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1.
用基因枪法将人工雄性不育基因导入小麦的研究初报   总被引:52,自引:1,他引:51  
傅荣昭  陈占宽 《遗传学报》1997,24(4):358-361
利用PDS1000/氦气基因枪将人工构建的雄性不育基因(TA29-Barnase基因)导入小麦栽培品种豫责18号的幼胚细胞。然后在含有10~20mg/L除草剂Basta的培养基础上筛选与分化。从170个幼胚中获得6株绿苗,对照的70个幼胚中未得到绿苗。对其中3株已生根且长势好的绿苗进行Southem杂交分析,结果表明,这3株绿苗皆为转基因植株,转化效率达1.8%。  相似文献   

2.
利用激光微束穿刺法将外源基因导入小麦的研究   总被引:21,自引:1,他引:20  
用激光微束穿刺法将携带有新霉素磷酸转移酶(NPT-Ⅱ)基因的质粒pJIT101导入京花1号小麦幼胚细胞。方法是将小麦幼胚细胞进行高渗缓冲液预处理,用微米级的激光微束处理,然后在卡那霉素培养基上筛选出具抗性的愈伤组织及绿色小植株。第一年,从150个小麦幼胚中,在卡那霉素培养基上筛选出4株绿苗,取两株进行NPT-Ⅱ酶活性分析,测到了NPT-Ⅱ酶的活性。第二年重复实验,从245个小麦幼胚中,经筛选获得1株绿苗,进行了叶片DNAPCR扩增检测,转化的绿色小苗扩增出所导入的NPT-Ⅱ基因编码的片段。结果表明,外源NPT-Ⅱ基因已导入了小麦,并实现了整合表达。  相似文献   

3.
雪花莲外源凝集素基因转化番茄   总被引:20,自引:0,他引:20  
将含有雪花莲外源凝集素基因(GNA)的质粒pRSSGNA1通过冻融法转化到根癌土壤杆菌9Agrobacterium tumefaciens(Smith et Townsend)Conn)菌株LBA4404中。采用叶盘法转化番茄(Lycopersicon esculentum Mill.)栽培品种“C8”、“A39”和“A53”,获得了含GNA基因的43株转化植物株。通过卡那霉素抗性鉴定、NPTⅡ基  相似文献   

4.
通过基因枪轰击转化获得转基因小麦植株的研究   总被引:13,自引:1,他引:12  
利用JQ-700型高速基因枪将pDM302质粒DNA上的bar基因即PAT酶基因导入了冬小麦品种“农大146”的幼胚中。经过在含有的选择培养基上筛选,得到了9块具有ppt抗性的愈伤组织。PCR电泳检测与PCR-Southern发要交结果显示,外源bar基因已转化进了由其中4块愈伤组织再生出的转基因的小麦植株中。  相似文献   

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黄瓜花叶病毒(CMV)运动蛋白基因介导的抗病性   总被引:3,自引:0,他引:3  
利用Fny_CMV株系RNA3cDNA克隆,构建了含有全长和编码区缺失501个核苷酸的运动蛋白(MP)基因植物表达载体pBMPR和pBMPK。在土壤农杆菌(Agrobacteriumtumefaciens(SmithetTownsend)Conn)LBA4404介导下转化烟草(NicotianatabacumL.)品种“NC89”,分别经Southernbloting、RT_PCR或Westernbloting分析,外源基因已整合到再生植株中并得到表达。抗病性分析表明,含有缺失型MP基因的R0代转基因植株抗性较好,接种50d后,10株转化植株中仍有5株不表现症状。在自然发病条件下,这5个含有缺失型MP基因转基因株系在R1代都表现了一定的抗病性。抗性主要表现为症状出现推迟,严重度减轻。利用PCR筛选、种子卡那霉素抗性试验和温室抗病性测定等方法,初步认为R2代转基因烟草K_6_5株系为转基因抗病纯合系。而含有全长MP基因的R0代转化植株,前期没有表现明显的抗病性,但在接种40d后部分发病植株有恢复健康的趋势。  相似文献   

