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1.
红麻基因组总DNA的提取纯度,关系到新型分子标记ISSR、SRAP的PCR扩增效果。经反复实验,对CTAB法提取红麻总DNA技术进行了改进,即采集足量的红麻嫩叶、提取时加入β-巯基乙醇、提取过程中使用2.5%CTAB、最后用预冷的无水乙醇沉淀DNA。OD260/OD280比值检测结果为平均1.824,而且DNA得率也较高,证明用此法提取的DNA能完全满足ISSR和SRAP分析的要求和获得较理想的效果,本文还讨论了高质量DNA提取应注意的有关技术环节。  相似文献   

2.
剑麻核酸的高效提取及应用   总被引:1,自引:0,他引:1  
针对剑麻组织中多糖、多酚和次生物质含量高的特点,对其总RNA和基因组DNA提取进行研究。比较了改良SDS法、改良CTAB法提取总RNA和SDS法、CTAB法提取基因组DNA的效果,结果表明:改良CTAB法提取总RNA效果较佳,所得RNA OD260/OD280高于1.8,OD260/OD230大于2.0,而CTAB法提取基因组DNA纯度和完整性较好。基于RNA和DNA水平,分别对剑麻的苯丙氨酸解氨酶基因(PAL)RT—PCR和RAPD反应体系进行优化,均得到理想条带。此方法提取剑麻的核酸可运用于后续分子生物学研究,为其它材料核酸的分离提供参考依据。  相似文献   

3.
红麻处于光合高峰期的成熟叶片中的次生物质含量很高,给DNA的提取造成很大的困难。CTAB法能有效去除多糖杂质,但普通CTAB法用于红麻的DNA提取效果很不理想,为使CTAB法能更好地用于红麻的DNA提取,我们对此方法进行了改进,通过前处理除去大部分的杂质,减小了后续操作的难度。为了验证前处理是否会损失DNA,研究了液氮研磨对材料细胞的破碎程度问题,还讨论了DNA提取中的其它一些相关问题。  相似文献   

4.
改良CTAB法用于提取红麻成熟叶片高质量DNA的研究   总被引:6,自引:0,他引:6  
红麻处于光合高峰期的成熟叶片中的次生物质含量很高,给DNA的提取造成很大的困难.CTAB法能有效去除多糖杂质,但普通CTAB法用于红麻的DNA提取效果很不理想,为使CTAB法能更好地用于红麻的DNA提取,我们对此方法进行了改进,通过前处理除去大部分的杂质,减小了后续操作的难度.为了验证前处理是否会损失DNA,研究了液氮研磨对材料细胞的破碎程度问题,还讨论了DNA提取中的其它一些相关问题.  相似文献   

5.
CTAB法提取花生总DNA在SSR和SRAP中的扩增效果   总被引:9,自引:0,他引:9  
本试验采用相对简单快捷的CTAB法提取花生总DNA,并应用于SSR和SRAP扩增分析,结果表明,所提取花生DNA中RNA含量高,但完全可以满足SSR标记和SRAP标记分析要求.  相似文献   

6.
蓖麻DNA的提取及SRAP反应体系的建立   总被引:6,自引:2,他引:4  
为建立蓖麻快速有效的DNA提取方法和SRAP标记的技术体系,本研究采用SDS法和CTAB法对蓖麻不同叶片(新鲜嫩叶、老叶和陈旧的嫩叶、老叶)的基因组DNA进行提取,并以DNA模板用量、Mg2+浓度、引物浓度和Taq酶用量4个因素的不同水平配制了10个反应体系对蓖麻SRAP反应体系进行优化。结果表明,SDS法和CTAB法均可获得较高质量的DNA,多数样品的A260/A280值介于1.7~1.9之间,但以CTAB法更为省工省时,不同叶片DNA的提取效果,以嫩叶效果最佳;设计的10个反应体系中,以反应体系A(总体积为15µL,40ng DNA模板,1.5mmol/L Mg2+浓度,0.2mmol/L dNTPs,0.3µmol/L引物浓度和1Unit Taq酶)最为经济适用,将该体系用于蓖麻的SRAP扩增能获得稳定、清晰的多态性条带。本实验为蓖麻的遗传多样性研究及目标性状的分析定位提供了技术支撑。  相似文献   

