新细胞系──粘虫胚胎细胞系的建立(英文)

一株新细胞系-粘虫（Mythimnaseparata）胚胎细胞系（NEAU－Ms－927311简称Ms937311）被建立。该细胞系多为梭形细胞,少数圆形,贴壁性强。细胞群体倍增时间为58．8h;接种3天后细胞增长速度加快,第5天达到最大值;染色体分析结果表明：染色体聚集成簇,棒球状,呈典型的鳞翅目昆虫细胞核型;酯酶同工酶分析有两条明显的谱带;该细胞系可被同源核型多角体病毒（MsNPV）侵染并形成多角体。     

NEW CELL LINE FROM EMBRYOS OF MYTHIMNA SEPARATA (LEPIDOPTERA: NOCTUIDAE)

Abstract  A new cell line was established from 2-d-old embryonated eggs of Mythimna separata and has been designated as NEAU-Ms-927311. This cell line consists of a mixture of cell types, including spherical and spindle-shaped cells. The cell line has a population doubling time of 58. 8h. Chromosome analysis revealed its typical lepidopteran karyology. Isozyme characterization of esterase showed that the band pattern was different from that of other two cell lines (Xc-920730, and SF-21AE). Virus infectivity tests revealed this cell line can support replication of M. separata nuclear polyhedrosis virus.     

人工饲料饲养的粘虫对杀虫剂的敏感性

本文介绍了粘虫Mythimna separata的一种新人工饲料,并比较了用其饲养的粘虫和用玉米叶饲养的粘虫对杀虫剂的敏感性及对乙酰胆碱酯酶的抑制作用.结果表明,两种饲料饲养的粘虫对大多数杀虫剂的敏感性及胆碱酯酶的抑制作用没有明显区别.     

两株粘虫核型多角体病毒理化性质的比较研究

电镜观察粘虫(Leucania separala)核型多角体病毒的不同株系内蒙株(简称N株)和阜阳株(简称F株)的多角体、病毒粒子及核衣壳在形态大小方面都没有很大差别,DNA分子大小也相近。但在SDS-PAGE多肽图谱及DNA酶切图谱上则表现出一定的差异。经SDS-PAGE所得病毒粒子多肽图谱N株有17条多肽,F株有19条多肽。两株病毒DNA经BamHI酶切N株获得7条带,F株则有8条带,说明DNA酶切位点不尽相同。据此,生化分析能更快速、准确地进行株系分类。     

Differences between the pathogenic processes induced by Steinernema and Heterorhabditis (Nemata: Rhabditida) in Pseudaletia unipuncta (Insecta: Lepidoptera)

Larvae of Pseudaletia unipuncta are moderately susceptible to infections caused by entomopathogenic nematodes, being a desirable host to study pathogenic processes caused by Heterorhabditis bacteriophora, Steinernema carpocapsae, and Steinernema glaseri and their associated bacteria. The ability of the infective stage of these nematodes to invade hosts is quite different. S. carpocapsae invades the highest number of insects and presents the highest penetration rate, followed by H. bacteriophora. Regression analysis between the number of insects parasitized and the number of IJs counted per insect, over time, showed a high correlation for S. carpocapsae whereas for H. bacteriophora it was low. Dose-response was most evident at a concentration below 100 IJs per insect on H. bacteriophora, whereas on S. carpocapsae it was found for doses ranging from 100 to 2,000 IJs. Student's t test analysis of dose-response showed parallel, yet unequal, slopes for both strains of H. bacteriophora, whereas distinct regressions were obtained for S. carpocapsae and S. glaseri, thus, evidencing each species develop a distinct pathogenic process. Insects injected with Photorhabdus luminescens died within 50 h after injection, whereas those treated with X. nematophila died much later. Moreover, the mortality in insects exposed to H. bacteriophora complex and injected with P. luminescens was close, but insects injected with bacteria died faster. Insect mortality in treatments with complexes S. carpocapsae and S. glaseri was significantly higher than that which was observed in insects injected with symbiotic bacteria.     

