转基因稻及其非转基因亲本对照在间隔种植条件下的转基因漂移

随着转基因技术的迅速发展, 越来越多的转基因作物被培育出来。转基因作物的外源转基因通过花粉传播向非转基因作物的漂移, 会影响非转基因作物品种的种子纯度, 从而可能导致一系列生物安全问题。为了研究转基因栽培水稻(Oryza sativa)中的外源转基因通过花粉介导向非转基因水稻品种逃逸的可能性及其频率, 我们选用3个含双价抗虫基因(Bt/CpTI)的转基因水稻品系及其相对应的非转基因水稻亲本品种(近等基因系)进行了转基因漂移的实验。为了获取在近距离状况下转基因水稻与非转基因水稻品种之间的转基因漂移频率, 采用了转基因与非转基因水稻品种间隔种植的栽培方式, 分别在福建省福州市和海南省三亚市的转基因环境安全实验地进行实验, 并利用潮霉素抗性筛选标记基因来鉴定转基因和非转基因稻的杂种。共检测了从非转基因水稻品种随机收获的70,056颗种子, 以此计算转基因漂移频率。结果表明, 在相邻种植的情况下, 由这3个转基因水稻向对应的非转基因水稻品种的转基因漂移的频率比较低(0.275–0.832％)。如此近距离条件下获得的低转基因漂移频率表明, 对于严格自花授粉的水稻而言, 通过一定的隔离措施, 能有效地降低由花粉介导的转基因漂移导致的非转基因种子混杂。     

Bt/CpTI转基因稻及其非转基因亲本对照在间隔种植条件下的转基因漂移

随着转基因技术的迅速发展,越来越多的转基因作物被培育出来。转基因作物的外源转基因通过花粉传播向非转基因作物的漂移,会影响非转基因作物品种的种子纯度,从而可能导致一系列生物安全问题。为了研究转基因栽培水稻(Oryzasativa)中的外源转基因通过花粉介导向非转基因水稻品种逃逸的可能性及其频率,我们选用3个含双价抗虫基因(Bt/CpTI)的转基因水稻品系及其相对应的非转基因水稻亲本品种(近等基因系)进行了转基因漂移的实验。为了获取在近距离状况下转基因水稻与非转基因水稻品种之间的转基因漂移频率,采用了转基因与非转基因水稻品种间隔种植的栽培方式,分别在福建省福州市和海南省三亚市的转基因环境安全实验地进行实验,并利用潮霉素抗性筛选标记基因来鉴定转基因和非转基因稻的杂种。共检测了从非转基因水稻品种随机收获的70,056颗种子,以此计算转基因漂移频率。结果表明,在相邻种植的情况下,由这3个转基因水稻向对应的非转基因水稻品种的转基因漂移的频率比较低(0.275–0.832%)。如此近距离条件下获得的低转基因漂移频率表明,对于严格自花授粉的水稻而言,通过一定的隔离措施,能有效地降低由花粉介导的转基因漂移导致的非转基因种子混杂。     

基因枪法转基因水稻中hpt基因稳定遗传

基因枪转化将潮霉素磷酸转移酶基因（ｈｐｔ）导入粳稻品种７７１７０,获得可育的转基因植株,研究外源基因遗传的稳定性。自交后代（Ｔ１和Ｔ２）经潮霉素筛选获得抗性植株和敏感植株,分子鉴定结果表明抗性植株带有ｈｐｔ基因,而敏感植株中没有ｈｐｔ基因存在。Ｔ１和Ｔ２代中潮霉素抗性表现为显性单基因位点的遗传方式,符合孟德尔分离规律,并得到分子鉴定结果的证实。Ｓｏｕｔｈｅｒｎ杂交结果显示,ｈｐｔ基因多拷贝整合在水     

转基因植物外源基因逃逸的途径

随着转基因植物商品化进程的加快,对其进行生态风险性评价也日益引起许多学者和科学家的重视,人们对转基因逃逸到其它亲缘物种中能否产生超级杂草,昆虫是否会产生耐受性及转基因植物对生物多样性的影响等问题进行了广泛的研究,本文对转基因植物外中源基因逃逸的几种主要途径作了综述,并对研究方法与手段进行了简要讨论,希望能为有关决策和科研部门提供一些思路。     