7.
锥栗总RNA的提取质量是后期锥栗空棚分子机理研究的关键。以锥栗花序为材料,采用2种植物RNA快速提取试剂盒、改良Trizol法和改良CTAB法提取锥栗总RNA,并采用多种方法检测提取出的总RNA的质量。结果表明:采用改良CTAB法获得的总RNA可明显看到28S和18S条带,且亮度比约为2∶1,DNA基本无残留,点样孔无杂质,OD260/OD280均接近2,而OD260/OD230均超过2,经逆转录可作为模板,获得LFY基因。改良后CTAB法最合适,所得的总RNA产量高、比较纯净、完整,可用于后续的逆转录并进行基因片段的扩增。  相似文献   

8.
红麻SRAP、ISSR遗传连锁图构建的初步研究   总被引:10,自引:0,他引:10  
应用SRAP、ISSR标记构建红麻分子遗传连锁图,以半野生种Ga42(源自加纳)和栽培种阿联红麻(源自埃及)杂交产生的203株F2代分离群体作为作图群体。筛选出多态性好的27对SRAP引物组合和5个ISSR引物,对F2作图群体进行PCR扩增,共产生108个多态性条带,平均每个引物组合产生3.4个多态性条带。最多的可产生8条多态性条带。应用MAPMAKER/EXP3.0软件对108个多态性条带进行遗传连锁分析,被分为16个连锁群(LOD≥3.0),初步构建了首张红麻遗传连锁图谱。该图谱总长为1336.6cM,平均两个标记位点间距为17.82cM。本文还讨论了图谱构建存在的偏分离有关问题与对策。可为进一步构建较高密度、分布均匀的遗传图谱及基因定位奠定了良好的基础。  相似文献   

9.
分别采用TRIzol法和改良的CTAB法提取阔叶薰衣草叶片的RNA,并用紫外光谱和琼脂糖凝胶电泳对其提取效果进行鉴定。实验结果表明:TRIzol法提取的总RNA浓度高,但杂质较多且有部分降解、稳定性差;改良的CTAB法提取得到的总RNA的纯度较高,OD260/OD280值为1.909,效果稳定,有良好的可重复性。因此,改良的CTAB法是一种较为简便、经济的提取阔叶薰衣草叶片总RNA的方法。  相似文献   

10.
为了获取高质量的油梨果肉DNA,以油梨果肉为试材,比较改良SDS法、改良CTAB法和试剂盒法对油梨果肉DNA的提取效果。结果表明:上述3种方法均可从油梨果肉中提取DNA,但改良SDS法与改良CTAB法所获得的DNA浓度都在600 μg/mL以上,纯度(OD260nm/OD280nm)均在1.8左右,纯度高且完整性较好,可用于未来分子标记分析。  相似文献   

11.
Sorghum and millet phenols and antioxidants   总被引:5,自引:2,他引:5  
Sorghum is a good source of phenolic compounds with a variety of genetically dependent types and levels including phenolic acids, flavonoids, and condensed tannins. Most sorghums do not contain condensed tannins, but all contain phenolic acids. Pigmented sorghums contain unique anthocyanins that could be potential food colorants. Some sorghums have a prominent pigmented testa that contains condensed tannins composed of flavan-3-ols with variable length. Flavan-3-ols of up to 8–10 units have been separated and quantitatively analyzed. These tannin sorghums are excellent antioxidants, which slow hydrolysis in foods, produce naturally dark-colored products and increase the dietary fiber levels of food products. Sorghums have high concentration of 3-deoxyanthocyanins (i.e. luteolinidin and apigenidin) that give stable pigments at high pH. Pigmented and tannin sorghum varieties have high antioxidant levels that are comparable to fruits and vegetables. Finger millet has tannins in some varieties that contain a red testa. There are limited data on the phenolic compounds in millets; only phenolic acids and flavones have been identified.  相似文献   

12.
Summary Between 1993 and 1998 205 different potato cultivars and 1220 accessions/genotypes of wild and cultivated potato species from the IPK Genebank Gatersleben were evaluated. Parameters interesting for starch isolation and especially for the use of starch were determined. Altogether, there was a higher variability in wild potato species than in cultivated potatoes for all characteristics investigated: dry matter content, starch content, protein content, amylose content and mean particle diameter of starch granules.  相似文献   

