Imidacloprid‐induced transference effect on some elements in rice plants and the brown planthopper Nilaparvata lugens (Hemiptera: Delphacidae)

Abstract The widespread use of imidacloprid against insect pests has not only increased the rate of the development of target pest resistance but has also resulted in various negative effects on rice plants and Nilaparvata lugens resurgence. However, the effect of imidacloprid on elements in rice plants and the transference of these element changes between rice and N. lugens are currently poorly understood. The present study investigated changes of Cu, Fe, Mn, Zn, Ca, K, Mg and Na contents in rice plants following imidacloprid foliar sprays in the adult female of N. lugens that develops from nymphs that feed on treated plants and honeydew produced by females. The results indicated that imidacloprid foliar spray significantly increased Fe and K contents in leaf sheaths. Generally, Fe, Mn, K and Na contents in leaf blades were noticeably decreased, but Ca contents in leaf blades for 10 and 30 mg/kg imidacloprid treatments were significantly increased. The contents of most elements except K and Mg in the adult females and honeydew were significantly elevated. Multivariate statistical analysis showed that Fe, Mn and Na in leaf blades and Fe and Mn in leaf sheaths could be proportionally transferred to N. lugens. The relationship between most elements in adult female bodies and in the honeydew showed a positive correlation coefficient. There were significant differences in the contents of some elements in rice plants and N. lugens from different regions.     

AChE在水稻抗性诱导的褐飞虱凋亡中肠细胞中的定位及表达

【目的】褐飞虱Nilaparvata lugens是水稻的重要害虫之一。本研究旨在了解水稻品种抗性诱导的褐飞虱中肠细胞凋亡与细胞中乙酰胆碱酯酶(acetylcholinesterase,ACh E)的关系。【方法】从4龄Ⅰ型褐飞虱若虫中肠组织分离原代细胞,用不同抗性的水稻幼苗汁液处理第一代继代细胞,利用TUNEL染色法检测细胞的凋亡情况,再分别利用免疫组织化学和荧光定量PCR技术对ACh E进行亚细胞定位和检测其表达水平的变化。【结果】在对照组(未处理)细胞中,检测不到代表凋亡细胞核的绿色荧光,而免疫组化后阳性反应颜色很浅,表明细胞中存在ACh E的本底水平表达。感虫水稻品种TN1幼苗汁液处理细胞中,细胞的凋亡率为8%,抗性水稻B5幼苗汁液处理的细胞中有65%的细胞发生凋亡;而抗性水稻TKM-6幼苗汁液处理的细胞中则有85%的细胞发生凋亡。免疫组化的检测结果表明,所有凋亡的细胞中都存在ACh E的积累,且主要分布在细胞质中;ACh E在凋亡后期并不迁入凋亡小体中,而是集中在细胞核附近。荧光定量PCR检测表明,TKM-6,B5和TN1幼苗汁液处理的细胞中ACh E的表达量分别为对照细胞的29.9,18.4和8倍,该结果与细胞水平上免疫组化的检测结果一致。【结论】本研究结果证实了水稻品种抗性与其诱导的中肠细胞凋亡率呈正相关,且凋亡细胞的细胞质中存在ACh E的积累。这些发现为揭示ACh E在褐飞虱与水稻的互作中的功能、促进抗性水稻新品种的选育及开发新的褐飞虱防治措施提供了一定的参考信息。     

