甘肃省苹果蠹蛾不同地理种群遗传多样性分析

为探明苹果蠹蛾不同地理种群的遗传变异、种群分化及传播扩散情况,利用9对微卫星引物对甘肃9个不同苹果蠹蛾地理种群的遗传多样性进行了研究。应用POPGENE 1.32和NTSYSpc 2.1软件分析表明:9对引物共检测到21个多样性位点,多态性百分比为95.45%;9个不同地理种群的观测等位基因为1.954 5,有效等位基因为1.741 3,Nei氏指数为0.411 3,Shannon指数为0.592 7±0.147 2;遗传相似度在0.325 8~0.817 0范围内。UPGMA聚类分析表明甘州、金塔种群聚为一支,与肃州种群合为一支;临泽种群与总寨种群聚为一支;七里河与敦煌种群聚为一支;高台与民勤种群聚为一支,各分支间的遗传分化系数Fst分别为0.795 0、0.531 4、0.857 9、0.598 0。Mantel检测表明,种群间的遗传距离与地理距离无显著相关性。研究表明,甘肃苹果蠹蛾种群的遗传多样性较低,遗传变异主要来自种群内部,种群之间的基因交流较少,在甘肃省的传播以人为被动传播扩散为主。     

中国疫区内苹果蠹蛾微卫星位点的扩增稳定性及遗传多样性

为揭示欧洲苹果蠹蛾微卫星位点在中国苹果蠹蛾群体遗传学研究中的有效性,以12对微卫星引物对采自中国主要疫区新疆、甘肃、黑龙江的8个苹果蠹蛾地理种群的120头个体在各位点的遗传多样性及扩增稳定性进行研究。所选取的12个微卫星位点中,有8个能够稳定扩增,各位点在大多数种群中均显示偏离哈迪-温伯格平衡,各位点上的平均等位基因数量为3.750～12.500,平均观察杂合度Ho为0.025～0.783,平均期望杂合度He为0.284～0.892。位点Cp2.P和Cp4.56分别具有较低的观察杂合度(0.109、0.025)和较高的近交系数Fis(0.806、0.954),说明这2个位点上的杂合子非常缺乏,其余6个位点均具有较高的杂合度水平和等位基因数量,适用于中国疫区内苹果蠹蛾微卫星分子标记研究。     

不同温度下苹果蠹蛾实验种群生命表

在15、20、25、28、31、34℃的恒温条件下,测定了苹果蠹蛾Cydiapomonella（L．）各发育历期、存活率及产卵量,组建了苹果蠹蛾实验种群生命表,以探讨温度对苹果蠹蛾生长发育和繁殖的影响。在15～31℃范围内,苹果蠹蛾各阶段发育历期随温度的升高而缩短,世代平均发育历期在15、20、25、28、31℃时分别为112．0、46．8、33．3、29．6、26．2天;在34℃时,苹果蠹蛾发育受阻,世代平均发育历期延长为31．4天。成虫寿命亦随温度的升高而缩短;单雌平均产卵量在25℃时最高,为92．7粒;在15和34℃条件下,没有有效卵产生。生命表分析显示,种群趋势指数（I）在20～31℃时均大于1,25℃时苹果蠹蛾种群内禀增长率（rm）和种群趋势指数最高,分别为0．0972和39．70。研究表明,25℃左右是苹果蠹蛾生长发育和繁殖的适宜温度,温度过高或偏低均不利于苹果蠹蛾的生长繁殖。     

亚洲玉米螟Bt抗性种群的遗传多样性分析

近20年来,在北美、西欧、南美、菲律宾等20多个国家大面积商业化应用Bt玉米防治欧洲玉米螟Ostrinia nubilalis(Hübner)等鳞翅目害虫的实践证明,其不可避免地胁迫靶标害虫产生遗传分化,出现新的抗性种群。研究精准有效的抗性检测方法,将为抗性监测和抗性治理策略的有效性提供科学依据。本研究基于高通量测序获得的亚洲玉米螟Ostrinia furnacalis(Guenée)转录组数据,应用MISA (MicroSAtellite)软件搜索SSR位点,从61 622条EST(Expressed Sequence Tag)序列中获得了3 467个SSR位点。通过设计、筛选,共获得3 316对特异性引物,从中挑选了150对引物进行PCR扩增,共有51对扩增出目的条带,对亚洲玉米螟敏感种群(ACB-BtS)及5个Bt毒素抗性种群(ACB-AbR、ACB-AcR、ACB-AhR、ACB-FR、ACB-IeR)进行多态性检测,最终得到20条高多态性引物。利用这20对微卫星引物共检测到126个等位基因,平均每个位点6.3个。不同Bt抗性种群间产生了一定程度的遗传分化,种群间的平均遗传分化系数(F_(st))为0.195 9,即说明种群间的遗传变异为19.6%。根据遗传距离建立了UPGMA系统发育树,显示6个种群的相似度,即ACB-AbR与ACB-AcR相似度高。种群变异相似度规律与已报道的亚洲玉米螟对各Bt毒素的交互抗性规律相一致。本研究发现的SSR位点可作为亚洲玉米螟不同Bt毒素抗性种群的分子检测方法。     

