丙型肝炎病毒E2基因在转基因番茄植株中的表达

以T-E1E2为模板扩增得到丙型肝炎病毒包膜蛋白基因E2,构建该基因的植物表达载体p35s-E2．通过农杆菌介导的叶盘法转化番茄子叶．转基因番茄植株叶片总DNA的PCR、Southern blot检测结果表明,E2基因已整合进了转基因番茄植株基因组中;RT-PCR,Western blot分析证实E2基因在转基因番茄植株叶片中表达．     

转基因植物及其安全性问题

介绍了转基因植物在全球和我国发展概况,介绍了转基因植物的几种目的基因:抗除草剂基因、抗病虫基因、改良作物品质基因,及其在农业上的应用,从转基因植物食品的安全性和转基因生态环境的安全性讨论了正确对待转基因技术的安全性问题。     

简便实用的转抗除草剂基因后代植株鉴定方法--种子萌发测定法

种子萌发法是检测外源抗性基因在转基因植物中的表达和分析外源抗性基因在转基因植物中遗传行为的简单而直接的方法,以转ｂａｒ基因燕麦后代Ｒ２种子为材料,用不同体积分数的除草剂Ｃｈａｌｌｅｎｇｅ处理种子,萌发的幼苗经ＰＣＲ和Ｓｏｕｔｈｅｒｎ ｄｏｔ ｂｌｏｔ鉴定,发现在０．０３％的除草剂上萌发的幼苗,部分不含ｂａｒ基因,而在０．０７％和０．０９％的除草剂上萌发的幼苗,均含ｂａｒ基因。用０．０７％的除草剂测     

植物基因工程：希望与风险并存

如果说,1985年获得的抗除草剂转基因烟草为植物基因工程拉开了序幕,那么,10多年来,植物的基因工程已取得了令人震惊的成果。一批具有抗虫、抗病毒、抗除草剂和其他特殊性状的转基因植物相继问世,玉米、棉花、大豆等主要作物的转基因品种,已从实验室和试验田走向了日常耕作的大田。科学家们预言,植物的基因工程必将带来农业领域的“第二次绿色革命”。然而,和许多科学技术成果一样,转基因植物在给人类带来喜悦的同时,也使人们产生了忧虑。今年2月28日～3月1日,世界经济合作发展组织在英国爱丁堡召开的基因改性食品安全国际论坛上,与会各国的专     

植物转基因沉默的发生机理和应用

随着转基因技术在农业生产等领域的深入发展,转基因沉默现象已引起越来越多人的关注.基因沉默的机制是多方面的,包括转基因的拷贝数和构型、在植物上的整合位点、转基因的转录水平等.研究基因沉默的原因并寻找相应的方法来进行控制,对于植物基因工程的发展以及转基因沉默的应用有着重要的意义.     

转基因玉米遗传转化的研究进展

转基因玉米是目前植物基因工程的研究热点之一,已有转基因抗虫玉米、抗除草剂玉米进入商品化生产。通过对转基因玉米遗传转化方法的研究,概括了目前在转基因玉米的遗传转化中所用的各种方法、及其优缺点,以及已取得的成果和遗传转化的检测分析方法的研究进展。     

基因工程与农业（一）

80年代起,基因工程陆续在培育抗病毒植物、抗虫植物、抗除草剂植物,创造植物的雄性不育、培育生产医用蛋白的植物和其他有重要价值的蛋白质的植物以及改变植物的质量性状等方面初露头角,为高产、优质、高效农业的发展展示了诱人的前景。 时至今日,通过向植物转入人类所需要的基因(转基因),得到了一系列符合人类要求的植物(转基因植物)。 一、转基因植物抗病毒 病毒病是植物病害中的一类严重病害,不仅会造成产量大幅度下降,也会影响质量。1986年,美国科学家Beachy等从烟草花叶病毒(TMVUI)株中分离出病毒的外壳蛋白基因,当把这     

基因工程在番茄改良方面的研究进展

近十多年来,植物基因工程取得了辉煌的成就,许多大田作物的转基因产品已进入商品化生产阶段。文章就基因工程在番茄改良方面的研究作一简要综述,主要包括番茄抗病虫性、抗除草剂、抗逆性、耐贮藏性及品质改良等方面的研究进展。     

转基因植物对环境微生物的影响及其检测技术的研究进展

为了解转基因植物及其表达产物环境释放的生态影响,归纳和总结了转基因植物对微生物的影响及其检测技术的应用现状.从转基因植物对土壤微生物多样性的影响及其检测的一般方法和分子生物学方法,转基因植物中抗性标记基因对微生物影响和微生物在转基因植物产品毒理学检测等方面进行了综述,以期为转基因植物的应用和健康发展提供依据,同时也为转...     

