苏云金芽孢杆菌杀虫剂的研究进展

综述了苏云金芽孢杆菌的毒性作用,它的杀虫晶体蛋白基因及基因产物的特性,将杀虫晶体蛋白基因转移到植物上获得抗虫植物以及如何防止昆虫对该杀虫剂产生抗性的问题.     

甲壳质/壳聚糖特性及其在农业中的应用(综述)

综述了壳聚糖促进作物生长、杀菌、杀虫、易于成膜等特殊功能及其在农业中的应用进展情况。     

苏云金杆菌杀虫晶体蛋白的crylC基因

目前,有8种crylC类基因已被克隆,其主要分布在杀虫亚种和鲇泽亚种.本文对杀虫晶体蛋白CrylC的结构与功能、作用方式、抗性问题及基因改造等方面作一综述,还就存在的问题与前景进行探讨.     

致病杆菌功能基因的分子生物学研究进展

致病杆菌是与斯氏线虫共生的肠杆菌科细菌,存在于昆虫病原线虫肠道内,可随线虫的侵染进入寄主昆虫血腔,迅速繁殖并产生多种代谢产物,抑制其他杂菌的污染,保护线虫繁殖的环境。多年研究发现,该菌的代谢产物具有广泛的杀虫、抑菌、抗癌等生物活性。本文介绍了致病杆菌外膜蛋白基因、菌毛蛋白基因、共生作用相关基因、溶血素相关基因、杀虫毒素蛋白基因、抗生素相关基因以及专利申请基因的研究进展。     

芬兰人的环保生意经 访芬兰环境科技中国项目委员会总裁Mari Pantsar-Kallio女士、副总裁Ari Makkonen先生及在华首席代表杨凤辉先生

《科学》2008年9月19日卷321 no.5896 Bt抗虫棉抑制中国多个农作物地区的棉铃虫转基因棉花已投入生产的杀虫毒素苏云金杆菌(Bt)和抵御虫害棉铃虫(棉铃虫)已被广泛种植在亚洲。从1992至2007年在中国的棉铃虫数量动态分析了,标注的区域爆发这种虫害的多个作物与种植转Bt基因棉花的关系。     

杀虫微生物的研究应用方兴未艾：记全国第三届杀虫微生物学术研讨会

1990年10月17日在武汉召开了全国杀虫微生物学术研讨会,来自全国21个省市、自治区的大专院校、科研、管理、出版、生产等74个单位,200多位代表,交流了141篇论文,充分地反映了活跃在我国生物防治领域中杀虫微生物的研究和应用的新进展。其中有高层次的基础研究,例如转Bt杀虫基因的植物基因工程研究、昆虫病毒分子生物学的研究、昆虫病原细菌的致病机理的研究以     

苏云金芽胞杆菌的研究进展

围绕国内外对苏云金芽胞杆菌(Bacillus thuringiensis)研究的新进展,分别从转苏云金芽胞杆菌作物的安全性、苏云金芽胞杆菌的酶学研究、苏云金芽胞杆菌的基因学研究、苏云金芽胞杆菌的蛋白质学研究四个方面进行论述,为进一步预防昆虫对苏云金芽胞杆菌植物产生抗性、防止苏云金芽胞杆菌基因飘逸、构建新型苏云金芽胞杆菌载体及生产出杀虫毒性更高、专一性更强的苏云金芽胞杆菌植物提供了新的思路.     

植物抗虫基因工程研究与应用

近年来,植物抗虫基因工程研究及应用进展喜人.本文综述了利用Bt杀虫蛋白基因、豇豆蛋白酶抑制素基因、昆虫特异性神经毒素基因以及外源凝集素基因等抗虫基因开展植物抗虫基因工程研究的概况,并探讨了植物抗虫基因工程中存在的主要问题及解决的途径.     

苏云金芽孢杆菌营养期杀虫蛋白基因的克隆、表达及杀虫活性分析

选择本实验室分离的野生型苏云金芽孢杆菌菌株WY-197为出发菌株，用全长PCR方法从此菌株中克隆了2．3kb大小的vip3A基因,DNA序列比较发现所克隆的基因vip3A-197与已知的营养期杀虫蛋白基因存在很高的同源性．将基因vip3A-197亚克隆至原核表达载体pET33b构建了原核表达质粒pEVip，转化大肠杆菌BL21，转化子经IPTG诱导后可表达88kD大小的蛋白．该蛋白对甜菜夜蛾(Spodoptera exigua)棉铃虫(Helicoverpa armigera)的初孵幼虫进行生物测定，结果表明，营养期杀虫蛋白vip3A-197对夜蛾科害虫具有一定的杀虫活性。     

