兰州熊蜂气味受体家族鉴定及分析

【目的】了解兰州熊蜂(Bombus lantschouensis)基因组中气味受体基因(odorant receptors,Ors)情况,为进一步分析气味受体功能提供信息,从而为研究该基因家族在熊蜂觅食、交尾、通讯等行为中的重要功能打下基础。【方法】提取兰州熊蜂胸部基因组DNA,进行Illumina高通量测序,对测序所得原始序列进行质控并拼接获得基因组序列,然后使用本地blast 2.2.28+对构建兰州熊蜂基因组本地数据库,使用已知的地熊蜂(Bombus terrestris)和意大利蜜蜂(Apis mellifera ligustica)的气味受体蛋白序列作为种子序列搜索数据库获得兰州熊蜂的气味受体序列;使用EMBOSS1.5对气味受体蛋白序列进行基本理化分析,利用GSDS2.0对家族序列的内含子与外显子位置进行分析,然后使用MEME 4.11.2对氨基酸序列进行保守基序分析,最后利用Clusta W 2.1、Trim Al 1.2、Phy ML3.0对兰州熊蜂、地熊蜂和意大利蜜蜂气味受体家族序列进行系统进化分析。【结果】共获得165个兰州熊蜂气味受体基因,包括1个非典型气味受体(olfactory receptor co-receptor,Orco)、5个假基因和159个气味受体。基因结构分析发现,气味受体家族外显子数目从4个到9个不等。其中Or 47—57的序列中外显子数量最少,为4个;Or 128—161中外显子的数量最多,为9个。根据基因结构的不同,将气味受体分为10大类型,每个类型中的序列都有相似的外显子长度与数量。Or 1—38、Or 69—85、Or 128—164这3大类所包含成员数较多(分别为38、17、37),其他7类包含成员个数都约10个,且每个类型中序列在染色体上都呈串联排布,其中Or 1—38中都包含一个较长的第一外显子。保守基序分析发现,在检索的10个保守基序中,除基序5为未知基序外,其他9个基序均包含在其保守结构域(7tm_6 domain)中。Or1—38与Or 39—46多数成员包含全部10个保守基序,基序2、3、4、9广泛存在于该家族成员序列中,这4个基序可能为该家族关键的功能区域。系统发育分析将Ors分为5个亚家族(I—V),其中亚家族II包含2种基因结构类型序列(Bl Or 97—100与Bl Or 69—85),亚家族V包含4种(Bl Or 1—46、47—57、86—95和101—107)。Bl Or 150—155与Am Or 122—139分别聚为两个分支,Bl Or 47—57与Am Or 63—65也发现类似的聚类,这表明在进化中,蜜蜂与熊蜂的Ors出现了特异性的扩张与缺失。在进化树中发现Or 115较早与其他成员分离,位于树的基部,推测该序列可能更接近气味受体家族的祖先序列。【结论】探明了兰州熊蜂的Or基因数量、基因结构及其进化关系;该基因家族在进化过程中部分成员保守基序丢失,这些结果为进一步了解Or基因功能打下了基础。     

The damage risk evaluation of Aphis gossypii on wheat by host shift and fitness comparison in wheat and cotton

Aphids are considered as one of the key pests for wheat production worldwide. Major aphid species that infest wheat in China include Sitobion avenae, Rhopalosiphum padi, Schizaphis graminum and Metopolophium dirhodum. However, during our wheat field survey in Wenshang County of Shangdong Province, China, we observed that Aphis gossypii can feed on wheat. The damage risk of A. gossypii on wheat was assessed using host shift method. A population of A. gossypii collected from a wheat field in 2015 and another population reared on cotton under laboratory conditions for a decade without exposure to insecticides were used in the study. The results of host shift demonstrated that the A. gossypii colony from wheat has not yet developed wheat specialization. Moreover, the assessment of A. gossypii fitness on wheat and cotton showed that fecundity and net reproductive rate of A. gossypii population fed on wheat was significantly higher comparing to the population fed on cotton, whether the initial host of A. gossypii population was wheat or cotton. This study raises a warning that the cotton aphid has potential to establish well on wheat and it may cause significant effects under specific circumstances. Therefore, future studies are required to evaluate the effects of A. gossypii on wheat production.     

