番茄植株中生物碱类对菜青虫(Pieris rapae L.)活性组分的分离和提纯

为寻找和开发不污染环境、不伤害天敌而对菜青虫有活性的行为物质,本研究对番茄植株中的生物碱类进行提取、分离和部分纯化,得到了极性和非极性生物碱类以及番茄苷(tomatine)纯品,与标样对照薄层试验吻合。将提取物对5龄菜青虫进行试验表明:生物碱类及其中的番茄苷对菜青虫有明显拒食作用;番茄苷对菜青虫有抑制生长发育的作用。

家蚕微孢子虫(Nosema bombycis)感染菜粉蝶(Pieris rapae)幼虫初探

采用不同浓度的家蚕微孢子虫感染三峡库区菜粉蝶幼虫—菜青虫,结果表明:家蚕微孢子虫对菜青虫感染性强,致死率较高,其半致死浓度(LC50)为:1.46×104/mL。当微孢子虫浓度为1.0×107/mL时,菜青虫死亡率高达88.9%,校正死亡率也高达87.2%,显著体现了家蚕微孢子虫对菜青虫幼虫的致病能力,由此探索了家蚕微孢子虫做为生物农药的可行性。

番茄植株中黄酮类对菜青虫(Pieris rapae L.)的活性组分及其对截形叶螨(Tetranychus truncalus Ehara)的毒效

本研究对番茄(Lycopersicon esculentum)植株中黄酮类进行了提取、分离和部分纯化,从中得到一种黄酮醇,经红外光谱、紫外光谱、质谱、核磁共振等分析,并与标准图谱对照,确定其分子结构为芦丁。生物测定结果表明,黄酮类对菜青虫有一定拒食作用,但未达到显著标准;黄酮类及芦丁有抑制菜青虫生长发育的作用;芦丁对截形叶螨有毒效,LC_(50)为94.00 μg·g^(-1)。

菜粉蝶Pieris rapae(L)的人工饲养及幼虫取食量的测定与防治适期

通过对菜粉蝶的人工饲养，测定各龄期在甘蓝上的取食量，累计取食量，观察其生物学特性，结果显示其取食量从１—５龄逐步递增，在菜青虫卵孵化后８天左右，累计取食量进入激增，１５天左右达到高逢。此为制订菜青虫化防适期提供科学依据。

家蚕微孢子虫(Nosema bombycis)对三峡库区菜青虫(Pieris rapae)的感染性研究

本试验用不同浓度的家蚕微孢子虫感染三峡库区菜青虫,结果表明:家蚕微孢子虫对菜青虫感染性较强,致死率高,其致死中量(ID50)为:1.5×104个/m l。当微孢子虫浓度为1.0×107个/m l时,菜青虫死亡率高达87.8%,校正死亡率也高达86.4%,体现了家蚕微孢子虫对菜青虫的显著致病能力,由此探索了家蚕微孢子虫作为生物农药的可行性。

Antifeedant Activity of Three Plant Extracts Including P. acinosa against Pieris rapae

[Objective] The research aimed to study antifeedant activity of Phytolacca acinosa Roxb., Setaria viridis （L.） Beauv and Viola yedoensis Makino extracts against Pieris rapae. [Method] Activity material was extracted from S. viridis （L.）, P. acinosa and V. yedoensis using acetone cold soak method, and non-selective antifeedant activity of extracts to Pieris rapae larva was determined by using lobular plate addition method. [Result] The results showed that the acetone leaching agent of P. acinosa had most obvious antifeedant effects on Pieris rapae. The antifeedant rate were 74.53% and 82.34% at 24 and 48 h respectively. With the concentration increasing, the antifeedant effect of P. acinosa extracts increased. The antifeedant rate of 0.050 g/ml treatment was the highest, being 74.53% and 82.34% at 24 and 48 h. [Conclusion] P. acinosa could be studied and utilized as potential botanical insecticide.