Bt玉米(MG95)对粘虫的抗性和拒食作用

在室内以我国自主培育的转Bt基因玉米 (简称 :Bt玉米 )为材料 ,测定了Bt玉米对不同龄期 ( 1～ 4龄 )粘虫PseudaletiaseparataWalker的影响。测定指标包括 :幼虫体重、幼虫发育历期、幼虫死亡率、化蛹率、蛹重、蛹历期、羽化率、产卵量、成虫历期、卵孵化率。结果显示 :用Bt玉米喂养的 1～ 4龄幼虫在处理 6d及 1～ 3龄幼虫在处理 9d时的体重与对照相比均差异显著 ;除 1龄幼虫外 ,2～ 4龄幼虫处理 6d及 2～ 3龄幼虫处理 9d后的死亡率差异不显著 ;用Bt玉米喂养的各龄幼虫发育历期延长 ,与对照相比差异显著 ;1、2龄幼虫的化蛹率与对照相比有显著差异 ,3、4龄则差异不显著 ;1～ 4龄幼虫的蛹重、蛹历期及羽化率均差异显著 ;成虫历期仅 1龄幼虫差异显著 ;与对照相比 ,各龄粘虫的产卵数、卵孵化率均无显著差异。结果表明 ,1龄幼虫对Bt玉米最为敏感。另外 ,拒食实验结果显示 ,在实验开始后的 8,1 6,2 4,3 2及 48h ,非选择性测定的拒食率为 63 2 0 ,64 60 ,48 99,2 3 3 5和 0 5 4% ,选择性测定的拒食率为98 3 3 ,82 43 ,71 5 4,40 1 6和 0 2 7% ,总体趋势是拒食率随时间延长而降低 ,48h后Bt玉米对粘虫幼虫无拒食活性。     

Primary and continuous midgut cell cultures from Pseudaletia unipuncta (Lepidoptera: Noctuidae)

Midgut epithelial cells were isolated from fifth-instar Pseudaletia unipuncta larvae by collagenase treatment of midgut tissue, and cultured in TNM-FH medium. Long-term continuous culture and maintenance of midgut cells were achieved with P. unipuncta armyworm intestinal cells. Several cells lines were obtained from these P. unipuncta primary cultures, and they have been subcultured and maintained for over 24 mo. The three major midgut cell types were present in the cultures, including stem (regenerative), columnar, and goblet cells. In vitro morphogenesis and differentiation of columnar and goblet cells from stem cells were observed. There appeared to be a cycle of cell death of goblet and columnar cells followed by their replacement from stem cells every 7-8 wk. After approximately six passages, the cell density in T-flasks appeared to be somewhat constant, reaching 10(3)-10(4) cells per milliliter of medium. The columnar cells are round to rectangular in shape and possess a brush border, while the goblet cells have a classic flask-like shape with a central cavity. Peritrophic membrane-like secretions were observed in all the culture flasks. Infection of these cells with multiply embedded nucleopolyhedrovirus was confirmed, and we conclude that these midgut cells can be used as an in vitro model system to study early events in baculovirus infection.     

粘虫下唇须陷窝器及其感器的形态、类型与分布

【目的】明确我国重要农业害虫粘虫Mythimna separata Walker下唇须和陷窝器形态结构以及陷窝器内部感器的形态、类型与分布。【方法】利用光学显微镜观察粘虫成虫下唇须及陷窝器形态结构,利用扫描电子显微镜观察陷窝器内部的感器。【结果】结果表明,粘虫成虫下唇须似管状,具3节,各节形态和长度不同,其中第2节最长。粘虫下唇须长度雌雄异形,雌性的下唇须长度为2 463.50±143.65 μm,显著短于雄性的（2 566.11±70.28 μm）（t=2.722, df=34, P=0.012）。陷窝器凹坑深约280 μm,开口处直径约50 μm,内部直径约32 μm,雌雄间无显著差异。陷窝器内部的感器主要包括毛形感器和棒状感器2种类型。毛形感器位于陷窝器凹坑的上半部分,而棒状感器分布在陷窝器凹坑的下半部分。雌性毛形感器长为18.20±0.84 μm,显著短于雄性的21.24±0.34 μm（t=3.335, df=30, P=0.003）。而雌性的棒状感器长为14.69±0.48 μm,显著长于雄性的12.31±0.49 μm（t=3.452, df=21, P=0.002）。【结论】粘虫下唇须陷窝器感器分属2大类型,分布于陷窝器内不同的区域,其长度具有性别差异性。本文报道的这些形态学观察结果为进一步研究粘虫下唇须陷窝器感器生理和功能奠定了基础。     

粘虫不同发育期体内储存蛋白的检测及苦皮藤素Ⅴ对其含量的影响

采用PAGE和SDS-PAGE以及Western blot 的方法,分析了粘虫Mythimna separata幼虫、蛹及成虫体内的储存蛋白。结果表明,粘虫体内存在两种储存蛋白,其中一种为SP-1,即幼虫特异性储存蛋白,从6龄粘虫幼虫的2日龄开始出现在血淋巴中,到末日龄时达到峰值,停止取食后从血淋巴中消失;另一种为SP-3,在化蛹时开始出现在脂肪体中,一直到成虫期仍可持续表达,因此属于持续性储存蛋白。SP-1为分子量约94 kD和100 kD的2种亚基组成的蛋白质,而SP-3为分子量约94 kD的1种亚基组成的蛋白质。SP-1含8.16%的芳香类氨基酸,3.06%的甲硫氨酸。经苦皮藤素Ⅴ亚致死剂量处理5龄粘虫幼虫后的6龄2、3、4日龄粘虫幼虫体内储存蛋白的含量明显低于对照组,对5日龄后粘虫处理组和对照组体内储存蛋白的含量及雌性成虫产卵量没有明显影响。