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1.
应用重组PCR技术构建大豆脂肪氧化酶基因RNA干扰表达载体 总被引:3,自引:0,他引:3
RNA干扰是近年来生物技术领域新兴的研究热点之一。在作物的研究中,RNA干扰被认为在作物品质改良中具有重要的应用价值,并已在水稻、小麦、玉米等作物中得到成功应用,但在大豆的研究中尚鲜有报道。在研究植物RNA干扰的实验中,构建相应的RNA干扰表达载体是常用手段,但传统构建方法较费时费力。重组PCR是一种利用重叠延伸的方法进行高效和快速的体外基因片段拼接的PCR技术。研究将重组PCR技术应用于构建大豆脂肪氧化酶基因ihpRNA干扰表达载体,一方面探索一条更简便、快捷、适应范围更广泛的大豆基因RNA干扰表达载体构建方法,另一方面所构建的载体为进一步研究RNA干扰在大豆脂肪氧化酶改良中的应用奠定了基础。结果显示应用重组PCR技术构建大豆脂肪氧化酶基因RNA干扰表达载体是可行的,相比与常规方法此方法更简便、快捷、效率更高。 相似文献
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《中国油料作物学报》2015,(5)
为幼龄动物提供优质的大豆饲料,同时有效抑制α'亚基和β亚基基因表达量,根据dsRNAi原理,以α'亚基基因RNAi重组植物表达载体p CAMBIA3301-PFNZ-BADH为基础,构建以BADH安全标记为筛选标记的α'亚基和β亚基基因双价RNAi植物表达载体。用根癌农杆菌介导法转化大豆子叶节,并对再生植株进行分子生物学检测。结果表明:成功构建β-伴大豆球蛋白α'亚基和β亚基基因双价RNAi表达载体,并获得大豆转基因植株。T1转基因大豆植株经PCR检测得到11株阳性转化株,经Southern杂交检测证明构建的双价植物表达载体已经以1~2个拷贝整合到大豆基因组中,实时荧光定量PCR检测表明β-伴大豆球蛋白α'亚基和β亚基基因的表达被明显抑制。 相似文献
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HSSP基因植物表达载体构建及其对大豆的遗传转化 总被引:1,自引:0,他引:1
大豆中含硫氨基酸匮乏,限制了大豆的营养价值和商业价值。为了提高大豆的含硫氨基酸含量,利用人工设计合成的高含硫氨基酸贮藏蛋白基因(HSSP)转化大豆。以载体p TF101.1为骨架,构建植物表达载体p TFGS,利用农杆菌介导法转化大豆品种东农50,经PCR及试纸条检测,获得转基因大豆16株,基因的转化效率为1.94%。对转基因大豆株系GSDL5进行组织特异性分析,结果表明,HSSP基因在种子中超量表达,而在其它组织部位仅有微量表达。采用接头PCR方法对株系GSDL5基因组插入位点侧翼序列进行克隆,获得与大豆基因组序列匹配的436 bp侧翼序列,HSSP基因插入2号染色体基因的非编码区。经研究,获得了遗传背景明确的,仅在大豆种子中超量表达HSSP的转基因株系GSDL5。 相似文献
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利用冻融法将大豆11S球蛋白GY1基因RNA干扰表达载体转入农杆菌EH105中,以大豆"吉农28"为受体,通过农杆菌介导的大豆子叶节转化法导入大豆,获得T1代转基因苗12株,并对得到的转基因植株进行分子生物学鉴定。PCR和Southern杂交检测表明,RNAi植物表达载体p3301-Gy1已成功插入到转基因大豆植株的基因组DNA中;RT-PCR和SDS-PAGE检测结果表明RNA干扰在转录后水平发挥了作用,11S球蛋白表达含量降低;利用BUCHI N-500型近红外谷物分析仪对转化的大豆蛋白质和脂肪含量进行检测,结果转化植株的籽粒蛋白质含量(36.07%)平均降低1.43个百分点,脂肪含量(21.28%)平均提高0.76个百分点。因此,利用RNA干扰技术提高大豆脂肪含量是可行的。 相似文献
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大豆的真菌性病害已经成为了限制大豆产量增长及品质提高的主要原因,利用基因工程技术培育出具有广谱性抗病能力的大豆新品种,已成为目前提高大豆抗性的有效途径之一。本研究构建了含2种广谱抗病基因chi和hrp Zpsta的双价植物表达载体,利用农杆菌介导技术导入大豆中,获得了能够在转录水平上表达2种外源抗病性基因的转基因大豆。对经抗性筛选得到的阳性苗及后代植株进行PCR检测、Southern杂交、荧光定量PCR检测,共得到T0代阳性植株7株,T1代阳性植株27株。转化植株的基因组在整合外源基因时出现不同情况,chi-hrp Zpsta双价载体分别以单价和双价两种形式随机整合到受体大豆基因组中并且能稳定遗传,同时在转录水平上亦有不同。 相似文献
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Joseph E. Laferriere Charles W. Weber Edwin A. Kohlhepp 《Plant foods for human nutrition (Dordrecht, Netherlands)》1991,41(3):277-282
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
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Lakshman Lal 《Potato Research》1987,30(2):329-334
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
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《Journal of Crop Improvement》2013,27(2):81-114
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
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Tatiana G. Polotow Sandra C. Poppe Cristina V. Vardaris Douglas Ganini Maísa Guariroba Rita Mattei Elaine Hatanaka Maria F. Martins Eduardo F. Bondan Marcelo P. Barros 《Marine drugs》2015,13(10):6117-6137
Health authorities worldwide have consistently recommended the regular consumption of marine fishes and seafood to preserve memory, sustain cognitive functions, and prevent neurodegenerative processes in humans. Shrimp, crabs, lobster, and salmon are of particular interest in the human diet due to their substantial provision of omega-3 fatty acids (n-3/PUFAs) and the antioxidant carotenoid astaxanthin (ASTA). However, the optimal ratio between these nutraceuticals in natural sources is apparently the key factor for maximum protection against most neuro-motor disorders. Therefore, we aimed here to investigate the effects of a long-term supplementation with (n-3)/PUFAs-rich fish oil, ASTA-rich algal biomass, the combination of them, or krill oil (a natural combination of both nutrients) on baseline redox balance and neuro-inflammation indexes in cerebellum and motor cortex of Wistar rats. Significant changes in redox metabolism were only observed upon ASTA supplementation, which reinforce its antioxidant properties with a putative mitochondrial-centered action in rat brain. Krill oil imposed mild astrocyte activation in motor cortex of Wistar rats, although no redox or inflammatory index was concomitantly altered. In summary, there is no experimental evidence that krill oil, fish oil, oralgal biomass (minor variation), drastically change the baseline oxidative conditions or the neuro-inflammatory scenario in neuromotor-associated rat brain regions. 相似文献
