共查询到19条相似文献,搜索用时 109 毫秒
1.
GNK2-1为一种来自银杏(Ginkgo biloba)种仁的新型抗真菌蛋白,具有较强的真菌抗性且性质稳定。序列分析表明,其结构与所有已知的抗真菌蛋白不同,而与富含半胱氨酸的植物类受体激酶的胞外结构域相似。为探索GNK2-1基因在黄瓜(Cucumis sativus)抗病反应中的作用,利用基因重组技术构建了GNK2-1的高效组成型表达载体,并利用根癌农杆菌(Agrobacterium tumefaciens)介导转入黄瓜栽培品种农城3号(Cucumis sativus' Nongcheng No.3')基因组中。通过对获得的抗性植株进行PCR、RT-PCR和Western blot检测分析,结果表明GNK2-1基因可在T0代转基因植株中转录表达,并能在T1代转基因黄瓜中稳定遗传。离体枯萎病抗性鉴定结果表明,转GNK2-1基因的黄瓜对枯萎病的抗性增强,GNK2-1可以作为黄瓜抗病性改良的潜在基因资源。 相似文献
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以黄瓜无菌苗子叶切段为外植体 ,通过叶盘转化法与根瘤农杆菌进行共培养建立了黄瓜的转基因系统。农杆菌菌株为LBA44 0 4,内含双元载体pBPMWMV。该质粒载体带有一个npt Ⅱ基因 (筛选具有卡那霉素抗性的植株 )和一个WMV 2CP基因。抗卡那霉素 (Kanr)的黄瓜植株经DNA分子点杂交、PCR检测以及Southernblot证实 ,外源的WMV 2CP基因确实已导入黄瓜细胞且能稳定地遗传到子一代。对WMV 2CP基因在子一代的分离进行了统计。获得的转基因子一代植株对WMV 2表现出较强的抗性 ,可以延迟发病时间 ,减轻发病程度 相似文献
3.
《生物技术通报》2016,(6)
采用抗生素标记法,对海洋多黏类芽孢杆菌L1-9菌株进行标记,抗性菌株L1-9Str,rif对链霉素和利福平的抗性浓度分别为160μg/m L和20μg/m L。双抗菌株L1-9Str,rif的抑菌特性及其对链霉素和利福平的抗性经多次传代仍比较稳定。盆栽试验表明,该双抗菌株能在黄瓜根部土壤及根组织、茎基部、子叶和真叶组织中定殖。菌株L1-9Str,rif在黄瓜外根际、根际和根表土壤及黄瓜组织中的定殖动态基本一致,初期黄瓜组织中L1-9Str,rif菌量较少,随着时间的延长,菌量逐渐增加,达到高峰后逐渐减少。菌株L1-9Str,rif在根表土壤中菌量最多(1.76×109 CFU/g),其次是根际土壤,外根际土壤中菌量较少;在黄瓜组织中,菌株L1-9Str,rif其在子叶中的定殖能力最强(5.63×104 CFU/g),其次是根和茎基部(0-2 cm);调查至第26 d时在根部土壤中的含菌量仍保持在稳定的水平,其中根表土壤中含菌量最高(2.41×107 CFU/g),在黄瓜组织样品中,子叶中的含菌量最高(4.15×104 CFU/g);温室防病实验结果表明,菌株L1-9和L1-9Str,rif菌株对黄瓜枯萎病具有良好的防治效果,不同时期防效均达70%以上。上述结果表明来自海洋的多黏类芽孢杆菌L1-9菌株能在黄瓜根部土壤及幼苗组织中定殖,是一株有潜力的黄瓜枯萎病生防菌株。 相似文献
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《生物技术》2014,(2)
目的:克隆海岛棉GbNPR1基因并研究该基因在抗病及抗病信号传导中的功能。方法:根据已报道的棉花NPR1基因序列设计1对特异引物,利用RT-PCR从新海21号克隆GbNPR1基因并对其序列进行生物信息学分析;通过实时定量PCR技术检测海岛棉经枯萎病菌侵染、水杨酸和乙烯诱导后该基因的表达特性。结果:GbNPR1基因编码587个氨基酸,与可可树中NPR1基因编码的氨基酸同源性最高(89.61%),其蛋白含有保守的BTB/POZ、ANK和NPR1_like_C结构域;枯萎病菌侵染,水杨酸和乙烯诱导均可使GbNPR1明显上调表达,枯萎病菌侵染3h,水杨酸和乙烯诱导2h表达量最高。结论:GbNPR1基因在棉花抵御枯萎病菌侵染及植物抗病信号转导过程中起一定作用。 相似文献
5.