应用Bt防治菜青虫(pieris rapae linne)试验研究

应用 Bt 菌悬液对菜青虫进行了防治试验,防治效果为87.5～97.4％。应用适宜浓度为300～500倍。对人畜无害,是值得推广应用的生物制剂。

番茄植株中挥发性组分的鉴定及其对菜粉蝶(Pieris rapae L.)的触角电位活性

采用水蒸汽蒸馏法，从番茄（Lycopersiconesculentum）植株中分离到2个纯化合物，经光谱法鉴定为二苯胺和N－苯基－β萘胺；番茄植株的粗提物经GC－MS测定，除获得上述2化合物外，还得到2，6－二叔丁基－对－甲基苯酚。经触角电位（EAG）测定表明，二苯胺、N－苯基－β－萘胺与番茄植株粗提物对菜粉蝶均有较强的生物活性。

Control Efficiency of Four Biological Pesticides against Pieris rapae Linne and Their Impacts on Yield and Benefit of Chinese Cabbage

[ Objective] The paper was to screen out ideal biological pesticides, in order to provide guidance for pollution-free control against Pieris rapae Linne. [Method] Taking Chinese cabbage variety Taiyuan Erqing as the test material, the field control efficiencies of four pesticides including PrGV · Bt WP 1 000 times dilution, NPV · Bt SC 750 times dilution, 0.5% azadirachtin EC 750 times dilution, and 0.3% matrine AS 500 times dilution against P. rapae were studied, and their impacts on yield and planting benefits of Chinese cabbage were also determined. Using foliar spray method, the pesticides were sprayed for the first time when the second or third instar larvae of P. rapae first occurred in fields, and sprayed for the second time with the interval of 15 d. The fields sprayed with beta-cypermethrin EC （organic chemical pesticide） and water were set as control. [ Result] NPV · Bt SC 750 times dilution had the best effect after spraying for two times： the control efficiency against P. rapae at 15 d after spraying was 90.11% ; the damage rate of Chinese cabbage was only 0.21%, while the commodity rate reached 100% ; compared with chemical pesticide spraying, the commodity yield （177 262.5 kg/hm2 ） and the income after deducting spraying cost （48 858.5 yuan/hm2 ） were increased by 14.7% and 13.75%, respectively. [ Conclusion] Although biological pesticides are more expensive, they have long persistence and good control effect, resulting in green and safe Chinese cabbage with high commodity rate and yield, and higher eventual economic benefit after deducting spraying cost.

上海地区蔬菜害虫研究 Ⅳ菜粉蝶Pieris rapae L.生物学特性观察

菜粉蝶每年发生多代,以往资料上海地区均以每年发生5～6代为依据。为模清实际发生世代数和田间虫口消长、为害情况,自1980～1982年经过田间观察圃内定点、定期系统调查和养虫笼内饲养的结果如下:

A Method for Preparing Insecticide of Cabbage Worm Pieris rapae and Its Control Effects

[Objective]The paper was to study the field control effects of different formulae of plant extracts against cabbage worm Pieris rapae,and to provide a basis for promotion and application of insecticides in production.[Method]With the leaching solution of radix anemones rivularis,Artemisia carvifolia and Nicotiana rustica,and lime and salt water as the materials,field efficacy test was conducted for consecutive two years.[Method]When the mass ratios of radix anemones rivularis mother liquid,A.carvifolia mother liquid,N.rustica mother liquid,lime water and salt water were 4:2:2:1:1:6,4:2:0:1:1:8,4:0:2:1:1:8 and 0:4:2:1:1:8,the control effects were 82.7%-89.8%,38.3%-42.0%,41.0%-45.4%and 52.9%-56.4%,respectively.The insecticide showed the best control effect when the mass ratio of radix anemones rivularis mother liquid,A.carvifolia mother liquid,N.rustica mother liquid,lime water and salt water was 4:2:2:1:1:6.[Conclusion]The optimal formula has good control effect against P.rapae,and the technology has obtained national invention patent.

