转基因技术及其应用

本文对近20多年来转基因技术的研究及其应用进展情况进行了简要的概述。     

Selection of Homozygous Cotton Lines Transformed with Two Insect-Resistant Genes

A plant expression vector containing a chimeric Bt29K gene coding for the activated Cry1Ac protein and the arrowhead proteinase inhibitior gene API-B were introduced into the cotton cultivar Jihe321 mediated by Agrobactertium tumefaciens. Based on the results of kanamycin resistant testing, PCR detection for both foreign genes and insect bioassay using Heliethis armigera, nine transgenic homozygous cotton lines with insect-resistance of more than 90% and better agronomic traits were bred through six generations from the original transgenic plants. Results from insect bioassay and sequence analysis of the PCR products of plants from some homozygous lines indicated that the chimeric Bt29K gene was stably inherited in these transgenic cotton lines. The main agronomic characters of these homozygous cotton lines, such as boll productivity and fibre strength, were better than that of the original cotton cv. Jihe321.     

转甜菜碱醛脱氢酶基因水稻种子萌发和苗期耐盐性的研究

用0.0、3.0、5.0g/L NaCl对2个转甜菜碱醛脱氢酶(BADH)基因水稻51-22、52-7与受体亲本中花8进行萌发期盐胁迫试验.结果表明,盐胁迫下转基因材料受抑程度小于受体亲本,尤以52-7表现明显.用0.0、3.0、5.0g/L NaCl对51-22、52-7和中花8在幼苗期进行14 d盐胁迫试验.结果表明,低盐胁迫时幼苗受盐害较小,高盐胁迫时转基因材料耐盐性优于受体亲本;在盐胁迫7 d后,转基因材料体内脯氨酸含量明显高于受体亲本,同一品种随盐度增大,脯氨酸含量升高.     

转基因植物的研究与应用

介绍了转基因植物的主要研究方法及其在各个领域中的应用现状 ,并对其今后的应用前景加以展望。     

Persistence of an oilseed rape transgene in the environment

Transgenic bromoxynil (BX)-resistant spring oilseed rape (Brassica napus L.) was grown commercially in Canada only for two crop seasons – 2000 and 2001. We investigated the persistence of suspected BX-resistant oilseed rape volunteers in a 64-ha wheat field in Saskatchewan, Canada in 2007, 7 years after the BX-resistant cultivar BX Armor was grown. A small oilseed rape volunteer population, estimated at less than 100 plants, was observed in three main areas or patches in the field. These patches were located in low-lying areas that were too wet to plant or spray with herbicides in 2007. Viable seed was collected from 35 mature volunteers and F₁ progeny screened with BX at 280 g ai/ha in the greenhouse. Progeny of all of the volunteers were visually rated as BX-resistant; the presence of the Oxy 235 transgene in leaf tissue of progeny of all volunteers was confirmed by PCR analysis. This study has documented the longest persistence of oilseed rape volunteers in Canada. Volunteers were not observed in 2008 or 2009, because of drought conditions in spring of both years. Results support the findings from previous studies that persistence of volunteer oilseed rape populations in western Canada is generally ephemeral or transitory in the absence of seed bank immigration. However, this study shows that oilseed rape transgenes can persist in the environment for a number of years even after all cultivars with the conferred trait are removed from the market.     

转抗虫基因Cry1Iem大豆的分子鉴定与功能验证

通过转抗虫基因来提高作物的抗虫性是一条有效途径,以此来提高作物的产量和品质。本研究以转抗虫基因Cry1Iem大豆的T2、T3株系作为试验材料,利用常规PCR检测、Southern杂交、荧光定量PCR技术对转抗虫基因Cry1Iem株系进行分子鉴定;同时对40个转基因阳性株系进行室内人工接虫、200个转基因阳性株系室外网室接虫并进行抗虫性鉴定。选取其中10个株系进行常规PCR检测,结果在T2、T3当中都检测到了Cry1Iem、Bar、启动子CaMV35S、Nos目标片段,表明目的基因已被顺利导入受体JN28大豆植株当中并得到了稳定遗传;选取其中5个T2、T3转基因株系进行Southern杂交,结果显示:目的基因以单拷贝的形式整合到了大豆基因组当中;荧光定量PCR检测结果表明,Cry1Iem基因在选取的3个转基因株系中均得到了表达,而在非转化的受体JN28当中未检测到荧光信号。室内抗虫性鉴定结果显示40个转Cry1Iem基因株系豆荚内活虫的成活数量明显低于受体材料JN28大豆豆荚内的活虫数量,表明转基因材料具有显著的抗虫性。室外抗虫性鉴定结果表明:JN28大豆受体植株的虫食率为6.72%,属于感虫品种;而200个转Cry1Iem基因株系的平均虫食率为3.81%,属于抗虫品系。本研究结果为中国转基因抗虫大豆新品种的选育提供部分参考数据。     

现代分子生物学技术在植物抗病毒育种中的作用

概述了近年来利用分子生物学技术进行抗病毒育种的几种主要方法及其研究进展 ,主要内容有外壳蛋白介导的抗性、利用缺损的复制酶、干扰运动蛋白、RNA介导的抗性、卫星RNA介导的抗性、反义RNA介导的抗性、中和抗体基因等     

转基因玉米双抗12-5对根际土壤酶活性及 微生物多样性的影响

采用田间试验研究了转基因玉米瑞丰1号-双抗-12-5(RF1-12-5)种植对根际土壤酶活性、微生物群落的影响,可为RF1-12-5的环境释放和商业化应用提供科学的安全性评价数据支持。研究结果表明:①在5个生育期内,RF1-12-5与其非转基因对照品种瑞丰1号(RF1)根际土壤碱性蛋白酶、脲酶和酸性转化酶活性均没有显著性差异;RF1-12-5根际土壤过氧化氢酶活性在收获期、碱性磷酸酶活性在乳熟期显著低于RF1,其他生育期差异不显著。②在5个生育期内,RF1-12-5与RF1根际土壤微生物群落的Shannon多样性指数、McIntosh均匀度指数和Simpson优势度指数均不存在显著性差异,主成分分析未发现RF1-12-5与RF1根际微生物功能多样性存在规律性差异。     

转基因抗虫水稻中BT蛋白表达量的研究(英文)

对转基因抗虫水稻中Bt蛋白表达量进行研究。[方法]应用酶联免疫吸附测定法(ELISA)定量检测转基因抗虫水稻生同生长期不同部位的Bt蛋自表达量。[结果]转基因水稻灌浆期不同组织中Bt蛋白表达绝对含量的高低顺序为:叶片>未成熟种子及颖壳>根>茎杆;在水稻不同的生长发育(分蘖期、抽穗期和灌浆期)阶段,转基因Bt水稻中Bt蛋白的含量有一些变化;一般在水稻生长后期Bt蛋白的浓度有所下降,但幅度不大,对其抗性不会造成太大影响。[结论]该试验对田间害虫的防治以及转基因水稻的育种都具有重要意义。     

转基因抗虫水稻研究进展

介绍了水稻害虫及其化学防治产生的问题,综述了转基因抗虫水稻的研究进展,分析了转基因抗虫水稻的安全性,并展望了抗虫水稻转基因的发展方向。