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1.
以基因枪法将Bt杀虫蛋白基因导入常规玉米自交系的研究   总被引:1,自引:2,他引:1  
以生产上大面积使用的优良玉米自交系7922为材料,以来源于其幼胚的Ⅱ型胚性愈伤为受体,经预实验选择标准制弹用量一半的金粉,通过基因枪轰击法,将Bt杀虫蛋白基因转入其细胞中,并再生了大量的转基因植株.抗性植株再生频率为78.40%,PCR分子检测证明目的基因已转入玉米中,转化频率为3.09%.优良自交系的转化成功为进一步培育适合我国农业生产的转基因玉米奠定了基础。  相似文献   

2.
为了探索小麦成熟胚基因枪转化的影响因素,对Z50、郑麦366、Bobwhite和西农529四个小麦基因型的成熟胚进行了愈伤组织的诱导及植株分化再生研究,同时以郑麦366的成熟胚为外植体分析了不同渗透处理剂、每枪金粉用量和轰击距离对基因枪转化过程的影响。结果表明,Z50、郑麦366、Bobwhite三个基因型的出愈率及胚性愈伤率差异不大,均在85%以上,西农529的出愈率较低,胚性愈伤率与Z50、郑麦366和Bobwhite差异显著。不同小麦基因型分化率差异显著,其中郑麦366分化率最高(39.26%)。基因枪转化结果显示,渗透剂对小麦愈伤组织生长均有一定的抑制作用,蔗糖的抑制作用比甘露醇弱;每枪金粉用量100μg的分化率较高(45.57%),但金粉用量100μg时的GUS瞬时表达量与金粉用量60μg时差异不显著,轰击距离为9cm时的分化率和GUS瞬时表达量均比6cm和12cm时高;蔗糖用量0.4mol、金粉用量60μg、轰击距离9cm时GUS瞬时表达量最高,该条件下小麦成熟胚转化频率达0.67%,已初步优化了基因枪介导的小麦成熟胚遗传转化体系。  相似文献   

3.
以玉米自交系E28的胚性愈伤组织为受体,分别利用农杆菌介导法和基因枪轰击法对携带有GUS基因和Bar基因的质粒进行转化,借助GUS基因的瞬时表达率和抗性愈伤组织比例建立体系,并对两种方法进行比较。农杆菌介导法以OD6000.6、浸染30 min、25℃共培养3 d、加入0.01%的silwet L-77为最佳参数;基因枪轰击法为4.5 MPa、11 cm轰击距离最佳参数。农杆菌介导法与基因枪轰击法的GUS瞬时表达率没有显著差异,基因枪法的抗性愈伤组织比例显著高于农杆菌介导法,基因枪轰击法较农杆菌介导法更有利于获得转基因后代。  相似文献   

4.
利用基因枪介导法将含有Cry1Ac基因的质粒转入玉米愈伤组织,获得转Bt基因玉米植株,对所获得的转基因植株进行PCR分析和金标免疫试纸鉴定。结果表明,部分转化植株呈阳性,说明目的基因已经整合到玉米基因组中。将转基因植株自交获得T1代种子,对T1代植株进行田间抗虫鉴定,结果表明,与对照相比转基因玉米植株的抗虫活性明显强于非转基因植株。  相似文献   

5.
转GNA基因小麦新株系的分子检测和抗蚜虫性鉴定   总被引:5,自引:1,他引:5       下载免费PDF全文
为了进行抗蚜虫转基因小麦的研究,将人工合成的雪花莲凝结素(Galanthus nivlis agglutinin.GNA)基因通过基因枪转入优质小麦品种郑州9405的幼胚愈伤组织,经过在选择培养基上多次筛选,从360块被轰击的愈伤组织中再生到1株Bialaphos(除草剂)抗性植株,命名为G郑州9405。通过连续2代对转化株系进行PCR、Southern检测和蚜虫抗性鉴定,证明GNA基因已经整合到了郑州9405基因组中。目前已经获得了纯合稳定的转基因株系,并在河南农业科学院小麦所进行了环境释放试验。  相似文献   

