褐飞虱胁迫下两种水稻不同生育期玉米素核苷含量动态

应用酶联免疫吸附法（enzyme-linked immunosorbent assay, ELISA）研究了在褐飞虱胁迫下镇稻2号（粳稻）和协优63（籼稻）抽穗期和灌浆期水稻叶片和根中玉米素核苷（zeatin riboside, ZR）含量变化情况。结果表明, 镇稻2号抽穗期ZR含量变化比灌浆期对褐飞虱侵害更为敏感,密度分别为15、30、60、120头/株的褐飞虱侵害水稻抽穗期3、6、9天后,叶片和根中ZR含量显著下降;灌浆期除各个褐飞虱密度侵害9天叶片中ZR含量和120头/株褐飞虱密度侵害9天根部ZR含量显著下降外,其他处理根和叶片中ZR含量下降不明显。协优63抽穗期受褐飞虱侵害后体内ZR的变化不同于镇稻2号,密度分别为15、30、60头/株的褐飞虱侵害3天后,叶片中ZR含量明显升高;在灌浆期,除30、60、120头/株褐飞虱密度侵害6天和120头/株褐飞虱密度侵害9天叶片中ZR含量有显著增加外,其他处理变化不明显。表明不同品种水稻在不同密度褐飞虱侵害下对根和叶片中ZR含量有不同的影响。     

Photosynthesis and translocation of assimilates in rice plants following phloem feeding by the planthopper Nilaparvata lugens (Homoptera: Delphacidae)

Experiments were conducted to measure the effect of feeding by the planthopper Nilaparvata lugens (St?l) on photosynthesis and the translocation of assimilates in rice plants, Oryza sativa L. We used mature japonica rice plants and applied the 13CO2 feeding method to evaluate those physiological effects. The photosynthetic rate was suppressed by N. lugens infestation, especially at the lower leaf position, with rates 30% lower than that of control plants at the booting stage. Leaf nitrogen concentration in infested plants was also lower than that in control plants. After flowering, the chlorophyll content and total plant dry weight were reduced by N. lugens. Stem and sheath dry weights were significantly reduced by N. lugens infestation, whereas panicle dry weight was not affected. Little effect was found on disruption in translocation of assimilates, even when 13CO2 was supplied to the infested leaves. Results suggested that removal of assimilates and reduction in photosynthesis by N. lugens have the greatest effect on growth and yield of rice plants as compared with the disruption in the translocation of assimilates. Plant death can occur by N. lugens infestation if the amount of energy supplied is less than that required for tissue maintenance.     

褐飞虱侵害后不同水稻品种根及叶片脱落酸含量的变化

为了解褐飞虱Nilaparvata lugens （Stål）侵害后水稻耐虫性与植物体内源激素关系,应用酶联免疫吸附法（enzyme-linked immunosorbent assay, ELISA）研究褐飞虱若虫侵害分蘖期超级培矮64S/E32和TN1,灌浆期协优963和TN1后根及叶片脱落酸（abscisic acid, ABA）含量变化.结果表明：褐飞虱侵害分蘖期超级培矮64S/E32和TN1后3 d,叶片ABA含量显著上升,ABA含量根冠比（根ABA/叶片ABA）显著下降;侵害后6 d,超级培矮64S/E32叶片ABA含量显著下降,根冠比显著上升;但TN1叶片ABA含量在褐飞虱侵害后3 d和6 d显著上升,根冠比显著下降.褐飞虱侵害灌浆期协优963与分蘖期超级培矮64S/E32变化一致,TN1在褐飞虱侵害后3 d叶片ABA含量显著上升,根冠比显著下降;侵害后6 d,叶片ABA含量、ABA含量根冠比均显著上升.由ABA含量变化百分比可见,分蘖期ABA含量变化幅度较灌浆期大;耐虫品种变化幅度较感虫品种大,持续期较感虫品种短;叶片变化幅度较根部大.褐飞虱侵害后,两种不同生育期两种抗性不同的水稻品种比较,耐虫品种叶片ABA含量先上升（3 d）后下降（6 d）,ABA含量根冠比先下降（3 d）后上升（6 d）;感虫品种叶片ABA含量持续上升（3 d和6 d）,分蘖期ABA含量根冠比持续下降（3 d和6 d）,灌浆期ABA含量根冠比先下降（3 d）后上升（6 d）;耐、感虫水稻品种根部变化规律不明显.这些差别表明不同水稻（耐虫和感虫）品种受褐飞虱侵害后体内ABA含量变化规律不同.本研究结果对深入阐明水稻耐虫品种的机制具有重要参考价值.     