褐飞虱核糖体小亚基蛋白S8基因NlRPS8的克隆及其表达分析

【目的】核糖体蛋白在生物体内具有重要作用,本研究旨在探明核糖体蛋白S8在褐飞虱Nilaparvata lugens生长发育过程中的表达规律和功能。【方法】本文根据褐飞虱基因表达谱提供的差异表达基因信息及转录组提供的基因核心序列,结合褐飞虱基因组比对分析,对褐飞虱核糖体S8基因进行了预测,并通过RT-PCR获得了褐飞虱核糖体小亚基蛋白S8的全长c DNA序列,命名为NlRPS8(Gen Bank登录号:KU341337)。【结果】NlRPS8基因全长627 bp,编码208个氨基酸。进化分析表明,褐飞虱与桑粉介壳虫Maconellicoccus hirsutus亲缘关系最为接近。应用荧光定量PCR分析了基因NlRPS8的表达规律,结果表明,NlRPS8基因在褐飞虱怀卵雌虫中表达量最高,而在雄成虫、初羽化雌成虫与1~5龄若虫表达量相对较低;在褐飞虱体内,NlRPS8基因在卵巢中表达量最高,中肠次之,在其他部位表达量较低。在抗性品种ASD7和RHT上,NlRPS8基因表达量分别为感性品种TN1上的1.99倍和2.14倍。NlRPS8基因在经过饥饿处理1 d的褐飞虱组中表达量为正常组的1.6倍。【结论】研究结果显示NlRPS8基因在褐飞虱生长、繁殖中发挥作用,为进一步研究NlRPS8基因在褐飞虱生长发育和对抗性品种适应中的功能提供了线索。     

惠州地区褐飞虱对几种药剂的抗药性监测

2009年采用稻茎浸渍法测定广东省惠州地区褐飞虱Nilaparvata lugens(Stl)种群对吡虫啉、噻嗪酮、异丙威、丁烯氟虫腈、烯定虫胺和毒死蜱等杀虫剂的敏感性,测定结果表明:当地褐飞虱种群对吡虫啉产生了极高水平抗性(抗性倍数为422.2倍),对噻嗪酮、异丙威产生了中等水平抗性(抗性倍数分别为11.0和14.0倍),对丁烯氟虫腈仍处于敏感性降低(抗性倍数为3.7倍),对烯定虫胺和毒死蜱敏感(抗性倍数<3倍)。基于褐飞虱对这6种药剂抗性的明显差异,对田间治理褐飞虱合理使用药剂进行了讨论。     

中国和越南褐飞虱抗药性研究

2010年用稻茎浸渍法监测了我国和越南共14个褐飞虱Nilaparvata lugens(St(a)l)田间种群对5种杀虫剂的抗性,结果表明:14个褐飞虱田间种群对吡虫啉、噻嗪酮、氟虫腈、吡蚜酮和叶蝉散的LC50值分别在9.5287～46 6716、1.6621～17.8785、0.9818～ 8.4084、0.331...     

水稻活性成分的分离纯化及其对褐飞虱若虫的活性测定

水稻植株内的抗性次生物质是影响水稻生化抗性的主要因子之一。运用硅藻土系统溶剂分离法和硅胶柱层析分离方法分离纯化手段,从水稻(Oryza sativa L.)抗虫品种IR36甲醇粗提物中得到一种针状晶体,初步认为该晶体为一种糖甙黄酮类物质,通过该物质对褐飞虱Nilaparvata lugens(Stal)生物型Ⅱ若虫的活性测定,表明该种晶体对褐飞虱若虫的存活率、体重增加和羽化均具有显著的不良影响,不利于褐飞虱若虫的生长发育。     

中晚粳稻区褐飞虱防治策略的研究

为了解二代褐飞虱对粳稻的为害损失,确定合理的防治指标和防治策略,为科学使用噻嗪酮、吡虫啉等药剂防治稻飞虱提供依据,作者在武育粳３号上,通过接不同的虫量,并进行笼罩的方法,研究了二代褐飞虱的增殖规律和为害损失及防治指标。并进行了锐劲特防治不同世代褐飞虱策略试验。结果表明,不同水稻生育期,不同虫口密度的褐飞虱的增长速度不一样;在水稻同一生育期,低密度的增殖速度要大于高密度,但总虫量高密度区显著高于低密度区。二代褐飞虱百丛虫量与水稻产量损失呈极显著正相关（ ｒ＝ ０．９５７６**）,关系式为 ｙ＝－ １８．３３０２＋ １０． ２８９２ｌｇｘ（ｘ为 ８月上旬褐飞虱百丛虫量）,防治指标为 １１０－１２０头。因此,在早发年,迁入量较大的年份必须采取“治二压三控四”的策略,１９９６－１９９７年全面推广这一策略,成效显著。     

几种虫生真菌菌株的培养性状及其对褐飞虱的毒力

从褐飞虱Nilaparvata lugens (St(a)l)罹病虫体上新分离出的一种黄绿绿僵菌菌株Metarhizium flavoviride (Mf82),与实验室保存的黄绿绿僵菌、金龟于绿僵菌和白僵菌3种菌种5个菌株作为对比,研究了在SDAY和PDA培养基上的培养性状,并测定了它们对褐飞虱成虫的毒力.结果表明:...     

