Meconopsis wilsonii subsp.wilsonii (Papaveraceae) Rediscovered

Field studies in the Lamagetou Nature Reserve,Mianning Xian,Sichuan Province,in the summer of2005revealed a particularlyrichand diverseflora.Thevegetationinthe area comprises thickets and forests ofmixed shrubs(Salix,Lonicera,Rhododendron)andtrees(Abies,P…     

濒危植物小勾儿茶伴生群落特征研究

从植物生活型、群落垂直结构、物种多样性以及群落稳定性4个方面研究小勾儿茶伴生群落特征,分析小勾儿茶在该生境幸存的原因。生活型谱反映出生境温热高湿且四季分明的亚热带气候特点;小勾儿茶处于群落顶层,能够获得充足的阳光;群落结构稳定、物种丰富。小勾儿茶的数量已极为稀少,应采取调查、保护和扩繁3种主要措施扩大其种群数量,即：(1)加大力度在相似的生境中寻找小勾儿茶;(2)对其生境加大保护力度;(3)开展种子生理方面的研究,探索有效的发芽途径,同时用组培的方法繁殖幼苗。     

青杄PSAK的克隆及生物信息学分析

目的:获取青杄的PSAK全长cDNA序列,对其进行生物信息学分析,并鉴定其在青杄各组织中相对表达量。方法:以多年生青杄(Picea wilsonii)的cDNA文库为模板,通过RACE PCR的方法获取PSAK的末端序列,经过与EST序列拼接得到PSAK基因的cDNA全长序列。通过特异引物将其克隆在pEASY-T1载体上,基于其氨基酸序列,利用DNAMAN、ExPASy、SWISS-MODEL等工具对其进行生物信息学分析,进行了半定量RT-PCR与RT-qPCR实验来检测其在mRNA水平的组织表达情况。结果:青杄PSAK基因编码140个氨基酸,蛋白分子量为14.23kDa,理论等电点为10.29。蛋白的C端有较为保守的结构域,与拟南芥、烟草等物种具有较高的相似性。PwPSAK主要在青杄的针叶与茎中进行表达。结论:成功获取了PwPSAK的单克隆并进行了生物信息学分析,为青杄中PSAK后续的功能研究提供理论依据。     

香圆种子萌发特征及催芽技术

以香圆（Citrus wilsonii Tanaka）种子为实验材料,通过测定种皮透水性、种皮机械障碍、发芽温度等对种子萌发的影响从而探讨种子萌发特征;通过切割种子、热水浸种、赤霉素浸种、低温层积等处理方式探索种子的催芽方法。结果显示：香圆种皮不存在透水性障碍,但其机械障碍对种子萌发具有抑制作用,且内种皮对萌发影响较大;种子在15℃、25℃、35℃恒温及15℃/30℃变温条件下均可萌发,其最适萌发温度为25℃;采用低温层积、热水浸种、切除1/3种子等方法均可一定程度上促进种子萌发。生产中宜采用始温60℃热水浸种24h的方法对香圆种子进行催芽。     

Rediscovery of Berchemiella wilsonii (Schneid.) Nakai

Berchemiella wilsonii (Schneid.) Nakai, an endangered species in the Rhamnaceae, was rediscovered in the Houhe National Nature Reserve, Wufeng County, Hubei Province, China, a place about 130 km away from its type locality, Xingshan County, where Wilson i     

Seed Dormancy,Germination and Storage Behavior of Magnolia wilsonii (Magnoliaceae), an Endangered Plant in China

In this paper, we measured the embryo morphology and size of Magnolia wilsonii seeds. And the effects of temperature, stratification, KNO3 and soaking on seed dormancy and germination were studied. Seeds were desiccated to different moisture content, and then stored at 4℃ and -20℃ for 100 d. The effects of desiccation and storage temperature on seed viability of M. wilsonii were analyzed. The results showed that the underdeveloped embryos of Mwilsonii seeds needed cold stratification to achieve physiological afterripening. The favorable temperature for germination of Mwilsonii seeds was 25/20℃. Cold stratification and GA3 could effectively overcome dormancy of Mwilsonii seeds. Therefore, Mwilsonii seeds have morphophysiological dormancy. About 5350% of seeds could survive in the moisture content of 539%. After 100 d storage at -20℃, all seeds in different moisture contents died. However, after 100 d storage at 4℃, 76% of seeds survived. Thus, Mwilsonii seeds exhibited intermediate seed storage behavior. The optimum storage environments was dry or moist storage at 4℃.     

