膜荚黄芪种子中2种几丁质酶的分离纯化及活性测定

运用传统的层析技术对膜荚黄芪种子中两种几丁质酶进行分离纯化,并对分离纯化中各个步骤得到的蛋白进行了活性研究.结果表明:(1)粗提液的硫酸铵沉淀经再生几丁质亲和柱和凝胶过滤层析Sephadex G-75得到几丁质酶A.(2)粗提液的硫酸铵沉淀经离子交换色谱DE-52、CM、凝胶过滤层析Sephadex G-75得到几丁质酶B.(3)几丁质酶A、B的比活性分别为35.6 U/mg和4.1 U/mg.(4)从膜荚黄芪种子中分离纯化的几丁质酶A和B均为糖蛋白,含糖量分别为6.1%和5.8%.(5) SDS-PAGE显示,几丁质酶A、B的分子量分别为35.5 ku、39.6 ku;凝胶过滤层析测定几丁质酶A、B的分子量分别为36.9 ku、40.8 ku;表明几丁质酶A、B均为单亚基蛋白.     

荔枝草花的形态发生

在体式显微镜系统观察的基础上,对唇形科鼠尾草属植物荔枝草(Salvia plebeiaR.Br.)的花发育过程进行了扫描电镜观察.发现荔枝草的轮伞花序由多数交互对生的聚伞花序单位组成,花器官发育形式为向心式",各部分花器官从外向内依次形成;共形成4个雄蕊原基,其中2个雄蕊原基在形成后不再发育,另2个雄蕊原基每个均发育出1可育药室和1不育药室,不育药室膨大连接,并在花成熟之前参与组成特殊杠杆结构;子房四深裂"的形成实际是由4个原基分别发育,而后相互靠拢而成.     

丹参雄性不育系Sh-B的鉴定与花粉发育过程的解剖学研究

在显微水平上对新发现的丹参雄性不育系Sh-B花药发育过程进行了解剖学观察,并对其花粉活力和结实率进行了鉴定。结果显示:根据花器官及花药的形态、大小以及花丝的长度,可以将Sh-B不育株分为3个不育类型,即Sh-B1、Sh-B2和Sh-B3。这3种不育类型均属于雄性不育,其花丝不到正常可育株的1/2,花药干瘪而瘦小,内无花粉粒或花粉无活力;其根、茎、叶以及种子形态结构与正常可育植株基本相似。产生雄性不育的主要原因有:花粉囊药室内壁纤维层加厚,影响花药壁开裂;小孢子母细胞周围不产生胼胝质或产生的胼胝质很少;绒毡层细胞延迟解体;花粉粒畸形。在其花药发育的小孢子母细胞时期、四分体形成前期、单核期、双核期均可能产生雄性不育的小孢子或花粉粒。     

Phylogenetic relationships of Salvia (Lamiaceae) in China: Evidence from DNA sequence datasets

With 84 native species, China is a center of distribution of the genus Salvia (Lamiaceae). These species are mainly distributed in Yunnan and Sichuan provinces (southwestern China), notably the Hengduan Mountain region. Traditionally, the Chinese Salvia has been classified into four subgenera, Salvia, Sclarea, Jungia, and Allagospadonopsis. We tested this classification using molecular phylogenetic analysis of 43 species of Salvia from China, six from Japan, and four introduced species. The nuclear ribosomal internal transcribed spacer region and three chloroplast regions (rbcL, matK, and trnH-psbA) were analyzed by maximum parsimony, maximum likelihood, and Bayesian methods. Our results showed that the Chinese (except Salvia deserta) and Japanese Salvia species formed a well-supported clade; S. deserta from Xinjiang grouped with Salvia officinalis of Europe. In addition, all introduced Salvia species in China were relatively distantly related to the native Chinese Salvia. Our results differed from the subgeneric and section classifications in Flora Reipublicae Popularis Sinicae. We suggested that sections Eusphace and Pleiphace should be united in a new subgenus and that sect. Notiosphace should be removed from subg. Sclarea and form a new subgenus. Our data could not distinguish a boundary between subg. Allagospadonopsis and sect. Drymosphace (subg. Sclarea); the latter should be reduced into the former. Further clarification of the phylogenetic relationships within Salvia and between Salvia and related genera will require broader taxonomic sampling and more molecular markers.     