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.

     

药用野生稻挥发油对褐飞虱的驱避作用与化学成分

用四臂嗅觉仪测定了药用野生稻(Oryza officinalis)挥发油对褐飞虱(Nilaparvata lugens)行为选择的干扰作用。结果表明,在100和200mL/min流量下,药用野生稻挥发油对褐飞虱具有显著的驱避作用,与对照相比,供试褐飞虱较少选择药物处理区,且在处理区的滞留时间明显缩短。进一步采用气相色谱-质谱联用技术(GC-MS)对药用野生稻挥发油进行了化学成份分析,表明该挥发油主要成份是羰基化合物,其中含量最高的为4-乙烯基-2-甲氧苯酚(22.99%)和2,3-苯并二氢呋喃(10.80%)。推测含量较高的2,3-苯并二氢呋喃和4-乙烯基-2-甲氧苯酚可能是药用野生稻对褐飞虱的驱避活性成份。     

十六个水稻品种(系)对褐飞虱的抗虫性评价

本文通过苗期抗性鉴定、田间抗虫性鉴定、稻田节肢动物功能团优势度比较及产量测定的方法来评价16个水稻品种(系)对褐飞虱Nilaparvata lugens(Stl)的抗性。结果表明,有12个品种(系)对褐飞虱表现为抗级,2个品系表现为中抗。这14个抗虫-中抗的品种(系)在成株期也表现一定的抗虫性,且与对照TN1之间差异显著。本试验条件下,广占63-3S/华恢15、广两优106、Y58S/华15、广两优476、广占63-4S/华恢15这5个品种(系)不仅抗虫性好,而且产量也较高,分别比TN1和主栽品种扬两优6号高63.49%～66.57%和3.47%～11.57%,具有广泛的开发应用前景。     

Effects on the feeding behaviour of Nilaparvata lugens (Stål) of sublethal concentrations of the foliarly applied nitromethylene heterocycle 2-nitromethylene-1, 3-thiazinan-3-yl-carbamaldehyde

Abstract. An electrical recording technique, combined with simultaneous videofilming, was used to investigate the detailed feeding behaviour of Nilaparvata lugens (Stål) (Homoptera: Delphacidae) on rice. Phases of brief probing, of non-feeding activities, such as walking and of sustained feeding were observed. Monitoring of electrical signals recorded during long, uninterrupted periods of contact between the insect and its host plant, revealed three pattern classes. Two clearly distinguishable wave-like patterns represent the hopper sucking from xylem and phloem tissues. The third represents a 'complex' of signals believed to be associated with stylet penetration, salivation and sucking activities in non-vascular tissues of the host plant. This recording method and simple counts of honeydew excretion activity were used to assess the effect on the feeding behaviour of N. lugens of a sublethal concentration of the nitromethylene heterocycle insecticide 2-nitromethylene-l, 3-thiazinan-3-yl-carbamaldehyde applied to rice. The treatment caused a general reduction in feeding activity. Analysis of the electrical trace revealed that phloem feeding was strongly inhibited. Since these effects were largely reversed on transferring hoppers from treated to untreated hosts, the test compound was considered to have an antifeedant rather than a conventional poisoning effect on the hoppers when applied at sublethal doses.     

