共查询到18条相似文献,搜索用时 62 毫秒
1.
2.
3.
4.
CTAB法高效提取苹果叶片DNA的研究 总被引:9,自引:0,他引:9
为研究CTAB法在正常条件和逆境条件下,从富含多酚和多糖等多种次生代谢物质的苹果树叶片中分离出高质量的基因组DNA,以抗旱能力较强的八棱海棠(Malus micromalusRehd. )和抗旱性较弱的平邑甜茶(Malus hupehensis pamp Reld.)幼苗为试验材料.用20%聚乙二醇(polyethylene glycol,PEG6000)模拟干旱,采取CTAB提取缓冲液,按一定比例加入PVP和β-巯基乙醇,用Tris平衡酚-氯仿-异戊醇除去蛋白质及杂质,在粗提时用异丙醇沉淀DNA,并用DNA洗液(10 mmol/L醋酸铵,75%乙醇)洗涤DNA,在纯化时用乙醇沉淀DNA,且没有加入RNase去除RNA的方法,分别提取2个品种正常和干旱胁迫的苹果属叶片总DNA,并对提取的DNA进行琼脂糖凝胶电泳和紫外分光光度计检测.结果表明:此方法可有效去除次生物质对DNA的干扰,得到高质量、特征带形清晰的大量DNA.并就提取过程中遇到的问题,提出了相应的解决办法. 相似文献
5.
6.
高质量枣基因组DNA提取方法 总被引:11,自引:2,他引:11
主要针对枣组织中多糖严重干扰基因组DNA提取质量的问题,比较了常规CTAB法、TE3D法和改良CTAB法3种方法的处理效果。结果表明:常规CTAB法提取DNA难以除去多糖类杂质;TE3D法提取的DNA多糖杂质少,但产率低,易降解,褐化严重;改良CTAB法提取DNA产率高,无明显降解,杂质少,D260nm/D280nm比值1.80左右,通过基因组DNA-AFLP指纹图谱分析,完全满足实验要求。并对改良CTAB法的关键步骤作了具体分析讨论。 相似文献
7.
食用菌DNA提取方法研究 总被引:16,自引:0,他引:16
DNA是最重要的遗传物质,因此,DNA的提取方法和技术是生物化学及分子生物学最基本的技术,也是生物工程最常用的技术,为了得到足够纯的食用菌DNA进行基因和遗传操作,已经报导了各种样的提取DNA的方法,但是都比较复杂。本论文通过对多种提取方法对多种食用菌的研究,得出一种操作简便,不需要昂贵仪器设备,用时短,产品纯度高,即适合大规模生产,又能体现食用菌生物学特性的食用菌DNA提取技术。 相似文献
8.
使用改良的SDS法提取银杏老枝、半木质化嫩枝韧皮部及叶子的基因组DNA,结果表明:从嫩技韧皮部和嫩叶提取DNA可获得较好的效果,但老叶提取DNA因杂质较多,需要多次纯化。 相似文献
9.
10.
以改良CTAB法、改良SDS法、试剂盒法提取青藏高原蕨麻总DNA;用紫外分光光度法测定提取的青藏高原蕨麻的总DNA浓度和质量;用琼脂糖凝胶电泳法、分光光度法测定提取的青藏高原蕨麻的总DNA质量,以期筛选出青藏高原蕨麻总DNA提取的最佳方法。结果表明:用改良SDS法提取的蛋白质含量最高,OD260/OD2801.61左右,用改良CTAB法提取的DNA的OD260/OD2801.76左右;用试剂盒法提取的DNA的OD260/OD2801.70左右;用改良CTAB法提取DNA的浓度在100μg/mL。 相似文献
11.
12.
