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1.
通过根癌农杆菌(Agrobactrium tumefaciens)介导转化法,将含有稻瘟菌蛋白激发子基因pemG1的植物表达载体pCAMBIA2300-Ubi-pemG1-Ocs转化三生烟(Nicotiana tobacum cv. Samsun NN),获得了转基因植株。用PCR检测抗卡那霉素烟苗确认阳性转化株,用Southern、Northern和Western杂交进一步证实了pemG1基因的整合、转录和表达。对T2代转基因阳性株进行烟草花叶病毒(Tobacco Mosaic Virus)接种试验。接种5 d后发现,与非转基因对照相比,表达pemG1的烟草叶片枯斑数量减少,表明稻瘟菌蛋白激发子基因pemG1的表达提高了转基因烟草对TMV的抗性。 相似文献
2.
蛋白激发子基因pemG1转化三生烟及其对TMV抗性的提高 总被引：2，自引：0，他引：2
通过根癌农杆菌（Agrobactrium tumefaciens）介导转化法,将含有稻瘟菌蛋白激发子基因pemGl的植物表达载体pCAMBIA2300-Ubi-pemG1-Ocs转化三生烟（Nicotiana tobacum cv．Samsun NN）,获得了转基因植株。用PCR检测抗卡那霉素烟苗确认阳性转化株,用Southern、Northern和Western杂交进一步证实了pemG1基因的整合、转录和表达。对T2代转基因阳性株进行烟草花叶病毒（Tobacco Mosaic Virus）接种试验。接种5d后发现,与非转基因对照相比,表达pemG1的烟草叶片枯斑数量减少,表明稻瘟菌蛋白激发子基因pemG1的表达提高了转基因烟草对TMV的抗性。 相似文献
3.
pCAMBIA2300-betA-BADH双价植物表达载体的构建 总被引：1，自引：0，他引：1
将载体pCAMBIA2300-35s-OCS进行改造,增加了酶切位点Sac Ⅰ,之后,PCR扩增编码胆碱脱氢酶(CDH)的基因并用Sma Ⅰ和Xba Ⅰ将其与改造后的pCAMBIA2300-35s-OCS连接,得到单价植物表达栽体p2300-gz-betA.再通过Sal Ⅰ、Hind Ⅲ、Sac Ⅰ等酶切位点将编码甜菜碱醛脱氧酶(BADH)的基因及其启动子和终止序列连接到栽体p2300-gz-betA上,最终得到pCAMBIA2300-betA-BADH双价植物表达载体,并将其导入根癌农杆菌GV3101和LBA4404.betA和BADH是编码甜菜碱合成途径的两个关键酶的基因(CDH和BADH),也是近年来研究较多的耐盐基因,其双价植物表达栽体应用于遗传转化后,有望进一步提高转基因植物的耐盐性. 相似文献
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5.
以茎尖分生组织为受体用农杆菌介导法转化激发子基因pemG1棉花植株的获得 总被引：1，自引：1，他引：0
为了解决棉花常规遗传育种周期长,农杆菌介导的转化下胚轴再生难,转化率低,胚变异率高等难题。通过改善茎尖的遗传转化,利用农杆菌介导法转化棉花茎尖,试图缩短转化周期,获得转化苗。构建了稻瘟菌激发子基因pemG1的植物表达载体pCAMBIA2300-pemG1,分别以‘珂字312’、‘中棉24’棉花茎尖分生组织作为外植体,摸索无菌苗的最佳培养方案,确定最佳共培养温度、共培养时间及卡那霉素筛选浓度。经过共培养,茎尖在诱导芽培养基上诱导生芽,从不同浓度的BAP和NAA生长激素的组合中,最终确定0.1 mg/L的BAP和NAA最适合芽的诱导;经过两个月的继代培养,在含有GA3的茎增殖培养基上完成茎的增殖和延伸;切取幼芽并使其在含有0.1 mg/L的IBA生根培养基中生根;生根的抗性植株移栽到温室中成苗,获得了T0代转基因株系4株。进行了转pemG1阳性植株的初步验证。 相似文献
6.
昆虫抗冻蛋白基因转化烟草的抗寒性 总被引：8，自引：0，他引：8
将准噶尔小胸鳖甲抗冻蛋白基因MPAFP149亚克隆至pCAMBIA1302中,构建成单价植物表达载体pCAMBIA1302-MPAFP149,转化至农杆菌EHA105中,利用叶圆盘法转化烟草。通过PCR、PCR-Southern及RT-PCR检测表明抗冻蛋白基因已整合至烟草基因组中,并发生了转录。对T0代转基因烟草以－1℃处理48 h,转基因烟草的相对电导率和表型明显优于野生型烟草。室温恢复试验验证转基因烟草可存活并恢复生长,而野生型烟草受到了不可逆的低温冻害。研究证明转化后携带昆虫抗冻蛋白基因的烟草比野生型烟草具有明显的抗寒能力,该结果为减轻冷敏感经济作物在春季遭受霜害提供了理论依据和应用基础。 相似文献
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8.
