Distribution and biosynthesis of 20-hydroxyecdysone in plants of Achyranthes japonica Nakai

There is increasing interest in phytoecdysteroids (PEs) because of their potential role in plant defense against insects. To understand the mechanism regulating their levels in plants, the fluctuation, distribution, and biosynthesis of PE 20-hydroxyecdysone (20E) examined in Achyranthes japonica. The total amount of 20E per individual plant initially remained at a constant level, and increased markedly after the first leaf pair (LP) stage, while the concentration of 20E in a given plant decreased rapidly during vegetative growth. In addition, the incorporation of [2-(14)C]-mevalonic acid into 20E did not differ significantly depending on plant organs and developmental stages, suggesting that biosynthesis of 20E is not restricted to particular organs or growth stages.     

Distribution and Biosynthesis of 20-Hydroxyecdysone in Plants of Achyranthes japonica Nakai

There is increasing interest in phytoecdysteroids (PEs) because of their potential role in plant defense against insects. To understand the mechanism regulating their levels in plants, the fluctuation, distribution, and biosynthesis of PE 20-hydroxyecdysone (20E) examined in Achyranthes japonica. The total amount of 20E per individual plant initially remained at a constant level, and increased markedly after the first leaf pair (LP) stage, while the concentration of 20E in a given plant decreased rapidly during vegetative growth. In addition, the incorporation of [2-¹⁴C]-mevalonic acid into 20E did not differ significantly depending on plant organs and developmental stages, suggesting that biosynthesis of 20E is not restricted to particular organs or growth stages.     

The botany,uses and production ofWasabia japonica (Miq.) (Cruciferae) Matsum

Wasabi (Wasabia japonica) is a unique native plant and a traditional condiment crop of Japan. It is used in traditional Japanese raw fish and noodle dishes and in several modern foods for its hot taste and tangy flavor. Japanese farmers grow the crop in wet upland orchard soils for leaves, petioles and small enlarged stems, and in flooded gravel and sand fields along streams or near springs to produce whole plants and large succulent green enlarged stems. Recent studies in Japan have demonstrated numerous enzymatic and biocidal properties of the plant. This review of Japanese and other literature details the history, uses, botany, cultivars, ecological requirements, production techniques, insect pests and diseases of wasabi.     

基于mt-16S rDNA和mt-COI基因的海月水母分子生物学鉴定方法和检测技术

以我国近海海域常见的海月水母为目标生物,提取其水母基因组DNA,PCR扩增mt-16S rDNA(650 bp)和mt-COI基因(709 bp)的部分序列,经纯化、克隆、测序后,将获得的序列进行BLASTn分析,筛选目的序列中与其他水母差异较大的区域,设计出8对分别针对海月水母mt-16S rDNA和mt-COI基因序列的特异性引物.经过实验室培养样品和模拟现场样品检测发现,针对mt-16S rDNA的引物AS3和mt-COI的引物AC3具有较好的特异性,可以从海蜇、沙海蜇、霞水母、端鞭水母和海月水母中快捷地检测出目标水母,从而初步建立了海月水母的分子生物学鉴定方法和检测技术.本技术摆脱了水母形态学鉴定方法存在的局限,可准确、快速地对样品进行定性分析.     

直立百部遗传多样性的ISSR分析

利用ISSR分子标记,对直立百部4个居群的84个样本进行遗传多样性研究,探讨山东百部种的地位。结果表明:(1)15个引物共检测到184条清晰的谱带,其中多样性条带153条;直立百部的遗传多样性水平较高,其物种水平多样性条带百分率(PPF)为83.15%,有效等位基因数(Ae)为1.425,Shannon多样性指数(I)为0.388,Nei’s多样性指数(Hpop)为0.254。(2)居群平均水平上多样性条带百分率为63.04%,有效等位基因数为1.351,Shannon多样性指数为0.310,Nei’s多样性指数为0.206。(3)AMOVA分析显示直立百部居群间遗传分化水平(ΦST)为0.126 3,POPGENE分析显示居群间的遗传分化系数(GST)为0.191 1,二者均表明居群内变异大于居群间变异;直立百部居群间的基因流(Nm)为2.116 6,说明居群间的基因交流较频繁。(4)聚类及主成分分析显示,直立百部4个居群聚为两支,其中泰山居群与其他居群关系较远,以蔓生百部和大百部为外类群分析的结果支持将山东百部作为直立百部的异名,Mental检测显示居群间遗传距离与地理距离间没有显著相关性。     

菠菜rDNA及端粒多色荧光原位杂交分析

以拟南芥25S rDNA、5S rDNA以及端粒序列为探针对菠菜的中期染色体进行了多色荧光原位杂交分析, 其中25S rDNA用生物素标记, 端粒序列用地高辛标记, 5S rDNA用生物素和地高辛共同标记。杂交结果显示, 菠菜中期染色体25S rDNA杂交位点为6个, 分别位于3号、5号和6号染色体的随体部位; 5S rDNA杂交位点为4个, 分别位于3号和5号染色体长臂, 端粒序列杂交位点位于6号染色体端部以及其余染色体的端部和着丝粒部位。     

