一种提取高质量红树植物总RNA的有效方法

为了从富含单宁和多糖的红树植物中获得高质量的RNA,在总结及改良前人工作的基础上,优化了提取缓冲液的条件,采用碱性条件、PVP及β-巯基乙醇抑制单宁的氧化;利用RNA与单宁、多糖在不同浓度的2-丁氧乙醇中溶解度不同的特性来去除单宁和多糖;同时利用LiCl来选择性沉淀RNA,获纯度较高的RNA可以直接用于cDNA合成、cDNA文库的构建以及RACE等分子生物学操作。     

一种小量提取植物总RNA的有效方法

以水稻、玉米、辣椒为材料 ,摸索出了一套改良的异硫氰酸胍提取植物总RNA的方法 ,该方法简单、快速、纯度高、完整性强 ,方便有效 ,值得推广。     

一种优化的植物总DNA提取方法

根据自己提取植物基因组DNA的实际经验,改进了Clark利用CTAB提取植物基因组DNA的方法。在试验过程中发现,通过在研磨材料时加入液氮、用剪去端部的吸头转移含有DNA的溶液,并且将加入RNase A消化RNA的步骤改在最后进行等一系列改进,可以获得高质量的DNA。本文同时就主要提取步骤进行了分析。     

一种丹参高质量总RNA的提取方法

高质量RNA的获得是开展丹参分子生物学研究的基础。采用异硫氰酸胍(Guanidine Thiocyanate,GT)法、尿素法、CTAB法、苯酚法和热硼酸改良法等五种方法．以丹参组织培养的幼苗为材料,进行丹参RNA的分离试验,发现所采用的几种方法获得的RNA都有不同程度的降解。分析可能是由于丹参中含有大量的多糖以及各种次生代谢成分造成的。在分析现有结果的基础上对GT法进行了改良,在第二次沉淀前附加低浓度乙醇(10％～20％)沉淀20min对于高质量总RNA的获得效果较好。琼脂糖凝胶电泳检测改良后的GT法无论对于组织培养中幼嫩的根、茎、叶还是大田两年生的丹参根、茎、叶和种子均具有很好的RNA分离效果。mRNA电泳检测发现所得mRNA集中分布在500b～3kb之间且质量较高,完全可以满足丹参各种RNA相关的分子生物学实验要求,为丹参RT—PCR、Northern杂交等分子生物学实验提供了良好的基础。     

一种快速提取小麦叶片总RNA的方法

从植物组织中提取高质量的RNA是进行植物分子生物学研究的必要前提和关键.同种植物不同器官的组织由于组成分的差异,提取RNA的方法也存在不同的难点.在苯酚法和氯化锂沉淀法的基础上,改进并提出了一种适合小麦叶片总RNA的快速提取方法,消除了蛋白质、DNA、多糖、多酚等污染.该方法提取的小麦叶片总RNA,完整性好、纯度高,可用于RT-PCR、N orthern杂交、RACE等实验操作,而且简单经济、快速、实验结果稳定,重复性好,还适合富含多糖和脂质的植物组织总RNA的提取.     

一种提取葡萄芽中总RNA的方法

以葡萄品种‘Perletts‘‘(Vitis vinifera)冬芽为材料,研究了提取葡萄芽总RNA的方法,同时指出提取RNA时的注意事项。该方法具有可靠性高,易于大量制备,所提取的RNA质量高等优点。     

富含多糖草莓果实总RNA提取方法的改进

以草莓果实为模式实验材料,将Chomczynski提出的常规RNA提取方法与Kenneth等提出的改进方法相结合,并做了进一步改进,建立了一种简单实用的从富舍多糖的植物材料中提取总RNA的方法。先利用冷的丙酮去除色素类物质,再利用乙二醇丁醚去除多糖,从而有效克服了从富含色素和多糖娄物质的植物材料中提取RNA的困难;获取的RNA样品在纯度和浓度上都可以满足PCR及Northern杂交等分子生物学实验的要求。     

