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1.
转BADH基因玉米的获得及其耐盐性   总被引:11,自引:0,他引:11  
利用花粉管通道技术将甜菜碱醛脱氢酶(BADH)基因导入玉米,以提高玉米的耐盐性.对1286株转化玉米进行PCR检测,共有16株呈阳性,转化率为1.2%.Southern Blot进一步证明BADH基因已经整合到玉米基因组.将阳性植株的后代(T3)用含1.2%NaCl的Hogland营养液每日1次浇灌。14d后对玉米叶片的相对电导率和叶绿素含量进行测定.结果表明在同样的盐胁迫条件下.转基因植株所受到的盐伤害明显较对照植株轻,说明转入的BADH基因可以提高玉米的耐盐能力.  相似文献   

2.
丙型肝炎病毒E2基因在转基因番茄植株中的表达   总被引:2,自引:1,他引:2  
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.  相似文献   

3.
根癌农杆菌介导大麦Mlo反义基因转化小麦   总被引:8,自引:0,他引:8  
应用含有大麦Mlo反义基因表达载体的根癌农杆菌菌株Agl Ⅰ,对3种基因型小麦(核生3号、烟优361、扬麦158)的愈伤组织进行了转化.从其中2种基因型获得23株转基因植株,有2株是白化苗,转化频率分别为1.9%(核生3号)和2.8%(扬麦158).PCR及Southem分析表明,外源基因已整合进植物基因组.实验结果表明,筛选压力的存在对转基因植株的获得至关重要,在无筛选压力下未获得转基因植株.7棵转基因植株移栽至大田后正常结实.  相似文献   

4.
建立了番茄子叶高频再生体系,优化了农杆菌介导的外源基因转化番茄的条件,将IPt和etrl-1双基因导入番茄中,以除草剂PPT作为筛选标记,得到3株转化再生植株.通过PCR,RT-PCR检测证明etrl-1和IPT基因已整合到转基因番茄植株的基因组中,并在转录水平有一定表达.  相似文献   

5.
为分析高山离子芥CbPLDβ基因的抗寒性价值,本实验构建了高山离子芥CbPLDβ的表达载体PBI121-CbPLDβ并转化烟草,经过筛选及PCR测序鉴定,获得转基因烟草植株.通过低温胁迫实验,分析了转基因烟草的抗寒性,结果表明:外源基因已经整合到烟草基因组中,在低温胁迫下转基因烟草植株的电解质渗透率普遍低于野生型烟草,而游离脯氨酸含量、可溶性糖和可溶性蛋白的含量均高于野生型烟草,说明CbPLDβ基因提高了烟草抵抗低温的能力.  相似文献   

6.
GRP1.8融合4CL1基因调控杨树木质素生物合成   总被引:1,自引:0,他引:1  
将形成层定位启动子GRP1.8与毛白杨的4-香豆酸:辅酶A连接酶基因融合,叶盘法转化毛白杨741,southern杂交检测表明融合基因已经整合到转基因杨树的基因组中.通过实时定量PCR技术测定mRNA的表达量,GC-MS分析4CL底物香豆酸的含量和定量硫酸木质素含量,分析了4CL基因的过量表达对转基因杨树生理生长的影响.实验结果表明,整合了正义4CL1的转基因植株在茎中的酶活性比野生植株增加了30%,木质素含量增加了40%.  相似文献   

7.
在获得转CBF4和bar基因蒙农杂种冰草植株的基础上,应用PCR和Southern杂交技术检测外源基因在转基因蒙农杂种冰草植株中的整合及拷贝数.确定外源基因CBF4和bar基因已经整合到冰草基因组中,并且是以多拷贝的整合方式插入受体细胞的染色体上.说明外源基因对蒙农杂种冰草成功地进行了转化,可以作为后期转基因冰草植株研究及培育转基因冰草新品种的材料.  相似文献   

8.
将从一株邻单胞菌中克隆到的一个新的邻苯二酚1,2-双加氧酶基因(tfd C)的起始密码子由GTG突变成ATG,并克隆到农杆菌双元载体pPZPY122中,利用农杆菌介导转化模式植物拟南芥,获得了转化植株,经过PCR,PCR-Southern和Southern dot blot方法检测证实,ftd C基因已经整合到拟南芥基因组中,邻苯二酚1,2-双加氧酶酶活性检测表明,转基因植株具有一定的酶活性,而未转化的植株则不具有酶活性。  相似文献   