Phytochemical and chemotaxonomic study on Berchemiella wilsonii (Schneid.) Nakai

Phytochemical investigation of the aerial parts of Berchemiella wilsonii (Schneid.) Nakai (Rhamnaceae) led to the isolation of four flavonoids (1–4), three phenolic acids (5–7), two megastigmane derivatives (8–9) and one triterpene (10). The structures of these compounds were elucidated as taxifolin (1), (−)-epicatechin (2), quercetin 3-O-a-l-arabinopyranoside (3), vitexin (4), methyl p-hydroxycinnamat (5), 3,4-dihydroxybenzoic acid (6), 2-hydroxy-4-methoxy-3,6-dimethyl benzoic acid (7), (3S,5R,6R,7E,9S)-3,5,6,9-tetrahydroxy-7-en-megastigmane (8), (6S,9R)-roseoside (9) and lupeol (10) on the basis of NMR spectral data and comparison with literature values. These results are the first chemical constituent data of the genus Berchemiella, and the chemotaxonomic significance of these compounds is discussed.     

Molecular reexamination of korean umbelliferae based on internal transcribed spacer sequences of rDNA:Ligusticum tenuissimum (Nakai) Kitagawa andLibanotis coreana (Wolff) kitagawa

The taxonomic status ofLigusticum tenuissimum (Nakai) Kitagawa andLibanotis coreana (Wolff) Kitagawa has been controversial in Korea. We examined their phylogenetic relationships by comparing the internal transcribed spacer (=ITS) sequences of the nuclear ribosomal RNA gene (=rDNA) with another seven species in the Korean Umbelliferae. Our results support the legitimacy of the nomenclatural combination withL. tenuissimum Kitagawa rather than withAngelica tenuissima Nakai. We also suggest usingL coreana (Wolff) Kitagawa instead ofSeseli coreanum Wolff becauseL. coreana has been completely separated from most taxa of the genusSeseli. In addition, we have confirmed at least three phylogenetic lines in the genusLigusticum. One is the core group ofLigusticum that comprisesL tenuissimum and four other species. This clade includes theLigusticum hultenii line andLigusticum scothicum, known as an anomaly ofLigusticum. The second clade is composed ofLigusticum physospermifolium withSeseli monatum, which indicates the polyphyly ofLigusticum. The third line isLigusticum acutilobum, clustered as a sister grade ofDystaenia.     

槲寄生的研究与开发利用

本文对槲寄生的生物学特性、化学成分、药理作用及开发利用等作了阐述。     

Detection and identification of mycoplasmas by amplification of rDNA

Alignment of published 16S rRNA sequences allowed the definition of a pair of oligonucleotides suitable for polymerase chain reaction (PCR). Using this pair of PCR primers, several mycoplasmas including the four human parasites Mycoplasma genitalium, M. hominis, M. salivarium and M. orale were detected. This DNA amplification was restricted to species of the genus Mycoplasma while no cross-reaction was observed with DNA from other bacteria and eukaryotic cells. Subsequent analysis of amplified products by either specific oligonucleotide hybridization or dideoxy sequencing specified the identity of the detected mycoplasmas. This method offers a highly discriminating and sensitive assay for the direct detection and identification of these microorganisms without the need for prior cultivation.     

短果茴芹的组织培养与快速繁殖

1植物名称短果茴芹[Pimpinella brachycarpa(Kom.)Nakai]. 2材料类别小苗茎尖. 3培养条件(1)芽诱导培养基:MS 6-BA 2 mg·L-1(单位下同) NAA 0.01 IBA 1 KT 2;(2)继代培养基:MS 6-BA 2 IBA 0.1;(3)生根培养基:1/2MS NAA 0.5 IBA 0.3.上述培养基均附加30 g·L-1蔗糖和6.8 g·L-1琼脂,pH 5.8～6.0.室温20～25℃,自然光照10～12 h·d-1.     

九节茶的组织培养和快速繁殖

1植物名称 九节茶[Sarcandra glabra（Thunb．）Nakai],又名草珊瑚、肿节风、接骨莲。 2材料类别带节茎段。     

Comparative analysis of five varieties in <Emphasis Type="Italic">Perilla frutescens</Emphasis> (L.) Britton by 45S rDNA FISH and 5S rDNA sequences

In this study, the relationships of five varieties in Perilla frutescens (L.) Britton were analyzed. All the varieties showed the same chromosome number 2n = 40, and the banding patterns of PI-DAPI staining and the FISH results of 45S rDNA probe were on the same site of the chromosome pair. The 5S rRNA gene spacers were ranged from 413 bp in P. frutescens var. purpurascens and 408 bp in var. frutescens. The varieties analyzed here were clustered into two clades according to the phylogenetic analysis. The sequence identity of 96% between var. auriculato-dentata and var. arguta suggested they could be considered the same variety. The article is published in the original.     