《基因组学与应用生物学》2015,(9)
本研究利用人工接种及BSA法验证甜瓜抗枯萎病基因Fom-2的连锁分子标记SSR430和STS296应用于节瓜抗枯萎病鉴定的通用性。结果表明:(1)在12份抗病材料和27份感病材料的分子鉴定中,抗病材料均能找到SSR430标记,与高抗母本B-4带型一致,感病材料均无标记,准确率为100.00%,能用于节瓜抗枯萎病分子标记辅助育种;(2)利用STS296进行分子标记鉴定,只有B-4和3个抗性子代含有STS296标记,所有感病材料均不含此标记,表明STS296有可能可作为节瓜枯萎病的抗性标记。 相似文献
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转WMV—2CP基因黄瓜植株的再生 总被引:15,自引:0,他引:15
以黄瓜无菌苗子叶切段为外植体,通过叶盘转移化法与根瘤农杆菌进行共培养建立了黄瓜的转其因系统。农杆菌菌株为LBA4404,内含双元载体pBPMWMV。该质粒载体带有一个npt-Ⅱ基因(筛选具有卡那霉素抗性的植株)和一个WMV-2CP基因。抗卡那霉素(Kan^r)的黄瓜植株经DNA分子点杂交、PCR检测以及Southern blot证实,外源的WMV-2CP基因确实已导人瓜细胞且能稳定地遗传到子一代。 相似文献
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9.
JTV1基因的转录物编码p38蛋白质,也称AMIP2.p38/JTV1是人类tRNA合成酶复合物的支架蛋白质,是氨酰tRNA连接酶复合物的核心分子,对复合物的组装起到重要的作用.p38/JTV1的结构中包含了谷胱甘肤S-转移酶(GST)结构域,可以与损害的DNA结合,起到分子伴侣的作用;p38/JTV-1可以通过与FBP相互作用而下调c-myc,从而促进细胞的凋亡;AIMP2/p38是p53的正性调节因子,缺失AIMP2/p38,可增加DNA损伤引起的凋亡,因为JTV1拥有上述这些活性,并且人类肿瘤组织经常发生突变,提示JTV1可能作为一种新的肿瘤抑制剂. 相似文献
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11.
Chitinases, -1,3-glucanases, and ribosome-inactivating proteins are reported to have antifungal activity in plants. With the aim of producing fungus-resistant transgenic plants, we co-expressed a modified maize ribosome-inactivating protein gene, MOD1, and a rice basic chitinase gene, RCH10, in transgenic rice plants. A construct containing MOD1 and RCH10 under the control of the rice rbcS and Act1 promoters, respectively, was co-transformed with a plasmid containing the herbicide-resistance gene bar as a selection marker into rice by particle bombardment. Several transformants analyzed by genomic Southern-blot hybridization demonstrated integration of multiple copies of the foreign gene into rice chromosomes. Immunoblot experiments showed that MOD1 formed approximately 0.5% of the total soluble protein in transgenic leaves. RCH10 expression was examined using the native polyacrylamide-overlay gel method, and high RCH10 activity was observed in leaf tissues where endogenous RCH10 is not expressed. R1 plants were analyzed in a similar way, and the Southern-blot patterns and levels of transgene expression remained the same as in the parental line. Analysis of the response of R2 plants to three fungal pathogens of rice, Rhizoctonia solani, Bipolaris oryzae, and Magnaporthe grisea, indicated statistically significant symptom reduction only in the case of R. solani (sheath blight). The increased resistance co-segregated with herbicide tolerance, reflecting a correlation between the resistance phenotype and transgene expression. 相似文献
12.
转WMV-2外壳蛋白基因西瓜植株的病毒抗性 总被引:17,自引:0,他引:17
西瓜是夏季的重要水果,病毒病是影响其品质和产量的重要原因之一。植物基因工程的发展为抗病育种提供了新途径。利用外壳蛋白(coat protein)基因转化高等植物,赋予转基因植物以相应抗病性的成功例子已很多。本文报道WMV-2CP基因在自交子一代的分离符合孟德尔3:1的分离比。经过连续4代的选择鉴定,已从T7、T11和T323个独立转化子的后代中筛选获得8个转基因纯合株系,性状表现整齐一致。Western blot结果表明,R4T7-1、T4T11-3以及R4T32-73个不同来源的株系均能表达产生外壳蛋白。转基因纯合株系WMV-2感染后的病毒抗性实验表明,与未转基因对照相比,转基因株系可以推迟发病时间,减轻发病程度。实验筛选获得的转基因株系R4T32-7表现出对WMV-2的高度抗性,为利用植物转基因技术选育抗病新品种奠定了基础。 相似文献
13.