Electroantennogram responses of Nilaparvata lugens (Homoptera: Delphacidae) to plant volatile compounds

Electroantennogram (EAG) recordings were made from the plaque organ receptors of the rice brown plant hopper, Nilaparvata lugens (St?l). The receptors of the distal plaque organ of both male and female hoppers responded to the air from above chopped rice plants, and those of females gave a larger response. Female hoppers, tested with 27 plant volatiles, gave dose-related responses to 16 of the compounds, including a range of green leaf volatiles. The aliphatic aldehydes hexanal and (E)-2-hexenal elicited a larger response than a range of green leaf alcohols. Of the remaining compounds, acetophenone, benzaldehyde, ethyl and methyl benzoate, and amyl and isoamyl acetate were among the most effective in eliciting responses. Receptor recovery times showed some variation between compounds. A comparison of the dose-related response of 16 compounds in paraffin oil and in pentane showed that considerably larger responses were recorded when the highly volatile solvent pentane was used.     

Bacterial Symbionts of the Brown Planthopper,Nilaparvata lugens (Homoptera: Delphacidae)

The brown planthopper (Nilaparvata lugens Stål), the most destructive pest of rice, has been identified, including biotypes with high virulence towards previously resistant rice varieties. There have also been many reports of a yeast-like symbiont of N. lugens, but little is known about the bacterial microbes. In this study, we examined the bacterial microbes in N. lugens and identified a total of 18 operational taxonomic units (OTUs) representing four phyla (Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes) by sequencing and analyzing 16S rRNA gene libraries obtained from three populations of N. lugens, which were maintained on the rice varieties TN1, Mudgo, and ASD7. Several of the OTUs were similar to previously reported secondary symbionts of other insects, including an endosymbiont of the psyllid Glycapsis brimblecombei, an Asaia sp. found in the mosquito Anopheles stephensi, and Wolbachia, found in the mite Metaseiulus occidentalis. However, the species and numbers of the detected OTUs differed substantially among the N. lugens populations. Further, in situ hybridization analysis using digoxigenin-labeled probes indicated that OTU 1 was located in hypogastrium tissues near the ovipositor and ovary in biotype 1 insects, while OTU 2 was located in the front of the ovipositor sheath in biotype 2 insects. In addition, masses of bacterium-like organisms were observed in the tubes of salivary sheaths in rice plant tissues that the insects had fed upon. The results provide indications of the diversity of the bacterial microbes harbored by the brown planthopper and of possible associations between specific bacterial microbes and biotypes of N. lugens.Close associations between insects and the microbes they harbor appear to be common. Symbionts have been found to contribute to the nutrition, development, reproduction, speciation, and defense against natural enemies of their host insects (1, 11, 18, 30, 39). The small brown planthopper (Laodelphax striatellus) and the white-backed planthopper (Sogatella furcifrea) also reportedly harbor an alphaproteobacterial Wolbachia symbiont (29) that can be transferred horizontally between different insect species and that affects its hosts'' sexual reproduction, cytoplasmic incompatibility, and immune responses (21, 38, 39).The brown planthopper, Nilaparvata lugens Stål (Homoptera: Delphacidae), is a monophagous insect herbivore of rice (13) that feeds on rice phloem and causes serious damage to rice crops. N. lugens reportedly harbors an intracellular, eukaryotic “yeast-like symbiont” (YLS) in the fat body, which plays a key role in recycling uric acid (3, 33). However, little is known about bacterial symbionts in N. lugens.It has been well recognized that diversity exists within insect species and that “biotypes” or populations that are adapted to or that prefer a particular host can frequently develop (10, 12). The behavioral and physiological responses during insect establishment on plants are feeding, metabolism of ingested food, growth, adult survival, egg production, and oviposition (34). In N. lugens, the biotype is assigned to a population with the ability to damage varieties of rice that carry resistance genes and that were previously resistant to it (5). It has been claimed that some biotypes of N. lugens differ in small morphological features, isozymes, and DNA polymorphisms (6, 25, 36). However, the precise nature of the virulence-conferring mechanisms in N. lugens biotypes (and their modes and stability of inheritance) is not clear. It is interesting to survey symbionts in different biotype populations of N. lugens.Generally, the 16S rRNA gene has been used as a molecular marker enabling the detection of as-yet-uncultured microbes, and it facilitates a profound investigation of microbial diversity (2, 22, 44). We initiated a study using molecular methods to investigate the bacterial symbionts of N. lugens. The major objective of this study was to identify bacterial microbes in N. lugens. The identified bacterial microbes appeared to be associated with different populations of N. lugens and in some cases were located in specific tissues, according to in situ hybridization (ISH) analyses.     

