榆黄毛萤叶甲生物学习性及发生动态研究

【目的】榆黄毛萤叶甲Pyrrhalta maculicollis(Mots.)是近年来在新疆新发现的榆树食叶害虫,本文经对其生物学习性的研究,为该害虫的有效控制提供科学依据。【方法】经室内外连续2年的观察,较系统研究了榆黄毛萤叶甲在吐鲁番市的年生活史、习性及其发生动态。【结果】榆黄毛萤叶甲在吐鲁番市一年发生4代,以成虫在老树干基部树缝内、干基部土缝内群集越冬。翌年3月底开始出蛰,并以榆树嫩芽补充营养,4月上旬开始交尾产卵。各代幼虫分别于4月中旬、6月上旬、7月下旬和9月中旬发生,各代间世代重叠。5种颜色的光源对榆黄毛萤叶甲诱集试验结果表明,榆黄毛萤叶甲有较强的趋光性,蓝光对其诱集效果最好。【结论】榆黄毛萤叶甲在吐鲁番市3月至10月间,以成虫和幼虫对榆树不间断为害,且有世代重叠现象,4月、5月为该害虫防治关键时期。     

榆黄毛萤叶甲的发育起点温度与有效积温的测定

【目的】有效防控榆黄毛萤叶甲Pyrrhalta maculicollis(Mots.)在吐鲁番地区的危害。【方法】本研究在室内设置22、25、28、31和34℃5个温度梯度,分别测定榆黄毛萤叶甲各虫态的发育历期,并以有效积温法则和"最小二乘法"计算出榆黄毛萤叶甲各虫态的发育起点温度和有效积温。【结果】卵、幼虫、蛹、产卵前期及全世代的发育起点温度分别为13.83、12.64、12.48、14.63和14.01℃,有效积温分别为83.99、186.32、121.50、185.42和550.54日·度。榆黄毛萤叶甲发育速率与温度的关系分别采用线性日度模型式和Logistic曲线模型式进行拟合,建立各虫态发育速率与温度关系模型。由Lagrange中值定理求出各虫态的发育最适温度、适宜温区,其全世代的分别为:Tmid为28.23℃、Tmin为14.36℃、Tmax为42.11℃。【结论】明确了榆黄毛萤叶甲各虫态的发育起点温度及有效积温,计算出了各虫态的理论发育最适温度及适宜温区,为准确预报榆黄毛萤叶甲发生期及有效防治提供理论依据。     

沙葱萤叶甲化学感受蛋白基因的鉴定及表达谱分析（英文）

【目的】沙葱萤叶甲Galeruca daurica是一种在内蒙古草原上爆发成灾的新害虫。昆虫的化学感受蛋白(CSPs)是一类水溶性小分子蛋白,其主要功能是识别和传导环境中的化学刺激至受体,参与众多昆虫行为。本研究旨在鉴定沙葱萤叶甲化学感受蛋白基因,并对其表达谱进行分析。【方法】通过筛选本实验室组装的沙葱萤叶甲转录组鉴定沙葱萤叶甲化学感受蛋白基因;采用实时荧光定量PCR方法分析其在沙葱萤叶甲不同发育阶段(卵、1-3龄幼虫、蛹和成虫)和成虫组织[触角、头(去触角)、胸、腹、足和翅]中的表达水平。【结果】鉴定出10个化学感受蛋白基因,将其命名为Gdau CSP1-10(Gen Bank登录号:KY885471-KY885480)。10条编码蛋白的氨基酸序列一致性范围为17.27%~62.79%,彼此间分化程度较高。通过NCBI的Blast比对结果显示,Gdau CSPs与榆黄毛萤叶甲Pyrrhalta maculicollis的Pmac CSP氨基酸序列一致性最高,为90%。系统发育分析表明,4种Gdau CSPs首先与榆黄毛萤叶甲的Pmac CSPs聚为一支。实时荧光定量PCR结果显示,Gd CSPs在不同发育阶段中的表达量有显著差异,Gdau CSP4-5,Gdau CSP7-8和Gdau CSP10 5个Gd CSPs在成虫中的表达量显著高于其他发育阶段,而Gd CSP2在卵中的表达量显著高于其他发育阶段;Gd CSPs不仅表达于成虫触角中,在头(去触角)、胸、腹、足、翅等其他部位也有表达,且10个Gd CSPs在沙葱萤叶甲成虫各组织中有着不同的表达谱,其中Gdau CSP2,Gdau CSP4,Gdau CSP5,Gdau CSP8和Gdau CSP9 5个Gd CSPs在雌成虫触角中的表达量显著高于其他组织。【结论】结果提示化学感受蛋白在沙葱萤叶甲的生长发育和化学感受过程中可能起着不同的作用。本研究为进一步研究沙葱萤叶甲化学感受蛋白的生理功能及化学通讯的分子机理奠定了必要的基础。     