转Bt棉不同生长期及不同器官杀虫蛋白表达量的免疫学方法测定

转苏云金杆菌(Bacillus thuringensis)杀虫蛋白基因棉花(下称Bt棉)在不同生长期及不同器官上对棉铃虫杀虫活性存在明显差异。目前国内普遍使用生物测试法检测Bt棉对棉铃虫的杀虫活性,但无法定量检测Bt棉中杀虫蛋白的表达量。本实验室建立的酶联免疫学测定方法(ELISA),可定量检测棉器官内杀虫蛋白的含量。它具有检测周期短,操作简便的优点。用ELISA方法检测Bt棉杀虫蛋白的含量,结果表明,棉叶和花瓣杀虫蛋白含量最高,铃和蕾次之。在棉株生长的后期,各器官杀虫蛋白的含量则有大幅度的下降。     

壳聚糖杀虫与壳低聚糖抑菌活性研究

对以壳聚糖及壳低聚糖为有效成分的天然农药 ,分别进行室内杀虫和抑菌活性实验。结果表明，壳聚糖对鳞翅目和同翅目害虫均存在一定的杀虫活性。在相同浓度下，对小菜蛾的杀虫活性高于棉铃虫；对不同蚜虫的杀虫活性，一般在60%～80%之间，最高可达99%。并用室内离体平皿法检验测定了以壳低聚糖为有效成分的药剂对17种植物病原菌生长的抑制作用。结果表明，壳低聚糖对其中16种病原菌都有抑制作用，且随着浓度提高作用增强。药剂对芦笋茎枯病原菌的最佳抑制率可达95.1%，对黄瓜枯萎病、水稻纹枯病及棉花枯萎病等重要植物病害的病原菌也可达80%以上的抑制效果。     

壳聚糖在作物生产上的应用进展(综述)

介绍了地球上第二大可再生自然资源壳聚糖(Chitosan)作为丰富的无毒、无污染的生防农药,在抗病诱导、杀菌杀虫、抵御逆境、种子包衣、农药增效、土壤改良、地膜降解、促进生长、提高产量、改善品质等方面的研究进展及应用前景。     

Construction of a new plant expression vector containing two insect resistant genes and its expression in transgenic tobacco plants

A new plant expression vector (pBS29K-BA) containing two insect resistant genes, a synthetic chimeric gene BtS29K encoding the activated insecticidal protein Cry1Ac and a gene API-BA encoding the arrowhead (Sagittaria sagittifolia L.) proteinase inhibitor (API) A and B, is constructed. Transgenic tobacco plants expressing these two genes are obtained through Agrobacterium-mediated transformation of tobacco leaf discs. The average expression levels of Cry1Ac and API-BA proteins in transgenic plants are of 3.2 μg and 4.9 μg per gram fresh leaf respectively. The results of insecticidal assay of transgenic plants indicate that the pBS29K-BA transformed plants are more resistant to insect damage than the plants expressing the Cry1Ac gene or API-BA gene alone.     

Evaluation of transgenic tobacco expressing two insecticidal genes to delay resistance development ofHelicoverpa armigera

The resistance ratio ofHelicoverpa armigera to Cry1 Ac insecticidal protein fromBacillus thuringiensis (Bt) is 13.1- and 3.02-fold after 18 generations of selection by transgenic tobacco expressing Bt or two (Bt and CpTI) insecticidal protein genes, in which the average corrected mortality for each selection treatments is about 60%. The mortality of selected population by transgenic Bt gene tobacco is significantly lower than the control strain when fed on transgenic tobacco plants. The mortaltty of the selected population by transgenic two genes tobacco was not significantly different from the control strain. This is the first experiment under laboratory condition which has proved that transgenic two genes tobacco could significantly delay resistance development ofH. armigera compared with one gene.     

Ubi1 intron-mediated enhancement of the expression of Bt cry1Ah gene in transgenic maize (Zea mays L.)

The cry1Ah gene was one of novel insecticidal genes cloned from Bacillus thuringiensis isolate BT8. Two plant expression vectors containing cry1Ah gene were constructed. The first intron of maize ubiqutin1 gene was inserted between the maize Ubiquitin promoter and cry1Ah gene in one of the plant expressing vectors (pUUOAH). The two vectors were introduced into maize immature embryonic calli by microprojectile bombardment, and the reproductively plants were acquired. PCR and Southern blot analysis showed that foreign genes had been integrated into maize genome and inherited to the next generation stably. The ELISA assay to T1 and T2 generation plants showed that the expression of Cry1Ah protein in the construct containing the ubi1 intron (pUUOAH) was 20% higher than that of the intronless construct (pUOAH). Bioassay results showed that the transgenic maize harboring cry1Ah gene had high resistance to the Asian corn borers and the insecticidal activity of the transgenic maize containing the ubi1 intron was higher than that of the intronless construct. These results indicated that the maize ubi1 intron can enhance the expression of the Bt cry1Ah gene in transgenic maize efficiently     