Flight duration of the brown planthopper, Nilaparvata lugens (Homoptera: Delphacidae)

ABSTRACT.

• 1 The brown planthopper, Nilaparvata lugens (Stål), is a major pest of rice in Asia. It is known to make wind-assisted migratory flights each year to colonize the summer rice growing areas of China, Japan and Korea.
• 2 Modelling windborne displacements between rice growing areas in Asia requires migratory behaviour and flight duration to be established for this insect.
• 3 Field and laboratory observations suggest that N. lugens take-off at dusk and that some continue flying for up to 24–26 h if the temperature is ≥ 17°C.
• 4 Trajectories for 10 m above ground level and 1.5 km above mean sea level are used to identify possible sources and, hence, to estimate the flight times of N.lugens caught in nets on ships on the East China Sea in 1973 and 1981.
• 5 Estimated flight times between the sources and the ships ranged from about 9 to 30 h.
• 6 Results suggest that long-distance migration can occur in surface winds, when they are strong, but that long-distance migration is more likely at 1.5 km.
• 7 When simulating windborne displacements of N.lugens, it can be assumed that in areas and at heights where the temperature is ≥ 17°C, some migrants will fly downwind for up to 30 h after a dusk take-off. Others will fly for shorter periods, giving the population as a whole the opportunity to colonize all the rice crops flown over.

     

褐飞虱侵害后不同水稻品种根及地上部氮、磷、钾含量的变化

利用浓硫酸-过氧化氢消煮法,研究了不同耐、感虫品种水稻分蘖期在褐飞虱侵害胁迫后根及地上部间营养成分含量的变化情况。结果表明：褐飞虱侵害协优963后3天,根及地上部N、P、K含量、6天根及地上部N含量、9天地上部N含量在60、120头/株侵害后变化不明显;6天根及地上部P含量、6天根K含量、9天地上部P含量、9天根及地上部K含量在120头/株侵害后显著下降,60头/株侵害后变化不明显;6天地上部K含量、9天根N含量在60、120头/株侵害后均显著下降。对于协优63,3天地上部N含量、3天根及地上部P含量、6天及9天地上部N、P含量在60、120头/株侵害后变化不明显;6天根N、P含量、根及地上部K含量、9天根N含量在120头/株侵害后显著下降,60头/株侵害后下降不明显;3天、9天根及地上部K含量、9天根部P含量在60、120头/株侵害后均显著下降。表明不同水稻品种体内不同营养物质含量在褐飞虱侵害后变化不同,协优63较协优963敏感;K含量变化最明显,其次为P,最后为N;而且根比地上部对褐飞虱的反应敏感。     