北美车前种群内遗传多样性分析

目的:研究外来入侵种北美车前种群内遗传多样性水平,以期揭示其较强环境适应性在分子水平的根源。方法:用60条10碱基随机引物对生长于浙江师范大学校园内北美车前的9个不同样本进行了PCR扩增,筛选出7条有较高多态性检测能力的引物,用这7条随机引物对北美车前的RAPD指纹图谱进行了研究。结果:(1)共扩增到82条带,每条引物扩增带数在6~17条,平均带数为11.71条,其中60条带具有遗传多态性,约占总数的73.17%。(2)浙师大校园种群内的北美车前不同样本之间DNA指纹存在着较明显的差异,这种差异主要是受到了相对空间位置和极端土壤环境条件的影响。结论:(1)北美车前在种群内有较高的遗传多样性;(2)北美车前对极端土壤环境有较强适应性。     

野燕麦种群遗传多样性的聚类分析研究

野燕麦ＡｖｅｎａｆａｔｕａＬ．分布极为广泛,因各地自然地理、气候和作物栽培制度不同,不同地区的野燕麦通过长期的适应和进化,在形态特征和生物学特性等方面产生了明显的差异,形成了不同的表现型［１～３］。这些类型的确认和划分,对于野燕麦的生物学特性、危害以...     

苹果蠹蛾的发生现状、监测技术及防治方法研究进展

苹果蠹蛾(Cydia pomonella L.)是一种严重危害苹果、梨、桃、杏的检疫性害虫,目前在我国局部分布,严重威胁我国苹果主产区。本文综述了苹果蠹蛾的发生现状、化学生态学研究进展、常用的监测技术方法以及主要防治方法,对苹果蠹蛾综合防控存在的问题及改进方向进行了讨论,以期为减缓苹果蠹蛾的扩散传播提供理论基础。     

苹果蠹蛾在中国的适生性分析

苹果蠹蛾是严重危害全球水果生产的一种重要害虫。本研究根据中国760个气象站点的气象数据和苹果蠹蛾生物学数据,综合运用CLMEX模型和ArcGIS分析相结合的方法,对苹果蠹蛾在中国的适生性进行分析。分析结果表明:苹果蠹蛾在我国的适生区域较为广泛。中高度适生区主要包括黑龙江、内蒙古、山西、宁夏、甘肃,吉林、北京、陕西、新疆、西藏的大部分地区,辽宁西部、河北西部和北部、青海北部、云南北部、四川西部、贵州西部及山东沿海地区。     

中国不同地区亚洲韧皮杆菌遗传多样性分析

 柑桔黄龙病是最严重的柑桔病害之一,现已威胁世界柑桔产业。已被证实在中国南部该病害通过嫁接和木虱传播。因此黄龙病的流行病学研究就显得尤为重要。本研究中,基于外膜蛋白(omp)基因的PCR-RFLP的方法被用来研究中国南部各柑桔主产区的亚洲韧皮部杆菌的遗传多样性。研究采用不同的限制性内切酶消化各地区的亚洲韧皮部杆菌的omp基因产生了不同的RFLP指纹图谱,并对各地亚洲韧皮部杆菌的omp基因进行克隆测序然后构建系统发育树。结果显示:中国不同地理区域的亚洲韧皮部杆菌具有一定的遗传多样性,即便在某一特定的地区也有一定的差异。序列分析显示中国地区的亚洲韧皮部杆菌有很高的同源性。这些结果对揭示中国柑桔黄龙病的流行病学及制定科学有效的防治策略具有一定的指导意义。     

Insecticide resistance status of the codling moth Cydia pomonella (Lepidoptera: Tortricidae) from Greece