Cloning and Sequence Analysis of lef-3 Gene from Pieris rapae Granulovirus

To better understand the origination and evolution of lef-3 gene from baculovirus genome and determine the relationships between various viruses at molecular level, late expression factor 3 gene （lef-3） fragments of Pieris rapae granulovirus were obtained through conventional PCR method and sequenced after cloned into T-vector. Then, bioinformatics analysis on lef-3 gene and its encoding sequences were conducted by using bid-softs. Four mutations were appeared in the ORF of cloned lef-3 gene, which did not alter the characteristics of amino acids. It was inferred that PiraGV LEF-3 protein contained 399 amino acids, the molecular weight of which was 3.99 kD. Prediction of the LEF-3 advanced structure and homology comparison between other LEF-3 from various baculoviruses showed that the lef-3 gene might encode the single-stranded DNA-binding protein. The result of BLAST revealed that the lef-3 gene only existed in Lepidoptera host for the baculovirus genome, and the evolution analysis illustrated that lef-3 gene could be divided into 3 groups including one granulovirus （GV） group and 2 nucleopolyhedrovirus （NPV） groups. The selection pressure analysis of GV lef-3 gene coding region showed that the majority of lef-3 genes performed negative selection, while the Ka/Ks differed from different lef-3 gene, to some extent, which also performed positive selection. The origination analysis revealed that lef-3 gene of baculovirus might derive from bacteria. The lef-3 gene of PiraGV was cloned successfully and the possible patterns of origination and evolution were speculated through bioinformatics analysis.

Field Efficacy Trials of Different Pesticides against Pieris rapae and Plutella xylostella

Field efficacy trials of different pesticides against Pieris rapae and Plutella xylostella showed that seven pesticides had certain control effects against P.rapae and P. xylostella. 240 g/L Chlorfenapyr SC had the most ideal control effect,with quick effect and long persistence,and the control effects against P. rapae and P. xylostella were 91. 96% and 95. 73% after application for 7 d,respectively. 25% Thiamethoxam WDG 3 000 times dilution had the poorest control effect,and quick effect and persistence were not ideal; the control effects against P. rapae and P. xylostella were 49. 21% and 57. 20% after application for 7 d,respectively. The remaining pesticides had good control effect against both P. rapae and P. xylostella,with certain persistence. Slight injury such as yellowing tender leaves appeared in the area treated with 50% thiocyclam SP,although the injury was reversible,it was still not recommended to use; no other treatments had adverse effects on growth of cabbage.

Enzymatic properties of phenoloxidase from Pieris rapae （Lepidoptera） larvae

The kinetic parameters of partially purified phenoloxidase （PO, EC. 1.14.18.1） from the 5th instar larvae of Pieris rapae （Lepidoptera） were determined, using L-3, 4- dihydroxyphenylalanine （L-DOPA） as substrate. The optimal pH and temperature of the enzyme for the oxidation of L-DOPA were determined to be at pH 7.0 and at 42℃, respectively. The enzyme was stable between pH 6.5 and 7.4 and at temperatures lower than 37℃. At pH 6.8 and 37℃, the Michaelis constant （Kin） and maximal velocity （V） of the enzyme for the oxidation of L-DOPA were determined to be 0.80 mmol/L and 1.84 μmol/ L/min, respectively. Tetra-hexylresorcinol and 4-dodecylresorcinol effectively inhibited activity of phenoloxidase and this inhibition was reversible and competitive, with the IC50 of 1.50 and 1.12 μmol/L, respectively. The inhibition constants were estimated to be 0.50 and 0.47μmol/L, respectively.

