紫花含笑(♀)与灰岩含笑(♂)及其杂种F1代叶表皮微形态和叶结构的比较观察

通过比较观察紫花含笑(♀)和灰岩含笑(♂)及其杂种F₁代叶表皮微形态和叶结构发现,紫花含笑和灰岩含笑在叶表皮微形态及叶解剖结构方面有很大差异。杂种F₁代个体间在叶表皮微形态及叶解剖结构方面变异很大,为连续的数量遗传;其中57.7%以上的杂种F₁代气孔密度高于父母本。杂种F₁代矮化型植株叶片气孔密度较小,推测其抗寒性较强,可以通过进一步杂交改良,获得矮化型的盆栽含笑新品种。观察结果可为深入探讨含笑属种间杂种F₁代的遗传变异,并从这些杂种F₁代中选育观赏新品种积累科学资料。     

Germination studies on three critically endangered endemic angiosperm species of the Kashmir Himalaya,India

The critically endangered and perennial alpine endemic angiosperms, namely, Aquilegia nivalis, Lagotis cashmeriana and Meconopsis latifolia inhabit such habitats in the Kashmir Himalaya that are characterised by short growing season and heavy snow cover for about 3–4 months during winter season. The seeds of these species under natural conditions experience a long period of pre-chilling during winter prior to their germination in following spring season. Taking cue from such a requirement, present study investigated the effect of chilling and exogenous application of growth hormones, NO₃ ⁻ and NH₄ ⁺ on total percent germination of otherwise deep-dormant seeds of these species, under alternate light/dark and continuous dark light regimes. Prolonged pre-chilling followed by treatment of seeds with different doses of GA₃ had a pronounced stimulatory effect on the total germination percentage in all the three species. In fact, highest germination percentage in A. nivalis was recorded only when pre-chilled seeds were treated with 1.5 mM GA₃ under alternate light/dark conditions. Likewise, germination of pre-chilled seeds of L. cashmeriana with ruptured seed coats was improved when treated with various concentrations of GA₃. Seed germination in M. latifolia was also favourably influenced by treatment of pre-chilled seeds with GA₃ or nitrogen applied either as NO₃ ⁻ or as NH₄ ⁺ under alternate light/dark conditions. Treatment of seeds with kinetin (6-furfuryl-aminopurine) had no significant influence on germination percentage in any of the three species. Thus, prolonged chilling of seeds followed by their treatment with GA₃ under alternate light/dark conditions are the requirements necessary for seed germination in these species.     

工程菌种子制备方法对rEPA表达量影响的研究

由重组E.colirPE553D所表达的基因缺失突变脱毒的重组铜绿假单胞菌外毒素A(rEPA),目前大量以载体蛋白用于多种细菌多糖蛋白结合疫苗研究。rEPA的表达量受众多因素的影响,种子的制备方式就是重要因素之一。本文用长期冷冻保存的和新鲜提取的质粒分别转化宿主菌E.coliBL21(λDE3)后制备种子,并将它们和冻干保存的E.colirPE553D在相同条件下培养和诱导,经SDS-PAGE分析表明长期保存的质粒转化宿主后的重组工程菌生长速度较慢,但rEPA的表达量和新鲜质粒转化制备的工程菌基本相同,而冻干保存的工程菌E.colirPE553D几乎丧失了表达rEPA的能力。     

濒危植物水青树种子的生物学特性

通过连续三年（2006～2008）对水青树种子（Tetracentron sinense）的形态特征和生理特性等指标进行检测,探讨了不同储藏温度、时间和不同基质对水青树种子萌发的影响,分析了与种子生物学特性相关的濒危机制。水青树种子很小,其形态特征、千粒重、含水量和饱满度在不同年限间差异不显著,而吸水速率却差异显著,其中2007年种子的吸水速率最快。TTC染色测得3年水青树种子的活力几乎为0,但其活力指数分别为0、0.41和0.27,表明TTC染色法不适于水青树种子活力的估算。低温储藏后,3年种子的萌发率分别为1.25%、96.5%、70.8%,发芽势分别为1%、59.8%、41.8%。随储藏时间的增加,2007年种子活力降低的速度都慢于其它2年,低温储藏的种子活力降低速度慢于常温储藏的种子。不同基质对水青树种子的萌发和幼苗的生长有一定影响。结果表明,大年水青树种子的活力明显强于小年,随储藏时间的增加其活力的丧失速度也慢于小年;低温储藏有利于种子活力的保持。种子生物学特性方面导致水青树更新困难的原因可能有以下几点：（1）长期进化形成的小种子虽有利于传播和散布,但不利于种子和幼苗的存活;（2）长期低温不适于种子萌发,难以形成应有的幼苗格局;（3）长期湿润环境不适于种子安全度过寒冷期。     

The Occurrence of <Emphasis Type="Italic">p</Emphasis>-coumaric Acid and Ferulic Acid in Fossil Plant Materials and their Use as UV-proxy

The applicability of p-coumaric acid and ferulic acid concentrations or ratios in (sub)fossil plant remnant as UV-B proxies relies on various aspects, which are discussed in this paper and will be illustrated with some experimental data. A newly developed THM-micropyrolysis–gas chromatography–mass spectrometry method was tested on various spores, pollen and other plant remains, which were analysed for the presence of the UV-absorbing compounds p-coumaric acid and ferulic acid. This revealed that these supposed building-blocks of sporopollenin appear to be present in pollen of many plant species but also in moss spores. The development of this micropyrolysis method paved the way for the quantitative analysis of UV-absorbing compounds in case only a small amount of analyte is available, for example for fossil pollen and spores but also other small palynomorphs and plant fossils. The use of this technique will provide a better insight in the plant responses to UV-radiation, the chemistry of pollen and spores, their fossil counterparts and furthermore the means for a further development of a proxy for the reconstruction of past UV-B radiation.     