The codling moth Cydia pomonella L. is controlled mostly with chemical insecticides in Greece and control failures have been reported. However, there are no insecticide resistance studies in the country as yet. We examined the insecticide resistance status of 33 and 38 populations of fifth-instar non-diapausing and diapausing larvae, respectively by applying bioassays, biochemical and DNA diagnostics. Diagnostic concentrations of azinphos-methyl, phosalone, deltamethrin, thiacloprid, fenoxycarb, tebufenozide, methoxyfenozide and diflubenzuron were used in bioassays. Almost all populations showed reduced susceptibility to at least one insecticide and approximately half of them to all insecticides examined compared to a laboratory susceptible strain used as reference. However, only one out of six populations tested showed reduced susceptibility in ovicidal tests with fenoxycarb. Cross-resistances were observed among most insecticides, except from the pairs fenoxycarb–phosalone and thiacloprid–phosalone, in non-diapausing larvae. The more obvious biochemical marker associated with the reduced susceptibility observed in both larval instars was elevated cytochrome P₄₅₀ polysubstrate monooxygenases activity, followed by elevated glutathione-S-transferase activity and reduced carboxylesterases activity. Neither sodium channel nor AChE known resistance mutations were found in any of the approximately 1000 individuals of each larval instar screened with diagnostic PCR. Actions for Integrated Resistance Management and application of alternative control methods are discussed.     

苹果蠹蛾半人工饲料与饲养温度的优化

为实现苹果蠹蛾Cydia pomonella(L.)的人工大规模饲养,对国外学者已经报道的半人工饲料配方进行改良,比较了取食5种饲料对苹果蠹蛾生长发育和繁殖的影响,并组建苹果蠹蛾在20、24、28、32℃条件下取食最优配方时的实验种群生命表。结果表明,来自我国西北疫区的苹果蠹蛾,在以豆粕、蔗糖、面粉、小麦胚芽为主要成分并添加酵母粉配制成的配方B饲喂后,与传统饲料配方相比显著影响苹果蠹蛾的发育历期和种群参数,提高存活率至50.24%、缩短幼虫发育历期至20.18 d、增加蛹重至26.54 mg,并提高单雌产卵量至85.30粒。生命表结果显示,24℃时种群趋势指数最高为16.42、单雌产卵量最大为62.44粒,28℃内禀增长率最高为0.08、雌性比例最大为0.55。表明我国西北疫区的苹果蠹蛾在24~28℃条件下,取食优化的半人工饲料利于其种群数量增加。     

苹果蠹蛾对高效氯氟氰菊酯的抗性现状、机制及治理策略

苹果蠹蛾Cydia pomonella属鳞翅目卷蛾科,是全球仁果类水果种植地区最重要的果树害虫之一,也是我国重大农业入侵物种,对全球水果生产造成严重威胁。当前主要采用高效氯氟氯氰菊酯等杀虫剂对苹果蠹蛾进行防治,杀虫剂的频繁使用导致其对高效氯氟氰菊酯产生了抗性。针对苹果蠹蛾对高效氯氟氰菊酯的抗性问题,该文综述了全球苹果蠹蛾种群对高效氯氟氰菊酯的抗性现状,指出由编码细胞色素P450（cytochrome P450,P450）、羧酸酯酶（carboxylesterase,CarE）和谷胱甘肽S-转移酶（glutathione S-transferase,GST）的解毒酶基因过表达导致代谢能力增强是其最普遍的抗性机制,提出合理的抗性治理策略,并对苹果蠹蛾的绿色防控进行展望。     

光周期和温度对苹果蠹蛾滞育诱导的影响

为探讨光周期和温度对苹果蠹蛾Cydia pomonella滞育诱导的影响,通过人工诱导的方法对其滞育的光周期、温度诱导条件及滞育的敏感光照虫期进行了研究。结果表明:20℃时,光照时间短于12 h,苹果蠹蛾的滞育率可达到100%;25℃时,光周期为L 8∶D 16时滞育率最高,为98.3%;30℃时,光照时间少于10 h,苹果蠹蛾的滞育率可达85%以上。在20、25和30℃条件下,幼虫滞育的临界日长分别为14.21、13.68和13.73 h。苹果蠹蛾对光照反应有2个最敏感时期,即幼虫初孵化阶段和预蛹期阶段,卵期对光照反应不敏感。表明苹果蠹蛾是一种典型的短日照滞育型种类,光周期、温度及其交互作用均对其滞育诱导具有重要影响,其中光周期占主导作用,温度随着光周期起作用,低温对滞育诱导有很好的促进作用。     

Azinphos-methyl and carbaryl resistance in adults of the codling moth (Cydia pomonella (L.), Lepidoptera: Tortricidae) from Northeastern Spain

The resistance of Cydia pomonella (L.) to organophosphates is widespread throughout the pome fruit growing areas. The lethal effects of two insecticides inhibitors of the acetylcholine esterase, azinphos-methyl and carbaryl, were evaluated in adults of five and four field populations of the codling moth, respectively. The lethal concentrations (LC₅₀ and LC₉₀) of these insecticides were determined in a susceptible strain from Spain (S_Spain). Topical bioassays using the approximate LC₉₀ values (3000 mg (a.i.)/L of carbaryl and 2000 mg (a.i.)/L of azinphos-methyl) that were obtained in S_Spain were tested as diagnostic concentrations. The enzymatic activities of mixed-function oxidases (MFO), glutathione S-transferases (GST) and esterases (EST) were measured to investigate their potential role in the detoxification of these insecticides.Carbaryl and azinphos-methyl caused ?53% and ?39% corrected mortality, respectively, in field populations, although the diagnostic concentrations applied were twofold and fourfold higher than the maximum concentration registered in Spain, respectively. The activities of MFO and GST were 7.3- to 16.1-fold higher and 2.5- to 3.7-fold higher in all the field populations compared to those in S_Spain, respectively.     

