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1.
根癌农杆菌介导的水稻转化及转基因R1代植株表型特征   总被引:14,自引:0,他引:14  
用根癌农杆菌介导的方法对2个粳稻品种和4个籼稻品种进行了转化。粳稻成熟胚和籼稻幼胚来源的愈伤组织用携带质粒pGIH的农杆菌EHA101感染,对所有品种均获得较高的愈伤转化频率(20.83%~62.32%)。粳稻“申香粳4号”植株转化频率为17.39%,“秋丰”为9.21%。4个品种籼稻中仅“超丰早1号”获得1株转化植株。Southern杂交分析、GUS染色、T-DNA整合边界序列分析等结果表明外源  相似文献   

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农杆菌转化获得转B.t.基因水稻及其生物学鉴定   总被引:8,自引:0,他引:8  
黄健秋  卫志明 《植物生理学报》2000,26(6):519-524,T002
以水稻主栽品种“秀水11”和“春江11”预培养4d未成熟胚为转化受体,经农杆菌LBA4404/pGBI4A2B(含B.t.基因)感染后,筛选出抗性愈伤组织并获得转化植株。其中抗性愈伤组织产生频率达44% ̄70%,转化植株产生频率达27% ̄64%。转基因植物总DNA经PCR和Southern blot分子杂交试验表明,T-DNA上的外源基因已整合到水稻的基因组里。对转B.t.基因水稻植株进行了两种水  相似文献   

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以粳稻品种广陵香糯为材料,用携带有双元载体pBB的根癌农杆菌EHA105为载体,研究了根癌农杆菌介导转化粳稻成熟胚愈伤组织的几个影响因素,建立了合适的粳稻成熟胚愈伤组织转化系统。一种基于MS为基本培养基的商品培养基(HRM)较适合于作为粳稻成熟胚愈伤组织诱导培养基,合适的愈伤组织诱导培养天数为7~8 d,合适的筛选培养基为CC培养基。在此基础上,将Metr基因导入粳稻品种广陵香糯中,获得了一批转基因水稻植株,PCR分析表明外源基因已经整合进了水稻的基因组中。部分转基因植株后代遗传分析表明外源基因的分离符合3∶1的理论比例。  相似文献   

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Transgeni根癌农杆菌介导的小麦转基因植株再生(英文)   总被引:1,自引:0,他引:1  
根癌农杆菌菌株Agl Ⅰ的Ti 质粒pUNN-2 带有Ubi1 启动子驱动的npt Ⅱ基因。7 种基因型小麦幼胚或胚性愈伤组织用于农杆菌介导的转化实验。经过不同浓度巴龙霉素的筛选,3 种基因型小麦产生抗性愈伤组织并再生植株。再生植株经PCR 和Southern 杂交鉴定为转基因植株,转化频率( 再生转基因植株的小麦愈伤组织数/ 用于转化实验的愈伤组织数) 为3.7% ~5 .9% 。小麦基因型及转化材料的起始生理状态是影响TDNA转移的重要因素。  相似文献   

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根癌农杆菌介导的小麦转基因植株再生   总被引:19,自引:0,他引:19  
根癌农杆菌菌株Agl Ⅰ的Ti质粒p^UNN-2带有Ubi1启动子驱动的nptⅡ基因。7种基因型小麦幼胚或胚性愈伤组织用于农杆菌介导的转化实验。经过不同家度巴龙霉素的筛选,3种基因型小麦产生抗性愈伤组织并再生植株。再生植株经PCR和Southern杂交鉴定为转基因植株,转化频率为3.7%-5.9%。小麦基因型及转化材料的起始生理状态的影响T-DNA转移的重要因素。  相似文献   

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农杆菌介导籼稻优良恢复系bar基因的遗传转化研究   总被引:2,自引:0,他引:2  
应用农杆菌介导转化体系,成功地将含有CaMv35s启动子启动的bar基因导入籼稻幼胚来源的愈伤组织,获得籼稻优良恢复系T461、R402和752三个品种(系)共47个抗除草剂Basta的转基因株系,Southem分析结果表明,转基因植株基因组中检测到bar基因的整合,转基因植株自交后代Basta除草剂抗性鉴定表现出分离,且大多数为1-2个整合位点的孟德尔方式遗传。结果表明,根癌农杆菌介导法可以有效且可靠地转化籼稻。  相似文献   

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简单高效的根癌农杆菌介导的水稻基因转化方法   总被引:2,自引:0,他引:2  
本文在研究影响农杆菌介导的水稻转化的主要因素基础上,建立了一套简单、高效的水稻转基因系统。将水稻成熟胚来源的愈伤组织用农杆菌EHA101/pHQ9,EHA101/pHQ10,EHA101/pHQT3感染后,筛选抗性愈伤,经分化获得转化株。抗性愈伤的平均得率为约100个愈伤/g愈伤外植体,抗性愈伤的分化频率平均高达85%。转基因植株的GUS染色、Southern杂交结果表明,T-DNA上的外源基因已整合进转基因植物的基因组中。转基因植株T1代对潮霉素的抗性表明,多数转基因株系符合孟德尔分离比3:1。该系统的建立将有助于应用T—DNA标签法和基因打靶法进行水稻功能基因组的研究。  相似文献   