以前曾报道用RNA介导的抗病毒策略,获得了高度抗病的表达马铃薯Y病毒坏死株系外壳蛋白基因(PVY^N CP)的转基因烟草,并对T1、T2代转基因植株进行了遗传和抗病性分析。此次以T,代转基因植株为试验材料,在筛选高度抗病植株并证明其抗病性是基于转基因沉默的基础上,采用Northern杂交的方法,证明CMV侵染抑制了转基因植株中PVY^N CP基因的沉默,而且CMV对PVY^N CP基因沉默的抑制部位是发生在接种后的新生叶上,接种叶及其下部叶片中PVY^N CP基因沉默则未受到影响。采用ELISA方法对CMV PVY^N复合接种的转基因植株进行PVY^N检测,结果表明,接种叶及下部叶没有检测到PVY^N,植株叶片对PVY^N表现为抗病。而在CMV接种后植株新生叶中则检测出了高滴度的PVY^N,植株叶片对PVY^N表现为感病。该文报道了在表达PVY^N CP基因的RNA介导抗性转基因植株中,异源病毒侵染抑制了转基因的沉默,并导致转基因植株的抗病性丧失。 相似文献
14.
Resistance against Fusarium Head Blight in Transgenic Wheat Plants Expressing the ScNPR1 gene 总被引:1,自引:0,他引:1
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Guihong Yu Xu Zhang Jingbao Yao MiaoPing Zhou Hongxiang Ma 《Journal of Phytopathology》2017,165(4):223-231
Fusarium head blight (FHB) is a severe global wheat disease that may cause severe yield losses, especially during epidemic years. Transforming the regulatory genes in the metabolic pathways of disease resistance into wheat via transgenic methods is one way to improve resistance to FHB. ScNPR1 (Secale cereale‐NPR1), a regulatory gene for systemic acquired resistance (SAR), was isolated from S. cereale cv Jingzhouheimai and transformed into the moderately FHB‐susceptible wheat variety Ningmai 13. RT‐PCR analysis indicated that the ScNPR1 gene was stably expressed in transgenic plants. An evaluation of the resistance to FHB revealed that six ScNPR1 transgenic lines (NP1, NP2, NP3, NP4, NP5 and NP6) exhibited significantly higher FHB resistance than the wild‐type wheat Ningmai 13 and the null‐segregated plants. The expression of pathogenesis‐related (PR) genes after Fusarium graminearum inoculation was earlier or higher than those in the wild‐type variety Ningmai 13. The high expression in the early stages of PR genes should account for the enhanced FHB resistance in the transgenic lines. Our results suggest that overexpression of ScNPR1 could be used to improve FHB resistance in wheat. 相似文献
15.
以黄瓜子房 (幼果 )RNA为模板 ,应用逆转录 聚合酶链式反应 (RT PCR) ,首次扩增出黄瓜生长素结合蛋白基因 (ABP1)cDNA片段 ,并进行测序和同源性分析。对ABP1基因在黄瓜子房 (幼果 )中的mRNA表达水平作了初步探讨 ,结果表明 ,该基因在开花前 1d的子房中表达信号较弱 ,在授粉后 2、4和 6d的幼果中表达增强 ;开花后 2d未经授粉的子房中 ,绿而膨大、能形成单性结实果者信号较强 ,黄而萎蔫、不能形成果实者信号较弱。Southern杂交结果表明 ,黄瓜生长素结合蛋白为小基因家族编码 相似文献
16.