RNAi介导的GdHsp60和GdHsp70基因沉默对沙葱萤叶甲幼虫抗寒性的影响

【目的】本研究旨在比较不同RNAi方法对沙葱萤叶甲Galeruca daurica幼虫热激蛋白基因(GdHsp60和GdHsp70)的沉默效率,以选择一种可高效降低靶基因表达水平的研究方法,明确这两个热激蛋白在沙葱萤叶甲幼虫抗寒性中的作用。【方法】分别通过饲喂法和显微注射法进行RNAi沉默沙葱萤叶甲1和2龄幼虫的GdHsp60和GdHsp70,采用qPCR检测GdHsp60和GdHsp70的沉默效率;通过显微注射法分别 对GdHsp60和GdHsp70进行RNAi后24 h,用热电偶法测定沙葱萤叶甲2龄幼虫过冷却点和结冰点,生物测定沙葱萤叶甲2龄幼虫暴露于不同低温条件下(-6~-14℃) 2 h的半致死温度(Ltemp₅₀)以及在-5℃低温条件下处理不同时间后的半致死时间(Ltime₅₀)。【结果】用饲喂法和显微注射法进行RNAi均可以降低GdHsp60和GdHsp70的表达水平,但显微注射法的沉默效率更高。与对照组(显微注射dsGFP)相比,沙葱萤叶甲2龄幼虫分别显微注射dsGdHsp60和dsGdHsp70 24 h后,GdHsp60和GdHsp70的表达水平均降至最低,分别降低了84.15%和92.38%。在沙葱萤叶甲2龄幼虫中,显微注射dsGdHsp60 24 h后其过冷却点、结冰点、Ltemp₅₀及Ltime₅₀值分别-10.56±0.42℃,-7.66±0.56℃,-8.33℃和49.25 h,显微注射dsGdHsp70 24 h后其过冷却点、结冰点、Ltemp₅₀及Ltime₅₀值分别为-9.08±0.23℃,-6.09±0.28℃, -8.20℃和52.21 h,而对照组的分别为-14.71±0.11℃,-13.94±0.09℃,-10.63℃和87.13 h。与对 照组(显微注射dsGFP)相比,在沙葱萤叶甲2龄幼虫分别显微注射dsGdHsp60和dsGdHsp70 24 h后过冷却点 、结冰点和Ltemp₅₀显著上升,而Ltime50值显著缩短。【结论】显微注射法可作为沙葱萤叶甲Hsp相关基 因RNAi的主要干扰方法;沉默GdHsp60和GdHsp70基因均会显著降低了沙葱萤叶甲幼虫的抗寒能力。     