小菜蛾成虫对性信息素与寄主挥发性物质的触角电位反应

采用电生理方法测定小菜蛾雌、雄成虫对雌蛾性信息素（8expheromone,SP）两种主要组分顺-11-十六碳烯醛（Z11-16：Aid）和顺-11-十六碳烯乙酸酯（Z11-16：Ac）,以及性信息索与寄主植物5种挥发性物质混合物的触角电位反应．结果表明：当供试浓度为20.0g·L^-1时,雄蛾对Z11-16：Aid的EAG反应相对值显著高于Z11-16：Ac．将Z11-16：Aid和Z11-16：Ae按照质量比5：5和7：3进行混配,当供试浓度为20.0g·L^-1时,雄蛾对5：5配比的EAG反应相对值显著高于7：3配比．部分供试植物挥发性物质能增强小菜蛾对性信息素的EAG反应,但会因植物挥发性物质种类和小菜蛾性别而存在差异．雄蛾对反-2-己烯醛与SP混合物以及正己醛与SP混合物的EAG反应相对值显著高于其它供试混合物,其中对反-2-烯醛与SP混合物的EAG反应相对值显著高于雌蛾,而雌蛾对顺-3-己烯醇与SP混合物、异硫氰酸丙烯酯与SP混合物以及正己醛与SP混合物的EAG反应相对值显著高于其它供试混合物,其中对青叶醇与SP混合物、异硫氰酸丙烯酯与SP混合物的EAG反应相对值要显著高于雄蛾．因此,小菜蛾性信息素主要成分与植物挥发性物质合理混配可以降低性诱剂成本,提高诱蛾效率,特别是具有提高雌蛾引诱比例的潜能．     

The biotypes and host shifts of cotton-melon aphids Aphis gossypii in northern China

Aphis gossypii is a globally distributed species and therefore has a highly variable life cycle. Populations of A. gossypii in northern China exhibit greater genotypic diversity and a broader host range, yet the details of life cycles of different biotypes is still unclear. In this study, the Cytb and 16S gene regions of A.gossypii collected from 5 common summer hosts and 4 primary hosts were analyzed. A total of 57 haplotypes were obtained from 1 046 individual A.gossypii sequences. The sequence included 44 variable sites, 27 of which were parsimony informative sites and 17 of which were singleton variable sites. The most frequent 3 haplotypes were found in 896 individuals, representing a total of 85.7% of all individuals and 36 haplotypes were found in 1 individual. A neighbor-joining tree was constructed using 21 haplotypes that were found in more than 2 individuals. Considering the individual host plant, 5 biotypes were identified. Type 1 corresponded exactly to the cucurbit host-race and the other 4 biotypes were found as cotton host-races. Type 3 was the most abundant biotype in cotton fields in northern China.     

小菜蛾成虫对十字花科9种植物挥发性物质的触角电生理反应

采用昆虫触角电位测定方法,以十字花科9种植物挥发性物质为对象,研究小菜蛾雌、雄成虫对不同浓度植物挥发性物质的触角电位反应.试验结果表明:六碳醇或六碳醛均能引起较强的EAG反应,如正己醛、反-2-己烯醛、正己醇,而萜烯类化合物如(1R) -(+)-α-蒎烯、α-萜品烯,(+)-3-莰烯、桉树脑和D-柠檬烯激发的EAG相对...     

不同寄主植物对棉蚜酯酶活性及杀虫剂敏感性的影响

通过分析研究取食5种寄主植物的棉蚜对杀虫剂的抗药性及其α-乙酸萘酯酯酶对杀虫剂不敏感性的频率分布,为棉蚜抗药性的治理提供理论依据。结果表明,取食不同寄主植物的棉蚜对杀虫剂的敏感性存在一定的差异。取食不同寄主植物棉蚜的酯酶活力及杀虫剂对棉蚜酯酶的抑制作用均不相同。其中,棉花棉蚜种群中具有高酯酶活力的个体多于其它4种寄主植物棉蚜,杀虫剂对棉花棉蚜酯酶的抑制率也小于其它寄主植物棉蚜。表明取食寄主植物不同,可影响棉蚜解毒酶活力及其对杀虫剂的耐药性。     

Including predator presence in a refined model for assessing resistance of alfalfa cultivar to aphids

