CTAB法抽提香榧种子胚乳DNA的改进

香榧种子胚乳DNA抽提是RAPD和AFLP分子标记构建遗传图谱的基础性工作。经多次实验，我们对CTAB法抽提香榧种子胚乳DNA进行了改进，即先将胚乳冷冻，抽提时加入β－巯基乙醇，抽提过程中用高盐TE重悬DNA－CTAB沉淀，最后用异丙醇沉淀DNA。OD260／OD280比值检测结果为平均1．824，而且DNA得率也较高，证明用此法抽提的DNA能完全满足RAPD和AFLP的分析的要求。     

山茶属植物叶片DNA抽提

对植物中ＤＮＡ 抽提是在分子水平上对植物进行系统分析与研究的基础性工作．经多次实验,使用改良的ＣＴＡＢ法抽提山茶属植物中总ＤＮＡ,即在抽提前加入抗氧化剂β－巯基乙醇,并使用冷冻的异丙醇来沉淀ＤＮＡ, 获得了较好的效果,其纯度和得率完全能满足常规分子生物学操作的要求．     

不同提取方法的铁线莲属叶片总DNA提取效果

以铁线莲属植物的叶片为材料,采用CTAB-SDS法、简易CTAB法、改良CTAB法等3种提取方法,比较耗时、纯度、得率、浓度和质量等指标,以期为铁线莲属植物叶片总DNA的提取筛选适宜方法.结果表明,在纯度和浓度方面CTAB-SDS法＞简易CTAB法＞改良CTAB法,在节省提取时间方面改良CTAB法＞CTAB-SDS法＞简易CTAB法.以CTAB-SDS法所提取的9种铁线莲DNA为模板,进行PCR扩增,CTAB-SDS法提取的DNA得率高、质量好,PCR扩增成功率也较高.综合比较,采用CTAB-SDS法进行铁线莲属叶片总DNA提取效果更佳.     

不同处理方法对榆属植物叶片总DNA提取效果的影响

为探明榆属植物基因组DNA的有效提取方法,以春榆无性系、白榆无性系、金叶榆无性系1号和2号的不同成熟度叶片为材料,并对其做不同预处理,然后采用CTAB法提取榆树基因组DNA,结果表明:嫩叶提取的DNA得率是老叶的1.4~3.0倍,嫩叶的A260/A280值在1.82~1.87之间,老叶的A260/A280值大于1.90;鲜叶片提取的DNA得率是干叶片的1.0~2.0倍,鲜叶片的A260/A280值在1.86~1.94之间,干叶片的A260/A280值在1.90~1.98之间;用低浓度的乙醇(50%~75%)浸泡新鲜嫩叶12~24h可以有效减少DNA的降解,得率在498.62~1 295.97ng/μL,A260/A280值在1.84~1.94之间;预处理液中加入3g的PVP具有较好除酚去多糖的效果,所提取的DNA得率在862.51~1 791.44ng/μL,A260/A280值在1.92~1.98之间。     

山茶属植物油茶组与金花茶组的分子分类

以山茶属5种油茶组植物和20种金花茶组植物的叶片为材料抽提DNA,利用筛选出的16种多态性随机引物进行PCR扩增,将扩增出来的多态型谱带转化为0-1数据,利用0-1型数据聚类软件进行RAPD聚类分析,将5种油茶组植物分为3大类,20种金花茶组植物分为4大类,分类结果与张宏达分类系统大致相同,并对差异很小的一些物种提出了合并建议.     

常山胡柚叶片DNA的抽提与分析

经反复实验，采用在抽提前加入一定量的抗氧化剂β－巯基乙醇，用冰冻的异丙醇来沉淀DNA，并增加氯仿／异戊醇抽提次数的所谓改良CTAB法，对常山胡柚叶片DNA抽提与分析，不仅成功提取到常山胡柚叶片DNA，而且所得的DNA浓度高、纯度好，20个试样的DNA苹均质量浓度为272.86μg/mL，平均每克鲜叶片可获DNA约82μg，纯度基本上都在1.4-1.8范围内，并经RAPD反应验证，完全能满足常规分子生物学分析的要求。     

油菜线粒体DNA提取纯化方法研究

以油菜为实验材料,探讨了植物线粒体DNA提取纯化过程中的影响因素,构建了一套快捷、经济、高效的植物线粒体DNA提取纯化方法.该方法结合差速离心和密度梯度离心分离得到线粒体,酶消化处理去除其他基因组的污染,用SDS与蛋白酶裂解线粒体,通过高盐沉淀,有机溶剂抽提,最后再用CTAB纯化得到mtDNA.用该方法提取的油菜线粒体DNA,经紫外分光光度计测定其纯度、限制性内切酶进行酶切.结果表明,此提取方法得到的mtDNA,其纯度可以满足后续遗传学分析.该法适用于不同植物材料mtDNA的提取纯化.     