Control of the codling moth (cydia pomonella) in apple and pear orchards in Israel by mating disruption

Control of the codling moth,Cydia pomonella (L.) (Lepidoptera: Tortricidae), by pheromone-mediated mating disruption, was studied in Israel using two types of dispensers: Shin-Etsu twist-tie (normal and long-life) and TNO matrix formulation (square wafers). Factors affecting mating disruption: initial pest population density, timing of the pheromone application, early season insecticide treatments and monitoring of the pest, were evaluated. Efficacy of mating disruption was assessed by comparing catch of males in pheromone traps located in pheromone-treated and chemically sprayed control plots. Fruit infestation during the season and at picking time was the major criterion to assess mating disruption. The preferred time for pheromone application was before the first flight of the moth. When pheromones were applied at a later stage, one or two early insecticide sprays had to be used to reduce the initial level of the codling moth population. In all tests, trap catch in pheromone-treated plots was drastically reduced as compared with respective control plots. In pheromone-treated plots where total seasonal trap catch (after pheromone application) was less than 12 males/trap, fruit infestation was less than 1.5%; zero trap catch resulted in 0.1-0.3% infestation. At high population levels, mating disruption was ineffective, with the rate of infestation at picking-time being similar in pheromone-treated and control plots. All the field tests indicated that mating disruption with either Shin-Etsu ropes or TNO squares can control codling moth infestations in apple and pear orchards, at relatively low densities of the pest, as efficiently as commercial insecticide sprays.     

Prospects of monosodium glutamate use for enhancement of spinosad toxicity against codling moth neonates

It was demonstrated that neonates of the codling moth,Cydia pomonella (L.), feed on ‘Red Delicious’ apple leaves and successfully molt to the second instar. Next, using a non-choice bioassay, we targeted codling moth neonates feeding on apple leaves, with standard concentrations of a culinary taste enhancer, monosodium glutamate (MSG), and Success®, which contains 22.8% spinosad as its active ingredient. The addition of 25 ppm MSG increased feeding by 20–30%. Stimulatory properties of MSG were preserved in the presence of 12.5 ppm Success, and mortality from a 12.5 ppm Success + 25 ppm MSG combination increased by factors of 3.1–1.6 compared with Success alone. In a field experiment without rain, MSG maintained its stimulatory properties for 24 h, increasing feeding by 37%. Consistently, without rain, MSG increased the toxicity of Success in the field by a factor of × 3.5. However, the stimulatory properties of MSG dropped to 19% with 4.3 mm of rain, and to zero with 9.6 mm of rain. Increased Success toxicity by MSG was reduced to × 1.6 with 4.3 mm of rain, and dropped to zero after 9.6 mm of rain. It is concluded that MSG seems to be a promising feeding stimulant, enhancing the toxic properties of Success which itself is a good candidate for codling moth control. However, field persistence of MSG needs to be improved, either by formulating the Success + MSG combination into some field-stable matrix, or by employing a sparingly water-soluble substance mimicking MSG’s action as a feeding stimulant in codling moth neonates.     

苹果蠹蛾在辽宁省彰武县梨树上的发生规律

为明确辽宁省苹果蠹蛾Cydia pomonella(Linnaeus)的发生规律,于2014—2016年连续3年对辽宁省彰武县哈尔套镇10个村59个不同梨园采用三角屋胶粘式诱捕器进行了雄成虫数量消长动态的监测,并对幼虫蛀果率、老熟幼虫数量及化蛹行径、蛹期进行调查。结果表明,在辽宁省彰武县,从4月下旬至9月下旬,苹果蠹蛾成虫共有2次发生高峰:第1次为5月下旬,第2次为7月中下旬;苹果蠹蛾在2016年全年内有3次明显的蛀果高峰期:第1次为6月下旬至7月上旬,第2次为8月上旬,第3次为8月下旬。老熟幼虫更趋向于脱果后直接掉落到地上再从地上沿着树干往上爬并在老翘皮下寻找合适位置化蛹;苹果蠹蛾经人工饲养的蛹期为9.95 d。研究表明,苹果蠹蛾在辽宁省彰武县1年发生2代,第1代从5月中旬到9月下旬,越冬代从7月中旬到翌年6月下旬。