6.
为验证小麦病程相关蛋白基因TaPR-1的功能,利用gus基因瞬间表达技术对基因枪法转化小麦幼胚的两个重要参数金粉用量和轰击距离进行了优化.研究结果表明,金粉用量为250.0 μg,轰击距离9cm的轰击参数最为理想,GUS瞬间表达率和平均蓝点数分别可达89.8%和33.7个.采用此轰击参数,将TaPR-1基因导入扬麦158幼胚,经3~5 mg/L Bialaphos筛选获得49株抗性再生植株,经PCR、Southern杂交分析,其中2株为阳性植株,转化率为0.11%.对转基因小麦植株进行苗期离体叶段和田间成株白粉病接种鉴定,初步结果表明,转基因植株对白粉病抗性与对照相比无明显差别.  相似文献   

7.
以幼胚来源的荔枝EC作为受体材料,采用基因枪轰击的方法将外源gus基因转入荔枝,并对不同条件下的GUS瞬时表达进行了详细研究。结果表明:在其它参数不变的条件下,轰击距离、真空度、可裂膜片压力、金粉用量、质粒DNA用量等理化参数显著影响GUS瞬时表达;渗透处理可显著促进GUS瞬时表达。因此,荔枝EC基因枪转化的适宜条件为在轰击距离6 cm、真空度84.66 k Pa、可裂膜片压力7 584.23 k Pa、轰击1次的情况下,每次轰击用1μg质粒DNA包裹600μg金粉对事先用0.25 mol/L前处理4 h的荔枝EC轰击,并在轰击后渗透处理20 h。另外,经50 mg/L潮霉素筛选得到荔枝抗性愈伤组织,并通过体胚发生途径获再生植株,且GUS染色显示获得了稳定表达GUS蛋白的细胞系和植株。  相似文献   

8.
口蹄疫病毒结构蛋白P1基因转化玉米的初步研究   总被引:4,自引:0,他引:4  
余云舟  王罡  金宁一  季静  王萍 《玉米科学》2004,12(3):022-025
实验在构建口蹄疫病毒(Foot and Mouth Disease Virus,FMDV)结构蛋白P1基因植物双元表达载体(pBI131SP1、pBI21P1和pBI121AP1)的基础上,以玉米自交系8902、340、4112的Ⅱ型胚性愈伤组织为受体,用农杆菌介导法和基因枪轰击法转化玉米,获得31株抗性再生植株.GUS染色证明外源目的基因在玉米细胞和组织中得到了表达.PCR检测、Southem blotting鉴定证实目的基因P1已整合到再生植株的基因组中,共获得了13株转基因玉米植株,首次进行了FMDV抗原基因转化玉米的研究。  相似文献   

9.
为验证小麦病程相关蛋白基因TαPR-1的功能,利用gus基因瞬间表达技术对基因枪法转化小麦幼胚的两个重要参数金粉用量和轰击距离进行了优化。研究结果表明,金粉用量为250.0μg.轰击距离9cm的轰击参数最为理想,GUS瞬间表达率和平均蓝点数分别可达89.8%和33.7个。采用此轰击参数.将TαPR-1基因导入扬麦158幼胚.经3~5mg/L Bialaphos筛选获得49株抗性再生植株.经PCR、Southern杂交分析.其中2株为阳性植株.转化率为0.11%。对转基因小麦植株进行苗期离体叶段和田间成株白粉病接种鉴定.初步结果表明,转基因植株对白粉病抗性与对照相比无明显差别。  相似文献   

10.
洁净DNA转化是基因枪介导外源基因表达框导入植物的转化技术,能从根本上消除载体框架序列对转基因植株的不利影响。2mG2-epsps基因是具有重要育种价值的草甘膦除草剂抗性基因。以日本晴为材料,研究了草甘膦对水稻愈伤组织生长及分化的影响,采用洁净DNA转化技术将2mG2-epsps基因表达框导入水稻。结果表明:1)草甘膦对水稻愈伤组织的生长及分化有明显的抑制作用,当草甘膦浓度为2mmol/L时,愈伤组织绿苗分化率为18.97%,较对照71.67%显著降低。2)基因枪介导2mG2-epsps基因表达框转化水稻时,经草甘膦筛选获得抗性愈伤后,在植株再生培养基中去除筛选剂利于抗性愈伤的分化,转化率为17.20%。经Southern杂交分析,2mG2-epsps基因表达框均以单拷贝整合到受体基因组。52.17%(12/23)转基因株系可耐受12~50mmol/L的草甘膦。  相似文献   