国产18种鳞盖蕨属植物叶表皮微形态特征及其系统学意义

鳞盖蕨属是一个自然类群,由于各类群之间的形态差异以及种内变异比较大,因此其分类一直存在争议。该研究利用光学显微镜(LM)和扫描电子显微镜(SEM)对鳞盖蕨属18种植物(含3变种)的叶表皮微形态进行了观察和比较,并结合形态学、孢粉学和分子系统学探讨其系统学意义。结果表明:在光学显微镜下,18种鳞盖蕨属植物叶上、下表皮的脉上均有毛,叶表皮细胞都为不规则形,垂周壁为深波状或波状; 气孔都分布在下表皮,气孔器类型主要有极细胞型和腋下细胞型两种,叶表皮微形态特征支持鳞盖蕨属是碗蕨科中的一个单系类群; 叶上、下表皮脉间毛的变异特征支持光叶鳞盖蕨、毛叶边缘鳞盖蕨和二回边缘鳞盖蕨处理为边缘鳞盖蕨变种; 在扫描电镜下,叶上表皮角质层多为脊状凸起,多数还具丝状纹饰,角质膜特征与叶片回数有一定的相关性; 共环极细胞型、聚腋下细胞型、不等细胞型和不规则四细胞型只在少数种出现,气孔外拱盖多数凹陷,光滑或有颗粒; 气孔外拱盖内缘平滑、浅波状或齿状; 保卫细胞两极大多数有“T”型加厚,不同种间叶表皮微形态特征表现出一定差异。该研究结果为其分类、演化、系统位置等进一步研究提供了参考。     

Purification and characterization of phytocystatins from kiwifruit cortex and seeds

Kiwifruit cysteine proteinase inhibitors (KCPIs) were purified from the cortex and seeds of kiwifruit after inactivation of the abundant cortex cysteine proteinase actinidain. One major (KCPI1) and four minor cystatins were identified from Actinidia deliciosa ripe mature kiwifruit cortex as well as a seed KCPI from A. chinensis. The predominant cortex cystatin, KCPI1, inhibited clan CA, family C1 (papain family) cysteine proteinases (papain, chymopapain, bromelain, ficin, human cathepsins B, H and L, actinidain and the house dust mite endopeptidase 1), while cysteine proteinases belonging to other families, [clostripain (C11), streptopain (C10) and calpain (C2)] were not inhibited. Inhibition constants (K(I)) ranged between 0.001 nM for cathepsin L and 0.98 nM for endopeptidase 1. The K(I) (14 nM) for KCPI1 inhibiting actinidain is at least 2 orders of magnitude higher than for other plant proteinases measured. The cortex KCPI1 and a seed KCPI purified from seeds had the same N-terminal sequence (VAAGGWRPIESLNSAEVQDV). BLAST-matching the peptide sequence against an in-house generated Actinidia EST database, identified 81 cDNAs that exactly matched the measured KCPI1 peptide sequence. Peptide sequences of two other cortex KCPIs each exactly matched a predicted peptide sequence of a cDNA from kiwifruit. The predicted peptide sequence of KCPI1 of 116 amino acids encodes a signal peptide and does not contain cysteine. Without the signal peptide (mature protein), KCPI1 has a molecular mass of approximately 11 kDa, possesses the consensus sequence characteristic for the phytocystatins and shows the highest homology to a cystatin from Citrusxparadisi (52% identity). This is the first report of phytocystatins from the Ericales.     

小叶栒子的分类学研究

据有关模式和产地标本的研究，结合野外调查、叶表皮微形态观察结果，对小叶栒子（Cotoneaster microphyllus Wall.ex Lindl.)进行了分类学修订，将分属 ３系（Series Procumbens Klotz; Series Alpigeni和Series Marginati Klotz）的１５个名称归并作小叶栒子的同物异名。     

Regulation of the maize ubiquitin (Ubi-1) promoter in developing maize (Zea mays L.) seeds examined using transient gene expression in kernels grown in vitro

Kernel culture was assessed for evaluating novel gene expression in developing maize (Zea mays L.) seeds by comparing the transient expression of maize ubiquitin (Ubi-1) promoter-driven β-glucuronidase (GUS) delivered by particle bombardment in kernels grown in culture with those grown in planta. With kernels from either source, GUS expression, as determined by histochemical staining, was widespread in young, actively growing kernels, but it diminished with kernel age and by 25 days after pollination was found only in the embryo. Transient expression of Ubi-1 in kernels grown in vitro was not affected by wounding, ethylene treatment, pathogen invasion, or heat shock. In contrast, the plant hormones indole-3-acetic acid and kinetin both stimulated transient Ubi-1 expression in the endosperm, particularly at the periphery. Transient gene expression in developing maize seeds grown in vitro should allow for facile and rapid evaluation of the tissue-specificity and environmental responses of novel gene constructs in developing maize seeds. Received: 31 July 1997 / Revision received: 13 October 1997 / Accepted: 31 October 1997     

Plant tRNA nucleotidyltransferase

A method for purification of tRNA nucleotidyltransferase from Lupinus luteus L. seeds on a preparative scale is presented. This involved ammonium sulfate fractionation and chromatography on DEAE-cellulose, hydroxylapatite, and QAE-Sephadex A-50. The final enzyme prepartion was apparently homogeneous and a 2000-fold purification was achieved. Data presented suggest the existence of only one form of tRNA nucleotidyltransferase in dry lupin seeds.