濒危植物毛柄小勾儿茶生存群落的数量分类

应用除趋势对应分析(DCA)与双向指示种分析(TWINSPAN)对濒危植物毛柄小勾儿茶(Berchemiella wilsoniivar.pubipetiolata)69块样地进行了排序与分类,共划分9个群落类型。结果表明,毛柄小勾儿茶生境特异性高。毛柄小勾儿茶在枫香(Liquidambar formosana)林、马尾松(Pinusmassoniana) 杉木(Cunninghamia lanceolata) 青冈栎(Quercus glauca)针阔混交林、杂木林和青榨槭(Acer davidii) 大果山胡椒(Lindera praecox)林中生长较好。这4个群落林下层优势种具有相似性:草本层的优势种中均有求米草(Oplismenus undulatifolius),常有三脉紫菀(Aster ageratoides);灌木层优势种常有大果山胡椒。毛柄小勾儿茶分布于海拔500~900 m的低山区,多分布于湿润、排水良好的沟谷边,生境坡向多为阴坡。     

青杄PsbO基因cDNA序列克隆及生物信息学分析

利用RACE技术从已有的青杄均一化cDNA文库中克隆得到青杄(Picea wilsonii)PsbO基因cDNA的全长,并用生物信息学的方法对该cDNA的核苷酸序列、氨基酸序列的相似性,及编码蛋白PwPsbO的理化性质、亲水性/疏水性、二级和三级结构,PwPsbO基因进化树等进行了预测和分析.采用实时荧光定量PCR(RT-qPCR)技术,测定了青杄不同组织中的PsbO基因的相对表达水平.结果发现:青杄PsbO基因编码346个氨基酸,预测相对分子质量为36.6 kD,理论pI值为5.29,属于可溶性蛋白.二级和三级结构预测表明其含有较多的α螺旋、β折叠和随机卷曲结构.RT-qPCR发现该基因在青杄针叶和茎中表达量最高.这些结果为青杄中PsbO基因的初步功能研究奠定了基础.     

青杄均一化cDNA文库构建及EST序列分析

以青杄花粉和针叶为材料,将青杄全长cDNA与Gateway供体载体pDONR222重组,构建了其非剪切型全长cDNA原始文库,利用基因组DNA饱和杂交技术对原始cDNA文库进行均一化处理,构建青杄的均一化全长cDNA文库。文库的总库容量为1.1×106CFU/mL,平均插入片段长度大于1.0 kb,重组率大于95%。定量RT-PCR检测表明,青杄高丰度表达基因EF1-α在均一化cDNA文库中的表达量下降了约41倍。接着对文库中随机的5 144个克隆进行了测序,获得高质量的有效EST(expressedsequence tag)序列为5 144条,经拼接共获得单一基因(unigene)为2 717个,其中包括片段重叠群(contig)628个和单一EST序列(singlet)2 089个。NCBI同源比对分析表明,其中1 887个序列unigenes获得分子功能注释,这些EST涉及细胞生长、信号转导、转录、抗逆、能量代谢等功能。这些数据有助于对青杄的相关功能蛋白及分子机制开展进一步的研究。     

Observation on Differentiation Potential and Chromosome Stability of Callus in Subcultures of Picea wilsonii

Embryogenic calli of Picea wilsonii Mast. cultured on modified 59 medium supplemented with 1 ppm 2, 4-D and 1 ppm Kt still remained high reproductive potential after sequential monthly subcultures for 3 years. When the callus were transferred to 1/2 strength modification 59 medium with 1 ppm ABA, a lot of somatic embryos were produced after culturing about 3 monthes. The frequency of callus embryogenesis was above 90 %. Chromosome counting showed that the chromosome ploidy of callus cells of P. wilsonii was stable in subcultures for 3 years. The chromosome number and karyotype formula is 2n= 24= 16m (6sc) +8sm+2B. This result agrees with that form root-tip squash of seedling.     