转基因小黑杨对土壤微生物群落结构的影响

为评价转基因林木环境释放可能引起的生态风险,以及转基因林木在生产应用中的可行性和可靠性,以实验室种植的转Bt-蜘蛛神经毒肽重组基因的小黑杨为材料,对转基因与非转基因植株根际(通常为几毫米至几厘米)土壤细菌、放线菌、真菌数量和基因水平转移情况进行检测,结果表明:转基因小黑杨种植一段时间后其根系附近的微生物数量略高于非转基因小黑杨。利用卡那霉素选择培养基分离根系周围细菌菌落发现转基因植株周围的土样细菌中,Kan^R抗性细菌菌落数占总细菌菌落的比例在11.58%～13.00%之间,而非转基因植株根际周围和空白土壤细菌菌落数则在5.73%～6.40%之间,表明转基因小黑杨根系对土壤细菌的抗生素抗性产生一定的影响。利用转基因植株的目的基因和抗性基因设计引物,以土壤中细菌基因组DNA为模板进行PCR扩增,转基因植株种植7个月后,根际土壤细菌菌落中含有抗性基因的菌落数量明显增加。虽然在种植1个月的转基因植株根际土壤土样细菌中未检出目的基因,但在种植7个月后的土样中检验到了很少的目的基因阳性细菌,阳性细菌菌落比例为2%～5%,检测结果表明可能有极少数Bt-蜘蛛神经毒肽重组基因发生了水平转移。     

Xenorhabdus nematophila BP品系xptB1基因的克隆和序列分析

采用PCR方法,从该室构建的Xenorhabdus nematophila BP品系粘粒文库的一个对棉铃虫有强口服杀虫活性的粘粒cos83中扩增出全长的xptB1基因,将扩增片段回收纯化后,连接到pMD-18T载体,转化大肠杆菌TG1后进行序列测定和分析。结果显示,该基因全长为3051bp,编码1016个氨基酸,其编码的毒素BP XptB1与X.nematophila PMF1296品系的XptB1的氨基酸序列同源性平均为95.8%,两侧高度一致,中间第607—738位差异明显。结构分析显示,两者在跨膜区,α螺旋等方面也有差异。BP XptB1与Photorhabdus luminescens的TccC毒素,Serratia entomophila的SepC,以及Yersinia pseudotubercusis和Y.pestis等的杀虫毒素均有58%以上的同源性。分析结果预示了XptB1可能是一个杀虫毒素。     

苏云金芽孢杆菌工程菌BM B_(20-4)的杀虫晶体蛋白检测及毒力测定

苏云金芽孢杆菌的杀虫晶体蛋白由单基因编码,经过遗传操作,构建成BM B 20-4工程菌.本研究对工程菌BM B 20-4进行了杀虫晶体蛋白检测和发酵液毒力测定.结果表明:工程菌BM B 20-4杀虫晶体蛋白质含量比天然菌株提高44%,毒力效价显著提高,对小菜蛾提高了0.72倍,对棉铃虫提高了0.75倍.     

Commercial production of transgenic Bt insect-resistant cotton varieties and the resistance management for bollworm (Helicoverpa armigera Hubner)

There are currently three kinds of transgenic Bt insect-resistant cotton germplasm lines, Shanxi 94-24, Zhongxin 94 and R19, in China. They showed high resistance to the neonate larvae of bollworm (Helicoverpa armigera). Transgenic Bt insect-resistant cotton varieties or hybrids have been bred using the three kinds of germplasm lines as parents. Our researches reveal that there exist different expressions in resistant level at different developmental stages in the three categories of germplasm lines. When neonate larvae are fed with leaves of cotton plant at the seeding stage with less than 10 leaves on the main stem, the mortality of the neonate larvae is 100%, but the resistance level will decline at later season. When Bt gene has been transferred to the cotton genome, it can be steadily transferred to the progeny, the level of resistance to bollworm keeps fundamentally uniform. Such insects as tobacco budworm (Heliothis virencens) in laboratory directive selection are very apt to produce resistance to the Bt insecticidal crystal protein. From the present crop system of cotton region in the Yangtze and Yellow River Valleys, and the expression characteristic of transgenic Bt resistant cotton, we suggest that the resistance to toxin protein in bollworm is not apt to be produced if the transgenic Bt insect-resistant cotton varieties are released and grown in the regions except in the Xinjiang cotton region. The managing strategies to delay or retard the resistance are discussed.