6.
小麦抗白粉病相关基因的转化   总被引:7,自引:0,他引:7  
王华忠  邢丽萍  陈佩度 《遗传》2007,29(2):243-249
利用玉米花青素苷合成调节基因C1-Lc作为报告基因, 通过瞬间表达后愈伤组织表面红色斑点的统计分析, 优化了小麦幼胚愈伤组织的基因枪转化参数。小麦Beclin1类似基因TaTBL和硫代硫酸硫转移酶基因TaTST是2个在白粉菌诱导条件下具有增强表达特性的抗病相关基因。本实验进一步利用基因枪将ubi强启动子控制下的2个基因导入到小麦品种扬麦158的幼胚愈伤组织细胞中, 使用除草剂经两轮选择培养基上的筛选和再生获得抗性植株, 进一步通过抗性植株的PCR分析获得转TaTBL基因植株5株, 转TaTST基因植株6株。转基因植株离体叶片的人工接种实验表明, 外源基因的导入不同程度上增强了植株的白粉病抗性, 表现为延缓了白粉菌的发育。利用玉米花青素苷合成调节基因C1-Lc作为报告基因,通过瞬间表达后愈伤组织表面红色斑点的统计分析,优化了小麦幼胚愈伤组织的基因枪转化参数。小麦Beclin1类似基因TaTBL和硫代硫酸硫转移酶基因TaTST是两个在白粉菌诱导条件下具有增强表达特性的抗病相关基因。本实验进一步利用基因枪将ubi强启动子控制下的两个基因导入到小麦品种扬麦158的幼胚愈伤组织细胞中,使用除草剂经两轮选择培养基上的筛选和再生获得抗性植株,进一步通过抗性植株的PCR分析获得转TaTBL基因植株5株,转TaTST基因植株6株。转基因植株离体叶片的人工接种实验表明,外源基因的导入不同程度上增强了植株的白粉病抗性,表现为延缓了白粉菌的发育。  相似文献   

7.
表达barstar基因及bar基因的转基因油菜的研究   总被引:30,自引:0,他引:30  
从细菌Bacilusamyloliquefaciens染色体DNA中克隆了barnase抑制剂barstar的基因,构建了带有TA-29基因5′调控区(-1300-+3)与barstar基因编码区、CaMV35S启动子与除草剂抗性基因bar两个表达框架的植物表达质粒pBBS。以“双低”油菜“5-4”的子叶柄为受体,通过农杆菌介导的遗传转化,获得了在含有10mg/L卡那霉素和20mg/LPPT的筛选培养基上再生的转基因植株。PCR分析结果表明,barstar基因已整合到油菜染色体上;Northernblot检测表明,barstar基因及bar基因在转基因植物中得到了正确的调控与表达。以转基因油菜“5-4”为父本授粉给表达barnase基因的雄性不育植株,不育株能正常结实。  相似文献   

8.
日本Hiroshima大学和Kagawa大学的M.Nishihara和H.Morikawa及其同事报道,他们从用粒子轰击法转化的花粉第一次获得了稳定的转基因单倍体植株。Nishihara等人用含有新霉素转移酶Ⅱ(NPTⅡ)和β-葡糖苷酸酶(GUS)基因的质粒DNA包被的金粒,转化烟草属Nicotianarusticana的未成熟花粉。从被轰击的花粉再生了卡那霉素抗性花粉胚胎,并从总数为2×106个被轰击细胞中获得两个独立的转基因株系。一个同时具有NPTⅡ和GUS活性,另一个只有NPTⅡ活性。两个…  相似文献   

9.
抗除草剂草甘膦EPSPs基因在小麦中的转化   总被引:22,自引:1,他引:21  
陈梁鸿  张文俊 《遗传学报》1999,26(3):239-243
通过基因枪法,用抗除草剂草甘膦的EPSPs基因转化小麦京花1号的幼穗约1000个,及幼胚约800个,经草甘膦选择后分别获得38株和4株再生植株。这些再生植株经PCR和(或)Southern杂交证明,其中有部分再生植株的基因组中稳定整合了外源EPSPs基因,并且转化体中部分是可育的。首次用实验证明,抗除草剂草甘膦的EPSPs基因作为单子叶禾谷类作物小麦基因转化的选择标记是行之有效的。  相似文献   

10.
植物抗毒素转化水稻和转基因植株的生物鉴定   总被引:15,自引:0,他引:15  
用基因枪法转化了水稻(OryzasativaL.)6个材料的未成熟胚、成熟胚及胚性愈伤组织。质粒pSSVst1和pVE5+是由葡萄中分离出的编码芪类合成酶的植物抗毒素基因与35S或它自己的启动子组成。G418(100~150mg/L)或潮霉素(50mg/L)筛选后,经PCR、Southernblot或Dotblot分析证明的转基因植株共54株。对转基因植株及其后代进行了稻瘟病和白叶枯病的抗性鉴定。初步结果表明,芪类合成酶基因可以提高转基因植株及后代的抗性。  相似文献   

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Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.  相似文献   

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Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.  相似文献   

17.
Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.  相似文献   

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正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases  相似文献   

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