9.
将本室克隆的编码复制酶基因3′端约1/2序列及其3′端非编码区的苜蓿花叶病毒中国分离株(AIMV-Ch)RNA:3′端cDNA,重组到植物表达载体pROKⅡ中,通过致瘤农杆菌(Agrobacterium tumefaciens)介导,以叶圆片为转化材料,转化普通烟草,并获得了转基因植株.经卡那霉素抗性选择、PCR检测目的基因证明.AIMV RNA:3′端基因已整合到转基因烟草的基因组DNA中.转基因植物的攻毒试验表明转基因植株对苜蓿花叶病毒产生高水平的抗性.  相似文献   

10.
利用基因枪法,以bar基因转化高羊茅(Festuca arundinacea Schreb.)的下胚轴愈伤组织,对获得的再生植株进行的PCR检测和点杂交分析表明,外源bar基因片段已经整合至高羊茅基因组中.同时,对转基因植株进行的草丁膦涂抹实验发现,转基因植株对除草剂的耐性提高.  相似文献   

11.
摘 要 为研究人参亲还素基因的抗盐活性,为该基因在人参抗逆育种方面的应用提供参考,通过植物转基因技术和外施不同浓度NaCl的方法获得阳性拟南芥植株,研究了不同植株类型不同盐浓度下的种子萌发率、植株生存率、植株主根长、植株分支数等相关指标。结果表明:在盐胁迫作用下转基因拟南芥种子萌发率高于野生型拟南芥;在盐胁迫作用下转基因拟南芥植株生存率显著高于野生型拟南芥;在盐胁迫作用下转基因拟南芥植株主根长大于野生型拟南芥;在盐胁迫作用下转基因拟南芥植株分支数与野生型拟南芥植株分支数没有明显差异。可见人参亲还素基因提高了转基因拟南芥抵御高盐胁迫的能力。  相似文献   

12.
转蔗糖: 蔗糖-1-果糖基转移酶基因提高烟草的耐旱性   总被引:4,自引:0,他引:4  
蔗糖: 蔗糖-1-果糖基转移酶(sucrose: sucrose 1-fructosyltransferase, 1-SST)以蔗糖为底物催化生成蔗果三糖等低聚合度的果聚糖.将从莴苣中克隆的1-SST基因重组到pCAMBIA1300-als中,构建了在CaMV 35S启动子调控下的植物表达载体,利用农杆菌介导的叶盘转化法将1-SST基因导入烟草中,PCR和Southern杂交检测表明获得了转基因植株,RT-PCR结果表明该基因在烟草中正常表达. 对T0代转基因烟草进行的耐旱性分析结果表明,干旱胁迫6d的转基因植株丙二醛含量和电解质渗漏率显著低于未转基因对照,叶片相对含水量下降速度也明显比对照慢. 对转基因植株叶片糖分分析表明,转基因烟草植株积累果聚糖,并在干旱胁迫后含量明显增加,而未转基因对照植株不积累果聚糖. 在14%PEG溶液中未转基因烟草种子的萌发率仅为转基因烟草种子的一半;在附加200mmol/L甘露醇的培养基中未转基因烟草种子根的生长明显受到抑制,而转基因烟草根的生长发育正常. 以上研究结果表明,转1-SST基因烟草植株耐旱性的提高可能与该基因的表达有关.  相似文献   

13.
It was found that the supplement of 10 -4 mol/L AgNO3 in N6 medium enhanced the induction of type Ⅰ embryogenic calli from immature embryos of maize inbred P9-10. After high-osmotic treatment, the induced calli was taken as transformation recipient to be bombarded with plasmid pMG6 carrying synthetic Bt gene by PDS-1000/He genegun. A total of 14 resistant calli were obtained after screening subsequently on the mediums with gradually increasing selective pressure. 10 plants were regenerated from the resistant calli on 3 different induction media, and 8 out of the 10 regenerated plants were confirmed to be integrated with Bt gene by PCR and Southern blotting analysis. Results of ELISA showed that the Bt-protein content in the leaves of the transgenic plants varied between 20-200 ng/g fresh weight.  相似文献   