唇形科鼠尾草属的物种多样性与分布

为全面了解唇形科鼠尾草属(Salvia)植物的多样性和分布格局及其形成机制, 作者查阅了国际权威生物多样性信息网站(GBIF, The Plant List)、中国数字植物标本馆(CVH)、教学标本资源共享平台和中国自然保护区标本资源共享平台中该属物种名称及标本采集信息, 以及国内32家标本馆的标本, 分析并绘制其物种分布图。结果显示, 具有明确地理坐标的世界和中国分布信息分别有57,674条和11,596条, 已接受种名952个。在世界范围内, 以中南美洲(510种)物种数量最多, 其次是西亚(270种)、欧洲(117种)、东亚(97种)和北美(94种); 在国家尺度上, 以墨西哥物种数量最多(322种), 其次是俄罗斯(109种)、土耳其(88种)、美国(85种)和中国(82种)。在中国, 以云南和四川省鼠尾草种数最多(合计占全国的63%), 两省分布最多的县域地区分别是玉龙县(23种)、香格里拉县(20种)、大理市(13种)和木里县(17种)、宝兴县(13种)、马边县(13种)。在自然地理区域上, 以横断山区最为丰富, 占该属全国物种总数的52.8%, 特有种达23种; 广布种以荔枝草(S. plebeia)分布的县域数量最多(395县), 其次是鼠尾草(S. japonica) (199)、丹参(S. miltiorrhiza) (192)、贵州鼠尾草(S. cavaleriei) (173)、华鼠尾草(S. chinensis) (153)和粘毛鼠尾草(S. roborowskii) (100)。鼠尾草属主要分布于北半球温带及亚热带高海拔地区, 中国是东亚的多样性分布中心, 代表性广布种及狭域特有种均有分布, 尤以云南、四川以及横断山区的物种多样性和特有种比例最高。     

Encapsulation-Dehydration and Encapsulation-Vitrification Based Cryopreservation of in vitro-Grown Shoot-Tips of Medicinal Plant Astragalus membranaceus

Shoot tips of Amembranaceus excised from in vitro grown axillary bud were encapsulated in calcium alginate beads. Subsequently, shoot tips were precultured in liquid MS medium enriched with 075mol·L-1 sucrose for 5d at 25℃ and then desiccated aseptically on dried silica gel for 5h to a water content of 231% (fresh weight basis) prior to immersion in liquid nitrogen (LN) for 1d. After rewarming at a 40℃ water bath for 2-3min and transferred to solid culture medium for shoot tip recovery. About 50% of cryopreserved shoot tips grew into shoots within 2 weeks after plating. Cryopreservation of Astragalus membranaceus (Fisch.) Bge. shoot tips by encapsulation vitrification has also been developed. Excised shoot tips were firstly encapsulated into alginate gel beads and then precultured in liquid MS medium containing 1mg·L-1 6 BA, 005mg·L-1 NAA and 075mol·L-1 sucrose at 25℃ for 3d. After loading for 90min with a mixture of 2mol·L-1 glycerol and 04mol·L-1 sucrose at 25℃, shoot tips were dehydrated with PVS2 for 120min at 0℃ prior to direct immersion in liquid nitrogen for 1d. After rapidly thawing at a 37℃ water bath for 2-3min, shoot tips were washed for 10min with liquid MS medium supplemented with 1mg·L-1 6 BA, 005mg·L-1 NAA and 12mol·L-1 sucrose at 25℃ and then post cultured on solid MS medium supplemented with 2mg·L-1 6 BA, 005mg·L-1 NAA. The regeneration rate of shoot tips amounted to nearly 80%. Both of plantlets regenerated from cryopreserved shoot tips were morphologically uniform, which both showed as that of control plants. Thus, this encapsulation dehydration and encapsulation vitrification technique appears promising as a routine method for the cryopreservation of shoot tips of Amembranaceus.     

The anatomical and micromorphological properties of three endemic and medicinal Salvia species (Lamiaceae) in Erzincan (Turkey)

In this study, we comparatively investigated three endemic Salvia species spreading in Erzincan (Turkey) in terms of anatomy and micromorphology. For anatomical investigation, cross sections taken from stems and leaves of the species were examined under light microscope. For micromorphological investigation, epidermal surface and nutlet structure were examined using scanning electron microscopy. The S. euphratica and S. divaricata species were examined anatomically and micromorphologically for the first time, and S. hypargea was examined micromorphologically for the first time. In anatomical examinations, it was seen that stem and leaf structures of the species were similar. In micromorphological analyses, it has been seen that hairiness of the nutlet surface and nutlet ornamentations (verrucate and rugose type) created a difference between the species.     