《Scientia Horticulturae》2002,95(3):213-221
Multiple shoots were generated via direct organogenesis on hypocotyl segments excised from in vitro germinated seedlings (45-day-old) of Psidium guajava L. cv. Allahabad Safeda. Modified basal Murashige and Skoog (MMS) medium supplemented with 6-benzylaminopurine (BAP), zeatin or thidiazuron with or without α-naphthalene acetic acid (NAA) were tried. Thidiazuron (0.1 μM) along with 0.54 μM NAA gave the highest response (44.6%) with the regeneration of 3.6 shoots per original explants. These shoots upon subculture gave rise to about 5.0 shootlets per explant in shoot proliferation medium, i.e. MMS supplemented with 2.22 μM BAP + 2.32 μM kinetin. The regenerated microshoots were elongated using a quick dip of gibberellic acid (GA3; 1.44 mM) prior to culture on MMS medium supplemented with 0.88 μM BAP and adenine sulphate (54.29 μM) for 2 weeks. Rooting of microshoots was achieved best on half strength MMS medium supplemented with 4.90 μM indole-3-butyric acid along with 100 mg l−1 activated charcoal. 相似文献
13.
Akhtar Nasim 《The Journal of Horticultural Science and Biotechnology》2013,88(6):556-562
SummaryAn embryogenic protocol for plant regeneration of guava (Psidium guajava L.) was established using 10-week post-anthesis, zygotic embryo explants. Somatic embryogenesis was induced on Murashige and Skoog medium (MS) containing 3% (w/v) sucrose, 0.8% (w/v) agar and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) by continuous treatment of the zygotic embryo explants. Somatic embryos appeared as globular structures at the end of the third week from culture initiation, and heart-shaped, cotyledonary-stage, and torpedo-stage embryos appeared within the next few weeks. The development of somatic embryos was asynchronous and showed five-to-seven discernible stages. Depending upon the response of the somatic embryos during their maturation, germination, acclimatisation, and encapsulation, they were grouped into one of three categories. The preferred type of somatic embryos (≥ 1.5 mm) were called the “elongated torpedo” (ET) category. The slightly less-preferred type of stomatic embryos (from 1.0 – 1.5 mm) were termed the “short torpedo” (ST) category. The least preferred types of somatic embryos, at the cotyledonary, heart-shaped, and/or globular stages of development (< 1.0 mm), were grouped into a third category designated “CHG”. The suitability and efficacy of various growth regulators and other treatments were assessed based on six different embryogenic parameters: (i) the frequency of embryogenesis; (ii) the intensity of embryogenesis, defined as the average number of somatic embryos produced per culture (“ANEPC”); (iii) the frequency of ET somatic embryos; (iv) the frequency of ST somatic embryos; (v) the frequency of CHG somatic embryos; and (vi) the overall efficiency of embryogenesis, defined as the potential of a treament to produce somatic embryos at the ET or ST stages, or at both stages of development, that could be converted into plantlets. In the present report, we found that 0.01 mg l–1 2,4-D gave the maximum frequency and intensity of embryogenesis. But the highest frequencies of ET and ST somatic embryos were produced on MS medium containing 3% (w/v) sucrose and 0.001 mg l–1 2,4-D, while CHG embryos were produced at the highest frequency on the same medium, but containing 0.5 mg l–1 2,4-D. It was difficult to calculate the most effective concentration of 2,4-D for somatic embryogenesis based on parameters (i) – (v) above. Hence, quantitative estimations of the efficiency of embryogenesis (sixth parameter) were imperative in order to analyse the potential of the different treatments. The highest efficiency of somatic embryogenesis was achieved by continuous treatment of 10-week post-anthesis, zygotic embryo explants with 0.01 mg l–1 2,4-D on full-strength MS agar medium containing 3% (w/v) sucrose. These somatic embryos matured normally on the same medium, and germinated well both on half-strength solid and in half-strength liquid MS medium containing 3% (w/v) sucrose. They grew in full-strength liquid MS medium with 3% (w/v) sucrose and showed maximum survival upon transfer to soil and hardening. Evaluations of the efficiency of somatic embryogenesis in guava, based on the six parameters defined above, have also helped us to understand and evaluate processes for high efficiency micropropagation in other species. 相似文献
14.
15.
以番石榴无菌茎段为材料,研究了生长调节剂及MS强度对番石榴无菌茎段芽萌发和茎段生根的影响.结果表明,以MS为基本培养基时,最有利于番石榴无菌苗生长的NAA浓度为0.4 mg·L-1,叶片数(7.59片)、节数(3.95节)、苗高(5.10cm)和节间长度(1.41 cm)最大;6-BA浓度为5.0 mg·L-1时,茎段具有最高的芽诱导率(100%)和诱较高的芽诱导数(2.44个);1/2 MS+NAA 0.05 mg·L-1最有利于番石榴茎段的生根,生根率100%,根数2.30条,根长4.64 cm.在番石榴组织培养中,NAA能够促进番石榴茎段的生长,但会抑制芽的诱导. 相似文献
16.