变温和恒温对沙葱萤叶甲发育速率的影响

沙葱萤叶甲为近年来在内蒙古草原猖獗成灾的新害虫,为明确温度对其发育速率的影响,分别设置5个变温组合(8/20℃,11/23℃,14/26℃,17/29℃和20/32℃)和6个恒温(13℃,17℃,21℃,25℃,29℃和33℃),比较了变温和恒温对沙葱萤叶甲幼虫和蛹发育速率的影响。结果表明,不同变温组合和恒温对沙葱萤叶甲幼虫和蛹的发育速率有显著的影响。发育历期随温度的升高而缩短,在变温条件下,1龄幼虫期、2龄幼虫期、3龄幼虫期、总幼虫期和蛹期分别从最低温度组合(8/20℃,平均15℃)的11.00,13.44,23.18,46.42和16.89 d,缩短至最高温度组合(20/32℃,平均27℃)的4.92,4.63,9.17,17.83和5.83 d;在恒温条件下,13℃下幼虫不能发育和存活,1龄幼虫期、2龄幼虫期、3龄幼虫期、总幼虫期和蛹期分别从17℃的14.50,10.75,20.63,45.50和11.00 d,缩短至33℃的6.10,5.47,10.60,22.17和5.33 d。在变温条件下,幼虫和蛹的发育起点温度分别为7.44℃和8.48℃,有效积温分别为344.82日度和113.52日度;在恒温条件下,幼虫和蛹的发育起点温度分别为0.64℃和5.11℃,有效积温分别为714.28日度和147.06日度。变温促进了沙葱萤叶甲幼虫和蛹的发育,本研究结果为沙葱萤叶甲的预测预报及综合防控提供了科学依据。     

豚草条纹萤叶甲各虫态的空间分布型

豚草条纹萤叶甲Ophraella communa LeSage是恶性入侵豚草(Ambrosia artemisiifolia L.)的天敌,用频次分布拟合和多种聚集指数测度等方法对该叶甲的成虫、幼虫和蛹、卵的空间分布型进行研究。结果表明,豚草条纹萤叶甲卵、幼虫+蛹、成虫的空间分布符合负二项分布,种群个体的空间分布为聚集分布。用几种衡量聚集度的指标,对上述各虫态分布的聚集程度进行测定;然后计算出各虫态田间最适理论抽样数。     

榆绿毛萤叶甲寄生微粒子虫新种记述：微孢子门：微粒子科

本文记述寄生于榆绿毛萤叶甲的一种微粒子虫新种的光匀和电镜形态特征及寄生习性。并讨论了新种鉴别特性。     

沙葱萤叶甲丝氨酸蛋白酶基因GdSP的克隆及对温度胁迫的响应

【目的】丝氨酸蛋白酶(serine protease,SP)是一类以丝氨酸为活性中心的重要的蛋白水解酶。本研究旨在克隆获得沙葱萤叶甲Galeruca daurica丝氨酸蛋白酶基因,分析其对温度胁迫的响应,以期为进一步揭示沙葱萤叶甲耐温性的调控机制及其他生理功能奠定基础。【方法】根据沙葱萤叶甲2龄幼虫转录组数据,采用RACE技术克隆得到沙葱萤叶甲丝氨酸蛋白酶基因的cDNA全长序列,并进行生物信息学分析;应用qPCR技术检测其在不同温度(-10,-5,0,5,25和35℃)下处理1 h后及25℃下恢复30 min后在沙葱萤叶甲2龄幼虫中的表达量变化。【结果】自沙葱萤叶甲克隆获得一个丝氨酸蛋白酶基因,命名为GdSP(GenBank登录号:MG797556)。该基因全长1 110 bp,开放阅读框969 bp,编码322个氨基酸;蛋白预测分子量35.41 kD,等电点5.61;编码蛋白具有丝氨酸蛋白酶的典型特征,具有一个跨膜结构,无信号肽。同源序列比对和系统发育分析表明,GdSP与光肩星天牛Anoplophora glabripennis SP的同源性最高,氨基酸序列一致性为30.53%。qPCR测定结果表明,不同温度处理间2龄幼虫中GdSP表达量差异不显著,但对各高低温(-10℃除外)胁迫处理回温后GdSP表达量显著上升。【结论】快速冷驯化对沙葱萤叶甲丝氨酸蛋白酶基因表达无显著影响,而回温可诱导其上调表达。     

