A Study on Pollen Morphology of Eucommia ulmoides Oliver

Eucommia ulmoides Oliver is endemic to China. The pollen morphology and exine ultrastructure were examined under LM, SEM and TEM. Pollen grains are prolate, polar axis 30.5-54.8 μm long, with the average 32.7 μm, equatorial axis 27.8-31.3 μm long, with the average 29.2 μm, tricolporoidate, but sometimes the outline of ora could be observed and elliptic in shape. Colpi are narrow, uequal in length, often two long and one short or two short and one long, sometimes rather irregularly arranged, with indistinct and thin colpus membrance. Exine psilate under LM, granulate under SEM, and shortly baculate under TEM. Tectum is thin with dense and small granules, columellae layer consists of short bacules, and foot layer very thick. Some taxonomists (Cronquist, 1968) consider that Eucommiaceae is related to Hamamelidales, but others (Takhtajan, 1969) to Urticales. The Urticales is of the porate type of pollen grains, while Eucommiaceae of tricolporoidate type, and thus the former is more advanced than the latter. Pollen grains of some members of Hamamelidales, tricolporoidate, are similar to those of Eucommiaceae. We therefore consider that Eucommiaceae is related to Hama-melidales.     

产杜仲黄酮内生真菌的初步研究

由杜仲植株的根、茎、叶分离到内生真菌,分别对各菌株进行液体培养,筛选能产生与杜仲药材相同或相似黄酮类化合物的内生真菌。将培养物适当处理后,根据黄酮类化合物所特有的颜色反应进行初筛,然后根据TLC分析和紫外-可见光分光光度法测定结果进行复筛,结果从杜仲植株共分离得到44株内生真菌,其中1株(DZY5)能产出与其宿主相同或相似的黄酮类物质。     

Abnormal Development of Regenerated Bark in Eucommia ulmoides

After the girdled trunk of Eucommia ulmoides Oliv. was locally damaged usually no bark was regenerated at the injured area. However, abnormal structures were observed in the newly-formed bark in the adjacent region. Similar results also occurred in simulated experiments. After girdling, a strip about 10×2 cm2 and 1 mm in depth was scraped in the exposed area ensuring no bark regeneration in this artificially ruined area. Hence one month after the treatment it was seen that new vascular cambium developed in the regenerated bark around the destroyed area. However, the newly-formed bark above the harmed area was about two-fold thicker than that below the area. The differentiation of cambium and starch distribution were quite different on both sides.     

Microsatellite genetic variation in the Chinese endemic Eucommia ulmoides (Eucommiaceae): implications for conservation

Eucommia ulmoides, a traditional Chinese medicinal plant, is endangered as a consequence of long‐term and widespread harvest in the late 20th century. It has been widely cultivated as a source of herbal medicine and for use in the organic chemical industry in China. In this study, eight microsatellite markers were applied to investigate genetic diversity in E. ulmoides. Three hundred individuals from one semi‐wild population and nine cultivated populations across its main production area were collected. A high level of genetic diversity at population levels (H_E = 0.716) was observed. The highly outcrossed mating system, high longevity of E. ulmoides and seed admixture may be responsible for high genetic variation within populations. A genetic bottleneck was observed in one population. Populations were only slightly differentiated from one another (F_ST = 0.063); this was also supported by AMOVA, which revealed that 94.05% of the total variation resided within populations. This is probably attributable to long‐distance gene flow mediated by the exchange of seeds by local farmers. Implications of these results for the conservation of genetic resources of E. ulmoides are discussed. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 775–785.     

一株具有特异AFLP指纹图谱的杜仲古树

杜仲(Eucomm ia ulm oidesO liver)是原产于我国的传统药用植物,虽然目前杜仲被广泛栽培,但野生杜仲已非常稀少,几乎难以见到。作者在利用AFLP分子标记技术研究杜仲的遗传多样性和居群遗传结构时,发现一株采自神农架自然保护区的树龄200多年的雄性杜仲古树(编号为SNJ5)的AFLP指纹图谱与其它582株树龄为10~52年的栽培杜仲的指纹图谱有显著的不同。在总计为191条多态带中,有44条为高频率显性等位基因带(h ighfrequency dom inant allele bands,HFDAB,即583个个体中只有1~3个个体缺乏的带)。SNJ5缺乏23条HFDAB,其中有12条HFDAB是SNJ5单独缺乏的,与其他582个个体差异显著(p<0.05);另外,SNJ5还具有一条特有带(E ACG/M CTG引物组合的217 bp带)。据此推测这一杜仲古树是原始野生植株,是近年来众多学者认为野生杜仲灭绝后发现的非常重要的种质资源,应加以大力保护。     

