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1.
为了提取高质量的割手密叶绿体DNA(chloroplast DNA,cp DNA),本研究立足于高效、省时、低耗的原则,以割手密幼叶为材料,采用改进的高盐-低p H法经差速离心分离叶绿体、DNaseⅠ处理、10%SDS和蛋白酶K裂解叶绿体、DNA萃取、乙醇沉淀等一系列操作。所提取的割手密cp DNA经显微观察、紫外吸收光度测定、琼脂糖凝胶电泳、叶绿体基因特异引物PCR扩增等进行检测,结果表明该提取方法简单高效,A260/A280的比值介于1.9~2.0,cp DNA纯度高,浓度为2.90μg/μL,适用于基因扩增等分子操作。研究结果为进一步开展割手密叶绿体基因功能、系统发育、遗传多样性等方面的研究奠定了基础。  相似文献   

2.
提取高质量的线粒体DNA,对植物细胞质雄性不育的研究起着关键性的作用。以胡萝卜叶片为材料,通过高盐差速离心和蔗糖沉淀差速离心2种方法提取线粒体DNA。通过改变分离和沉淀线粒体离心力大小、使用DNaseⅠ处理得到了无核DNA污染的线粒体。用SDS和蛋白酶K裂解线粒体经酚/氯仿/异戊醇抽提除去蛋白,并用RNaseA消化从而得到单纯线粒体DNA。还特别设计了叶绿体特,异性引物检测线粒体DNA纯度。结果表明:利用优化后的蔗糖沉淀差速离心法提取胡萝卜线粒体DNA,不仅操作简单,而且所得线粒体DNA纯度高、产量高,每克叶片能够得到34.46μg线粒体DNA。  相似文献   

3.
胡萝卜叶片线粒体DNA提取方法研究   总被引:1,自引:0,他引:1  
本实验以胡萝卜叶片为材料,通过高盐差速离心和蔗糖沉淀差速离心两种方法提取线粒体DNA。通过改变分离和沉淀线粒体离心力大小、使用DNaseⅠ处理得到了无核DNA污染的线粒体。用SDS和蛋白酶K 裂解线粒体,经酚/氯仿/异戊醇抽提除去蛋白,并用RNase A 消化从而得到单纯线粒体DNA。本实验还特别设计了叶绿体特异性引物检测线粒体DNA纯度。结果表明:利用优化后的蔗糖沉淀差速离心法提取胡萝卜线粒体DNA,不仅操作简单,而且所得线粒体DNA纯度高、产量高,每克叶片能够得到34.46 μg 线粒体DNA。  相似文献   

4.
本研究以胡萝卜根为材料,通过高盐差速离心和蔗糖沉淀差速离心两种方法提取线粒体DNA。通过改变缓冲液、使用DNaseⅠ处理得到了无核DNA污染的线粒体。用SDS和蛋白酶K裂解线粒体,经酚:氯仿:异戊醇抽提除去蛋白,并用RNaseA消化从而得到单纯线粒体DNA。本实验还特别设计了叶绿体特异性引物检测线粒体DNA纯度。结果表明:利用优化后的蔗糖沉淀差速离心法提取胡萝卜线粒体DNA,所得线粒体DNA纯度高、产量高,每克肉质根能够得到27.60μg线粒体DNA。  相似文献   

5.
梭梭属2种植物线粒体基因组的提取   总被引:1,自引:1,他引:0  
为了分离提取高质量梭梭(Haloxylon ammodendron)、白梭梭(Haloxylon persicum)线粒体DNA,以藜科梭梭属植物梭梭、白梭梭黄化苗为材料,通过采用蔗糖密度梯度离心和差速离心相结合的方法,使用DNaseⅠ处理得到了无核DNA污染的线粒体,用3% CTAB裂解线粒体,经氯仿/异戊醇抽提除去蛋白,从而得到单纯线粒体DNA。结果表明,所提取的线粒体DNA经电泳检测条带清晰,无拖尾现象,且A260/A280≈1.8,说明纯度较高、质量好,可满足后续有关线粒体DNA的相关实验。梭梭、白梭梭高纯度线粒体DNA提取技术的建立,为开展梭梭、白梭梭线粒体基因组研究奠定了基础。  相似文献   

6.
甘蔗野生种割手密抗逆性强,在甘蔗抗逆育种中具有较好的应用前景;割手密SSR分子标记技术体系的建立及优化可为割手密指纹图谱绘制、种质鉴定、遗传多样性及亲缘关系分析提供技术支持。本研究以21份割手密材料用改良CTAB法提取DNA,采用降落PCR和正交试验设计对扩增反应体系主要成分的浓度(DNA模板,引物,Mg2+,d NTPs,Taq聚合酶)进行了优化。结果表明建立的割手密Touchdown SSR-PCR分子标记技术体系操作简便,扩增条带清晰且多态性丰富,有较强的稳定性和可重复性,可为割手密资源的相关研究提供技术支持。  相似文献   