14.
Soybean is one of the crops most difficult to be manipulated in vitro. Although several soybean marker genes, all the selectable markers used were from bacteria origin. To find suitable selectable marker gene from plant origin for soybean transformation, a mutant acetolactate synthase (ALS) gene from Arabidopsis thaliana was tested for Agrobacterium-mediated soybean embryo axis transformation with the herbicide Arsenal as the selective agent. Transgenic soybean plants were obtained after the herbicide se- lection and the To transgenic lines showed resistance to the herbicide at a concentration of 100 g/ha. ALS enzyme assay of To transgenic line also showed higher activity compared to the wild type control plant. PCR analysis of the T1 transgenic lines confirmed the integration and segregation of the transgene. Taken together, our results showed that the mutant ALS gene is a suitable selectable marker for soybean transformation.  相似文献   

15.
16.
The cry1Ah gene was one of novel insecticidal genes cloned from Bacillus thuringiensis isolate BT8. Two plant expression vectors containing cry1Ah gene were constructed. The first intron of maize ubiqutin1 gene was inserted between the maize Ubiquitin promoter and cry1Ah gene in one of the plant expressing vectors (pUUOAH). The two vectors were introduced into maize immature embryonic calli by microprojectile bombardment, and the reproductively plants were acquired. PCR and Southern blot analysis showed that foreign genes had been integrated into maize genome and inherited to the next generation stably. The ELISA assay to T1 and T2 generation plants showed that the expression of Cry1Ah protein in the construct containing the ubi1 intron (pUUOAH) was 20% higher than that of the intronless construct (pUOAH). Bioassay results showed that the transgenic maize harboring cry1Ah gene had high resistance to the Asian corn borers and the insecticidal activity of the transgenic maize containing the ubi1 intron was higher than that of the intronless construct. These results indicated that the maize ubi1 intron can enhance the expression of the Bt cry1Ah gene in transgenic maize efficiently  相似文献   

17.
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.  相似文献   

18.
转ChIFN-γ基因烟草抗虫机制研究   总被引:2,自引:0,他引:2  
为探索转ChIFN-γ基因烟草的抗虫作用机制,对虫体蛋白酶活性变化、烟草挥发性成分以及表皮腺毛密度进行了研究.结果表明,烟青虫进食转基因烟草量较少,导致其体内分泌蛋白酶量少、活力低;转基因烟草T0和T1代,黑松三烯和西柏三烯二醇相对含量较对照高;转基因烟草腺毛密度比对照高49.2%.ChIFN-γ基因激活的多种转录因子可能结合了腺毛发育相关基因的cis调节元件,上调了腺毛发育基因的表达,导致腺毛的数量增加,分泌的萜烯化合物增加从而具有显著的抗虫性.  相似文献   

19.
利用露天盆栽试验研究了不同浓度Cd胁迫下转PprI基因油菜和非转基因野生型油菜对Cd的吸收特性以及生理响应。结果表明:随着Cd胁迫浓度的升高,转基因和非转基因野生型油菜对Cd的吸收均显著增加,且二者地下部分的吸收都大于地上部分;在0.5 mg/kg Cd处理下,转基因和非转基因油菜对Cd的吸收差异不显著,但1和2 mg/kg处理下转基因油菜对Cd的吸收显著低于非转基因野生型油菜。两种油菜对Cd的富集系数和转运系数均随处理浓度的升高而降低;随着Cd浓度的增加,超氧化物歧化酶(SOD)活性呈现先上升后下降的趋势,且在相同处理浓度下,转基因油菜的SOD活性均高于非转基因野生型油菜;丙二醛(MDA)含量呈上升趋势,且其在转基因油菜中的含量均低于非转基因野生型油菜;叶绿素a和叶绿素b的含量随Cd浓度的升高而下降。因此PprI基因的导入,显著降低了油菜对Cd的吸收,提高了其可食部分安全性。  相似文献   

20.
转基因玉米遗传转化的研究进展   总被引:1,自引:0,他引:1  
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。  相似文献   

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