褐飞虱EST资源的微卫星信息分析

表达序列标签（expressed sequence tags,ESTs）是开发微卫星标记的一个重要的资源。褐飞虱Nilaparvata lugens （Stål） EST序列的公布为开发EST-SSRs提供了宝贵的数据资源,本研究利用生物信息学对NCBI公共数据库中的37 398条褐飞虱ESTs序列进行EST-SSRs特征分析,得到全长为7 619 324 kb的无冗余EST 9 852条。按照3个不同的查找标准在这些序列中搜索SSR。查找结果显示：褐飞虱EST-SSRs主要重复基元以1～3碱基为主,占总EST-SSR的95％以上。在单碱基重复基元中,A/T是占优势的重复基元,在二相重复类型中,AG/CT重复基元出现的频率最多,而AAG/CTT是三相重复中占绝对优势的重复基元。在褐飞虱EST-SSRs中未查找到GC重复基元。以100 bp为参照,在3种查找标准下含有SSR的EST序列中两端侧翼序列均≥100 bp的序列分别为738,89和42个。通过分析褐飞虱EST-SSRs标记可以为褐飞虱和近缘种的SSR标记的开发提供信息,同时通过分析褐飞虱EST-SSRs的分布频率和分布特征可以为昆虫EST-SSRs的研究提供借鉴和参考。     

褐飞虱卵黄蛋白的分离及其生化特性

电泳结合不同染色方法证实, 褐飞虱Nilaparvata lugens (Stål)卵黄蛋白为一种糖脂结合蛋白,其分子量约为314 kD,由148 kD、124.5 kD和39.6 kD 3个亚基组成。免疫反应证明,卵黄蛋白只存在于生育期的雌性褐飞虱成虫体内。褐飞虱卵黄蛋白具有种的特异性,其免疫血清与白背飞虱的卵黄蛋白无交叉反应。     

Preference and performance of Anagrus nilaparvatae (Hymenoptera: Mymaridae): effect of infestation duration and density by Nilaparvata lugens (Homoptera: Delphacidae)

The effect of Nilaparvata lugens (St?l) infestation duration and density on the host preference and performance of Anagrus nilaparvatae Pang et Wang, an egg parasitoid of rice planthoppers, was determined. The results showed that the parasitoid preferred N. lugens eggs on the plants infested with 10 gravid N. lugens females for 1 d to those plants infested with 10 gravid females for 2 or 3 d. It was also found to prefer N. lugens eggs on plants infested with 10 or 20 adult females after 24 h of infestation to those plants infested with 5 or 80 females. The parasitoid's offsprings had lower survival rates, fecundities, female ratios, indexes of capacity for population increase, and longer developmental durations on plants when they were infested with high N. lugens density (80 adult females per plant). However, the performance of the parasitoid on plants infested with low N. lugens density (5 female adults per plant) was similar to those on plants with intermediate N. lugens density (10 or 20 adult females per plant). Low preference of the parasitoid for N. lugens eggs on plants with heavy or light infestation levels may be correlated with low host suitability and detectability, respectively. The result implies an important role of herbivore-induced rice volatiles in the host preference of the parasitoid A. nilaparvatae, by which the parasitoid perceives the host and its suitability.     

利用抑制差减杂交技术分离受水稻抗性调控的褐飞虱基因

为分离受水稻抗性调控的褐飞虱Nilaparvata lugens基因, 以取食感虫水稻台中1号和高抗水稻B5的2叶1芯秧苗24 h的褐飞虱4龄若虫为起始材料, 采用抑制差减杂交技术构建了两个群体间的正反向差减cDNA文库。通过斑点杂交从差减文库中筛选代表受水稻抗性调控的基因的cDNA克隆, 进行测序和功能分析, 挑选具功能的基因进行Northern杂交验证。结果表明, 通过斑点杂交筛选到的98个阳性克隆代表92个互不重复的单基因, 其中25个与动物的已知蛋白基因存在较高的同源性。Northern杂交表明, 这25个基因有11个表达上调, 8个表达下调, 提示它们可能在褐飞虱适应抗性水稻过程中发挥了重要作用。本研究结果为克隆上述新基因的全长cDNA序列及进一步研究其在褐飞虱与水稻互作中的功能奠定了基础。     