无患子叶皂苷提取及结构表征

以无患子叶子为研究对象,分别考察了索氏抽提法与微波萃取法对皂苷提取得率的影响,确定了较优的提取工艺.结果表明,索氏抽提选择80％乙醇为提取剂,在液料比30∶1(g∶g,下同),提取温度95℃,抽提时间为3h,提取2次时皂苷元提取得率达到21.7％；微波萃取选择3 g原料,微波功率300 W,温度70℃,90％的乙醇,液料比20∶1,提取时间45 min,皂苷的提取得率达到22.3％.目标提取物经IT-IR、HPLC和1H NMR表征,确定其结构为常春藤皂苷元.     

桦木醇提取及纯化工艺的研究

以白桦树皮为原料,研究了桦木醇提取和纯化的工艺,确定了适宜的提取工艺条件:树皮原料30 g,树皮颗粒大小为0.42~1.16 mm,采用氯仿为溶剂,用索氏抽提器抽提7 h,得到粗桦木醇黄色固体,得率为38.5%。通过重结晶对桦木醇进行纯化,确定了适宜的重结晶条件:以异丙醇为重结晶用溶剂,粗提物与异丙醇的比为1∶20(g∶mL),结晶温度-5℃,结晶时间1 h,在此条件下桦木醇的结晶得率41.4%,产品熔点为245~254℃,纯度为95.4%。     

八角基因组DNA的提取方法

采用4种植物基因组DNA提取方法(CTAB法、SDS法、高盐法、试剂盒法)对八角基因组DNA进行提取试验.结果表明,对传统CTAB法稍加改良可以得到较为理想的提取效果,改良的内容包括①在加入CTAB提取缓冲液之前,先用CTAB-free缓冲液抽提1～2次;②将CTAB提取缓冲液中CTAB的用量从2％提高到3％;③提取缓冲液的用量由样品干重的10倍增加到60倍.用改良CTAB法提取的DNA经RAPD-PCR扩增,条带清晰,其质量能够满足分子标记的要求.     

Effect of border planting of Eucalyptus citriodora Hook. on the growth and biomass production performance of Cymbopogon winterianus Jowitt ex Bor Grass at Wondo Genet,Southern Ethiopia

The use of Eucalyptus trees, in association with other plants, has the potential to reduce the growth of neighbouring plants or grasses by competing for resources or releasing allelochemicals that may cause other plants to grow slowly. Our purpose here is to assess whether border planting of Eucalyptus is harmful to the establishment,growth, and production of Cymbopogon winterianus at various distances from tree lines in the Wondo Genet Agricultural Research Center in Southern Ethiopia. Several factors were studied: growth performances and yield of aromatic grasses; soil properties, including total nitrogen,organic carbon, and moisture content; and plant leaf nutrient content(NPK) at various distances away from the Eucalyptus citriodora border line were studied. Chemical compositions of hydro-distilled essential oils obtained from selected C. winterianus grasses leaves were analysed by gas chromatography. Plant height, biomass yield, amount of essential oil yield, and essential oil composition all were significantly affected due to Eucalyptus effect in the first10-m distance as compared to beyond 10 m from the tree border line. There were increases in soil total nitrogen,organic carbon, and moisture content with decreasing distances to the Eucalyptus tree border. The result of plant leaf nutrient content(NPK) differed across treatment and distance. The pot experiment(15 cm 9 20 cm), shoot length,and number of leaves of aromatic plants varied in the degree of inhibition based on the concentration of the aqueous extract. Significantly higher inhibitions of the shoot length and number of leaf parameters were observed when using 25% fresh leaf extract compared to other extract levels.     