Bt转基因水稻对抗生素反应的初步研究（简报）

以相同浓度卡那霉素处理的Bt转基因水稻,其发芽率,根系生长力和叶色变化与对照品种无显著差异;用潮霉素处理时,对照秀水11对50mg.L^-1潮霉素浸种处理十分敏感,发芽率降为0,而Bt转基因水稻则未受到明显的影响,据此可以认为,浸种催芽阶段用潮霉素处理可快速 有效地筛选Bt转基因水稻植株。     

hpt与bar基因作为水稻转基因筛选标记的比较研究

标记基因的选择是影响植物遗传转化和转基因后代筛选成败的关键因素。hpt与bar作为两种常用的水稻转化筛选标记被广泛应用于水稻的转化。为比较两者在实际应用中的效果, 文章首先对比了在潮霉素和除草剂(Bialaphos)两种筛选剂下水稻遗传转化的情况。研究表明, hpt基因的转化筛选体系相对于bar基因在转化效率上提高近两倍, 转化周期提前至少10 d, 且插入基因拷贝数更低。随后, 文章分析了利用潮霉素浸种法在田间筛选转基因水稻的可行性, 研究显示当潮霉素浓度大于167 mg/L时, 可以对以水稻品种kitaake为亲本的转基因材料进行有效筛选, 达到常规除草剂的筛选效果。但与除草剂相比较, 潮霉素的田间筛选成本却处于劣势。文章研究和讨论了hpt和bar基因在遗传转化和后代田间筛选中的优缺点, 并提供了一种利用潮霉素浸种法筛选转基因后代阳性植株的手段, 为将来在水稻转基因研究工作中根据实际需求选择合适的遗传转化、筛选体系提供参考。     

农杆菌介导将白叶枯病抗性基因Xa21导入水稻获得转基因植株

利用农杆菌介导的高效遗传转化系统,将白叶枯病抗性基因Xa21转入黄淮稻区主栽品种豫粳6号的胚性愈伤组织,获得转基因植株,GUS染色和PCR分析证明Xa21基因已整合到水稻基因组中,其自交T1代植株经GUS染色和白叶枯病接种鉴定呈现3：1分离,研究为培育抗白叶枯病水稻品种奠定了基础。     

转基因水稻中外源基因的荧光原位杂交（FISH）分析

Using fluorescence in situ hybridization (FISH) with metaphase preparations, we localized a transferred barnase-psl DNA sequence onto chromosomes in 8 rice transgenic plants. All the tested rice transgenic lines showed hybridization signals on the middle and terminal regions of chromosome arms except for those close to centromeres. In two lines, two different integration sites were identified, and the other lines showed only one integration site. With the aid of Southern analysis and expression detection, we found that the barnase tended to show a higher level expression in the lines whose integration sites near the distal regions of chromosomes, while the expression level became lower in the lines whose integration sites near the centromeres. This result suggested a possible relationship between chromosomal location of transgenes and the expression level. However it showed no obvious relationship between copy numbers and expression levels. In most cases, the results of multi-color FISH showed that barnase-ps1 always integrated at the same position on the chrmosome as the reporter genes(pHctinG).     

农杆菌介导的苏云金杆菌抗虫基因cryIA（b）和cryIA（c）在水稻中 …

通过农杆菌介导法用含有抗潮霉素和ＧＵＳ基因的双元载体将杀虫结晶蛋白基因ｃｒｙＩＡ（ｂ）和ｃｒｙＩＡ（ｃ）导入到籼,粳稻幼穗愈伤组织中,然后经过在含有不同浓度潮霉素的２在上进行数次筛选,获得一批Ｂｔ转基因株。经ＰＣＲ,Ｓｏｕｔｈｅｒｎ杂交及ＳＷｅｓｔｅｒｎ印迹分析证实此二基因已整合进水稻中,饲虫试验结果表明,转基因株具有１００％杀虫率。     

家蝇（Musca domestica）对三氟氯氰菊酯的抗药性遗传

研究了家蝇对三氟氯氰菊酯抗性的遗传,通过对抗性品系和敏感品系杂交后代的抗性遗传分析发现家蝇对三氟氯氰菊酯的抗性受多因子控制,抗性显性率为－0．102,为不完全隐性。其对三氟氯氰菊酯的抗性现实遗传力为0．120。     

Low frequency of transgene flow from Bt/CpTI rice to its non-transgenic counterparts planted at close spacing

Crop-to-crop transgene flow will affect seed purity of non-GM rice varieties, leading to unwanted consequences. To assess the maximum probability of transgene outflow in rice (Oryza sativa), gene flow experiments were conducted with three cultivation patterns with different mixed-planting proportions of adjacent GM and non-GM rice at two sites in Fujian and Hainan Provinces of China. Three GM rice lines containing two insect-resistance genes (Bt/CpTI) and their non-GM counterparts were used in the experiments to allow natural hybridization to occur. A hygromycin resistance gene was used as a selective marker for identifying hybrids. Based on the examination of > 645 700 geminated seeds, the result showed low frequencies (0.05-0.79%) of transgene flow from GM to non-GM rice at close spacing, although with significant variation among mixed-planting proportions. It is concluded that rice transgene flow will occur at a very low frequency (< 1.0%), even if the GM rice is planted at close spacing with non-GM rice, and high densities of GM rice cultivated in the neighborhood of non-GM rice will increase the probability of outcrossing with the non-GM rice.     

