转基因甘蔗BtG-2的T-DNA侧翼序列分析及其转化事件特异性检测     

冯翠莲  万玥  冯小艳  王俊刚  赵婷婷  王文治  沈林波  张树珍 《热带作物学报》2021,42(9):2468-2477

转基因甘蔗BtG-2是利用农杆菌介导法把Cry1Ac-2A-gna融合抗虫基因导入‘新台糖22号’的转基因甘蔗株系,具有良好的抗虫特性和农艺性状。为了明确转基因甘蔗BtG-2的分子特征及其检测方法,推进其生物安全性评价工作,以BtG-2的T2代为研究材料,利用Southern杂交检测外源基因在转基因甘蔗基因组内的拷贝数;利用染色体步移技术分离外源基因在甘蔗基因组中插入位点的侧翼序列,并建立了该转化体高效灵敏的特异性PCR检测方法。结果表明：Southern杂交检测证明外源T-DNA以单拷贝方式插入BtG-2株系;经过3次的热不对称巢式PCR扩增,获得外源基因T-DNA左边侧翼序列984 bp和右边侧翼序列705 bp;以这2个序列和相应的T-DNA的左右端序列分别设计3对检测引物对,建立了BtG-2株系的转化事件特异性PCR检测方法,扩增效率最高的引物对LS011/LA451和RS160/RA588分别扩增到440 bp和428 bp的特异片段。其中T-DNA左侧设计的LS011/LA451检测引物对扩增的灵敏度高、特异性强,能够在甘蔗BtG-2基因组DNA相对含量为0.1%的模板中检测出转基因目的成分,相当于9个单倍体基因组拷贝数。本研究完成了转基因株系BtG-2的分子特征及其转化事件特异性检测,为该转基因甘蔗及其衍生产品的检测和身份识别提供技术依据。  相似文献   

Correction to: Predation on glass eels of Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> in the Tone River Estuary,Japan     

Yoichi Miyake  Aigo Takeshige  Hikaru Itakura  Hajime Itoh  Hiroaki Onda  Akira Yamaguchi  Akihito Yoneta  Kohma Arai  Yulina V. Hane  Shingo Kimura 《Fisheries Science》2018,84(6):1015-1016

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Alessandro Bonanno and Lawrence Busch (eds): Handbook of the international political economy of agriculture and food     

Marie Louise Ryan 《Agriculture and Human Values》2018,35(3):731-732

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异硫氰酸丙烯酯毒杀松材线虫作用机制初步研究     

韩双双  谭周倩  黄晓德  钱骅  陈斌  赵伯涛  陈双林  闫淑珍 《农药学学报》2020,22(1):48-53

对异硫氰酸丙烯酯毒杀松材线虫作用机制进行了初步研究。测定了其对线虫虫体形态、运动行为、体内总糖和蛋白质含量的影响,以及处理前后线虫体内过氧化氢酶(CAT)、乙酰胆碱酯酶(AChE)和谷胱甘肽-S-转移酶(GST)活性变化。结果表明:经异硫氰酸丙烯酯处理后的松材线虫虫体形态严重扭曲,运动形态出现由正常的S形变化为多节弯曲和螺旋状卷曲等异常状况,线虫行动随时间的推移变得迟缓,最终呈C形、新月形或直接死亡。随着异硫氰酸丙烯酯处理质量浓度的增大,线虫体内总糖和蛋白质含量显著上升,糖和蛋白质的代谢发生紊乱;线虫体内CAT、AChE和GST活性显著降低,且均低于对照组,表明线虫体内氧化和抗氧化作用失衡;细胞受到极大的影响,神经系统遭到破坏,解毒能力明显降低。研究结果可为研究利用异硫氰酸丙烯酯作为植物源杀线虫剂提供依据。  相似文献   

动物细胞遗传学研究现状及其应用   总被引：1，自引：0，他引：1  

门正明  韩建林 《甘肃农业大学学报》1993,28(4):317-322

本文介绍了细胞遗传学在动物方面的研究现状及其在畜牧业上的应用。细胞遗传学的研究主要以染色体为对象。目前世界上现存的哺乳动物约4100多种,其中进行过染色体研究或核型分析的有1468种,我国已研究过的有242种。细胞遗传学在人类医学上的应用是众所周知的,在畜牧业上主要用于对种畜进行细胞学检查,以消除对家畜繁殖性能有影响的畸变染色本,以染色体标记来识别家畜品种,对环境污染的监测及遗传操作中的染色体工程。  相似文献   

杂种小麦产量和蛋白质含量的优势表现与优势环境分析(简报)     

刘录祥  黄铁城  张树榛  刘广田 《中国农业大学学报》1993,(4):107-108

许多研究表明,杂种小麦在产量和主要品质性状上存在显著的基因型与环境互作,杂种小麦比纯系品种更能适应变化的环境条件,表现出更好的稳定性。但是,关于杂种小麦优于纯系品种所表现出的杂种优势在不同环境中的表现则报道很少。正如一种性状的表现型由  相似文献   

‘Plantibodies’: a flexible approach to design resistance against pathogens     

A. Schots  J. De Boer  A. Schouten  J. Roosien  J. F. Zil Verentant  H. Pomp  L. Bouwman-Smits  H. Overmars  F. J. Gommers  B. Visser  W. J. Stiekema  J. Bakker 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(2):183-191

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.  相似文献   

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>     

Tsutomu MATSUMOTO  Yuichiro NARA  Hiromitsu FURUYA  Harumi TAKAHASHI  Kiichi TAIRAKO  Hideki YAMAMOTO 《Journal of General Plant Pathology》2002,68(4):382-384

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002  相似文献   

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields     

Shohei MATSUURA  Shigeru HOSHINO  Hideaki HAYASHI  Tetsuyuki KOHGUCHI  Kyoji HAGIWARA  Toshihiro OMURA 《Journal of General Plant Pathology》2002,68(1):99-102

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001  相似文献   

Distribution of citrus viroids and <Emphasis Type="Italic">Apple stem grooving virus</Emphasis> on citrus trees in Japan using multiplex reverse transcription polymerase chain reaction     

Takao Ito  Nobuyuki Namba  Tsutae Ito 《Journal of General Plant Pathology》2003,69(3):205-207

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