或总酚酸含量存在诱导滞后性,其酚酸种数只是高抗和中抗类丁香的1/2。此外,一些特定酚酸种类的有无及其含量的多少也与丁香的抗性相关,如：高抗类及中抗类丁香在5—8月份均检测到肉桂酸,易感和高感类丁香5—8月均未检测到肉桂酸。对不同危害期内13种丁香叶中的POD、SOD、CAT、PPO、 PAL、TI CI共7种防御蛋白活性的测定结果表明：危害时期、丁香种(品种)对丁香防御蛋白活力有极显著影响(p<0.01)。 POD活性,在危害盛期,高抗、中抗类丁香均显著升高(p<0.05),易感、高感类应激滞后,分别在危害弱期和危害末期升高。CAT在易感类和高感类丁香防御上起主导作用,其活性在危害盛期显著升高,中抗和高抗类未见明显变化。SOD活性,高抗类丁香危害盛期无显著性变化,而其余3类则显著降低。PPO活性,未危害期的高低与抗性相关,虫害盛期诱导表达的强弱也与抗性相关；PAL活性未能反映抗性品种间的差异。TI和CI活性,在未危害期的高低与其抗虫性无明显相关性,在危害盛期的升高幅度与抗虫性成正相关。综上所述,丁香中酚酸种类多、总酚酸或单宁含量高,或者其单宁和总酚酸含量能在虫害后迅速升高、增幅大的种类,对柳蛎盾蚧的抗性就强,反之则弱。PPO未危害期活性的高低可作为筛选丁香抗虫品种的指标,危害盛期POD、PPO、TI和CI活性升高的幅度与丁香抗虫性水平呈正相关。因此,在抗性品种的选育上丁香酚酸种类数、总酚酸和单宁含量、PPO活性可作为选育指标。更多还原

【Abstract】 Syringa plants are excellent landscaping trees and many species of them have good medicinal effects. The plants have long been considered less pests and resistant infertility. However, our survey found there were at least11different species of insects damaged Syringa plants and2-3species of them made trees weak growth, even death. Studying the rule of pests damage and insect-resistant mechanism are necessary prerequisite for protecting Syringa plants resources and further develop its using potential. For this purpose, the pest species during the period from clove leaves expansion to deciduous were detailed investigated and the resistance mechanism of Syringa spp. on Lepidosaphes salicina were analyzed from the point of view of the chemical ecology. Furthermore, the effects of cloves tannins, phenolic acids and a variety of defense protein on L. salicina were discussed. Our study will provide basic data for better protection and use of the cloves resources and breeding insect-resistant varieties, and also provide a theoretical basis for the prevention and treatment of L. salicina.Our survey found there were at least11different species of insects damaged Syringa plants in Harbin. Except phassus excrescens and Lepidosaphes salicina, all the other9species are reported for the first time. Thereinto, the trunk borer Phassus excrescens damaged nine variations, including Syringa oblata, Syringa villosa, Syringa emodi, Syringa microphylla, Syringa vulgaris., Syringa velutina, Syringa vulgaris’dahua’, Syringa reticulata var. mandshurica and Syringa dilatata. The hosts of this pest are in large scope, with the highest fatality rate to Syringa plants. The defoliator Psilogramma increta damaged Syringa oblata, Syringa villosa, Syringa emodi, Syringa microphylla, Syringa velutina and Syringa vulgaris ’zihong’. The Phloem-Sucking pests Coccurra ussuriensis was only found in Syringa reticulata var. mandshurica. Pseudococcus comstocki endangers Syringa reticulata var. mandshurica and Syringa microphylla. Lepidosaphes salicina greatly damaged Syringa oblata, Syringa villosa, Syringa emodi, Syringa oblata, Syringa dilatata, Syringa vulgaris and its variations, and occasionally endangered Syringa reticulata var. mandshurica, but not Syringa microphylla, Syringa velutina, Syringax chinensis f. chinensis and Syringa vulgaris’zihong’. Moreover, other insects were also found to damage Syringa spp., for instance, the Dendrothrips ornatus and two Lymantridae larvae that feed on Syringa oblate, and two thrips and one aphid that feed on Syringa reticulata var. mandshurica.On the basis of the population density of L. salicina on Syringa spp.,13species/cultivars of Syringa spp. were categorized into following four groups:highly-resistant group (e.g. Syringa chinensis et al), moderately-resistant group (e.g. S. emodi et al), susceptible group (e.g. S. oblate et al), highly-susceptible group (e.g.5. villosa), by cluster analysis. The contents of various tannins and phenolic acids in leaves of these13species/cultivars of Syringa from four sampling periods:the pre-infesting period (end of May), the peak-infesting period (end of June), the weakly-infesting period (end of July) and the late-infesting period (end of August), were measured with Vanillin-HCl and HPLC methods; and then their relationships with the susceptibility to L. salicina attacks were analyzed. The results showed that in the highly-resistant group, a full range/types of phenolic acids were detected in the uninfected leaves, and the total contents of phenolic acids were significantly higher