海岛棉GbRar1和GbSgt1基因的过量表达提高烟草离体叶片对赤星病的抗性

RAR1 和 SGT1 是两个植物抗病蛋白 (R) 防卫反应的信号元件,在 R 蛋白下游和活性氧产生之间发挥作用. 利用简并引物 PCR 及3′RACE 等技术,克隆了海岛棉 GbRAR1 和 GbSGT1的编码基因. Northern blot 表明,海岛棉GbRar1和GbSgt1基因在转录水平上受棉花黄萎病菌的诱导,诱导后 mRNA 表达量显著增加. 构建组成型植物表达载体 pRar 和 pSgt,分别转化烟草品种 NC89. 离体叶片抗病性鉴定表明,两类转基因烟草对赤星病的抗性明显提高,为植物防卫反应信号元件RAR1 和 SGT1在广谱抗病基因工程中的应用提供了实验证据.     

Transgenic rice homozygous lines expressing GNA showed enhanced resistance to rice brown planthopper

Mature seed-derived calli from two elite Chinese japonica rice (Oryza sativa L.) cultivars Eyi 105 and Ewan 5 were co-transformed with two plasmids, pWRG1515 and pRSSGNA1, containing the selectable marker hygromycin phosphotransferase gene (hpt), the reporter β-glucuronidase gene (gusA) and the snowdrop (Galanthus nivalis) lectin gene (gna) via particle bombardment. 61 independent transgenic rice plants were regenerated from 329 bombarded calli. 79% transgenic plants contained all the three genes, revealed by PCR/Southern blot analysis. Western blot analysis revealed that 36 out of 48 gna-containing transgenic plants expressed GNA (75%) at various levels with the highest expression being approximately 0.5% of total soluble protein. Genetic analysis confirmed Mendelian segregation of transgenes in progeny. From the R2 generations whose R1 parent plants showing 3:1 Mendelian segregation patterns, we identified five independent homozygous lines containing and expressing all the three transgenes. Insect bioassay and feeding tests showed that these homozygous lines had significant inhibition to rice brown planthopper (Nilaparvata lugens, BPH) by decreasing BPH survival and overall fecundity, retarding BPH development and declining BPH feeding. These BPH-resistant lines have been incorporated into rice insect resistance breeding program. This is the first report that homozygous transgenic rice lines expressing GNA, developed by genetic transformation and through genetic analysis-based selection, conferred enhanced resistance to BPH, one of the most damaging insect pests in rice.     

Obtaining the transgenic poplars with low lignin content through down-regulation of 4CL

The antisense 4CL (4-coumarate: CoA ligase)gene was transformed into triploid Chinese white poplar(Populus tomentosa) mediated by Agrobacterium tumefaciens.PCR and Southern blot analysis indicated that antisense 4CLgene had been integrated into the genome of the transgenicChinese white poplars. The antisense gene had also beenexpressed, which was indicated by RT-PCR and Westernanalysis. Klason lignin content assay showed that repressionof 4CL expression could result in remarkable reduction oflignin content in transgenic poplars, with most reduction of 41.73% compared with that of wild type in this paper. Butthere is no significant difference in holocellulose content be-tween trans- genic and wild poplars. We considered that 4CLmight not be the metabolism control point between ligninand carbohy- drate biosynthesis. The stems of transgenicpoplars displayed red-brown color with different levels afterthe bark was peeled, while those of untransformed plantswere white. No visible differences in growth and developmentwere observed between transgenic and wild plants. Wiesnerreaction analysis of the transgenic plant stems with reducedlignin content exhibited red color, while that of untrans-formed plant was typically purple-red.     

OsNHX1基因转化84K杨的研究

采用农杆菌介导的方法开展了水稻Na /H 逆向转运蛋白基因OsNHX1转化84K杨的研究.建立了84K杨的叶外植体高频再生系统,经诱导不定芽及诱导生根阶段卡那霉素连续筛选,获得了大量卡那霉素抗性转化植株.PCR检测和Southern杂交检测表明,OsNHX1基因已成功整合到84K杨基因组.耐盐实验表明,转基因植株能在200 mmol NaCl条件下正常生长.     