桉树体内的生根抑制物质研究综述*

对桉树内源生根抑制物Ｇ和“巨桉酚”，分别列出了化学结构和物理性质，分析对植物生根的抑制作用表明，随着茎叶组织成熟，Ｇ含量上升；不同种类桉树和不同成熟度的组织，Ｇ含量也不同，并有周期性的变化，在１０＾－４Ｍ时，抑制巨桉插条３０％－４０％发根，但在１０＾－５Ｍ时，不表现有抑制作用，巨桉的幼苗随着茎节数增多，Ｇ含量升高，在１２节数时，生根率几为０。两者的生理效应还有抑制种子萌发和叶片光合作用，具有生长素     

桉树不同成熟度叶片营养元素内循环特征

揭示桉树人工林不同成熟度的叶片器官所需营养元素变化以及土壤肥力对其影响,对于桉树营林和科学施肥具有重要的生产指导意义。选取广西隆安县小林镇和田东县思林镇的桉树2代萌芽林作为研究对象,测定两个人工林嫩叶、成熟叶、黄叶的C、N、P、K、B、Ca、Mg以及土壤C、N、P、K、Ca、Mg元素含量,并进行相关分析。结果表明,植物叶片的N、P、K、Mg营养元素转移规律是黄叶向成熟叶和嫩叶转移,但C、Ca、B营养元素则是随着叶片的生长发育逐渐积累,黄叶中含量最高,说明施肥时应关注Ca、B元素的持续和有效供给。两片人工林都表现出类似的规律,说明土壤肥力的差异不影响桉树人工林叶片营养元素的内循环规律。桉树嫩叶营养元素含量及其生态化学计量比学特征具有与成熟叶和黄叶不同的表征,在营养物质研究中应予以足够重视。     

粗皮桉再生系统的研究(英文)

该研究在国内外首次报导了粗皮桉的再生系统 .利用粗皮桉的外植体通过两种不同的再生途径建立了高效的再生系统 .首先探讨了粗皮桉的下胚轴和子叶的体细胞胚胎发生 .研究发现 ,下胚轴和子叶在含有 2 ,4 D(添加有BA或激动素 )的培养基上可诱导三种典型不同的愈伤类型 .其中 ,来源于下胚轴的颗粒状的、松脆的胚性愈伤在分化培养基上可获得高频率的再生小植株 .讨论了不同类型的愈伤组织和其对应的分化再生率之间的关系 .此外 ,本研究探讨了来源于粗皮桉的下胚轴、子叶及成熟叶片在添加有一系列BA和NAA的不同浓度组合的B5培养基上的器官发生途径 ,三种不同的外植体在不同的再生培养基上的最高再生频率不同 ,其中成熟叶片、子叶和下胚轴的最高再生率逐渐提高 ,分别为 4 1 6 7%、6 6 6 7%和 83 33% .该文还讨论了两种不同再生系统的意义     

铝处理对2个桉树无性系幼苗丙二醛含量的影响

采用室内水培试验法,研究了铝处理对广西区2个优良桉树无性系(Eucalyptus grandis×E.urophylla No.9和E.urophylla No.4,分别记为G9、G4)根系及叶片内MDA含量的影响。结果表明:2个桉树无性系根内MDA含量均比叶内高;铝处理下,2个桉树无性系幼苗的MDA含量明显升高,G4根中MDA含量最高,达11.49μmol·g-1,根中MDA含量比叶中高,桉树无性系幼苗根部受铝伤害较叶严重。     

Water and nutrient supply regimes and the water relations of juvenile leaves of Eucalyptus globulus

Pressure-volume analysis was used to study effects of irrigation and fertilization on the water relations of newly expanded juvenile leaves of Eucalyptus globulus Labill. seedlings growing in Portugal's Atlantic region. In May, at the beginning of the dry season, fertilization and irrigation treatments had no significant effects on the water relations parameters investigated. In September, at the end of the dry season, leaves from non-irrigated plants had a significantly higher apoplasmic water content and a higher dry weight/turgid weight ratio than leaves of similar physiological age from irrigated trees. The osmotic potential at full turgor and the water potential at the wilting point were lower in non-irrigated than in irrigated plants. Changes in osmotic potential at full turgor were negatively correlated with changes in dry weight/turgid weight ratio, suggesting that reductions in osmotic potential at full turgor were largely the result of decreases in cell size. Fertilization had no detectable effect on these variables or on leaf tissue bulk modulus of elasticity. Tissue elasticity was also unaffected by irrigation. Independently of water and nutrient supplies, leaf elasticity was higher and relative water content at the wilting point lower in leaves sampled in May than in leaves sampled in September. In non-irrigated plants, leaves sampled in September had a lower tissue elasticity and a lower osmotic potential at full turgor than leaves sampled in May, indicating that leaves produced at the end of the dry season generate lower water potentials as turgor is lost than leaves expanded early in the season.     