WANG Jing-ru ZHAO Jing-jing YE Chun-ling YE Kai-he ZHANG Xiao-qi WANG Ying YE Wen-cai 《园艺学报》2012,28(6):1109-1113
AIM: To investigate the effects of total triterpenoids from Psidium guajava leaves (TTPGL) on blood glucose and lipids in type 2 diabetic rats. METHODS: The diabetic rats were induced by intraperitoneal injection of streptozotocin at dose of 35 mg/kg and feeding with high-fat diet. The animals were divided into 5 groups: diabetic model control group (model), TTPGL treatment groups (with the doses of 60, 120 and 240 mg/kg, respectively) and rosiglitazone treatment group (3 mg/kg). Another 12 normal SD rats were used as the normal controls. The rats received daily treatment for 6 weeks, and then the levels of fasting blood glucose (FBG), fasting insulin (FINS), triglyceride (TG), total cholesterol (TCH), free fatty acid (FFA), glycosylated hemoglobin (GHb) and glycosylated serum proteins (GSP) were measured. The protein expression of peroxisome proliferator-activated receptor γ (PPARγ) in adipose tissues was detected by Western blotting. RESULTS: Compared with normal control group, the levels of FBG, GHb and blood lipids were increased in type 2 diabetic rats. The FINS, insulin sensitivity index, and the protein expression of PPARγ in adipose tissues were decreased. Compared with model group, the levels of FBG and GSP were decreased,and the FINS, insulin sensitivity index, and the protein expression of PPARγ in adipose tissues significantly increased in TTPGL treatment groups (with the doses of 120 and 240 mg/kg). The levels of serum TG,TCH and FFA were significantly lower in TTPGL treatment groups (P<0.01 or P<0.05) as compared with the model controls. CONCLUSION: TTPGL decreases the levels of blood glucose and lipids in diabetic rats. TTPGL also increases serum insulin level and improves insulin sensitivity. The action mechanism of TTPGL may be related to the increase in the protein expression of PPARγ. 相似文献
17.
不同方法提取牛心朴子基因组DNA效果的比较研究 总被引:1,自引:1,他引:1
以牛心朴子的叶片、茎段、根为试材,采用改良的CTAB法Ⅰ、CTAB法Ⅱ和SDS法对其进行了基因组DNA提取效果的比较分析.结果表明.CTAB法Ⅰ从3个部位都能提出较高浓度的DNA,电泳条带完整清晰;CTAB法Ⅱ从3个部位都能提出DNA,条带明亮,但有明显拖尾现象;SDS法从叶片中能提出较高的DNA,但从茎和根中所提的DNA含量较小,电泳条带暗;3种方法所提的DNA其RAPD扩增效果以CTAB法Ⅰ最好,CTAB法Ⅱ次之,SDS法最差.叶片提取的DNA平均纯度、浓度和得率为最高,RAPD扩增效果最好,茎次之,根最低.说明CTAB法工是牛心朴子DNA的高效提取方法,不同部位以牛心朴子叶子提取的DNA得率高,扩增效果最好. 相似文献
18.
葡萄总RNA提取方法的研究 总被引:63,自引:3,他引:63
针对葡萄组织中多酚、多糖类物质含量较高的特点,比较了改进SDS/酚法、异硫氰酸胍法和市售总RNA提取试剂盒等不同的提取方法,3种改进方法均能从葡萄幼嫩叶片中提取到总RNA。其中改进的SDS/酚法能有效抑制酚类物质和多糖对总RNA提取的影响,能够从成熟的叶片和完熟果实的果皮中获得质量高、完整性好的总RNA,28SrRNA亮度约为18SrRNA的2倍,A260/A280介于1.8~2.0之间,A260/A230大于2.0,且总RNA产率高,其中幼叶总RNA产率为261.20μg/g,成熟叶产率为191.40μg/g,完熟果皮产率为31.50μg/g,完全适于进行DDRT-PCR、Northernbolt和cDNA文库构建等研究。且该方法具有快速、简单、有效的特点。 相似文献