13.
种子加工、检验理论与技术现状及思考   总被引:3,自引:0,他引:3  
种子加工及检验是种子向市场流通的关键,是商业化育种中赋予种子商品属性不可或缺的环节。种子加工、检验理论与技术的研究是完善种业产业链、实现规模化商业育种、夯实“育繁推一体化”种业科学的重要一环。中国的种业科学技术体系正在形成与完善之中,受制于行业发展水平,种子加工及检验理论与技术相对薄弱。本文回顾了种子加工及检验理论与技术研究发展历程,提出了完善中国种业科学发展的策略与建议。围绕种业发展的需要,在做好品种优质化繁育的基础上,分析种子加工及检验理论与技术的新方向和新需求,加强种业应用性研究,建立先进的种子质量检验体系,研制适合现代种业发展需求的种子加工设备,打造种子加工产业标准化生产体系,完善种业科学技术学科建设,促进种业产业持续健康发展。  相似文献   

14.
Seed processing and testing hold the key to the market circulation of seeds, and are an indispensable link in commercial breeding to endow seeds with commodity attributes. The research of seed processing and testing theory and technology is an important link in improving the seed industry chain, realizing large-scale commercial breeding and consolidating the seed industry science of ‘integration of breeding, propagation and promotion’. The scientific and technological system of seed industry in China is being formed and perfected, which is subject to the development level of the industry. The theory and technology of seed processing and testing are relatively weak. We reviewed the development of seed processing and testing theory and technology, and put forward strategies and suggestions to improve the sound development of China’s seed industry. In order to meet the needs of seed industry development and on the basis of high quality breeding of varieties, the new directions and demands of seed processing and testing theory and technology were analyzed. We will work to strengthen the applied research of seed industry, establish advanced seed quality inspection system, improve seed processing equipment suitable for the development needs of modern seed industry, establish standardized production system of seed processing industry, develop the scientific and technological disciplines of seed industry, and promote the sustainable and healthy development of seed industry.  相似文献   

15.
Summary

Arbuscular mycorrhizae were inoculated into phosphorus-deficient soil fertilized with either organic or chemical fertilizer with cucumber (Cucumis sativus L.) as the first crop and lettuce (Lactu-ca sativa L.) as the second crop but without additional fertilization and AM inoculation. AM increased dry matter and fruit yield of cucumber significantly in the unfertilized, organic-fertilized and P-deficient plants compared with the fully chemical-fertilized plants. AM inoculation increased the available phosphorus in plant and soil by around 30% for all treatments except for those chemically-fertilized. The rate of AM infection did not differ significantly among the fertilization treatments, but the infection intensity was higher in unfertilized, organic-fertilized and phosphorus-deficient treatments than chemical-fertilized treatment. The residual effects of AM-inoculated to cucumber were evident for lettuce in all pre-treatments that were unfertilized and un-inoculated for the second cropping. Without P-fertilization, neither crop could grow optimally even when the soil was inoculated with AM, suggesting that AM could not serve as a substitute for phosphorus fertilizer. However, the other beneficial effects of AM on crop growth and yield could not be fulfilled with phosphorus fertilizer.  相似文献   

16.
种子加工及检验是种子向市场流通的关键,是商业化育种中赋予种子商品属性不可或缺的环节。种子加工、检验理论与技术的研究是完善种业产业链、实现规模化商业育种、夯实"育繁推一体化"种业科学的重要一环。中国的种业科学技术体系正在形成与完善之中,受制于行业发展水平,种子加工及检验理论与技术相对薄弱。本文回顾了种子加工及检验理论与技术研究发展历程,提出了完善中国种业科学发展的策略与建议。围绕种业发展的需要,在做好品种优质化繁育的基础上,分析种子加工及检验理论与技术的新方向和新需求,加强种业应用性研究,建立先进的种子质量检验体系,研制适合现代种业发展需求的种子加工设备,打造种子加工产业标准化生产体系,完善种业科学技术学科建设,促进种业产业持续健康发展。  相似文献   

