Introduction of new traits into cotton through genetic engineering: insect resistance as example

The main goal of gene transfer into cotton is the development of insect-resistant varieties. The stakes are important since cotton protection against insects uses almost 24% of the world's chemical insecticides market, which is not without consequences on the environment. The first approach was to introduce and express in the cotton genome, genes from the bacterium Bacillus thuringiensis (B.t.) which produces entomopathogenic toxins. The development of an efficient Agrobacterium tumefaciens mediated transformation system was the first step. The expression of B.t. genes was studied and synthetic genes more adapted to a plant genome have been constructed. Studies on their expression in cotton is underway. The second focus was to develop strategies that would minimize the risks of inducing insect resistance. The main approach is to associate several genes coding for entomopathogenic proteins with different modes of action. Genes encoding protease inhibitors were chosen. One possibility is to associate a B.t. gene and a gene encoding a protease inhibitor. Several protease inhibitors were tested in artificial diets on major pests of cotton. The corresponding genes have been introduced into the cotton genome. These various orientations of the research program will be presented. This revised version was published online in July 2006 with corrections to the Cover Date.     

Powdery mildew resistance in barley landrace material. I. Screening for resistance

A total of 4,681 accessions of Hordeum vulgare landrace material from Ethiopia, East Mediterranean, Near East, Nepal and China were sown in the field and subjected to the natural powdery mildew epidemic in Denmark. Apparently resistant accessions were selected. Selfed progeny from them were retested and reselected in subsequent years at four locations in Denmark. Finally, 16 promising donors of resistance were retained. They were characterized in the field and tested in the seedling stage for reaction to up to 72 different isolates of the powdery mildew fungus. The absence of the corresponding virulences in the Danish airborne powdery mildew population was ascertained in five years. The resistances in the 16 donors are apparently mutually different and from known sources of powdery mildew resistance in barley. This revised version was published online in July 2006 with corrections to the Cover Date.     

一种高产L-赖氨酸枯草芽孢杆菌的构建研究

为了选育一种非抗性、高产出、遗传稳定性好、低生产成本的L-赖氨酸生产菌株,以辣椒花药和枯草芽孢杆菌为试验材料,以穿梭表达载体pHT43和基因敲除载体pK18mobsacB为媒介,将辣椒花药中的高赖氨酸基因CFLR转化到枯草芽孢杆菌中。经过2次细胞内同源交换和培养基筛选,最终获得不带质粒抗性标记的高产L-赖氨酸枯草芽孢杆菌株CMCC(B)63501/pK18mobsacB-ΔALA::CFLR,该菌株连续传代20次后仍具有明显高于野生型的CFLR基因拷贝数,经HPLC验证,重组菌株L-赖氨酸含量比野生菌株提高了64.89%。结果表明,本研究所构建的非抗性高产L-赖氨酸枯草芽孢杆菌株产量高、遗传稳定性好、低纯化成本,具有良好的应用潜力。     

苏云金芽孢杆菌营养期杀虫蛋白Vip3的研究进展

为了更好地理解Vip3毒素的研究进展,本研究从Vip3毒素的类型、理化特性、功能、杀虫机理以及应用研究等方面做了详细的综述。并指出,尽管Vip3毒素的研究取得了很大的进展,但是还有一些重要的问题在后续研究中亟待解决。     

一株产蛋白酶海洋菌的筛选、鉴定及培养条件优化

为了获得高产蛋白酶的菌株,将其广泛应用于水产养殖中。从青岛市胶州湾海泥中分离得到一株产蛋白酶的海洋菌ZR-Pw。通过菌株ZR-Pw的菌落形态、菌体形状、革兰氏染色,结合16S rDNA基因序列,对菌种进行鉴定;将ZR-Pw接入发酵培养基中,测定该菌株的生长曲线、产酶曲线,确定其产酶的最适温度及pH。结果显示：菌株ZR-Pw为地衣芽孢杆菌(Bacillus licheniformis)。菌株在生长到18 h时,菌体生长量达到最大值。在菌株生长到24 h时,产酶量达到最大值。该菌株产酶最适温度为35℃,最适pH 8.0。试验为菌株ZR-Pw产蛋白酶的产业化开发提供依据。     

一株病原真菌拮抗菌的分离与鉴定研究

本实验室从食用菌的栽培料从分离得到一株产抗真菌物质的菌株MAF-1。经检测其对1株土壤未知真菌、一些炭疽病菌、木霉、梢腐病原菌具有良好的抑制效果。经生理生化实验及16S rDNA 同源性序列分析,鉴定该菌株为短小芽孢杆菌属(Bacillus pumilus)。     

芽胞杆菌菌体浓度的快速检测方法研究

为了快速检测芽胞杆菌的菌液浓度,采用分光光度法测定蜡状芽胞杆菌的菌液浓度,代替传统常规费时费力的生物细胞测定法。本研究以不同pH值下培养的模式菌株蜡状芽胞杆菌为例,通过可见光分光光度法测定不同pH值菌液的吸光度,分别以水和培养基作为空白对照,并与其对应的菌体浓度进行比较分析。利用生物统计软件DPS分析蜡状芽胞杆菌菌液浓度(Y)与其吸光度(X)（以水为空白对照）之间有着极好的相关性,其测定的吸光度可以作为快速地检测芽胞杆菌菌体浓度的指标。综上所述,本研究结果证明吸光度法可以代替细胞计数法作为芽胞杆菌的菌体浓度生长变化的指标,且此方法简便、快速、可靠。     

香蕉枯萎病菌4号小种致病力分化的初步研究

自2003年6月至2010年1月,我们通过采集或交流的方式共得到海南、广东、福建、云南和广西等省区的35个市县的56个Fusarium oxysporum f.sp. cubense Race 4菌株,并通过盆栽伤根淋灌法接种巴西蕉苗,系统性地评价了56个Race 4菌株的致病力。实验结果表明：供试的56个Race 4菌株表现出了很明显的致病力分化,其中,强致病力的菌株有29个、中致病力的有9个、弱致病力的有18个,分别占供试菌株的51.79%、16.07%、32.14%。     

氟磺胺草醚降解菌F-12的分离鉴定及降解特性,

为了研究氟磺胺草醚污染土壤的生物修复机理,利用富集培养技术从长期施用氟磺胺草醚的土壤中分离得到1株能够以氟磺胺草醚为唯一碳源生长的细菌,命名为F-12。通过菌落形态、生理生化特性和16SrDNA基因序列分析,初步鉴定菌株F-12为克雷伯氏菌属（Klebsiella sp.）。并分析了氟磺胺草醚的初始浓度、接种量、温度和pH值对菌株F-12降解氟磺胺草醚效果的影响,确定了最佳降解条件。结果显示,该菌在氟磺胺草醚浓度为100 mg/L、接种量为15%、pH为6.0、温度35℃条件下,培养2 d后对氟磺胺草醚的降解效率达到80%以上。具有应用到氟磺胺草醚污染土壤生物修复的能力。