青杆PwUSP1基因的克隆及表达模式分析

广泛逆境胁迫蛋白（universalstressprotein,USP）在非生物胁迫响应中起重要作用,但在植物中其功能还大部分未知。本研究通过BLAST分析青杆EST文库,得到职zP基因的EST序列,通过RACEPCR方法获取USP基因的末端序列,经过与EST序列拼接得到USP基因的cDNA全长序列,命名为PwUSP1。分析发现PwUSP1全长cDNA为1167bp,编码区为519bp,编码172个氨基酸残基。生物信息学分析显示,PwUSP1编码的蛋白相对分子质量为19．07kDa,理论等电点为6．38,为非跨膜的亲水性蛋白。PwUSP1具有USP家族典型的UspA结构域和ATP结合位点G-（2x）-G-（9x）-G（S／T）。半定量RT-PCR与RT-qPCR分析表明,PwUSP1在青杆的根、茎、针叶、花粉、种子中均有表达,在根和花粉中表达量较高。同时,PwUSP1受干旱和盐胁迫的诱导表达上调,均在处理6h后表达量较高,推测该基因可能在青杆逆境胁迫响应中发挥作用。     

濒危植物毛柄小勾儿茶种子休眠与萌发生理的初步研究

毛柄小勾儿茶（Berchemiella wilsonii （Schneid．）Nakai var．pubipetiolata H．Qian）为中国特有种,国家三级保护植物。TTC染色法测得毛柄小勾儿茶种子的生活力较低,仅为32％,空粒和无生活力的种子分别为28％和40％,说明毛柄小勾儿茶发育完全的种子较少。种子吸水速度试验证明毛柄小勾儿茶种皮透水性较差,不利于种子萌发时对水分的吸收,发芽抑制物的生物鉴定结果表明,毛柄小勾儿茶种子的不同部位都存在发芽抑制物,且种胚中的含量最高,不利于种子的萌发,使毛柄小勾儿茶种子处于休眠状态之中。几种不同浓度GA。溶液浸种24h的预处理可以不同程度地提高毛柄小勾儿茶种子的发芽率和发芽指数,其中300mg／L GA3的效果最显著,并且300mg／L GA3的预处理可以在萌发初期（萌发前3天）显著地提高毛柄小勾儿茶种子中过氧化物酶、过氧化氢酶和脱氢酶的活性。毛柄小勾儿茶种子生活力低,且处于休眠状态,再加上与种子发芽力有关的几种酶的活性较低,致使毛柄小勾儿茶种子的发芽率较低,同时这些也是从种子萌发生理方面导致毛柄小勾儿茶濒危的重要原因。     

珍稀濒危植物毛柄小勾儿茶的离体培养

以珍稀濒危植物毛柄小勾儿茶（Berchemiella wilsonii（Schneid．）Nakai var．pubipetiolata H．Qian）嫩梢为外植体进行组织培养实验,结果表明：以1／2 MS＋6-BA0．5mg／L＋NAA 0．05mg／L为芽诱导培养基、MS＋6-BA0．5mg／L＋NAA 0．05mg／L为增殖培养基,可以得到大量的毛柄小勾儿茶试管苗。     

Three-dimensional reconstruction of Picea wilsonii Mast. pollen grains using automated electron microscopy