Uniqueness of the Gossypium mustelinum Genome Revealed by GISH and 45S rDNA FISH

Gossypium mustelinum ((AD)₄) is one of five disomic species in Gossypium. Three 45S ribosomal DNA (rDNA) loci were detected in (AD)₄ with 45S rDNA as probe, and three pairs of brighter signals were detected with genomic DNA (gDNA) of Gossypium D genome species as probes. The size and the location of these brighter signals were the same as those detected with 45S rDNA as probe, and were named GISH-NOR. One of them was super-major, which accounted for the fact that about one-half of its chromosome at metaphase was located at chromosome 3, and other two were minor and located at chromosomes 5 and 9, respectively. All GISH-NORs were located in A sub-genome chromosomes, separate from the other four allopolyploid cotton species. GISH-NOR were detected with D genome species as probe, but not A. The greatly abnormal sizes and sites of (AD)₄ NORs or GISH-NORs indicate a possible mechanism for 45S rDNA diversification following (AD)₄ speciation. Comparisons of GISH intensities and GISH-NOR production with gDNA probes between A and D genomes show that the better relationship of (AD)₄ is with A genome. The shortest two chromosomes of A sub-genome of G. mustelinum were shorter than the longest chromosome of D sub-genome chromosomes. Therefore, the longest 13 chromosomes of tetraploid cotton being classified as A sub-genome, while the shorter 13 chromosomes being classified as D sub-genome in traditional cytogenetic and karyotype analyses may not be entirely correct.     

钩藤的组织培养与植株再生

1植物名称钩藤[Uncariarhynchophylla(Miq.)Jacks.]。2材料类别成熟种子。3培养条件以B5为基本培养基。(1)种子萌发培养基:加活性炭和不加活性炭的B5(不含任何激素);(2)增殖培养基:B5+6-BA2.0mg·L-1(单位下同)+NAA0.2+0.2%活性炭;(3)生根培养基:B5+NAA0.5+0.2%活性炭。上述培养基均附加3%蔗糖和0.8%琼脂粉,pH5.8。培养温度为(25±2)℃,光照时间12h·d-1,光强为30～40μmol·m-2·s-1。4生长及分化情况4.1无菌材料的获得参考张建康等(2005)的方法,选取成熟饱满、色泽好且未开裂的蒴果,先用70%的酒精进行表面消毒30s,再用0.1%的升汞溶…     

兰香草的组织培养与快速繁殖

1植物名称 兰香草[Caryopteris incana（Thunb．）Miq．]。 2材料类别 茎段。 3培养条件 以MS为基本培养基。芽诱导培养基：（1）MS＋6-BA0．5mg&#183;L^-1（单位下同）＋IBA0．2;（2）MS＋6-BA1．0＋IBA0．2。     

大麦45S和5S rDNA定位及5S rDNA伸展纤维的FISH分析

用荧光原位杂交技术对45S和5SrDNA在大麦(Hordeum vulgare L．)有丝分裂中期染色体进行了确定分析,较强的45SrDNA信号共有2对,分别分布在大麦的第1染色体的短臂和第2染色体的长臂。而5SrDNA则只有1对杂交信号,位于第3染色体的长臂,但信号较弱。用伸展DNA纤维的荧光原位杂交(Fiber—FISH)技术测定了5SrDNA在大麦的基因组中的拷贝数,计算出5SrDNA的拷贝数约为408～416。对大麦品种中rDNA位点数目的可变性进行了讨论。     

基于16S rDNA与Cytb序列的疏广蜡蝉属三个近似种的分子鉴定

本文通过PCR扩增首次获得了疏广蜡蝉属Euricania Melichar,1898 3个近似种:透明疏广蜡蝉E.clara Kato,1932、长刺疏广蜡蝉E.longa Xu,LiangJiang,2006和短刺疏广蜡蝉E.brevicula Xu,LiangJiang,2006的16S r DNA序列和Cytb序列;使用MEGA 6.0软件,分析了其序列组成及变异,计算了遗传距离,以宽广蜡蝉属Pochazia AmyotServille,1843的圆纹宽广蜡蝉P.guttifera Walker,1851和眼斑宽广蜡蝉P.discreta Melichar,1898为外群,利用邻接法(NJ)和最大简约法(MP)构建了系统发育树。结果显示:透明疏广蜡蝉、长刺疏广蜡蝉和短刺疏广蜡蝉的16S r DNA序列长度分别为414 bp,404 bp,435 bp,而Cytb基因序列长度分别为492 bp,468 bp,472 bp,三者的序列比对、碱基组成成分存在差异;2属5种广翅蜡蝉16S r DNA基因的种间遗传距离为0.008-0.098,属间遗传距离为0.122-0.197,而Cytb基因的种间遗传距离为0.038-0.055,属间遗传距离为0.181-0.188,表明16S r DNA和Cytb基因序列可作为DNA条形码候选基因片段,用于广翅蜡蝉近似属、种的分子鉴定。系统发育树的拓扑结构显示,3个近似种的亲缘关系基本一致,各自聚为一支,且置信值均达96%以上。本文亦结合了透明疏广蜡蝉、长刺疏广蜡蝉和短刺疏广蜡蝉形态学特征的差异对其进行了讨论和分析。