含硫氨基酸基因植物表达载体的构建及对百脉根的转化 总被引：2，自引：0，他引：2
在本研究中,用限制性内切酶Nco Ⅰ和Bgl Ⅱ从中间载体pMD18zeolin和pMD18zein 上切下zeolin基因(约1 600bp)和γ-zein基因(约720 bp),将其定向连接在经相同酶切的质粒载体pCAMBIA1302上,构建成植物表达载体pCBzeolin和pCBzein.采用冻融法将pCBzeolin和pCBzein导入根癌农杆菌(Agrobacterium tumefaciens)菌株LBA4404,利用该菌株转化百脉根(Lotus corniculatus L.).经过共培养、筛选分化、再生,得到抗件植株.对抗性植株进行了PCR、RT-PCR检测表明,γ-zein和zeolin基因已经整合到百脉根基因组中,在核酸水平得到了表达.含硫氨基酸数据分析表明,转zeolin基因植株含硫氨基酸含量极显著高于转γ-zein基因植株和对照植株的含量(P＜0.01),但转γ-zein基因植株与对照植株间差异不显著. 相似文献
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10.
优化的VHb基因和GFMcryIA基因构建的双价基因在烟草中的表达 总被引：1，自引：0，他引：1
利用人工合成的、密码子优化后的透明颤菌血红蛋白（VHb）基因与人工合成的GFMcryIA基因构建成双价基因植物高效表达载体。通过根癌农杆菌介导转化烟草,获得了36株卡那霉素抗性植株。经转基因植株的PCR及Southern blot检测,证实了双价基因在35株烟草基因组中的整合。Western blot检测证实了VHb基因的表达,杀虫实验证实GFMcryIA基因也表达出活性毒蛋白。且转基因烟草平均每株干重比非转基因烟草植株高7％。本研究结果表明转基因育种技术在培育出既高产又具抗虫性的作物或经济作物新品种方面具有良好的应用前景。 相似文献
11.
Jens Jensen 《Euphytica》1979,28(1):47-56
Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans. 相似文献
12.
Biological Activity and Quantification of Suspected Allelochemicals from Alfalfa Plant Parts 总被引：7，自引：0，他引：7
Autotoxicity restricts reseeding of alfalfa (Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments. A series of aqueous extracts from leaves, stems, roots and seeds of alfalfa ‘Vernal’ were bioassayed against alfalfa seedlings of the same cultivar to determine their autotoxicity. The highest inhibition was found in the extracts from the leaves. Extracts at 40 g dry tissue l?1 from alfalfa leaves were 15.4, 17.5 and 28.7 times more toxic to alfalfa root growth than were those from roots, stems and seeds, respectively. A high‐performance liquid chromatography (HPLC) analysis with nine standard compounds showed that the concentrations and compositions of allelopathic compounds depended on the plant parts. In leaf extracts that showed the most inhibitory effect on root growth, the highest amounts of allelochemicals were detected. Among nine phenolic compounds assayed for their phytotoxicity on root growth of alfalfa, coumarin, trans‐cinnamic acid and o‐coumaric acid at 10?3 m were most inhibitory. The type and amount of causative allelochemicals found in alfalfa plant parts were highly correlated with the results of the bioassay, indicating that the autotoxic effects of alfalfa plant parts significantly differed. 相似文献
13.
Hongyan WEI Jian ZHOU Jinguang LU Jingzheng SONG Qiongxi YU Zhihong JIANG 《Medicinal Plant》2019,(6):27-29
[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol... 相似文献
14.
C. J. Spurr D. A. Fulton P. H. Brown R. J. Clark 《Journal of Agronomy and Crop Science》2002,188(4):275-280
Development of onion (Allium cepa L., cv. ‘Early Cream Gold’) seed under cool climate conditions in Tasmania, Australia occurred over a longer duration than previously reported, but similar patterns of change in yield components were recorded. In contrast to previous studies, umbel moisture content declined from 85 to 67 % over 57 days while seed moisture content decreased from 85 to 31 %. Seed yield continued to increase over the duration of crop development, with increasing seed weight compensating for seed loss resulting from capsule dehiscence in the later stages of maturation. Germination percentage was high and did not vary significantly from 53 to 77 days after full bloom (DAF), but mean germination time declined and uniformity of germination increased significantly over the same time period. The percentage abnormal seedlings declined with later harvest date, resulting in highest seed quality at 77 DAF. The results of this study suggest that the decision to harvest cool climate onion seed crops before capsule dehiscence will result in a loss of potential seed yield and quality. 相似文献
15.
Chunli TANG Fengxian ZHAO Hongxia CHEN Jiabao MA Jiangcun WEI Meiyan QIU Zhen XIE 《Medicinal Plant》2019,(6):60-65
[Objectives]To optimize the water extraction process of Chinese Herbal Compound Man Gan Ning and establish a method for its extraction and content determination... 相似文献
16.