7.
黏类小麦细胞质雄性不育系mtDNA消减文库的构建   总被引:1,自引:0,他引:1  
本研究利用抑制性消减杂交技术,构建了黏类小麦细胞质雄性不育线粒体DNA的消减文库。分别提取相同细胞质背景下的不育和可育等基因系线粒体基因组DNA(mtDNA),用RsaⅠ酶切成大小不等的片段,并各自与不同的接头连接,连续经过两次消减杂交和两次PCR扩增,将PCR产物与克隆载体连接,转化为大肠杆菌感受态细胞DH5α,经蓝白斑筛选后,再用PCR方法插入片段筛选出阳性重组质粒,构建差异DNA消减文库。在构建的不育mtDNA文库中,将SSH文库的全部扩增产物与SSH文库中的单条扩增条带分别进行胶回收和克隆转化,分别挑取54个和6个阳性克隆进行测序分析和相关功能初步比对。结果表明,不育mtDNA的SSH文库差减杂交效率较高,质量较好;回收单条扩增条带分别进行克隆测序的结果准确率较高;对测序序列进行BLASTx比对及功能注释分析发现,约90%的序列来源于线粒体基因nad1和nad5的非编码区。  相似文献   

8.
芝麻高纯度线粒体DNA提取技术研究   总被引:3,自引:0,他引:3  
以2个芝麻品种黄化苗为试验材料,通过比较植物线粒体DNA(mtDNA)提取过程中的关键影响因素DNase处理时间,建立了一种经济、快速、高效的芝麻mtDNA提取方法.该方法分别通过匀浆化处理、2 400 g离心15min结合12 000 g离心12 min的差速离心方法分离获得粗制线粒体,再经过DNase酶切处理1.5...  相似文献   

9.
选择一种快速、简便、经济的从马血液中提取基因组DNA的方法。分别采用了常规酚—氯仿抽提法、简易酚—氯仿快速抽提法和V—gene Biotechnology Limited试剂盒三种方法从马血中提取DNA,并利用琼脂糖凝胶电泳技术和紫外分光光度计,比较各方法提出DNA的浓度和纯度,且利用PCR扩增技术检测DNA的实用性。结果表明,常规酚—氯仿抽提法适合从马血中大量提取DNA。  相似文献   

10.
为解决现有的橡胶树基因组DNA提取方法步骤繁琐、耗时长等问题,摸索出一种橡胶树叶片基因组DNA快速PCR扩增方法。通过比较分析快速提法与CTAB法,结果显示快速提取方法提取的基因组DNA完整性很高,同时也适用于RAPD分型分析。相比传统的基因组DNA提取、PCR扩增等其它分子生物学流程,快速PCR扩增方法操作简单、简单快速、成本低廉,尤其适用于大批量样品的基因分型分析。  相似文献   

11.
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   

12.
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   

13.
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   

14.
G. H. Kroon 《Euphytica》1994,76(1-2):125-125
Summary K x vadensis is a hybrid of K. blossfeldiana and K. marmorata obtained after doubling the number of chromosomes.  相似文献   

15.
[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   

16.
Summary Avoidance of rust fungi that was based on poor appressorium induction was previously found in Hordeum chilense. In the present study 95 accessions of Triticeae were screened for avoidance of Puccinia hordei. The percentage of appressorium formation per germinated spore ranged from 6 to 90%. On none of the 41 accessions of Aegilops, Agropyron, Elymus, Secale, Thinopyrum or Triticum studied was the rate of appressorium formation lower than 25%. Lower rates of appressorium formation were, however, found on accessions of wild barley species Hordeum brachyantherum, H. marinum, H. parodii and H. secalinum. Its implications in cereal breeding are discussed.  相似文献   

17.
P. A. York  R. Cook 《Euphytica》1989,43(1-2):135-141
Summary Reactions of 13 grasses to Meloidogyne naasi varied with species; ryegrasses, fescues and their hybrids were generally susceptible and cocksfoot and timothy resistant. Marked variation in host resistance levels occurred between genotypes within cultivars.Selection of single plants, followed by tests on replicate tillers, identified resistant and susceptible genotypes in both Italian and perennial ryegrass cultivars. Resistant plants had few nematode-induced galls and fewer females and eggs than susceptibles. There was more or less continuous variation, with many genotypes intermediate between extremes of resistance and susceptibility. Selected resistant and susceptible genotypes are of use in assessing variation in nematode populations and as controls for breeding and selection programmes.  相似文献   

18.
Lentil production is limited by lack of moisture and unfavorable temperatures throughout its distribution. Waterlogging and salinity are only locally important. Progress has been made in breeding for tolerance to drought through selection for an appropriate phenology and increased water use efficiency and in breeding for winter hardiness through selection for cold tolerance.The diseases rust, vascular wilt, and Ascochyta blight, caused by Uromyces viciae-fabae, Fusarium oxysporum f. sp. lentis, and Ascochyta fabae f. sp. lentis, respectively, are the key fungal pathogens of lentil. Cultivars with resistance to rust and Ascochyta blight have been released in several countries and resistant sources to vascular wilt are being exploited. Sources of resistance to several other fungal and viral diseases of regional importance are known. In contrast, although the pea leaf weevil (Sitona spp.) and the parasitic weed broomrape (Orobanche spp.), and to a lesser extent the cyst nematode (Heterodera ciceri), are significant yield reducers of lentil, no sources of resistance to these biotic stresses have been found. Directions for future research in lentil on both biotic and abiotic stresses are discussed.  相似文献   

19.
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   

20.
[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   

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