than those species/varieties in other groups (p<0.05), except S. microphylla. Among the infested Syringa spp. within the highly-resistant group, the contents of tannins in S. dilatata leaves during the pre-infesting period were significantly higher than those in the other infested species within the same group (p<0.05); during the peak-infesting period, except that the increasing range of tannin is not significant in S. dilatata, the contents of tannin and total phenolic acids of the non-infested species were significantly higher (around81.75%to888.43%) than the infected Syringa spp. As for the moderately-resistant group, the increase range of total phenolic acids contents was significantly higher than that in the susceptible group and the highly-susceptible group during the peak-infesting period, and the tannin content was significantly higher than that in the susceptible group. In the susceptible group and highly-susceptible group, the contents of tannin and total phenolic acids were relatively low during the pre-infesting period, and there was a lag of induction in the total contents of tannins and phenolic acids during the peak-infesting period, and the types of phenolic acids were just half of the highly-resistant and moderately resistant groups. In addition, some specific kinds of phenolic acids and the changes of contents were related to the resistance of Syringa spp. eg. From May to August, the cinnamic acid was detected in all highly-resistant group and moderately-resistant group, whereas it wasn’t detected in the susceptible and the highly susceptible groups.We investigated the relationship between activities of various defensive plant proteins and pest-resistance to the L. salicina by testing seven kinds of defensive protein activities (POD, SOD, CAT, PPO, PAL, TI and CI) from the leaves of13Syringa species (seven species with high pest-resistance, three species with general resistance, two species with vulnerable type and one with very susceptible type) in different pest periods. The results showed highly significant difference in defensive proteins activities between different Syringa species (/?<0.01) in the pest period. POD activity of the species with high and general pest-resistance were significantly increased (p<0.05) in the peak-infesting period (end of June). In contrast, POD activity of the species with vulnerable and very susceptible pest-resistance was increased in the weak-infesting period (end of July) and the late-infesting period (end of August). CAT activity was significantly increased in the peak-infesting period; it may play a primary role in protection of the two types of species from pest damage. SOD activity was significantly decreased in all the tested species except the high resistance species in the peak-infesting period. The change of the PPO activity was positively correlated with the resistance in the pre-infesting period (end of May) and its increment also had correlation with pest resistance in the peak-infesting period. There were no differences in PAL activity between different resistance species. Similarly no significant correlation was found between the activities of TI and CI and the pest resistance in the pre-inf esting period. In summary, the level of PPO activity in the pre-infesting period is recommended as an index screening the resistant species.In summary, the resistance of Syringa spp. against L. salicina is related to its constitutive and induced defenses. The species that contain more types of phenolic acids and high contents of total phenolic acids and tannin, or can quickly respond to insect attacks by producing or increasing the contents of tannin or phenolic acids, might have a strong resistance potential against L. salicina, and vice versa. The activity level of the PPO non-harmful period can be used as index for screening cloves insect-resistant varieties. Increment of POD, PPO, TI and CI activities in the peak-infesting period were positively correlated with pest-resistance level of Syringa spp.. Therefore, on the breeding of resistant varieties, the species number of cloves phenolic acid, the total phenolic acid, and tannin contents, PPO activity can be used as breeding index.更多还原