柑橘叶片总RNA的两种提取方法比较

为了简便、快速提取高质量的柑橘(Citrus reticulata Banco)叶片总RNA,采用TRlzol法,对北京天根公司RNAplant Reagent和日本TaKaRa公司RNAiso Reagent两种RNA提取试剂进行比较并适当改进.结果表明:通过琼脂糖凝胶电泳,使用TaKaRa公司试剂提取的总RNA28S和18S条带清晰,紫外分光光度计检测分析A260/A280为1,820,A260,A230为2.088,RNA的浓度为2.840 μg μl-1,用于RT-PCR反应可成功克隆柑橘β-actin看家基因228 bp片段;采用天根公司试剂,琼脂糖凝胶电泳显示RNA带型较模糊,紫外分光光度计检测分析A260/A280为1.464,A260/A230为1.603,RNA的浓度为2.020 μg μl-1,达不到RNA的标准.TaKaRa公司RNAiso Reagent试剂提取的柑橘叶RNA纯度和完整性较好,能用于Northern杂交、cDNA文库的建立及基因克隆等分子生物学实验,为柑橘的进一步分子生物学研究奠定了基础.     

一种快速的胚胎组织总RNA的提取方法

简要介绍一种改进的提取人体器官组织总RNA的方法,该方法具有费用低,快速简便,重复性好的优点,提取的RNA无DNA等污染物,完全能满足基因表达研究的需要。     

茶树叶片总RNA提取方法的比较研究

针对茶树叶片中富含多酚类物质的特点,以陕西地方茶树代表种质紫阳槠叶种的叶片为材料,比较了Trizol法、异硫氰酸胍法和改良SDS-酸酚法三种不同的总RNA提取方法。结果表明改良SDS-酸酚法能够从新鲜叶片、冷冻叶片和干燥叶片中提取到质量高、完整性好的总RNA,经检测28SrRNA亮度为18SrRNA的2倍,A260/A280值介于1.83～2.01之间。以鲜叶、冻叶和干燥叶片的总RNA为材料,进一步用于DDRT-PCR分析,均可获得清晰稳定的多态性结果。说明改良的SDS-酸酚法能抑制茶树叶片中多酚类物质对总RNA提取的影响,是一种适于茶树叶片总RNA提取的有效方法。     

Gibberellins from mangrove plants

Gibberellins were isolated from three mangrove plants: A₁ and A₃ from Sonneratia apetala; A₃, A₅ and A₉ from Rhizophora mucranata and A₃, A₄ and A₇ from Bruguiera gymnorhiza. Biological activity of these gibberellins were examined using three bioassays.     

海南红树林群系及分子生物技术在其研究中的应用

红树林是生长在热带亚热带海岸潮间带的木本植物群落。海南红树林是中国种类最多,分布和保存面积最大的区域之一,具有极为重要的研究价值。综述了海南红树林在种类、群落组成及红树林资源引种恢复以及分子生物技术等方面的研究,重点综述了分子生物技术在海南红树林植物研究中的应用,包括等位酶的应用、随机扩增多态DNA技术(RAPD)的应用和红树林植物耐盐性研究。     

广西北部湾红树植物内生真菌多样性

【目的】研究广西北部湾地区红树植物内生真菌多样性,建立北部湾红树植物内生真菌种质资源库,为利用内生真菌生物技术促进农业可持续发展提供理论依据。【方法】从广西北部湾地区采集红树植物组织样本,采用表面消毒法分离真菌,通过测定分离菌株对宿主植物是否具有致病性来筛选内生真菌,结合形态学特征和分子生物学分析对内生真菌进行分类与鉴定。【结果】从60个红树植物样本中分离得到1 764个菌株,经过致病性测定筛选获得41株内生真菌,分离率为2.3%。其中从宿主植物红海榄分离得到15株内生真菌,占总菌株数的36.6%,比例最高。通过分析,发现这些内生真菌在ITS-NJ、NS-NJ两个系统发育树上各聚为7个大分支,分属8个科(目)。其中球腔菌属Mycosphaerella、德福里斯孢属Devriesia、假尾孢属Pseudocercospora、枝孢霉属Cladosporium、Pleosporales等属(科)真菌是广西红树林的优势菌。【结论】广西北部湾地区红树植物内生真菌菌种资源丰富。     

Use of a simple method to isolate intact RNA from partially hydratedSelaginella lepidophylla plants

Isolation of high-quality RNA is a necessary step in gene expression analysis. Although many methods can be used to isolate RNA from plants where contamination of preparations with complex carbohydrates or phenolic compounds is a problem, the application of these methods toSelaginella lepidophylla tissues has failed to obtain good-quality RNA. Here we introduce 2 modifications to the method developed by Chomczynski and Sacchi (1987), generating a simple and rapid method that allows the isolation of intact RNA fromS. lepidophylla-dehydrated tissues. Although the introduced modifications are not new, their addition proved to be decisive for success in RNA isolation. Quality of the RNA obtained was evaluated by electrophoresis in agarose and by 3 different PCR-based techniques—RT-PCR, RNA differential display, and synthesis of a cDNA library.     