沙葱萤叶甲的过冷却能力与抗寒性

【目的】沙葱萤叶甲Galeruca daurica Joannis于2009年开始在内蒙古草原暴发成灾, 发生地区不断扩大, 危害日趋严重, 严重影响内蒙古草原畜牧业的可持续发展和生态安全。低温是影响昆虫生长发育和存活的关键因子, 而昆虫对低温的耐受性决定了其越冬存活率。了解沙葱萤叶甲的过冷却点及抗寒能力有助于预测其分布范围及种群数量动态。【方法】采用热电偶法, 在室内测定了沙葱萤叶甲各发育阶段的过冷却点; 比较了幼虫在不同低温条件下（-6~-14℃）暴露2 h及在-5℃低温条件下暴露不同时间（0.5~8 d）的存活率。【结果】沙葱萤叶甲不同发育阶段的过冷却点存在显著差异, 从低到高依次为卵（-29.8℃）、 1龄幼虫（-14.6℃）、 2龄幼虫（-13.3℃）、 蛹（-12.1℃）、 3龄幼虫（-10.2℃）和成虫（-9.0℃）; 越冬卵12月和1月的过冷却点最低, 2月的过冷却点最高。随着处理温度的降低和处理时间的延长, 幼虫的存活率降低。1, 2和3龄幼虫在-5℃下的半致死时间（Ltime₅₀）分别为3.84, 3.80和2.28 d, 低温处理2 h后半致死温度（Ltemp₅₀）分别为-10.1, -9.1和-8.5℃, 高于其过冷却点。【结论】说明沙葱萤叶甲幼虫为不耐寒冷型(chill-intolerant)。     

双斑长跗萤叶甲在玉米田的种群消长规律

【目的】明确双斑长跗萤叶甲Monolepta hieroglyphica(Motschulsky)的发生规律,为双斑长跗萤叶甲的预测预报和综合治理提供科学依据。【方法】2011年通过田间系统调查对山西省忻州市和晋中市玉米田的双斑长跗萤叶甲的种群消长动态进行了系统研究。【结果】在山西省忻州市的玉米田,越冬卵在5月下旬开始孵化,幼虫以玉米根系为食,6月中旬为幼虫的发生高峰期,部分老熟幼虫开始化蛹,6月下旬成虫开始羽化出土,为害玉米叶片,8月初成虫种群数量达到最高峰,8月中旬以后随着玉米花丝大部分萎蔫,叶片开始衰老,玉米田间的成虫种群数量也急剧下降。10月中旬玉米田成虫基本消失,但是在杂草上还能发现少量的成虫。在山西省晋中市,玉米的生育期比忻州市晚10 d左右,双斑长跗萤叶甲的发生期也相对晚一些,但是种群数量的发生发展趋势基本是一样的。双斑长跗萤叶甲成虫在靠近杂草的玉米田边比在玉米田中部的发生更重,而在成虫的发生高峰期,田边和田中间的种群密度无显著性差异。【结论】双斑长跗萤叶甲在山西省1年发生1代,在山西省忻州市的发生期比晋中市的要早10 d左右,这与两地玉米田的不同的土壤性质、灌溉方式、玉米生育期等因素有关。     

Notes on black elytron species of Pyrrhalta Joannis and the description of a new species from China (Coleoptera,Chrysomelidae, Galerucinae)

Thirteen species of Pyrrhalta Joannis, 1865 with black elytron are reviewed. A key to species, photographs of aedeagus and habitus are provided. Pyrrhalta qianana sp. n. is described from Guizhou, China. Pyrrhalta martensi Medvedev & Sprecher-Uebersax, 1999 is newly recorded from China (Tibet).     

A Taxonomic Study on the Larvae of the Genus Pyrrhalta Joannis (Coleoptera: Chrysomelidae: Galerucinae) from Korea

Last instar larvae of Pyrrhalta annulicornis, P. fuscipennis, P. humeralis, P. lineola and P. maculicollis are described and illustrated for the first time in Korea. Their taxonomic remarks, tubercle patterns and a key are also given.     