Cultivation,identification and quantification of one species of yeast‐like symbiotes,Candida, in the rice brown planthopper,Nilaparvata lugens

Abstract The brown planthopper (BPH), Nilaparvata lugens Stål, which is one of the most destructive pests of rice, has been confirmed to harbor yeast‐like symbiotes (YLS) in the fat body. Several morphologically different YLS have been previously isolated and cultured in vitro from BPH, but direct evidence is lacking to further clarify whether the cultured YLS were from BPH. In this study, one species of YLS was successfully cultured in vitro and simultaneously verified to exist in the fat body of BPH by sequence analysis and nested polymerase chain reaction (PCR). The cultured YLS isolate in vitro was identified as a member of the genus Candida on the basis of 18S rDNA (ribosomal DNA) and 5.8S‐ITS (internal transcribed spacer) rDNA sequence and a phylogenetic analysis of ITS sequences from yeast. Therefore, this yeast isolate was named as Candida‐like symbiotes. Candida‐like symbiotes was found to exist in fat bodies, ovaries and newly laid eggs of the BPH, but not in the heads, thoraxes and mid‐guts. In addition, the number of Candida‐like symbiotes in 1 × 10⁶ of purified YLS from BPH fat bodies was speculated to be (5.32 ± 0.22) × 10⁴ on the basis of a quantitative PCR analysis.     

褐飞虱侵害对水稻叶片、根SOD活性及丙二醛含量的影响

以分蘖期水稻协优963、TN1为供试材料,研究褐飞虱Nilaparvata lugens(St(a)l)侵害后水稻叶片、根超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量变化情况.结果表明:协优963、TN1叶片SOD活性变化不明显;15和30头/株侵害协优963后3、6、9d根SOD活性分别显著下降了88.4%和7...     

Feeding‐induced changes in defence enzymes and PR proteins and their implications in host resistance to Nilaparvata lugens

Six rice genotypes showing susceptible and resistant reactions to brown planthopper (BPH), Nilaparvata lugens were studied for feeding‐induced changes in defence enzymes and pathogenesis‐related (PR) proteins. The high resistant genotypes PTB 33, ADT 45 and ASD 7 and moderately resistant genotypes CO 43 and KAU 1661 recorded the greater expression of defence enzymes peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, total phenol and β‐1,3 glucanase in response to N. lugens feeding at 1 day after infestation (DAI) compared with susceptible genotype TN1. The greater activity of chitinase was observed in resistant cultivars at 3 DAI and the activity was sustained for more than 1 week compared with susceptible TN1. In conclusion, the current study revealed that these defence enzymes and PR proteins might attribute to the resistance mechanisms in rice plants against BPH infestation.     

褐飞虱flightin的原核表达及其差异表达检测

flightin最早发现于果蝇Drosophila melanogaster的间接飞行肌中,并且定位于粗肌丝。这种蛋白对维持肌节的结构和功能起到了重要的作用,但其在具有长短翅型分化的褐飞虱Nilaparvata lugens Stl的不同翅型间差异并不清楚。本研究以长翅型雌虫褐飞虱cDNA为模板,通过PCR扩增得到褐飞虱flightin基因ORF全长,将其连接到表达载体pGEX-6P-1中以与谷胱甘肽S-转移酶(GST)融合表达。将表达载体转入大肠杆菌表达株Rosseta,在不同温度、不同浓度IPTG的条件下诱导表达flightin,得到了最优表达条件,获得了高水平可溶性表达。在用GST抗体进行Western blotting验证GST-flightin融合重组蛋白表达的正确性后,我们通过GST柱纯化了的GST-flightin,进而用纯化后的蛋白免疫新西兰兔制备了高特异性的多克隆抗体。最后,我们用制备的多克隆抗体检测了长、短翅型雌成虫和不同发育阶段的褐飞虱体内flightin的表达差异。结果显示,flightin仅在长翅型成虫中表达,在短翅型雌成虫中未检测到其明显表达,而且flightin只在成虫期表达。本研究为进一步研究褐飞虱的flightin与其它蛋白互作、翅肌发育和翅型分化打下了基础。     

Quantitative genetic analysis of biotypes of the brown planthopper Nilaparvata lugens: heritability of virulence to resistant rice varieties