D. A. Bond G. J. Jellis G. G. Rowland J. Le Guen L. D. Robertson S. A. Khalil L. Li-Juan 《Euphytica》1993,73(1-2):151-166
Progress is being made, mainly by ICARDA but also elsewhere, in breeding for resistance to Botrytis, AScochyta, Uromyces, and Orobanche; and some lines have resistance to more than one pathogen. The strategy is to extend multiple resistance but also to seek new and durable forms of resistance. Internationally coordinated programs are needed to maintain the momentum of this work.Tolerance of abiotic stresses leads to types suited to dry or cold environments rather than broad adaptability, but in this cross-pollinated species, the more hybrid vigor expressed by a cultivar, the more it is likely to tolerate various stresses. 相似文献
17.
Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed. 相似文献
18.
[Objectives] To determine the optimum extraction technology for total phenols of leaves in Acanthopanax giraldii Harms.[Methods]The single factor test and ortho... 相似文献
19.
E. Keep 《Euphytica》1986,35(3):843-855
Summary Cytoplasmic male sterility (cms) is described in the F1 hybrids Ribes × carrierei (R. glutinosum albidum × R. nigrum) and R. sanguineum × R. nigrum. In backcrosses to R. nigrum, progenies with R. glutinosum cytoplasm were either all male sterile, or segregated for full male fertility (F) and complete (S) and partial (I) male sterility. Ratios of F:I+S suggested that two linked genes controlled cms, F plants being dominant for one (Rf
1) and recessive for the other (Rf
2).Segregation for cms in relation to three linded genes, Ce (resistance to the gall mite, Cecidophyopsis ribes), Sph
3(resistance to American gooseberry mildew, Sphaerotheca mors-uvae) and Lf
1(one of two dominant additive genes controlling early season leafing out) indicated that Rf
1and Rf
2were in this linkage group. The gene order and approximate crossover values appeared to be: % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% aabaqaciGacaGaamqadaabaeaafaaakeaacaWGdbGaamyzamaamaaa% baGaaiiiaiaacccacaGGWaGaaiOlaiaacgdacaGG0aGaaiiiaiaacc% caaaGaaiiiaiaacccacaGGGaGaamOuaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaaccdacaGGUaGaaiOmaiaacs% dacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaaaacaWGsbGaamOzaSGa% aGOmaOWaaWaaaeaacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccaaaGaamitaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccacaGGGaaaaiaadofacaWGWbGaamiAaSGa% aG4maaaa!6E4D!\[Ce\underline { 0.14 } Rf1\underline { 0.24 } Rf2\underline { } Lf1\underline { } Sph3\]. Crossover values of 0.36 for Ce-Lf
1, and 0.15 for Lf
1-Sph
3were estimated from the relative mean differences in season of leafing out between seedlings dominant and recessive for Ce and Sph
3.It is suggested that competitive disadvantage of lf
1-carrying gametes and/or zygotes at low temperatures may be implicated in the almost invariable deficit of plants dominant for the closely linked mildew resistance allele Sph
3. Poor performance of lf
1- (and possibly lf
2-) carrying gametes and young zygotes during periods of low temperature at flowering might also account for the liability of some late season cultivars and selections to premature fruit drop (running off). 相似文献
20.
Parasitic angiosperms cause great losses in many important crops under different climatic conditions and soil types. The most widespread and important parasitic angiosperms belong to the genera Orobanche, Striga, and Cuscuta. The most important economical hosts belong to the Poaceae, Asteraceae, Solanaceae, Cucurbitaceae, and Fabaceae. Although some resistant cultivars have been identified in several crops, great gaps exist in our knowledge of the parasites and the genetic basis of the resistance, as well as the availability of in vitro screening techniques. Screening techniques are based on reactions of the host root or foliage. In vitro or greenhouse screening methods based on the reaction of root and/or foliar tissues are usually superior to field screenings and can be used with many species. To utilize them in plant breeding, it is necessary to demonstrate a strong correlation between in vitro and field data. The correlation should be calculated for every environment in which selection is practiced. Using biochemical analysis as a screening technique has had limited success. The reason seems to be the complex host-parasite interactions which lead to germination, rhizotropism, infection, and growth of the parasite. Germination results from chemicals produced by the host. Resistance is only available in a small group of crops. Resistance has been found in cultivated, primitive and wild forms, depending on the specific host-parasite system. An additional problem is the existence of pathotypes in the parasites. Inheritance of host resistance is usually polygenic and its transfer is slow and tedious. Molecular techniques have yet to be used to locate resistance to parasitic angiosperms. While intensifying the search for genes that control resistance to specific parasitic angiosperms, the best strategy to screen for resistance is to improve the already existing in vitro or greenhouse screening techniques. 相似文献