Mdogg1基因参与家蝇体内氧化还原平衡的维持

[目的]本研究旨在克隆鉴定家蝇Musca domestica中8-羟基鸟嘌吟糖苷酶1(8-hydroxyguanine glycosylase 1,OGG1)编码基因Mdogg1,明确其是否参与家蝇氧化应激调控.[方法]根据家蝇转录组数据,利用RT-PCR克隆Mdogg1基因cDNA序列,并进行生物信息学分析;利用qR...     

转基因抗草甘膦油菜的检测方法

目的:建立检测田间转基因抗草甘膦油菜的方法。方法:用CTAB法提取DNA,经PCR分别扩增内参基因BnACCg8,及抗草甘膦外源基因CP4-EPSPS、FMV 35S启动子和E9 3’终止子等4种基因的片段,以琼脂糖凝胶电泳对扩增产物进行分析比对。结果:抗草甘膦油菜中分别扩增出与外源基因FMV 35S启动子、E9 3’终止子和CP4-EPSPS大小一致的片段。结论:该方法能有效地用于转基因抗草甘膦油菜的检测。     

Detection of gene flow from GM to non-GM watermelon in a field trial

Gene flow from genetically modified (GM) crops to conventional non-GM crops is a serious concern for protection of conventional and organic farming. Gene flow from GM watermelon developed for rootstock use, containing cucumber green mottle mosaic virus (CGMMV)-coat protein (CP) gene, to a non-GM isogenic control variety “Clhalteok” and grafted watermelon “Keumcheon” was investigated in a small scale field trial as a pilot study. Hybrids between GM and non-GM watermelons were screened from 1304 “Chalteok” seeds and 856 “Keumcheon” seeds using the duplex PCR method targeting theCGMMV- CP gene as a marker. Hybrids were found in all pollen recipient plots. The gene flow frequencies were greater for “Chaiteok” than for “KeumcheonD; with 75% outcrossing in the “Chaiteok” plot at the closest distance (0.8 m) to the GM plot. A much larger scale field trial is necessary to identify the isolation distance between GM and non-GM watermelon, as the behaviors of insect pollinators needs to be clarified in Korea.     

STUDIES ON A CADMIUM-INDUCED METALLOTHIONEIN IN HOUSEFLY LARVAE, MUSCA DOMESTICA

Abstract  The induction of metallothionein (MT) was studied in the housefly larvae ( Musca domestica ). Upon dietary exposure to Cd, two Cd-binding proteins were isolated from the whole body homogenates, using gel filtration and ion exchange chromatography. Mass spectrometric measurement revealed that they have high purity and the molecular weight are 9045.9Da and 11560.2Da, respectively. Amino acid analysis showed that the content of cysteine is the highest, attaining to 18.2%. However, aromatic amino acid residues such as tyrosine (2.5%) and phenylalanine (3.1%) were also detected. In addition, MT from housefly larvae has strong heat stability. All these results suggest that the properties of MT isolated from housefly are very similar to that of mammalian MT, but some differences still exist.     

多寄主型寄生蜂蝇蛹俑小蜂对新寄主的选择与适应性

【目的】明确多寄主型寄生蜂蝇蛹俑小蜂Spalangia endius对不同类型新寄主的偏好选择与适应性。【方法】选择分别以瓜实蝇Zeugodacus cucurbitae、南瓜实蝇Z.tau和家蝇Musca domestica的蛹为寄主饲养的蝇蛹俑小蜂成蜂,比较研究其对上述3种寄主中其他2种非饲养寄主的选择行为、寄生选择及适应性。【结果】与家蝇蛹相比,以瓜实蝇蛹为饲养寄主的蝇蛹俑小蜂成蜂偏好选择和寄生南瓜实蝇蛹,且在南瓜实蝇蛹上产生更多的子代;以南瓜实蝇蛹为饲养寄主的蝇蛹俑小蜂成蜂偏好选择和寄生瓜实蝇蛹,且在瓜实蝇蛹上产生更多的子代;而以家蝇蛹为饲养寄主的蝇蛹俑小蜂成蜂对瓜实蝇蛹和南瓜实蝇蛹的选择数量以及在选择和非选择试验条件下的寄生率与子代数量上并无显著差异。此外,以瓜实蝇和南瓜实蝇蛹为饲养寄主的蝇蛹俑小蜂成蜂分别更容易适应南瓜实蝇和瓜实蝇蛹。【结论】基于3种蝇的生态位关系,推测蝇蛹俑小蜂偏好选择和更容易适应与其饲养寄主有重叠生态位的新寄主。