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91.
农杆菌介导霞多丽葡萄胚性细胞系遗传转化条件的优化 总被引:3,自引:1,他引:2
为建立葡萄(Vitis vinifera)遗传转化技术体系,以霞多丽葡萄(Chardonnay)胚性细胞系为靶组织,采用GUS检测法,对影响农杆菌介导葡萄遗传转化效率的主要因素进行了研究。结果表明,超声波处理时间长短对转化效率有较大影响,在所试的0.5、1、5、10 min 4种不同时间的超声波处理中,以5 min时转化效率较好,平均达到9.11个蓝色斑点;AS浓度对转化效率有明显差异,当浓度为50μmol/L时,平均蓝色斑点数为8.89个,100μmol/L时,达到12.44个;DTT质量浓度对转化效率也有较大影响,在所试的3种质量浓度(1,2,3 mg/L)中,以3 mg/L为最佳,有16.67个蓝色斑点。通过GUS瞬时表达检测,确立了农杆菌介导葡萄胚性细胞系遗传转化的几个最适影响因素,从而为葡萄遗传转化技术体系的建立奠定了基础。 相似文献
92.
影响根癌农杆菌介导香蕉胚性悬浮细胞遗传转化的因素 总被引:1,自引:0,他引:1
用含质粒载体pCAMBIA1301的根癌农杆菌EHA105转化贡蕉(Musa A A Pisang Mas cv. Mas)品种的胚性悬浮细胞,通过测定GUS基因瞬时表达率,对转化初期的主要影响因素进行了研究。结果表明,取处于对数期的胚性悬浮细胞和D600nm为0.2的菌液混合,在室温、8000r/min离心5min,然后在110r/min的旋转摇床上摇5min,在体细胞胚诱导培养基上共培养8d,瞬时表达率达到最高值25.30%。 相似文献
93.
94.
黄瓜棒孢叶斑病病原菌RFP标记转化株的构建 总被引:1,自引:0,他引:1
由多主棒孢(Corynespora cassiicola)侵染引起的黄瓜棒孢叶斑病已成为我国黄瓜生产上的重要新流行病害,目前尚缺乏抗性品种,病原菌侵染机制及与寄主的互作关系尚不清楚。为了开展多主棒孢的病原学研究,本试验采用农杆菌基因介导技术(ATMT),获得了红色荧光蛋白(RFP)标记的多主棒孢转化株,转化株在PDA培养基转接6次后仍能在菌丝和分生孢子上发出强烈的红色荧光,生物学测定和致病性测定结果表明该转化株与野生型菌株无显著差异。 相似文献
95.
96.
DREB2A基因对苜蓿遗传转化的研究 总被引:11,自引:4,他引:7
以黑龙江紫花苜蓿Medicago sativa主栽品种:肇东苜蓿、敖汉苜蓿、公农1号和公农2号为受体材料,系统地探讨了除菌剂种类和浓度以及卡那霉素筛选浓度等,建立了高效的苜蓿再生体系和遗传转化体系;分别构建了由诱导型启动子rd29A和组成型启动子E12调控的DREB2A基因的2个植物表达载体pB2A29A和pB2AE12;采用农杆菌介导法进行遗传转化,获得大量转基因抗性植株并进行了分子生物学(PCR和Southern blot)检测.结果表明,DREB2A基因已整合到苜蓿基因组中. 相似文献
97.
Inheritance and expression of transgenes in T2 and T3 generations of Lotus corniculatus transformed using Agrobacterium tumefaciens 总被引:2,自引:0,他引:2
K. Judith Webb Mervyn O. Humphreys Leif Skøt Margaret Gibbs John Gatehouse 《Euphytica》1999,108(3):169-179
The inheritance and expression of the reporter gene uidA, encoding β-glucuronidase (GUS), was previously analysed in the T1 generation of 25 independent transformed lines of Lotus corniculatus cv. Leo. In the work reported here, GUS activity in various tissues of seven of these lines was tested in the T2 generation. Four representative lines were chosen for more detailed study in the T3 generation. Lines 25 and 38 had multiple, independently segregating transgene inserts; lines 24 and 39 appeared to transmit one segregating transgene insert to their T1 progeny, although transgene expression was low and was detected in fewer seedlings than expected in line 39. The uidA gene was inherited and expressed in seedlings of T1, T2 and T3 generations of all four lines. In all lines, transgene expression varied between tissues, with more embryos than seedlings
having detectable GUS activity. Studies in the T2 generation showed that use of transgenic plants as female or male parents altered the frequency of expression of the transgene
in progeny. By contrast, in the T3 generation the use of transgenic plants as female or male parents did not effect either frequency of transmission, or expression
of the transgene, in any of the four lines. Transgene inheritance was also similar among individual pods within flower heads
and between individual flower heads.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
98.
豇豆胰蛋白酶抑制剂转基因棉花的获得 总被引:32,自引:10,他引:22
利用根癌农杆菌(Agrobacteriumtumefaciens)介导法将豇豆胰蛋白酶抑制剂(Cow-peaTrypsinInhibitor,CpTI)基因转移进入棉花(GosypiumhirsutumL.)。棉花幼苗的下胚轴与携带有CpTI基因和Npt-Ⅱ基因的根癌农杆菌共培养,在选择培养基上诱导出了卡那霉素抗性(Kmr)愈伤组织。选择Npt-Ⅱ阳性的胚性愈伤组织在胚状体诱导培养基上进行胚状体诱导,继而在分化培养基上进行植株分化,最终获得了再生棉花植株。经Npt-Ⅱ分析、PCR及Southern检测证明,外源CpTI基因和标记基因(Npt-Ⅱ基因)存在于转化棉株及其后代中。抗棉铃虫(Heliothisarmigera)生物活性检测表明转基因植株后代具有明显的抗棉铃虫能力。 相似文献
99.
Evidence of Migration and Endophytic Presence of Agrobacterium tumefaciens in Rose Plants 总被引:2,自引:0,他引:2
Rubén Martí Jaime Cubero Antonio Daza Jaime Piquer Carmen I. Salcedo Clara Morente María M. López 《European journal of plant pathology / European Foundation for Plant Pathology》1999,105(1):39-50
Agrobacterium tumefaciens was isolated from stem tumors of several rose cultivars showing that the bacterium is the causal agent of aerial galls in rose plants. No differences were observed in the characteristics of the Agrobacterium isolates from crown or aerial galls. Stem inoculation of ten rose cultivars showed that all of them were susceptible to A. tumefaciens but differences in the size of the resulting tumors were observed. The movement of A. tumefaciens in rose plants was demonstrated using two wild type strains and two antibiotic resistant mutants. Three months after inoculation, the inoculated strains were recovered in the roots, crown and below and above the inoculation site but low numbers of pathogenic Agrobacterium cells were isolated. New tumors appeared in 5% of the noninoculated wounds. A. tumefaciens was isolated from the stem at different distances from the tumor in naturally infected plants. In symptomless commercial plants, the isolation from the roots, crown and at different stem levels demonstrated the existence of systemic and latent infections in rose. Direct isolation using a nonselective and selective media with or without a previous enrichment step were efficient methods for isolating tumorigenic Agrobacterium from the different parts of rose plants. 相似文献
100.