Antimicrobial Activity and Mechanism of Essential Oil of Endemic Salvia hypargeia Finc. & Mey. in Turkey

In this study, the aerial parts of Salvia hypargeia were subjected to hydrodistillation and the resulting compounds were analyzed in GC–MS. Antimicrobial activity of S. hypargeia essential oil (EO) against A. baumannii, S. aureus, and C. tropicalis were determined by agar well diffusion assay and microdilution method. Antimicrobial mechanism of the EO were investigated based on change of TTC-dehydrogenase relative, leakages of intracellular protein, DNA and potassium ion (K +). The main components of the EO were β-pinene, 1,8-cineole, camphor, α-pinene, 4-terpineol, and 4-thujanol. The MICs of the EO against the microorganisms were 15.2 mg/mL for S. aureus, 17.5 mg/mL for C. tropicalis and 28.8 mg/mL for A. baumannii. The inhibition zones were 18.16 mm, 25.01 mm, and 27.01 mm for A. baumannii, S. aureus, and C. tropicalis, respectively (p < 0.05). An increase in DNA, protein and K⁺ leakage was observed when microorganisms were exposed to the EO. The TTC-DRA of the treated microorganism cells was also significantly decreased because of slowing the respiration. The present study provided an experimental basis of practical application of S. hypargeia EO as a natural agent.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12088-021-00939-1.     

Non-random organization of the Biomphalaria glabrata genome in interphase Bge cells and the spatial repositioning of activated genes in cells co-cultured with Schistosomamansoni

Biomphalaria glabrata is a major intermediate host for the parasitic trematode Schistosoma mansoni, a causative agent of human schistosomiasis. To decipher the molecular basis of this host-parasite interaction, the Bge embryonic cell line provides a unique in vitro model system to assess whether interactions between the snail and parasite affect the cell and genome biology in either organism. The organization of the B. glabrata genome in Bge cells was studied using image analysis through positioning territories of differently sized chromosomes within cell nuclei. The snail chromosome territories are similar in morphology as well as in non-random radial positioning as those found in other derived protostome and deuterostome organisms. Specific monitoring of four gene loci, piwi, BgPrx, actin and ferritin, revealed non-random radial positioning of the genome. This indicates that specific parts of the snail genome reside in reproducible nuclear addresses. To determine whether exposure to parasite is reflected in genome organization, the interphase spatial positioning of genes was assessed after co-culturing Bge cells with either normal or irradiation attenuated miracidia for 30 min to 24 h. The loci of actin and ferritin, genes that are up-regulated in the snail when subjected to infection, were visualized by fluorescence in situ hybridisation (FISH) and their radial nuclear positions i.e. their position in the interphase nucleus with respect to the nuclear edge/envelope, mapped. Interestingly, large scale gene repositioning correlated to temporal kinetics of gene expression levels in Bge cells co-cultured with normal miracidia while irradiated parasites failed to elicit similar gene expression or gene loci repositioning as demonstrated using the ferritin gene. This indicates that normal but not attenuated schistosomes provide stimuli that evoke host responses that are reflected in the host’s nuclear architecture. We believe that this is not only the first time that gene-repositioning studies have been attempted in a mollusc but also demonstrates a parasite influencing the interphase genome organization of its host.     

丹参异戊烯焦磷酸异构酶基因（SmIPI）的生物信息学及表达分析

异戊烯焦磷酸异构酶（IPI）是萜类合成途径的关键酶之一。本文在丹参转录组高通量数据分析的基础上,对丹参IPI基因（SmIPI）进行了克隆及序列分析。SmIPI脸长1234bp,包含681bp的开放读码框,编码226个氨基酸。生物信息学结构分析表明,SmIPI亲水性α／β蛋白,包含有IPI结构域,在序列组成、结构及活性位点等方面与其他植物的IPI均具有高度的相似性。实时荧光定量PCR分析结果表明,SmIPI在丹参生长的各个时期和不同组织器官中差异表达,其表达受病原菌和茉莉酸甲酯的诱导。