Inheritance and expression of multiple disease and insect resistance genes in transgenic rice

2—3 anti-fungal disease genes are coinserted with hygromycin phosphotransferase in the same vector. Two insecticidal genes and PPT acetyl transferase genes are placed in another one. The vectors are co-delivered to rice embryonic cellus tissue at a molar ratio of 1︰1 using the particle gun method. 55 independent regenerated lines have been obtained through screening for hygromycin resistance. Of these, 70% transgenic plants harbor 6—7 foreign genes. The genes on the same vectors are always co-delivered to rice plant. Northern blot analysis has indicated that the multiple foreign genes give stable expression. In the 6 transgenic plants carrying 6—7 foreign genes, multiple foreign genes tend to integrate in 1 or 2 genetic loci. Progeny segregation is consistent with Mendel’s 3︰1 segregation law. 8 homozygous R₁ transgenic plants harboring 2—3 anti-fungal and 2 insecticidal genes are selected from large number of transgenic progeny screening for hygromycin and Basta resistance.     

利用农杆菌介导法将抗虫基因导入油菜的研究

以油菜的子叶柄作为转化的受体材料,经农杆菌感染和共培养,在含卡那霉素的筛选培养上诱导产了大量的不定芽,经生根培养,获得了再生植株,通过PCR扩增和基因组Southern杂交分析,证明其中5株为转基因植株,对转基因植株进行抗虫性检测,有2株表现出较好的抗虫性。     

Expression of two insect-resistant genes crylA ( b&c )/GNA in transgenic tobacco plants results in added protection against both cotton bollworm and aphids

The synthesized Bacillus thuringiensis insecticidal protein gene crylA(b&c) and the synthesized gene GNA, (the mannose specific lectin from snowdrop ( Galanthus nivalis)), tumefaciens have been inserted into plant expression vector pGW4BAI. Leave stripes of Nico-tiana tabacum var. K326 have been transformed with Agrobacterium tumefaciens strain LBA4404 harboring the plant expression vector. 28 kanamycin resistant tobacco plants have been obtained. PCR and Southern blot analyses show that the foreign crylA and GNA genes have been inserted into the genome of transformed tobacco plants. Haemagglutination assays show that GNA has a functional activity. Leaf disc bioassays against cotton bollworm ( H. armigera) show that the transgenic tobacco plants have a high insecticidal activity. The inhibition of aphid population in leaf disc bioassays against Myzus persicae shows that the fecundity of aphid on transgenic plants is lower than that on untransformed plants; the aphid population on the transgenic tobacco plants is 25%-70% that on untransformed tobacco plants. ELISA analysis of CrylA protein in tobcco leaves provides similar data to bioassay results. Through the two bioassays against H. armigera and M. persicae, several transgenic tobacco plants showing high insect-resistant activities to both pests have been obtained.     

Salt tolerance of transgenic rice (Oryza sativa L.) with mtlD gene and gutD gene

Southern blot analysis indicated that mtlD gene (encoding mannitol-1-phosphate dehydrogenase) and gutD gene (encoding glucitol-6-phosphate dehydrogenase) had been integrated into the rice genome mediated by Agrobacterium tumefaciens LBA4404(pBIGM). The expression of the above two genes in transgenic rice plants was demonstrated by Northern blot analysis and enzymatic activity assay. Analysis of sugar alcohol showed that transgenic rice plants could produce and accumulate mannitol and sorbitol. The salt tolerance of transgenic plants was much higher than that of their controls.     

转几丁质酶基因烟草对三种真菌拮抗作用的研究

几丁质酶广泛存在于高等植物体内,它可以分解真菌细胞壁中的几丁质,破坏其细胞结构,从而获得对真菌的抗性.试验以转苦瓜几丁质酶基因烟草(T-Chit)为供试植物,测定植株根系和叶片内几丁质酶活性,选取了3种具有代表性的真菌:毛霉、木霉、禾谷镰刀霉,通过抑菌试验验证了T-Chit组织萃取液对真菌的抗性.结果表明:1)T-Chit几丁质酶活性显著高于非转基因烟草(Nt-X),且转基因烟草根系几丁质酶活性比叶片高86%;2)T-Chit组织萃取液对毛霉生长具有明显抑制作用,根系强于叶片;3)T-Chit对木霉和禾谷镰刀霉的抑制具有时空效应:抑菌效果与植株部位及作用时间有关.     

Efficient selection of homozygous lines of hybrid rice restorer with the transgeneXa21 using test cross and PCR

The homozygous restorer lines with a single copy of the transgene Xa21 have been obtained from the progenies of transgenic Minghui63 and Yanhui559 plants through PCR marker-assisted selection and test cross. These homozygous transgenic restorer lines can be used to breed hybrid rice with high resistance to bacterial leaf blight.