Oviposition strategy as a means of local adaptation to plant defence in native and invasive populations of the viburnum leaf beetle

Herbivores have been hypothesized to adapt locally to variation in plant defences and such adaptation could facilitate novel associations in the context of biological invasions. Here, we show that in the native range of the viburnum leaf beetle (VLB, Pyrrhalta viburni), two populations of geographically isolated hosts-Viburnum opulus and Viburnum tinus-have divergent defences against VLB oviposition: negative versus positive density-dependent egg-crushing wound responses, respectively. Populations of beetles coexisting with each host show an adaptive behavioural response: aggregative versus non-aggregative oviposition on V. opulus and V. tinus, respectively. In parallel, we show that in North America, where VLB is invasive, defences of three novel hosts are negatively density-dependent, and beetles' oviposition behaviour is aggregative. Thus, local adaptation to plant defences has the potential to facilitate the invasion of herbivores onto novel hosts.     

Patch-exploitation patterns in an egg parasite of elm leaf beetle

Eggs of elm beetle,Pyrrhalta luteola (Muller), normally occur in masses which can be viewed as discrete host patches in space and time. Analysis of>800 egg masses from 3 field sites in northern California revealed that the number of eggs/mass varied from 2 to>40 and that the imported egg parasiteTetrastichus gallerucae (Fonsc.) exploited a relatively large proported of the egg masses at certain times. The spatial relationship between parasitization and number of eggs/egg mass was assessed in 2 ways-i.e., for exploited masses only and for all masses combined (exploited+nonexploited). Percent parasitization was density independent in most cases for both data sets. It is suggested that both methods of analysis can provide patterns which are relevant to biological control of insect pests.      

Indirect effects of a non-target species, Pyrrhalta luteola (Chrysomelidae) on the biodeterioration of Brussels tapestries

Treatments to preserve artwork are usually designed to control for known target species. Nevertheless, indirect effects of non-target species may cause important biodeterioration. During the restoration of a series of Brussels tapestries preserved in the Abbey of Sacromonte (Granada, Spain), we studied one of the tapestries which presented a high degree of deterioration, principally large losses of material and color alterations. We found important biodeterioration effects produced by a non-target insect species, the beetle Pyrrhalta luteola (Chrysomelidae). We raised P. luteola beetles in the laboratory on different constituent materials to identify the effects of this insect on the tapestry. Results showed that beetle remains were mainly responsible for the chromatic changes. Although beetles did not consume textile materials, and had not direct effects in tapestry destruction, its presence indirectly affected the tapestry due to the higher mold-growth obtained in samples of materials with beetle remains. Based on these results, we propose some recommendations to consider control for these potential biodeterioration agents in artwork conservation programs.     

Resistance of temperate Chinese elms (Ulmus spp.) to feeding by the adult elm leaf beetle (Coleoptera: Chrysomelidae)

Newly acquired Ulmus species from the temperate regions of China growing at The Morton Arboretum, Lisle, IL, were evaluated in laboratory bioassays for feeding preference and suitability for larvae and adults of the elm leaf beetle, Pyrrhalta luteola (Müller). Larval and adult no-choice and adult multiple-choice feeding studies revealed that U changii, U. lanceaefolia, U. prunifolia, U. pseudopropinqua, and U. taihangshanensis were the least suitable for feeding and reproduction by the elm leaf beetle. Ulmus wallichiana and the highly preferred U pumila were more suitable for feeding and reproduction. Suitability had no significant effect on male and female longevity. The least suitable species of U. changii, U. lanceaefolia, U. prunifolia, U. pseudopropinqua, and U. taihangshanensis show promise for future elm breeding programs.     

Adaptation to different host plant ages facilitates insect divergence without a host shift

Host shifts and subsequent adaption to novel host plants are important drivers of speciation among phytophagous insects. However, there is considerably less evidence for host plant-mediated speciation in the absence of a host shift. Here, we investigated divergence of two sympatric sister elm leaf beetles, Pyrrhalta maculicollis and P. aenescens, which feed on different age classes of the elm Ulmus pumila L. (seedling versus adult trees). Using a field survey coupled with preference and performance trials, we show that these beetle species are highly divergent in both feeding and oviposition preference and specialize on either seedling or adult stages of their host plant. An experiment using artificial leaf discs painted with leaf surface wax extracts showed that host plant chemistry is a critical element that shapes preference. Specialization appears to be driven by adaptive divergence as there was also evidence of divergent selection; beetles had significantly higher survival and fecundity when reared on their natal host plant age class. Together, the results identify the first probable example of divergence induced by host plant age, thus extending how phytophagous insects might diversify in the absence of host shifts.     