Parasitism of Pieris rapae （Lepidoptera： Pieridae） by the endoparasitic wasp Pteromalus puparum （Hymenoptera： Pteromalidae）： Effects of parasitism on differential hemocyte counts, micro- and ultra-structures of host hemocytes

Parasitism by the endoparasitic wasp Pteromalus puparum （Hymenoptera： Pteromalidae） by using only its associated venom, can suppress the immunal responses of Pieris rapae （Lepidoptera： Pieridae）. However, up to now, current knowledge of the mech- anisms has been limited. The response of host hemocytes to parasitism was investigated using a combination of light and transmission electron microscopy （TEM）. Five hemocyte types, prohemocytes （PRs）, granulocytes （GRs）, plasmatocytes （PLs）, oenocytoids （OEs） and coagulocytes （COs）, were observed and characterized from both unparasitized and parasitized Pieris rapae pupae. Light microscopy showed that both GRs and PLs became more round and spread abnormally after parasitism, whereas the shape of other types of hemocytes remained unaffected. In addition, the size of PRs and PLs became larger while OEs became smaller. The proportion of PRs significantly increased after parasitism and that of PLs decreased by 43.9%, but there was no significant increase of GRs and OEs. TEM showed that all types of hemocytes except COs were damaged to various degrees after parasitism, especially resulting in electron opaque cytoplasm and nucleus, fewer cell organelles of rough endoplasmic reticulum, mitochondria and vesicles. Our results indicate that parasitism by P. puparum affects differential hemocyte counts and structures of host hemocytes, particularly for GRs and PLs, which may be the main cause of the parasitoid suppressing host cellular immune responses.

Proteome changes in the plasma of Pieris rapae parasitized by the endoparasitoid wasp Pteromalus puparum

Parasitism by the endoparasitoid wasp Pteromalus puparum causes alterations in the plasma proteins of Pieris rapae. Analysis of plasma proteins using a proteomic approach showed that seven proteins were differentially expressed in the host pupae after 24-h parasitism. They were masquerade-like serine proteinase homolog (MSPH), enolase (Eno), bilin-binding protein (BBP), imaginal disc growth factor (IDGF), ornithine decarboxylase (ODC), cellular retinoic acid binding protein (CRABP), and one unknown function protein. The full length cDNA sequences of MSPH, Eno, and BBP were successfully cloned using rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis indicated that the transcript levels of MSPH and BBP in the fat bodies of host pupae were inducible in response to the parasitism and their variations were consistent with translational changes of these genes after parasitism, while the transcript levels of Eno and IDGF were not affected by parasitism. This study will contribute to the better understanding of the molecular bases of parasitoid-induced host alterations associated with innate immune responses, detoxification, and energy metabolism.

cDNA of an arylphorin-type storage protein from Pieris rapae with parasitism inducible expression by the endoparasitoid wasp Pteromalus puparum

This report presents the cDNA cloning of a storage protein, PraAry, from Pieris rapae and investigates its expression regulated by parasitism of an endoparasitoid wasp Pteromalus puparum. The full-length eDNA of PraAry is 2 270 nucleotides and contains a 2 121 nucleotide open reading frame encoding 707 amino acids with calculated molecular weights of approximately 83 kDa. Analysis of the primary protein sequence revealed that it possesses a signal peptide of 16 amino acids at the N-terminus and contains two highly conserved storage protein signature motifs. According to both phylogenetic analysis and the criteria for amino acid composition, PraAry belongs to the subfamily of arylphorin-type storage protein （1.42% methionine and 18.82% aromatic amino acids）. Reverse transcription- polymerase chain reaction analysis indicated that the transcriptional level of PraAry mRNA in P. rapae pupae fat body is inducible in response to parasitism by P. puparum.

Inhibitory Kinetics of p-Substituted Benzaldehydes on Polyphenol Oxidase from the Fifth Instar of Pieris Rapae L.