Study of the relationship between the content of the rare earth element Eu in rice plants and in Nilaparvata lugens (Hemiptera: Delphacidae)

Abstract The current study investigated changes in the content of the rare earth element Europium (Eu) in roots, shoots and leaves of rice plants and in Nilaparvata lugens Stål (Hemiptera: Delphacidae) using an Eu marker and hydroponic culture. The results showed that N. lugens infestation significantly reduced Eu content in roots, shoots and leaves of two rice varieties, Shenyou 1 and Xieyou 963. The Eu content in roots, shoots and leaves of the susceptible variety, Shenyou 1, was significantly higher than that in the resistant variety, Xieyou 963. The Eu content of N. lugens fed on Shenyou 1 was significantly higher than of those fed on Xieyou 963. In addition, the Eu level was elevated at a higher density of N. lugens infestation. Eu content in the bodies of N. lugens was related to their weight and honeydew excretion, with a significant positive correlation. Thus, Eu content in the bodies of N. lugens can be considered an index of the amount of phloem sap taken in by N. lugens because the amount of honeydew excretion is proportional to the amount of phloem sap consumed. The ratios of Eu content in N. lugens to that in roots, shoots and leaves of rice plants were elevated at a higher N. lugens infestation density. That ratio was maximal in leaves, was intermediate in shoots and was minimal in roots. There was no significant difference in ratios between the two plant varieties. An Eu marker may be useful in the screening of resistant varieties and in the study of the mechanisms of resistance.     

褐飞虱侵害对不同抗性水平水稻根部吸收氮、磷、钾的影响

为了解褐飞虱Nilaparvata lugens侵害后水稻抗性水平与根系吸收氮（N）、磷（P）、钾（K）的关系,对不同抗性水平的水稻受褐飞虱侵害后的平均受害水平、植株功能损失指数及水稻吸收水培液营养元素N,P,K情况进行了研究。结果表明：不同水稻品种（TN1、协优63、协优963和超级培矮64S/E32）接种褐飞虱后,TN1受害最严重,协优63和超级培矮64S/E32 次之;受害最轻的是协优963,30头/株侵害后植株功能损失指数仅为0.661。40,60和80头/株侵害水稻后根系对K吸收下降程度最显著,其次为P,最后为N;且随褐飞虱侵害时间的延长（6 d, 9 d）影响愈显著。60头/株侵害TN1、协优63、协优963和超级培矮64S/E32后9 d,根系对K吸收下降率分别为164.11%,74.61%,55.16%和46.60%。由此可见,随水稻品种抗性水平的下降,接种褐飞虱后,水稻根部对水培液营养元素吸收下降程度愈显著。本研究结果可对深入阐明不同抗性水平水稻抗（耐）虫机制与根系吸收能力关系提供参考。     

Imidacloprid susceptibility survey and selection risk assessment in field populations of Nilaparvata lugens (Homoptera: Delphacidae)