Floral scent chemistry of mangrove plants

The flowers of mangrove plants are pollinated by a variety of pollinators including birds, bats, and insects. This study analyzed the floral scent chemistry of mangroves on Iriomote Island (located near Taiwan) including Bruguiera gymnorrhiza (L.) Lamk. (Rhizophoraceae), Kandelia candel (L.) Druce (Rhizophoraceae), Rhizophora stylosa Griff. (Rhizophoraceae), Sonneratia alba J. Smith (Sonneratiaceae), Nypa fruticans (Thunb.) Wurmb. (Palmae), Lumnitzera racemosa Willd. (Combretaceae), Avicennia marina (Forsk.) Vierh. (Avicenniaceae or Verbenaceae), and Pemphis acidula Forst. (Lythraceae). A total of 61 chemicals (fatty acid derivatives, terpenoids, carotenoid derivatives, benzenoids, nitrogen-containing compounds, 13 unknown chemicals) were detected in the floral scents of the various species. The species displayed a distinct chemical profile ranging from only two chemicals in the floral scent of Kandelia candel to more than 25 chemicals in the floral scent of Nypa fruticans. All of the identified chemicals have been found in the floral scents of other angiosperms. The chemical profile of some species can be correlated with their floral morphology and pollinators. Received: August 18, 2001 / Accepted: October 9, 2001     

A Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants

Introduction –  RNA quality and integrity are critical for many studies in plant molecular biology. High‐quality RNA extraction from grapevine and other woody plants is problematic due to the presence of polysaccharides, polyphenolics and other compounds that bind or co‐precipitate with the RNA. Objective  – To develop an optimised cetyltrimethylammonium bromide (CTAB)‐based protocol, to reduce the time and cost of extraction without reducing quality and yield of RNA extracted from polysaccharide‐rich tissues of several plants. Methodology  – Several changes were introduced to the original CTAB protocol. All centrifugation steps were carried out at 4°C, the sample weight was decreased and the concentrations of PVP‐40 and LiCl were increased reducing incubation time prior to RNA precipitation. This rapid CTAB protocol was compared with six different RNA extraction methods from three grapevine tissues, namely, in vitro plantlets, and leaves and mature canes from actively growing field vines. Results –  The rapid CTAB method gave high‐quality RNA in only 3 h at low cost with efficiency equal to or higher than that obtained with other time‐consuming and expensive protocols. The procedure was applied to RNA extraction from other grapevine tissues and other woody species including olive, lemon, poplar, chestnut, apple, pear, peach, cherry, apricot, plum and kiwi fruit. RNA of high quality could be isolated from all tissues and from all species. Conclusion –  The study has shown that the improvement of a CTAB‐based protocol allows the rapid isolation of high‐quality RNA from grapevine and many woody species. Copyright © 2008 John Wiley & Sons, Ltd.     

Soil salinity and pH in Japanese mangrove forests and growth of cultivated mangrove plants in different soil conditions

Soil conditions of mangrove forests in southern Japan were found to correlate largely with zonal distributions of the species.Kandelia candel grew in soils with low salinity and low pH,Avicennia marina, Rhizophora stylosa andSonneratia alba in soils with high salinity and high pH, andBruguiera gymnorrhiza in soil with a wide range of pH but limited range of salinity.Lumnitzera racemosa colonized soil with a wide range of pH and medium salinity. Seedlings ofKandelia candel, Bruguiera gymnorrhiza andRhizophora stylosa were planted in soils with differing salinity and pH. Optimum seedling growth ofKandelia, Bruguiera andRhizophora occurred when plants were cultivated in soils similar to those of their natural habitats, suggesting that growth of mangrove species and their zonal distributions were regulated by salinity and soil pH.