Polyphenol oxidase （PPO） is the enzyme responsible for enzymatic browning during the growth of insects. It is also involved in defense reactions and is related with immunities in insects. PPO a metalloenzyme oxidase, catalyzes the oxidation of o-diphenol to o-quinone. The present paper describes the effects of benzaldehyde and its p-substituted derivatives on the activity of PPO from the fifth instar of Pieris rapae L. PPO from the fifth instar of Pieris rapae L. was purified using ammonium sulfate fractionation and chromatography on Sephadex G-100. The enzyme kinetics was characterized using L-3,4-dihydroxyphenylalanine （L-DOPA） as substrate. The results show that benzaldehyde, phydroxybenzaldehyde, p-chlorobenzaldehyde, and p-cyanobenzaldehyde can inhibit the PPO activity for the oxidation of L-DOPA. The inhibitor concentration leading to 50% activity lost, IC50, was estimated to be 5.90, 5.62, 2.83, and 2.91 mmol/L for the four tested inhibitors, respectively. Kinetic analyses show that the inhibitory effects of these compounds are reversible. Benzaldehyde, phydroxybenzaldehyde, and p-chlorobenzaldehyde are noncompetitive inhibitors while p-cyanobenzaldehyde is a mixed-type inhibitor. The inhibition constants were determined for all four inhibitors. p-chlorobenzaldehyde and p-cyanobenzaldehyde were more potent inhibitors than the other compounds. These results provide a basis for developing PPO inhibition-based pesticides.

中国菜粉蝶线粒体基因组遗传多样性分析

菜粉蝶Pieris rapae是十字花科蔬菜重要的农业害虫。为了深入了解中国菜粉蝶的群体遗传特征,通过高通量测序获取了中国5个群体48头菜粉蝶的线粒体全基因组,并分析了不同群体间的遗传结构与多样性。结果表明:菜粉蝶群体间有较高的线粒体单倍型多样性;相对封闭的山区和盆地地区的核苷酸多样性较低;群体间的平均遗传分化指数(F_(ST))仅0.042,表明不同群体间的遗传分化水平较低,这也暗示了群体间基因交流频繁,推测这与甘蓝等蔬菜频繁运输有关;线粒体进化树和网络图均未形成明显的地理谱系,南北方群体之间没有明显的遗传结构差异;总群体的Tajima's D和Fu's Fs值均为负值,且达到了显著水平,这意味着中国地区的菜粉蝶可能经历过群体扩张事件。

菜粉蝶及其近缘种东方菜粉蝶全球潜在分布区预测

重要农业害虫菜粉蝶(Pieris rapae)及其近缘种东方菜粉蝶(Pieris canidia)幼虫主要危害十字花科的多种蔬菜和油料作物,对农业粮食安全造成重大威胁。收集到菜粉蝶和东方菜粉蝶分布数据共计248393个,其中野外调查收集361个。基于上述现存分布点,利用6种物种分布模型算法对历史、当代和未来气候条件下两种粉蝶在全球的潜在分布区进行了预测。结果表明:(1)基于去除空间自相关后的6个气候变量构建的13种算法的模型预测结果中,有6种算法的模型评估指标的值高于随机状态(0.5)。(2)当代气候条件下,菜粉蝶的潜在分布区主要包括北美洲、南美洲南部、欧洲西部、中国东南部以及澳大利亚南部;东方菜粉蝶主要分布于南美洲北部和亚洲南部等地区。相比于东方菜粉蝶,菜粉蝶在3个时期的潜在分布区面积都要更大,但是东方菜粉蝶在未来气候条件下潜在分布区增加的面积将会更大,具有更高的潜在入侵概率。(3)相比于作为入侵生物的菜粉蝶,其近缘种东方菜粉蝶更能适应高温多雨的气候,面对未来全球变暖的趋势,东方菜粉蝶很有可能成为入侵物种,有必要对其未来气候条件下的潜在分布区,加强虫害监测和防控预报。研究对重要农业害虫菜粉蝶和东方菜粉蝶在不同气候条件下的潜在分布区进行预测和比较,为其防治提供理论依据。