The virulence of Nilaparvata lugens Stål (Homoptera: Delphacidae) to a resistant variety of rice, Oryza sativa L., is suggested to be under polygenic control. To determine whether the virulence of N. lugens is a quantitative character that has continuous distribution or a threshold character that has a few discontinuous phenotypic forms and is determined by some underlying continuous variable, I examined the frequency distribution of honeydew excretion that has been used as a measure of ability of attacking the rice plants, and the relationship between honeydew excretion and the oviposition rate of N. lugens females using a japonica rice line Saikai 190 which has a resistance gene Bph 1. The frequency distribution in honeydew excretion significantly differed from the normal distribution, but two distributions below and above 10 mg honeydew excretion for two days did not significantly deviate from normality, suggesting a bimodal distribution. There were significant differences in the proportion of females ovipositing and the number of eggs deposited between the female groups that excreted 0–10 mg honeydew and more than 20 mg honeydew. Within these female groups, however, the reproductive performances were not different. Thus, virulence of N. lugens can be analyzed as a threshold character that has two distinct phenotypes, virulent and avirulent. I estimated the heritabilities of the virulence of N. lugens by parent-offspring regression using the percentage of virulent females in a full-sib family. The estimates of heritability were 0.41 on Saikai 190 and 0.55 on ASD7, carrying the bph 2 gene. These results show that the N. lugens population has substantial genetic variation in virulence. The regression coefficients on female and male parents were similar, suggesting similar genetic contributions by both parents. When the parental families were examined on Saikai 190 and the progeny on ASD7, the regression coefficient approached zero. There may be a low genetic correlation between virulences to the two varieties.     

水稻品种和稻虱缨小蜂对褐飞虱协同作用的模拟分析

利用我们先前的试验结果和文献资料,组建了水稻品种－褐飞虱－稻虱缨小蜂３者相互关系的模拟模型．模拟分析表明水稻品种能直接和间接地通过物理或化学特性影响稻虱缨小蜂的寄生作用,其中以影响褐飞虱卵垂直分布型、稻虱缨小蜂的迁入时间、未成熟期历期和怀印量等的水稻品种特性起主要作用．同时,模拟分析还表明中等抗性的水稻品种,配以增强稻虱缨小蜂寄生作用的品种特性,就能有效地控制褐飞虱的危害．文中还就协调水稻品种抗性与天敌协同控制褐飞虱的途径进行了讨论．     

The long-distance migration of Nilaparvata lugens (Stål) (Delphacidae) in China: radar observations of mass return flight in the autumn

Abstract.

• 1 A case study is presented of the autumn migration of the brown planthopper, Nilaparvata lugens (Stål), in the area of Nanjing in the People's Republic of China. The study was made using a high frequency (8 mm wavelength) radar and a net suspended from a kytoon.
• 2 The observations confirmed that long-distance return migrations occur in China in mid and late September, with N.lugens being carried on the prevailing north-easterly wind towards the autumn infestation and overwintering areas of the species.
• 3 After mass take-off in the late afternoon or at dusk, the migrants flew for several hours during the evening, often in a dense layer which formed at heights between about 400 and 1000m above ground. These layers often had well-defined ceilings corresponding to an air temperature of about 16°C. The migration height was above the top of the surface temperature inversion, i.e. the migrants did not fly at the height of the warmest air.
• 4 The dense layer concentrations overflying the radar were backtracked to source areas up to 240 km away in the north-east of Jiangsu Province. Planthoppers observed emigrating from the Nanjing area would reach areas in south Anhui Province or north Jiangxi Province if they flew for 12 h.
• 5 There was a second period of mass take-off at dawn. Insect layers sometimes formed but did not last longer than 1–2h.
• 6 The present results were strikingly different from those previously observed in the dry season in the Philippines, where migratory flight durations were largely confined to periods of about 30min at dusk and dawn.
• 7 Our observations are discussed in relation to the equator-wards return migrations undertaken in autumn by other insect species, and the importance of these migrations for the maintenance of long-flying genotypes in the overwintering populations is considered.