Fusarium wilt is caused by the soil-inhabiting fungus Fusarium oxysporum ff. spp. and is one of the most devastating plant diseases, resulting in losses and decreasing the quality and safety of agricultural crops. We recently reported the structures and biochemical properties of two biotin-binding proteins, streptavidin C1 and C2 (isolated from Streptomyces cinnamonensis strain KPP02129). In the present study, the potential of the biotin-binding proteins as antifungal agent for Fusarium wilt pathogens was investigated using recombinant streptavidin C1 and C2. The minimum inhibitory concentration of streptavidin C2 was found to be 16 µg ml–1 for inhibiting the mycelial growth of F. oxysporum f.sp. cucumerinum and F. oxysporum f.sp. lycopersici, while that of streptavidin C1 was found to be 64 µg ml–1. Compared with the nontreated control soil, the population density of F. oxysporum f.sp. lycopersici in the soil was reduced to 49·5% and 39·6% on treatment with streptavidin C1 (500 µg ml–1) and C2 (500 µg ml–1), respectively. A greenhouse experiment revealed that Fusarium wilt of tomato plants was completely inhibited on soil drenching using a 50-ml culture filtrate of the streptavidin-producing strain KPP02129. 相似文献
17.
Coca M Bortolotti C Rufat M Peñas G Eritja R Tharreau D del Pozo AM Messeguer J San Segundo B 《Plant molecular biology》2004,54(2):245-259
The Aspergillus giganteus antifungal protein (AFP), encoded by the afp gene, has been reported to possess in vitro antifungal activity against various economically important fungal pathogens, including the rice blast fungus Magnaporthe grisea. In this study, transgenic rice ( Oryza sativa ) constitutively expressing the afp gene was generated by Agrobacterium -mediated transformation. Two different DNA constructs containing either the afp cDNA sequence from Aspergillus or a chemically synthesized codon-optimized afp gene were introduced into rice plants. In both cases, the DNA region encoding the signal sequence from the tobacco AP24 gene was N-terminally fused to the coding sequence of the mature AFP protein. Transgenic rice plants showed stable integration and inheritance of the transgene. No effect on plant morphology was observed in the afp -expressing rice lines. The inhibitory activity of protein extracts prepared from leaves of afp plants on the in vitro growth of M. grisea indicated that the AFP protein produced by the trangenic rice plants was biologically active. Several of the T(2) homozygous afp lines were challenged with M. grisea in a detached leaf infection assay. Transformants exhibited resistance to rice blast at various levels. Altogether, the results presented here indicate that AFP can be functionally expressed in rice plants for protection against the rice blast fungus M. grisea. 相似文献
18.
Xiaoyan Zhao Xiaomeng Zhao Yanmin Wei Qiaoxia Shang Zhenping Liu 《Journal of Phytopathology》2013,161(1):43-48
Bacillus subtilis strain F3, isolated from peach rhizosphere soil, is an antifungal bacterium against many plant pathogens. In this study, the antifungal protein was isolated and purified by ammonium sulphate and chromatography, then identified by mass spectrum analysis. By sequential chromatography of Sephadex G‐50, DEAE‐Sephadex A‐25 anion exchange and Sephadex G‐100, a fraction designated as F3A was isolated to show a single protein band in SDS‐PAGE and be antagonistic towards Monilinia fructicola. The peptide mass fingerprinting of the protein band of F3A had high similarity with the amino acid sequences of several flagellin protein of B. subtilis. There were seven amino acid fragments matched with the protein having the highest score, and sequence coverage was 33%. F3A showed a strongly inhibitory effect to the growth and sporulation of M. fructicola. There were little aerial hyphae and conidia at the antifungal zone, and the hyphae were abnormal with some cell wall collapse and several vacuoles in cells. 相似文献
19.
天麻抗真菌蛋白基因(gafp)转化彩色棉的研究 总被引:8,自引:0,他引:8
天麻抗真菌蛋白(gastrodia antifungal protein简称GAFP)是从我国传统中药天麻(Gastrodia elata B1.)中分离到的一种具有广谱抗真菌活性的蛋白质,它对许多植物真菌病包括棉花枯萎病、黄萎病等的致病菌离体具有很强的抑制作用,因此,在植物抗真菌病基因工程上有很重要的应用价值。本研究通过花粉管通道法,将GAFP的基因.gafp转入3个新疆彩色棉品种中,通过田间抗病筛选和分子检测,得到了高抗黄萎病的转基因植株,两株Southem杂交阳性植株LB-5-8和ZB-1—49对黄萎病表现整株免疫。RT-PCR的结果显示,LB-5-8和ZB-1—49中均有gafp的正确转录;离体的抑菌实验也表明,它们的蛋白粗提物对棉花黄萎病致病菌离体有明显的抑制,表明了gafp在转基因植株中的正确表达,翻译的产物具有活性。经过进一步选育和扩繁,发现转基因彩色棉后代具有稳定的、较强的抗黄萎病能力,本研究为通过植物抗病基因工程的方法防治棉花黄萎病提供了一条新的途径。 相似文献