具有ACC脱氨酶活性的杜仲内生细菌的分离鉴定及其抗菌活性

采用富集筛选法从杜仲根中分离到5株具有ACC脱氨酶活性的内生细菌, 利用纸片法测定它们的抑菌活性, 通过形态特征、生理生化试验和16S rRNA序列分析对分离菌株进行鉴定。结果显示, 5株杜仲内生细菌均具有较高的ACC脱氨酶活性, 其中4株菌对大肠杆菌CGMCC1.1103和枯草芽孢杆菌CGMCC1.769均有较好的抑菌活性, 通过生理生化试验和16S rRNA序列分析, 将菌株JDM-2、JDM-8、JDM-11、JDM-14和JDM-19分别鉴定为Pseudomonas koreensis、肺炎克雷伯氏菌(Klebsiella pneumoniae)、路德维希肠杆菌(Enterobacter ludwigii)、变栖克雷伯氏菌(Klebsiella variicola)和阿氏肠杆菌(Enterobacter asburiae)。     

杜仲种质资源的现状及保护对策

在野外调查的基础上,根据杜仲的分布特点,划分了7个主要分布区,并对种内存在的形态差异进行了描述。在评述杜仲种质资源现状的基础上,对种质资源的破坏以及造成遗传流失的原因进行了分析。提出了有效保护杜仲种质资源的原理和方法,并强调了迁地保护的重要性。     

Chemical Constituents from Staminate Flowers of Eucommia ulmoides Oliver and Their Anti-Inflammation Activity in Vitro

Three new monoterpenoids, named eucomylides A−C ( 1 – 3 ), along with six known compounds ( 4 – 9 ) were isolated from the staminate flowers of Eucommia ulmoides Oliver. The structures were elucidated by extensive analyses of spectroscopic methods, and their absolute configurations were established by time-dependent density functional theory electronic circular dichroism (TDDFT ECD) calculation. All the compounds along with previously isolated components ( 10 – 14 ) were tested for their anti-inflammatory effects. Two iridoid glycosides ( 11 and 12 ) and a flavonoid glycoside ( 14 ) showed potent suppressive effects on nitric oxide (NO) production in RAW 264.7 cells, with IC₅₀ values ranging from 17.11 to 22.26 μM.     

杜仲花粉壁发育的超微结构观察

利用透射电子显微镜和场发射扫描显微镜对杜仲花粉壁结构和发育进行了研究.结果显示,杜仲花粉在四分体末期形成原外壁,质膜随之出现波状样式,原基粒棒的形成在波状质膜的凹陷处积累,同时细胞中的核质类核仁向核膜外扩散,并进一步分布在质膜下;单细胞花粉早期其外壁结构(覆盖层、柱状层、基足层和外壁内层)已经分化完全,之后花粉内壁由萌发孔起始发育,然后全面增厚,至花粉成熟时萌发孔处的内壁变得很厚,并形成具有丰富径向微通道的结构,此时外壁内层崩溃解体.重点讨论了花粉外壁模式决定以及类核仁和花粉外壁形成之间的关系.     

杜仲叶及提取物营养价值和药用成分研究

对不同产地和不同收获季节的杜仲叶常规营养成分、杜仲叶及其提取物中药用成分进行了研究 ,结果表明不同收获季节的杜仲叶营养有差异 ,杜仲叶及其提取物中含有较高的绿原酸和单宁。生产实践中 ,可利用秋末落叶作为很好的饲料资源     

杜仲橡胶合成相关酶基因HMGR的克隆与表达分析

为解析HMGR基因在杜仲橡胶生物合成中的作用,本研究以杜仲良种华仲6号叶片和果皮为材料,参考杜仲基因组和转录组数据,克隆了杜仲HMGR基因,命名为Eu HMGR,全长1770 bp,编码590个氨基酸。实时荧光定量PCR测定结果显示,EuHMGR基因在果皮中的表达量于5月中旬达到最大值,而在叶片中的最大值出现在7月中旬。通过索氏提取测定相应时期果皮和叶片的含胶量表明,果皮中的含胶量从4月中旬到5月下旬迅速增加,而叶片中的含胶量一直平稳增长。分析不同发育时期果皮和叶片Eu HMGR基因表达量和含胶增长速率相关性发现,果皮Eu HMGR基因在不同发育时期的相对表达量和含胶增长速率呈显著正相关,而二者在叶片中无明显相关性。因此,推测Eu HMGR基因的表达与杜仲果皮中橡胶生物合成相关。     

杜仲含胶细胞的形态学研究

通过变性处理、整体透明及离析等方法的综合运用,可见杜仲体内含胶细胞是一种细长、两端膨大的单细胞,含胶细胞的分枝比较常见,大多为二叉状分枝,罕见三叉状分枝。整体透明后可见含胶细胞在叶片和果皮中的分布状况,同时对含胶细胞的形态学指标进行了测量。     

Influences of Special Damage Upon Exposed Surface of Girdled Trunk in Eucommia ulmoides

Mechanical injury to the exposed surface of the girdled trunk of Eueommia ulmo- ides Oliv. was conducted by scraping frames 10×l0 cm2 in four varieties of shapes, i.e. ∪、| |、Π and □ The scraping edge was 2 cm in width and 1 mm in depth. One month after treatment, periderm was formed at the central portion of all the scraped areas, however, among them the thickness, degree of differentiation and starch distribution of the regenerated bark were different. The newly-formed bark of the ∪ -shaped area was the thickest, and vascular cambium-like meristem was formed in the central portion which later gave rise to vascular tissues. The thickness of the newly formed bark in Ii -shaped area was half as :thick as the former, although similar tissue differentiation was observed. The thick- ness of the newly-formed bark inΠ-shaped area was half of that in∪ -shaped area. The flat meristematic cells were only seen in the deeper layer, and no vascular tissues were visible. The thickness of the newly-formed bark in the □ -shaped area was only 1/4 of that in ∪ -shaped area. There were undifferentiated parenchyma cells below the periderm in which very few flat meristematic cells were formed later.     