11.
12.
Summary

The efficiency of N fertilizers is usually poor; often less than 50% of the applied N is taken up by the crop. This review focuses on various N fertilizers with respect to the significance of different N loss pathways, namely (i) ammonia volatilization, (ii) dinitrogen and nitrogen oxide emissions, and (iii) nitrate leaching. Further, the significance of biological N immobilization, ammonium fixation and, finally, the impact of nitrate vs. ammonium uptake on crop yield are also discussed. The reviewed literature shows that N fertilizers may differ markedly in their susceptibility to losses. There is, however, considerable scope to improve N efficiency of each N source by proper N management practices.  相似文献   

13.
Summary The effects of the leaves of five plant species, one from each of the generaAmbrosia, Anemone, Eupatorium, Eucalyptus andLantana, on potato tuber moth were investigated under indigenous storage conditions at the Central Potato Research Station, Shillong (1800 m above sea level). Their action was compared with that of a biological insecticide (spores ofBacillus thuringiensis), a chemical insecticide (carbaryl), and an untreated control. The data collected after six months storage on tuber damage, sprout damage and the rotting indicated that the leaves ofLantana aculeata provided most protection to the tubers, reducing damage from over 70% in the check to below 5%, and sprout damage from over 45% to below 3%. Next best wasEucalyptus globulus followed byB. thuringiensis. They may be used on tubers stored for table use or for seed as they had no adverse effect on germination or on the yield of a subsequent crop.  相似文献   

14.
15.
Teas of plant origin traditionally consumed by the Mountain Pima of Chihuahua, Mexico, were analyzed for mineral nutritional content. Fe, Cu, Zn, Ca, and Mg composition was determined for native teas made from shoots ofTagetes lucida, T. filifolia, Elytraria imbricata, andHolodiscus dumosus, and from root xylem ofCeanothus depressus andPhaseolus ritensis. Native uses of these teas are also described.  相似文献   

16.
Novel food and non-food uses for sorghum and millets   总被引:4,自引:3,他引:4  
Sorghum and millets have considerable potential in foods and beverages. As they are gluten-free they are suitable for coeliacs. Sorghum is also a potentially important source of nutraceuticals such antioxidant phenolics and cholesterol-lowering waxes. Cakes, cookies, pasta, a parboiled rice-like product and snack foods have been successfully produced from sorghum and, in some cases, millets. Wheat-free sorghum or millet bread remains the main challenge. Additives such as native and pre-gelatinised starches, hydrocolloids, fat, egg and rye pentosans improve bread quality. However, specific volumes are lower than those for wheat bread or gluten-free breads based on pure starches, and in many cases, breads tend to stale faster. Lager and stout beers with sorghum are brewed commercially. Sorghum's high-starch gelatinisation temperature and low beta-amylase activity remain problems with regard to complete substitution of barley malt with sorghum malt . The role of the sorghum endosperm matrix protein and cell wall components in limiting extract is a research focus. Brewing with millets is still at an experimental stage. Sorghum could be important for bioethanol and other bio-industrial products. Bioethanol research has focused on improving the economics of the process through cultivar selection, method development for low-quality grain and pre-processing to recover valuable by-products. Potential by-products such as the kafirin prolamin proteins and the pericarp wax have potential as bioplastic films and coatings for foods, primarily due to their hydrophobicity.  相似文献   

17.
Summary The in vitro antimicrobial activity of extracts from accessions ofSolanum commersonii Dun. collected in the south of Uruguay was investigated against five microorganisms including the pathogenRalstonia solanacearum. A total of 30 extracts corresponding to organic and aqueous extracts were studied. Interestingly, most of the positive results for growth inhibition were againstR. solanacearum. The extracts were also analyzed for the presence of glycoalkaloids and lectins. Six of the ten aqueous extracts showed lectin presence and a wide variation in the type and amounts of glycoalkaloids, was found. Results indicate that there is no clear relationship between the antimicrobial activity against the five microorganisms screened and the presence or amounts of lectins and glycoalkaloids, traditionally regarded as possible antimicrobial metabolites in theSolanum genus, which indicates the presence of as yet unidentified antimicrobial compounds.  相似文献   