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根癌农杆菌介导的水稻高效转化系统的建立   总被引:4,自引:0,他引:4  
比较了影响根癌农杆菌转化水稻的各种因素后,建立了农杆菌介导的水稻高效转基因实验体系。按该体系,水稻品种中花11号预培养4d的幼胚经农杆菌EHA105/pCAMBIA1301感染后,具有GUS基因瞬间表达的幼胚比例在50%以上,最高可达90%;按产生潮霉素抗性愈伤和转基因植株的比例计算,转化率分别达到87.6%和64.6%。转基因植株总DNA的Southern杂交分析表明T-DNA上的外源基因已整合进了水稻基因组,且在大多数转基因植株中表现为单拷贝插入;遗传分析证明T1代的表型分离符合孟德尔法则。此转化系统的建立为高效地将有用的外源DNA导入水稻植株奠定了基础。  相似文献   

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根癌农杆菌介导的水稻高效转化系统的建立   总被引:132,自引:5,他引:127  
比较了影响根癌农杆菌转化水稻的各种因素后,建立了农杆菌介导的水稻高效转基因实验体系。按该体系,水稻品种中花11号预培养4d的幼胚经农杆菌EHA105/pCAMBIA1301感染后,具有GUS基因瞬间表达的幼胚比例在50%以上,最高可达90%,按产生潮霉素抗性愈伤和转基因植株的比例计算,转化率分别达到了87.6%和64.6%。  相似文献   

10.
本文在研究影响农杆菌介导的水稻转化的主要因素基础上,建立了一套简单、高效的水稻转基因系统。将水稻成熟胚来源的愈伤组织用农杆菌EHA101/pHQ9,EHA 101/pHQ 10,EHA 101/pHQ T3感染后,筛选抗性愈伤,经分化获得转化株。抗性愈伤的平均得率为约100个愈伤/g愈伤外植体,抗性愈伤的分化频率平均高达85%。转基因植株的GUS染色、Southern杂交结果表明,T-DNA上的外源基因已整合进转基因植物的基因组中。转基因植株T1代对潮霉素的抗性表明,多数转基因株系符合孟德尔分离比3∶1。该系统的建立将有助于应用T-DNA标签法和基因打靶法进行水稻功能基因组的研究。  相似文献   

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K. Hausmann 《Protoplasma》1979,100(2):199-213
Summary The membranes of the pellicle of the ciliatePseudomicrothorax dubius are investigated using thin section electron microscopy and freeze-fracture replicas. The plasma membrane is covered by a surface coat and is connected to the outer alveolar membrane by short, sometimes branched, bridges. The inner alveolar membrane is coated on both sides. The epiplasm lies in intimate contact with the cytoplasmic surface of this membrane, and there is a corresponding deposit on the other surface. This deposit is regularly striated.The epiplasmic layer and the alveoli are interrupted at sites of cytotic activity,e.g., the attachment sites of trichocysts, the cytoproct, and the parasomal sacs. The striated deposit ends where the epiplasm ends, indicating a direct relationship between these two epimembranous layers.There is a deposit along the sides of the first part of the tip of the trichocysts, and in this region the trichocyst membrane is free of intramembranous particles.The membrane of the parasomal sacs has a coat on both surfaces. That on the extraplasmic surface is similar to the surface coat of the plasma membrane. The origin of the cytoplasmic coat is unknown. The cytotic activity of these sacs is indicated by their highly irregular profiles.  相似文献   

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Summary The differentiation of the spermatid, especially in reference to the formation of the flagellum, and transformation of the shape of the nucleus was investigated in the domestic fowl.In the early stage of the spermatid, a prominent Golgi apparatus appears around the centrioles. The Golgi vesicles then surround the axial-filament complex which develops from the distal centriole. These vesicles fuse to form continuous membrane at the earliest stage of flagellar formation, and in the succeeding stage Golgi lamellae are attached to the plasma membrane of the developing flagellum. From these observations, it is assumed that Golgi apparatus may be a source of the membrane system of the flagellum.The microtubules distributed around the nucleus form the circular manchette. The anterior region of the nucleus with the manchette is cylindrical in shape and the posterior region without it remains irregular in shape. When the circular manchette has been completed, the whole nucleus acquires a slender cylindrical shape. The circular manchette then changes into the longitudinal manchette. The nuclei of spermatids without a longitudinal manchette are abnormal in shape. In view of these observations it is assumed that the nuclear shaping of the spermatid may be accomplished by circular manchette and the maintenance of shape of the elongated nucleus by longitudinal manchette.The authors wish to thank Mr. Takayuki Mori for his helpful suggestions and technical advices  相似文献   

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Summary The choriocapillaris is a fenestrated capillary bed located posterior to the retinal pigment epithelium. It serves as the main source of supply to the photoreceptors, retinal pigment epithelium, and other cells of the outer retina. The permeability of these capillaries to intravenously injected ferritin (MW — approx. 480,000; mol. diam. 11 nm) was examined in the mouse, rabbit, and guinea pig, each of which is characterized by a different type of retinal vascularization. In all three species, the bulk of the ferritin remained in the capillary lumina, where it appeared to be blocked at the level of the diaphragmed fenestrae. Some ferritin was present in endothelial cell vacuoles. The results confirm previous work on the rat choriocapillaris and indicate that the barrier function of the choriocapillary endothelium is present even among species in which the retinal circulation differs significantly.Supported by NIH grant EY03418  相似文献   

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