Imidacloprid has been used for many years to control planthopper Nilaparvata lugens (St?l) (Homoptera: Delphacidae) in China. To provide resistance assessment for the national insecticide resistance management program, we collected a total of 42 samples of the planthoppers from 27 locations covering eight provinces to monitor their dose responses and susceptibility changes to imidacloprid over an 11-yr period (1996-2006). Results showed that most field populations maintained susceptibility from 1996 to 2003 except for a population from Guilin, Guangxi, in 1997, which showed a low level of resistance to imidacloprid. However, surveys conducted in 2005 indicated that 16 populations from six provinces quickly developed resistance with resistance ratios ranging from 79 to 811. The data collected in 2006 revealed that the resistance levels in 12 populations collected from seven different provinces decreased slightly (RR = 107-316), except the Tongzhou population (Jiangsu Province), which developed 625-fold resistance. Dominant and intensive use of imidacloprid in a wide range of rice, Oryza savita L., growing areas might be a driving force for the resistance development. Migration of the insect also significantly boosted the resistance levels due to extensive and intensive use of imidacloprid in emigrating areas and continuous postmigration sprays of the chemical. In addition, laboratory resistance selection using imidacloprid showed that resistance ratio increased to 14-fold after 27 generations, suggesting that quick resistance development might be associated with more frequent applications of the insecticide in recent years.     

The variability of mating signals and preferences in the brown planthopper,Nilaparvata lugens (Homoptera: Delphacidae)

Male and female brown planthoppers, Nilaparvata lugens(Stål) (Homoptera: Delphacidae), exchange substrate-transmitted signals prior to mating. The pulse repetition frequency of the male song is known to be involved in mate recognition and also to vary among geographical populations. Here the variability of male signals, female signals, and female preferences has been examined within a population. Female preference variation has been partitioned into variation in mean preference and variation in the window of preference of individuals. The genetic component of variation has been examined using isofemale lines. Male signal variation was limited (CV=8%) and was mainly within individuals. Female signal variation was greater (CV=15%). Female mean preference varied little (CV=10%) and was closely matched to the male signal mean, but the preference window was wide (> 4 male signal standard deviations on average) and variable (CV=56%). There was evidence for genetic variation only for preference window. These results are discussed in relation to theories of signal system evolution.     

Repeated copulation in the brown planthopper, Nilaparvata lugens Stál (Homoptera; Delphacidae)

Abstract. 1. Mating of the brown planthopper, Nilaparvata lugens , was investigated in relation to oviposition. Females became unreceptive immediately after mating and showed various types of repelling behaviour to courting males. As a result, females usually did not mate repeatedly in quick succession, but after ceasing to lay fertilized eggs they behaved as virgins and mated again before producing more fertilized eggs.
2. Copulation, followed by deposition of fertilized eggs, occurred twice, or in a few cases three times, throughout the adult stage. Copulation lasted about 2 min at the first mating and about 1 min at the second or third mating.
3. When the number of fertilized eggs began to decrease rapidly, oviposition rate also decreased, but it increased again immediately after re-mating. Repeated copulation was also related to the potential rate of population increase. In the present experiments, the estimates of net reproductive rate ( R ₀) in brachypterous females was 287.7 (first mating), 88.3 (second mating) and 7.2 (third mating) and in macropterous females 286.8, 120.6 and 3.6 respectively.
4. In a patchy environment, repeated copulation following an invasion may contribute to the increase of a population.     

褐飞虱体内类酵母共生菌与氨基酸营养的关系

利用全纯人工饲料饲喂技术,研究了缺失不同氨基酸对高温(35℃)处理后的缺菌褐飞虱Nilaparvata lugens Stål相对生长速度、体内共生菌数量的影响,发现10种必需氨基酸对缺菌褐飞虱生长的影响明显大于10种非必需氨基酸,饲料中必需氨基酸的缺少对褐飞虱（特别是高温处理褐飞虱）体内共生菌数量有一定的刺激作用。分析了缺菌试虫体内氨基酸组成和转氨酶活性的变化规律,发现在摄取的氨基酸营养相同的条件下（用全纯饲料D-97饲养）,高温处理试虫体内蛋白质氨基酸组成无明显变化,而游离氨基酸总量明显上升,且必需氨基酸所占比例显著下降,其中组氨酸（His）、异亮氨酸（Ile）、亮氨酸（Leu）、赖氨酸（Lys）、蛋氨酸（Met）和苯丙氨酸（Phe）摩尔百分含量均显著下降,表明必需氨基酸的相对缺乏可能是体内蛋白质合成受阻的一个重要原因,推测这可能是由于试虫体内共生菌数减少致使所合成的必需氨基酸减少而引起。处理试虫体内谷氨酰胺合成酶（GS）和丙氨酸氨基转移酶（ALT）活性明显提高,天冬氨酸氨基转移酶(AST)活性显著降低,结合游离氨基酸中谷氨酰胺（Gln）显著增多,推测类酵母共生菌可能利用谷氨酰胺等为原料进行必需氨基酸的合成。     