18.
Summary Clones derived from thirty-one different accessions (nineteen of Argentine origin) belonging to eightSolanum species were screened for resistance to infection by potato virus X strain cp (PVX cp) by mechanical inoculation of plantlets that had been micropropagated in vitro. Estimates of PVX multiplication obtained by enzyme linked immunosorbent assay and slot blot nucleic acid hybridization allowed the identification of resistant clones derived from five accessions belonging toS. commersonii S. oplocense, S. sparsipilum andS. tuberosum andigena. Resistant genotypes supported PVX concentrations 5 to 15 times smaller than did the susceptible control cultivar Spunta. Graft inoculation test confirmed the presence of extreme resistance similar to that conferred by the ‘immunity’ gene X1 (also called RXact).  相似文献   

19.
The loss of density and elasticity, the appearance of wrinkles and hyperpigmentation are among the first noticeable signs of skin aging. Beyond UV radiation and oxidative stress, matrix metalloproteinases (MMPs) assume a preponderant role in the process, since their deregulation results in the degradation of most extracellular matrix components. In this survey, four cyanobacteria strains were explored for their capacity to produce secondary metabolites with biotechnological potential for use in anti-aging formulations. Leptolyngbya boryana LEGE 15486 and Cephalothrix lacustris LEGE 15493 from freshwater ecosystems, and Leptolyngbya cf. ectocarpi LEGE 11479 and Nodosilinea nodulosa LEGE 06104 from marine habitats were sequentially extracted with acetone and water, and extracts were analyzed for their toxicity in cell lines with key roles in the skin context (HaCAT, 3T3L1, and hCMEC). The non-toxic extracts were chemically characterized in terms of proteins, carotenoids, phenols, and chlorophyll a, and their anti-aging potential was explored through their ability to scavenge the physiological free radical superoxide anion radical (O2•−), to reduce the activity of the MMPs elastase and hyaluronidase, to inhibit tyrosinase and thus avoid melanin production, and to block UV-B radiation (sun protection factor, SPF). Leptolyngbya species stood out for anti-aging purposes: L. boryana LEGE 15486 presented a remarkable SPF of 19 (at 200 µg/mL), being among the best species regarding O2•− scavenging, (IC50 = 99.50 µg/mL) and also being able to inhibit tyrosinase (IC25 = 784 µg/mL), proving to be promising against UV-induced skin-aging; L. ectocarpi LEGE 11479 was more efficient in inhibiting MMPs (hyaluronidase, IC50 = 863 µg/mL; elastase, IC50 = 391 µg/mL), thus being the choice to retard dermal density loss. Principal component analysis (PCA) of the data allowed the grouping of extracts into three groups, according to their chemical composition; the correlation of carotenoids and chlorophyll a with MMPs activity (p < 0.01), O2•− scavenging with phenolic compounds (p < 0.01), and phycocyanin and allophycocyanin with SPF, pointing to these compounds in particular as responsible for UV-B blockage. This original survey explores, for the first time, the biotechnological potential of these cyanobacteria strains in the field of skin aging, demonstrating the promising, innovative, and multifactorial nature of these microorganisms.  相似文献   

20.
The peer-reviewed marine pharmacology literature from 2009 to 2011 is presented in this review, following the format used in the 1998–2008 reviews of this series. The pharmacology of structurally-characterized compounds isolated from marine animals, algae, fungi and bacteria is discussed in a comprehensive manner. Antibacterial, antifungal, antiprotozoal, antituberculosis, and antiviral pharmacological activities were reported for 102 marine natural products. Additionally, 60 marine compounds were observed to affect the immune and nervous system as well as possess antidiabetic and anti-inflammatory effects. Finally, 68 marine metabolites were shown to interact with a variety of receptors and molecular targets, and thus will probably contribute to multiple pharmacological classes upon further mechanism of action studies. Marine pharmacology during 2009–2011 remained a global enterprise, with researchers from 35 countries, and the United States, contributing to the preclinical pharmacology of 262 marine compounds which are part of the preclinical pharmaceutical pipeline. Continued pharmacological research with marine natural products will contribute to enhance the marine pharmaceutical clinical pipeline, which in 2013 consisted of 17 marine natural products, analogs or derivatives targeting a limited number of disease categories.  相似文献   

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