The aphid quantity ratio(AQR) is defined as the number of aphids on each cultivar divided by the number of aphids on all cultivars. AQR is based on the correlation between aphid populations and their host plants and is an important tool that has been utilized in evaluating Medicago sativa(alfalfa) cultivar resistance to aphids. However, assessment of alfalfa resistance to aphids can be confused by the presence of aphid predators, causing the assessment of plant resistance to aphids to be based on incorrect aphid population data. To refine the AQR and account for the effect of predators on aphid population assessments, we introduced a parameter ‘α', corresponding to the predator quantity ratio, and used αAQR as the ratio to quantify aphid populations. Populations of both aphids(4 species) and their predators(12 species) occurring in 28 M. sativa cultivars were sampled over two years at a research station near Cangzhou, Hebei Province, China. Results showed that the most suitable evaluation period was from May to June, as the aphid population was stable during this period. Compared with the AQR method, the predator population numbers based on the αAQR had a significant inverse relationship with aphid population numbers and the 28 cultivars were clustered into three classes: the resistant class, tolerant class, and susceptible class. In addition, 17 cultivars were reassigned when evaluated using αAQR. All numerical values calculated by αAQR were displayed as a Gaussian distribution, which showed that the 28 cultivars could be clustered into nine groups using a median value(±SE) of 1±0.1. Hence, ongoing alfalfa breeding trials will be assessed using the αAQR to establish a robust system that includes agronomic performance parameters in order to generalize the new method for further studies.     

武安国家森林公园蚜虫物种多样性研究

根据对河北武安国家森林公园蚜虫的标本采集和文献查阅,对该森林公园蚜虫的物种多样性、区系成分和寄主植物多样性进行了分析研究。结果表明:该区已知蚜虫5科23属47种;武安地处古北区南端,与东洋区种类有较明显的交汇,故蚜虫区系以古北+东洋种和广布种为主;已知蚜虫的寄主植物29科,其中蚜科寄主植物最为丰富,斑蚜科次之。     

Physiology and defense responses of wheat to the infestation of different cereal aphids

Cereal aphids are major insect pests of wheat, which cause significant damages to wheat production. Previous studies mainly focused on the resistance of different wheat varieties to one specific aphid species. However, reports on the physiology and defense responses of wheat to different cereal aphids are basically lacking. In this work, we studied the feeding behavior of three cereal aphids: the grain aphid, Sitobion avenae(Fabricius), the greenbugs, Schizaphis graminum(Rondani), and the bird cherry-oat aphid, Rhopalosiphum padi(Linnaeus) on winter wheat, and the physiology and defense responses of wheat to the infestation of these cereal aphids with focus on how these cereal aphids utilize divergent strategies to optimize their nutrition requirement from wheat leaves. Our results indicated that S. graminum and R. padi were better adapted to penetrating phloem tissue and to collect more nutrition than S. avenae. The harm on wheat physiology committed by S. graminum and R. padi was severer than that by S. avenae, through reducing chlorophyll concentration and interfering metabolism genes. Furthermore, cereal aphids manipulated the plant nutrition metabolism by increasing the relative concentration of major amino acids and percentage of essential amino acids. In addition, different cereal aphids triggered specific defense response in wheat. All of these results suggested that different cereal aphids utilize divergent strategies to change the physiological and defense responses of their host plants in order to optimize their nutrition absorption and requirement. These findings not only extend our current knowledge on the insect–plant interactions but also provide useful clues to develop novel biotechnological strategies for enhancing the resistance and tolerance of crop plants against phloem-feeding insects.     

寄主植物对百合上桃蚜的影响研究

对广东百合种植区进行了系统调查,结果表明为害宜兴百合Lilium peciosum的蚜虫有桃蚜Myzus persicae和豆蚜Aphis craccivora,其中桃蚜发生的数量较多。在百合未出苗前,1-3月桃蚜主要在小白菜和芥菜为害,并在这些作物上越冬,转入春季后,当食料条件发生变化后,产生有翅蚜迁出,当年3—6月主要在百合上为害,当百合现蕾及成熟后,又产生有翅蚜迁出,在菊花和茄子等植物上为害,随后再转到小白菜和芥菜上为害。当地种植的小白菜Brassica campestris var．Chinensis、芥菜B．juncea等植物都属于桃蚜的嗜食寄主。通过组建不同寄主上实验种群生命表,表明桃蚜在百合和小白菜上种群趋势指数分别为28．512和27．027,最有利于桃蚜种群的增长。     

绿盲蝽对寄主植物花器挥发性成分的电生理和风洞行为反应

为了揭示绿盲蝽与寄主植物间的化学通讯机制,采用触角电位(EAG)和风洞技术,测定了绿盲蝽雌、雄成虫对寄主植物花器挥发物的电生理和风洞行为反应。结果表明:除间二甲苯外,绿盲蝽雌、雄成虫对a-蒎烯、乙酸叶醇酯和松油烯均产生明显的EAG反应;风洞试验结果表明,绿盲蝽雌、雄成虫对乙酸叶醇酯的定向行为反应均较强,且与对照化合物苯乙酮和顺-3-己烯醇无显著差异。乙酸叶醇酯等植物花器挥发性成分可能在绿盲蝽寄主定位过程中起着重要作用。     