Anatomical and nutritional factors associated with susceptibility of elms (Ulmus spp.) to the elm leaf beetle (Coleoptera: Chrysomelidae)

A wide range of susceptibility exists across elm (Ulmus) species and hybrids to the elm leaf beetle, Pyrrhalta luteola (Müller) (Coleoptera: Chrysomelidae). We evaluated various elm species, hybrids, or cultivars (taxa) growing in an experimental plantation in the city of Holbrook, AZ, for leaf anatomical (toughness and trichome density) and nutritional (minerals and sugars) traits that may be associated with host resistance. Leaf toughness and percentage of defoliation (susceptibility) were not correlated. However, we found weak negative correlations between percentage of defoliation and density of trichomes on the leaf abaxial surface. Of the 11 leaf nutrients examined, concentrations of iron and phosphorus correlated inversely with percentage of defoliation. The remaining nine traits did not show any correlation with percentage of defoliation. We concluded that individual anatomical and nutritional traits of elm species/hybrids do not seem to create a strong barrier to elm leaf beetle defoliation. However, the results from a stepwise multiple regression analysis indicated that collectively, these traits may play an important role in determining susceptibility.     

Ovipositional biology of Viburnum leaf beetle, Pyrrhalta viburni (Coleoptera: Chrysomelidae)

Pyrrhalta viburni Paykull, a new landscape pest in the United States, feeds in both the larval and adult stages on foliage of plants in the genus Viburnum. We measured lifetime oviposition capacity of mated and unmated females reared in the laboratory versus field-collected females, as well as ovipositional response to physical characteristics of the host plant. Both mated and unmated females produced eggs, but at different rates. Field-collected females and mated females reared in the laboratory laid similar numbers of egg masses containing similar numbers of eggs, but unmated females laid approximately one half as many eggs, the result primarily of smaller clutch size. Mated females reared in the laboratory had a preovipositional period of 11.4 +/- 1.7 versus 29 + 11.7 d for unmated females, and unmated females lived significantly longer than mated females. The angle and diameter of stems of V. trilobum, a very susceptible host, both greatly influenced oviposition; females laid most eggs on vertically oriented stems, and those of smallest diameter; when these factors were combined, stem diameter predominated. Females also had a very strong geotactic response, preferring to lay eggs on portions of stems toward gravity, even when stems were at fairly shallow angles.     

Effects of induced water stress on leaf trichome density and foliar nutrients of three elm (Ulmus) species: implications for resistance to the elm leaf beetle

Seedlings of three elm species with variable susceptibility to the elm leaf beetle (Pyrrhalta luteola Müller) (Coleoptera: Chrysomelidae) were subjected to three water stress treatments (no stress, low stress, and high stress) in a greenhouse experiment. The species tested were Ulmus pumila L. (Siberian elm = highly susceptible), U. parvifolia Jacq. (Chinese elm = resistant), and U. americana L. (American elm = intermediate). The seedlings were analyzed for changes in the levels of selected host traits (trichome density, foliar concentration of nitrogen [N], phosphorus [P], potassium [K], calcium [Ca], magnesium [Mg], iron [Fe], and manganese [Mn]), some of which had previously been implicated in resistance to the elm leaf beetle. Density of leaf abaxial surface trichomes (simple, bulbous, and total trichomes) and foliar Fe and Mg concentrations increased significantly in the highly susceptible Siberian elms under water stress. In contrast, stress reduced trichome density in the moderately susceptible American elms, but it had no effect on levels of foliar mineral nutrients. The stress treatments had no influence on host traits in the resistant Chinese elms. The results suggest that environmental stress can alter plant traits that are likely involved in determining resistance of elms to the elm leaf beetle.