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Youna Hemery Ulla Holopainen Anna-Maija Lampi Pekka Lehtinen Tanja Nurmi Vieno Piironen Minnamari Edelmann Xavier Rouau 《Journal of Cereal Science》2011
Wheat bran is a composite material made of several layers, such as pericarp, testa and aleurone. It could be fractionated into purified fractions, which might either be used as food ingredients, or serve as a starting material for extraction of bioactive compounds. The aim of this work was to evaluate the potential of using electrostatic separation as a way to obtain purified fractions from wheat bran. Ultrafine-ground bran obtained either by cryogenic grinding or by grinding at ambient temperature was used as starting material. The ultrafine bran was then charged by tribo-electrification and introduced in a chamber containing two high voltage electrodes, where bran particles were separated depending on their acquired charge, allowing positively and negatively charged fractions to be collected separately. The particle size distribution, microstructure and biochemical composition of the obtained fractions were studied. The charge of the particles was influenced by their biochemical composition: particles rich in highly branched and cross-linked arabinoxylans (pericarp) were separated from particles rich in β-glucan, ferulic acid and para-coumaric acid (aleurone cell walls). The testa and the intracellular compounds from aleurone were not highly charged, neither positively nor negatively. The most positively charged fraction represented 34% of the initial bran, and contained 62% of the ferulic acid present in the initial bran. The yield of the separation process was good (5.4% loss), and could be further increased. 相似文献
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M. I. Siri P. Villanueva M. J. Pianzzola L. Franco Fraguas G. Galván M. Acosta F. Ferreira 《Potato Research》2004,47(3-4):127-138
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
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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
20.
Tovar LR Olivos M Gutierrez ME 《Plant foods for human nutrition (Dordrecht, Netherlands)》2008,63(4):189-194
Pulque is made by fermenting the agave sap or aguamiel of Agave atrovirens with a whole array of microorganisms present in the environment including several lactic acid bacteria and yeasts such as
Saccharomyces cerevisiae. Ascorbic acid was determined in pulque and aguamiel, respectively. Phytase activity in lees, liquid and freeze-dried pulque was assayed by measuring the appearance of phosphate from phytate by a colorimetric method likewise phosphate from phytate
present in fresh corn tortilla was measured after in vitro incubation with pulque. Iron, zinc, calcium, magnesium and selenium contents were measured in pulque and corn tortilla as well as in nixtamalized corn flour (NCF), the latter is used to make instant tortilla, since corn provides
most of the energy as well as most of the phytate in the Mexican rural diet. Pulque showed phytase activity but much less ascorbic acid and iron than previously reported; additionally, phytase in pulque hydrolyzed most of phytate’s corn tortilla. Lees, which is mostly made of pulque’s microbiota, significantly accumulated iron and zinc but no selenium. NCF was fortified with iron by the manufacturers but
poorly blended. There were significant differences on selenium content between tortillas samples, apparently some soils in
central Mexico are selenium deficient. Moderate pulque intake appears to increase the bioavailability of iron and zinc bound by phytate in corn. 相似文献