Histological observations on transovarial transmission of a yeast-like symbiote in Nilaparvata lugens Stal (Homoptera, Delphacidae)

Transovarial transmission of a yeast-like symbiote (YLS) in the brown planthopper, Nilaparvata lugens Stal, was observed with light and electron microscopy. Light micrographs showed that there was no YLS in testes and spermathecae of the mated females, indicating that sperm is not involved in the transovarial transmission of the symbiote. Both light and electron micrographs showed the processes of YLS transmission from fat body to the oocyte. In females, the symbiotes in mycetocytes moved out of the syncytium, which is formed from a layer of fat body cells, by exocytosis, and released into hemocoel. Then, the free YLS in hemolymph approached to the ovarioles near pedicel and were enclosed by follicle cells. They entered the follicle cells around the primary oocyte by endocytosis at epithelial plug of the ovariole. The YLS aggregated at the posterior end of the mature egg after entering, and finally formed a symbiote ball.     

褐飞虱对噻嗪酮抗性的遗传分析

害虫的抗性遗传特性是影响其抗性发展的一个重要因子,也是制订抗性治理对策的重要依据。我们采用稻茎浸渍法测定了褐飞虱Nilaparvatalugens (Stal)抗性和敏感亲本、正反交（F₁、F₁₂、F'₂）及回交（BC）后代3龄若虫对噻嗪酮的剂量反应数据,研究了褐飞虱对噻嗪酮的抗性遗传特性。结果表明: 正反交后代的显性度分别为-0.3153（F₁）和-0.376 3（F'₁）,表明抗性遗传为常染色体的不完全隐性;将自交及回交后代的剂量反应数据进行单个主基因假设的卡方(χ²)检验,其卡方值分别为42.11（F₂）、5.44 （F'₂）及93.57（BC）,均大于χ^２0.05= 15.51(df=8),表明其抗性是多基因控制的。还讨论了褐飞虱对噻嗪酮的抗性治理策略。     

褐飞虱乙酰胆碱酯酶基因全长cDNA的克隆及序列分析

利用反转录聚合酶链式反应(RT_PCR)结合快速扩增cDNA末端(RACE)技术克隆了褐飞虱Nilaparvata lugens 乙酰胆碱酯酶基因cDNA。该cDNA全长2 467 bp,包含一个1 938 bp的开放阅读框(GenBank登录号AJ852420); 在推导出的646个氨基酸残基的前体蛋白中, N端的前30个氨基酸残基为信号肽,随后的616个氨基酸残基是成熟的乙酰胆碱酯酶序列,其预测的分子量为69 418 D。在一级结构中,形成催化活性中心的3个氨基酸残基(Ser242,Glu371和His485),以及在亚基内形成二硫键的6个半胱氨酸完全保守; 位于催化功能域的14个芳香族氨基酸中有10 个完全保守。该酶的氨基酸序列与黑尾叶蝉的同源性最高,达83%。对来自23种昆虫（包括褐飞虱）的30个乙酰胆碱酯酶的聚类分析显示,褐飞虱的乙酰胆碱酯酶与其中6个Ⅱ型乙酰胆碱酯酶（AChE2）同属一个支系; 此外,只存在于昆虫AChE2中的超变区及特异的氨基酸残基,也存在于褐飞虱的乙酰胆碱酯酶中。以上结果表明,所克隆的褐飞虱的乙酰胆碱酯酶基因是一个与黑腹果蝇的orthologous型基因同源的AChE2基因。