Studles on the Initiation and Development of Gutta-containing Cells of Eucommia ulmoides Oliv.

The gutta-containing structure in the cortex of a stem of Eucommia ulmoides is a filamentous, secretory cell. Observation of thin sections revealed that when the procambium of a stem differentiated into the earliest sieve elenents of protophloem, the initials cells appeared as daughter cells originated from a longitudinally equational or unequational divisions, or from the terminal cell formed by several transverse divisions of some cortical ground meristematic cells. Before the ceils of the cortical ground'meristem ceased to divide, the initial cells, in an arbitrary position of origin, develops continuously. These initial cells were able to be distinguished from the surrounding cells by their large length/width ratio, the presence of elliptical nuclei and dense cytoplasm, etc. Later, both ends of the initial cells extended rapidly through intrusive growth, forming the very long, thin and filamentous unicell with two dilated terminations. During development, the gutta particles were gradually synthesized and accumulated in the cytoplasm, where as the organelles degenerated progressively. In the muture gutta-containing cell, the cell cavity was filled with gutta particles, the nucleus as well as other organelles was disintegrated leaving an intact fibrous cell wall.     

杜仲剥皮再生对生长的影响

杜仲（Ｅｕｃｏｍｍｉａ ｕｌｍｏｉｄｅｓ Ｌｌｉｖ．）剥皮后每年７月测定剥皮株和照株的径向与高度生长量,每年秋季观测这些树的落叶情况,春季观测其萌芽情况,连续观察４年。同是对剥皮３年后在原剥皮部位作第二次剥皮的植株作了同样的３年观测。结果表明,剥皮当年树的落叶提前、发芽推迟,高度和树粗的生长明显减慢,剥皮后第二年其各方面的生长状况与对照株接近,第三年剥皮株的生长与对照株相似,但剥皮株的木材年生长量     

酶法提取杜仲叶中绿原酸工艺研究

采用纤维素酶法提取杜仲叶中的主要药效成分———苯丙素类的绿原酸(CHA),通过单因素试验、正交试验和方差分析确定了纤维素酶法提取杜仲叶中绿原酸的最佳操作条件。结果表明,加入550 U/g纤维素酶0.50%,pH 4.5,温度40℃,提取率最高可达到51.84 mg/g。     

聚乙二醇（PEG）对杜仲胚乳愈伤组织茎芽分化的影响

实验中发现,在培养基中加入适量PEG可以显著提高杜仲胚乳愈伤分化频率。PEG这种促进分化的效果既与PEG的分子量和所用的浓度有关,也与培养基中无机离子的强度和蔗糖浓度有关。效果最佳的培养基配方是：在激素组成为BA（2.0-2.75mg/L）＋NAA0.15mg/L）的基本培养基（MS无机盐＋B5有机物＋3％蔗糖）中,添加浓度为4％－6％PEG 4000或4％－5％PEG6000。在这种培养基上杜仲胚乳愈伤组织的分化频率均超过50％,最高可达70％以上,而在同样的条件下不加PEG时分化频率不到10％。然而,经PEG处理分化出来的胚乳再生植株中,部分苗玻璃化现象严重。     

杜仲叶中总黄酮物质超声提取的研究

采用醇提法、超声提取法探讨了从杜仲叶中提取总黄酮类物质的最佳工艺。试验结果表明：通过单因素试验和正交试验,超声提取效果最好。其最佳条件是：用浓度为400%、按料液比1：60的比例的乙醇浸泡24h,再用超声提取45min,其杜仲叶中总黄酮类物质的提出率可达25．43％。     

杜仲雌雄株树皮和叶片中氨基酸及其含量的研究

杜仲（ＥｕｃｏｍｍｉａｕｌｍｏｉｄｅｓＯｌｉｖ）是我国特有的雌雄异株植物,具有极高的药用价值、工业价值和观赏价值。主要依靠种子繁殖。为了有计划地进行定性栽培,就需要对幼龄植株进行早期性别鉴定。许多生理指标都可能作为性别鉴定的依据。通过对杜仲中氨基酸含量和组成的分析表明,不同性别植株树皮和叶片中均含有被分析的１７种氨基酸,其中胱氨酸和蛋氨酸在雌雄株间差异明显,雄株树皮和叶片中胱氨酸的含量分别高出雌株７６．９８％和４５．０４％;树皮和叶片中蛋氨酸含量雌株分别高出雄株２０．５０％和１８．７７％。这是否反映了杜仲雌雄的性别差异,值得探讨。必需氨基酸总量在树皮和叶片中均表现为雌株高于雄株,且叶片中必需氨基酸总量高于树皮中的含量。