17.
通过比较分析芸薹属多倍体复合种与其祖先二倍体基本种在花柱、柱头大小、角果长度、宽度、种子直径 及种子干重变化上的异同,发现多倍体复合种在这些发育性状上表现出处于两个祖先二倍体亲本之间,或只偏向 于其中一个祖先亲本。同一个多倍体品种在不同发育性状上具有不同的表现,即在某一发育性状上偏向于其祖先 亲本之一,而在另一发育性状上偏向其另一祖先亲本。  相似文献   

18.
Pea (Pisum sativum L.) and oat (Avena sativa L.) were grown as sole and mixed crops in various densities under two different tillage systems on a loess soil near Göttingen/Germany in a 2-year field experiment (2002/2003). In the conventional tillage system a mouldboard plough (CT) was used and in the minimum tillage system a rotary harrow (MT) was employed. The effect of crop density and tillage system on the grain dry matter and grain N yields, N2 fixation and soil N uptake were determined to address the following questions: (i) which mixture compositions exhibit the highest grain yields compared to the sole crops, (ii) which mixture compositions also fix a high level of N2 and leave low levels of residual inorganic soil N after harvest, and (iii) whether the intercrop advantage is influenced by the tillage system. For (i) the result in 2002 showed that the highest grain yields of both sole cropped pea and oat and intercropped pea and oat were achieved at the highest densities. In 2003, when the inorganic soil N content was higher and weather conditions were warmer and drier, grain yields were significantly higher than in 2002, but sole as well as intercropped pea and oat gave their highest grain yields at lower densities. For both years and tillage systems, the highest intercrop advantages were achieved in mixtures with densities above the optimal sole crop densities. The result for (ii) was that a distinctly higher proportion of nitrogen was derived from the atmosphere (Ndfa) by intercropped pea than by sole cropped pea. However, the uptake of soil N by intercropped pea and oat was not reduced in comparison with that of sole cropped oat as the decrease in the uptake of N from the soil by oat at lower oat densities in the mixture was compensated for by the soil N uptake of pea. Additionally, the Nmin-N content of the soil following the mixtures and sole cropped oat did not differ, especially in the deeper soil layers because oat in mixture was forced to take up more soil N from deeper layers. Therefore, the risk of soil N losses through leaching after mixtures was lower compared to sole cropped pea. The tillage system (iii) had no significant influence on grain yield and soil N uptake, but N2 fixation and the competitive ability of intercropped pea were higher under CT than with MT. An additional result was that intercropping led to a significantly increased grain N content of both pea and oat compared to the sole crops. The increase in grain N content from sole to intercrop was from 3.30 to 3.42% for pea and from 1.73 to 1.96% for oat as a mean for both years and tillage systems. The present study confirms that growing pea and oat as intercrops highlights potential economic and environmental benefits which still need to be understood in more detail in order to exploit intercropping to a greater extent.  相似文献   

19.
对云南保山、德宏、普洱等地咖啡病虫害开展调查研究,结果表明:咖啡锈病(Hemileia vastatrix Berk et Br.)、炭疽病(Colletotrichum coffeanum Noack)、褐斑病(Cercospora coffeicola Berk. et Cooke)、幼苗立枯病(Rhizoctonia solani Kuhn.)、旋皮天牛(Acalolepta cervina Hope)、灭字虎天牛(Xylo  相似文献   

20.
Soil sulphur deficiency, which is increasingly prevalent in Western Europe, lowers wheat yields, and also affects the gluten quality of the flour. Differences in S availability may change the proportion of S-poor to S-rich gliadins and glutenin subunits. This may cause unpredictable and unwanted variations in wheat quality. The combined effects of nitrogen (N) and sulphur (S) fertilisers and split application of S and N on wheat gluten quality and composition were investigated. The results revealed effects of S fertilisation on dough quality. At high N fertilisation levels significant responses to S fertilisation were found which emphasised the need for precision application of S in intensive wheat production systems. Protein fractionation by SE-FPLC showed that quality differences were associated with changing proportions of high Mr polymeric proteins. Changes in protein composition of salt soluble proteins were also confirmed by proteomics. Glyceraldehyde-3-phosphate dehydrogenase and one of the serpin protein spots increased at high N, combined with the lower S level. The enzymes also increased in samples with increased S fertilisation combined with low N, but was not changed at higher N levels. Furthermore, at high N the serpin protein spot, and also a 27 K protein and one unidentified protein spot decreased with increasing S.  相似文献   

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