西维因对几种抗性害虫的药效试验

室内毒力测定和田间药效试验证明，西维因对抗性棉蚜和棉铃虫有较好防效，其对棉铃虫的ＬＤ０为１．７５０２μｇ／头，较对照药剂甲胺磷、马拉硫磷和氯氰菊酯的毒力弱；对棉蚜的ＬＤ５０为０．３０９４μｇ／头，较对照药剂的毒力强，田间防治棉蚜及在陕西、河北两地对棉铃虫的药效均在６０％以上，同对照药剂的药效相当。     

Potato virus Y in the Russian Far East (epidemiology,strains, and damage)

Biological, physical, chemical, and antigenic characteristics of a potato virus Y are investigated. Significant strain diversity of this pathogen is found in the Far East of Russia. There are ten strains affecting potato and weeds. It is shown that PVYntm strain has a remote immunochemical affinity with common PVY strain. In the Far East of Russia, the most widespread aphid vectors of this virus are Myzus percicae Sulz., Aulacorthum solani Kalt., and Aphis gossypii Glov.     

Structure,Binding Characteristics,and 3D Model Prediction of a Newly Identified Odorant-Binding Protein from the Cotton Bollworm,Helicoverpa armigera (Hübner)

The full-length sequence of the odorant binding protein 5 gene, HarmOBP5, was obtained from an antennae cDNA library of cotton bollworm, Helicoverpa armigera (Hübner). The cDNA contains a 444 bp open reading frame, encoding a protein with 147 amino acids, namely HarmOBP5. HarmOBP5 was expressed in Escherichia coli and the recombinant protein was purified by affinity chromatography. SDS-PAGE and Western blot analysis demonstrated that the purified protein can be used for further investigation of its binding characteristics. Competitive binding assays with 113 odorant chemicals indicated that HarmOBP5 has strong affinity to some special plant volatiles, including (E)-ß-farnesene, ethyl butyrate, ethyl heptanoate, and acetic acid 2-methylbutyl ester. Based on three-dimensional (3D) model of AaegOBP1 from Aedes aegypti, a 3D model of HarmOBP5 was predicted. The model revealed that some key binding residues in HarmOBP5 may play important roles in odorant perception of H. armigera. This study provides clues for better understanding physiological functions of OBPs in H. armigera and other insects.     

Studies on the Host Biotypes and Its Cause of Cotton Aphid in Nanjing, China

The fitness of cotton aphid, Aphis gossypii Glover, on different host plants was studied. The results showed that the cotton aphid population in Nanjing formed host preference bio-types on cucumber and cotton. The aphids from cotton and cucumber could not inter-transplant successfully between the two host plants. The apterous and alate offspring of stem mother from hibiscus could not survive, reproduce and form population normally when they were transplanted to cucumber and water melon, but they could form normally when transplanted to cotton and muskmelon. There was no significant difference between the stylet length of cotton aphids from cucumber and cotton, so the stylet length was not the cause of the host preference. When the aphids originated on hibiscus were transplanted to cotton, water melon and muskmelon, the stylet could reach host phloem to absorb host plant‘s nutrients, but it couldn‘t when transplanted to cucumber. Cotton aphid can feed on muskmelon without secretion of water-soluble saliva. It could be concluded that the two host biotypes of cotton aphid came from different winter host plants.     

游离脯氨酸在棉花品种抗蚜性中的作用

棉株游离脯氨酸含量高低与棉花蚜景或蚜害指数成显著负相关。游离脯氨酸是棉花重要的抗蚜性物质之一,对棉蚜的寄主选择性、生长、发育及繁殖均有较强的制约作用。棉株受蚜虫刺吸胁迫后产生生理应激反应,可使游离脯氨酸含量大幅度升高,从而诱导提高棉花的总体抗蚜性强度。     

Application of natural aminopolysaccharide in seed film-coating for pest control and cotton growth

Cotton suffers attacks of various pests that result in a decreased yield. Until recently, the chemical control of pests was achieved through seed film-coating treatment with toxic insecticides. The natural aminopolysaccharide extracts from crab is a promising candidate as a repellent to protect cotton from Aphis gossypii Glover and Helicoverpa armigera Hubner. Experiments were conducted to study the effect of aminopolysaccharide on pest control, seed germination, plant growth and lint yield. Results indicated that all aminopolysaccharide treatments of different content repelled pests efficiently and increased significantly seed germination, plant growth and cotton yield. The natural aminopolysaccharide extracts from shrimp is a promising candidate as a repellent to protect cotton from Aphis gossypii Glover and Helicoverpa armigera Hubner. The application of the potential aminopolysaccharide by seed coating is an appropriate option for controlling pests and avoiding environmental pollution.