Influence of light availability on leaf structure and growth of two Eucalyptus globulus ssp. globulus provenances

Light availability strongly affects leaf structure of the distinctive ontogenetic leaf forms of Eucalyptus globulus Labill. ssp. globulus. Late-maturing plants from St. Marys, Tasmania and early maturing plants from Wilsons Promontory, Victoria (hereafter referred to as Wilsons Prom.) were grown for 9 months in 100, 50 or 10% sunlight. Growth, biomass and leaf area were significantly reduced when plants were grown in 10% sunlight. Provenance differences were minimal despite retention of the juvenile leaf form by the Tasmanian plants throughout the study. The time taken for initiation of vegetative phase change by the Wilsons Prom. saplings increased with decreasing light availability, but the nodal position of change on the main stem remained the same. Both juvenile and adult leaves remained horizontal in low light conditions, but became vertical with high irradiance. Leaf dimensions changed with ontogenetic development, but were unaffected by light availability. Juvenile leaves retained a dorsiventral anatomy and adult Wilsons Prom. leaves retained an isobilateral structure despite a tenfold difference in light availability. Stomatal density and distribution showed ontogenetic and treatment differences. At all irradiances, juvenile leaves produced the smallest stomata and adult leaves the largest stomata. Amphistomy decreased with decreasing irradiance. Detrended, correspondence analysis ordination highlighted the structural changes influenced by ontogenetic development and light availability. Adult leaves had characteristics similar to the xeromorphic, sun-leaf type found in arid, high-light conditions. Although juvenile leaves had characteristics typical of mesomorphic leaves, several structural features suggest that these leaves are more sun-adapted than adult leaves.     

Antibacterial sideroxylonals and loxophlebal A from Eucalyptus loxophleba foliage

Loxophlebal A, a new antibacterial formylated phloroglucinol was isolated from the mother liquor obtained after separation of sideroxylonals from the chloroform–methanol extract of leaves of Eucalyptus loxophleba ssp lissophloia. The structure of loxophlebal A was determined to be 3-desformyl sideroxylonal A by spectroscopic methods including 1D- and 2D-NMR. The stereochemistry of loxophlebal A was determined by chemical correlation with sideroxylonal A. This article also reports an efficient, simple and economic method for large scale isolation of sideroxylonals in a purity of > 90% from the leaves of Eucalyptus loxophleba ssp lissophloia.     

DNA Extraction from Eriocaulon Plants and Construction of RAPD System

There have been many arguments on the classification of Eriocaulon Linn. by morphology so far, and little is known about the use of molecular marker for genetic for genetic diversity of Eriocaulon plants. To apply the technique of molecular marker to the research of genetic diversity of Eriocaulon plants, the study of the extraction method of DNA from the Eriocaulon plants and the RAPD system are essential for researchers. In this paper, the extraction of genome DNA from the silica-gel-dried leaves of several species of Eriocaulon distributed in China was studied, and the best RAPD analysis technique condition of Eriocaulon plants was analyzed.     

Physiology and anatomy of lenticel-like structures on leaves of Eucalyptus nitens and Eucalyptus globulus seedlings

Intumescences or abnormal, non-pathogenic, blister-like protuberant growths, form on Eucalyptus globulus Labill. and, to a much lesser extent, Eucalyptus nitens (Deane and Maiden) Maiden leaves when plants are grown in a high relative humidity environment. We examined the histology of intumescences and their effects on leaf photosynthetic processes. Intumescences were induced by placing E. globulus and E. nitens seedlings in a relative humidity of 80% in a greenhouse for 5 days. Symptomatic and asymptomatic leaves of plants with intumescence development were compared with leaves of control plants. Light-saturated carbon dioxide (CO(2)) assimilation (A(max)) and responses of CO(2) assimilation (A) to varying intercellular CO(2) partial pressure (C(i)) were measured. Symptomatic and asymptomatic leaf samples were fixed and sectioned and cellular structure was examined. Intumescences greatly reduced the photosynthetic capacity of E. globulus leaves and were associated with reduced electron transport rate and ribulose bisphosphate (RuBP) regeneration capacity. Tissue necrotization and cellular collapse of the palisade mesophyll and deposition of phenolic compounds in the affected areas, probably reduced light penetration to photosynthesizing cells as well as reducing the amount of photosynthesizing tissue. Photosynthetic capacity of E. nitens was unaffected. The intumescences resembled simple lenticels, both morphologically and developmentally. To our knowledge, this is the first time that lenticel-like structures developed in response to environmental conditions have been described on leaves.