Three-dimensional electron microscopy(3 D-EM) has attracted considerable attention because of its ability to provide detailed information with respect to developmental analysis. However, large-scale high-resolution 3 D reconstruction of biological samples remains challenging. Herein, we present a 3 D view of a Picea wilsonii Mast. pollen grain with 100 nm axial and38.57 nm lateral resolution using AutoCUTS-SEM(automatic collector of ultrathin sections-scanning electron microscopy). We established a library of 3,127 100 nm thick serial sections of pollen grains for preservation and observation, demonstrating that the protocol can be used to analyze large-volume samples. After obtaining the SEM images, we reconstructed an entire pollen grain comprising 734 serial sections. The images produced by 3D reconstruction clearly revealed the main components of the P.wilsonii pollen grain, i.e., two sacci and pollen corpus, tube cell, generative cell, and two degenerated prothallial cells, and their internal organization. In addition, we performed a quantitative analysis of the different pollen grain cells, including sacci, and found that there were 202 connections within a saccus SEM image. Thus, for the first time, this study provided a global 3D view of the entire pollen grain, which will be useful for analyzing pollen development and growth.     

The block of intracellular calcium release affects the pollen tube development of Picea wilsonii by changing the deposition of cell wall components

Two potent drugs, neomycin and TMB-8, which can block intracellular calcium release, were used to investigate their influence on pollen tube growth and cell wall deposition in Picea wilsonii. Apart from inhibiting pollen germination and pollen tube growth, the two drugs largely influenced tube morphology. The drugs not only obviously disturbed the generation and maintenance of the tip-localized Ca(2+) gradient but also led to a heavy accumulation of callose at the tip region of P. wilsonii pollen tubes. Fourier transform infrared (FTIR) spectroscopy analysis showed that the deposition of cell wall components, such as carboxylic acid, pectins, and other polysaccharides, in pollen tubes was changed by the two drugs. The results obtained from immunolabeling with different pectin and arabinogalactan protein antibodies agreed well with the FTIR results and further demonstrated that the generation and maintenance of the gradient of cross-linked pectins, as well as the proportional distribution of arabinogalactan proteins in tube cell walls, are essential for pollen tube growth. These results strongly suggest that intracellular calcium release mediates the processes of pollen germination and pollen tube growth in P. wilsonii and its inhibition can lead to abnormal growth by disturbing the deposition of cell wall components in pollen tube tips.     

Somatic Embryogenesis and Plantlet Formation of Picea wilsonii Mast. in Ditterent Conditions

Embryogenic callus (translucent callus) was produced from immature zygotic embryos of Picea wilsonii Mast. Subsequently somatic embryogenesis occurred on the brown callus. The somatic embryos could be stimulated to developinto plantlets on the medium without hormone. Young somatic embryos were produced from embryogenic callus in liquid suspension culture, in which suspensor was several or more than ten times the size of the somatic embryo. The somatic embryo showed very similar to zygotic embryos in micro-section and living material.     

濒危物种小勾儿茶的重新发现

在距离濒危物种小勾儿茶模式标本产地约130 km处的湖北省五峰县后河国家自然保护区中重新发现了小勾儿茶Berchemiella wilsonii (Schneid.) Nakai。威尔逊1907年在湖北省兴山县采集到小勾儿茶的模式标本有花,无果实。我们在五峰县采集了小勾儿茶的花和果实标本并对其果实特征予以补充描述。     

青杄中sPPa1的cDNA序列克隆及其生物信息学分析

以青杄(Picea wilsonii)均一化cDNA文库为模板,通过RACE方法克隆得到青杄PPa1基因cDNA全长,对该cDNA序列、核苷酸序列的相似性、理化性质、疏水性、二级结构、三级结构及是否跨膜进行了分析预测;进行了多序列比对并构建了系统树,同时对PPa1在青杄各组织中的表达量进行了检测.结果表明:青杄PPa1基因共由216个氨基酸组成,分子量为24.55 kD,理论PI为5.83,属可溶性蛋白;二级结构主要由α-螺旋、不规则卷曲和β-折叠构成;PPa1在青杄花粉中表达量最高.研究为进一步研究青杄PPa1的功能奠定了基础.     

湖北川桂叶精油的化学成分

川桂 ( Cinnamomum wilsonii Gamble)是中国特有植物 ,主产湖北、四川和湖南 ,在广西、广东、江西和陕西等省区也有分布 ,资源贮量相当丰富。由于其各部位器官均含有挥发油 ,具有比较浓烈的药香气味 ,我国民间历来习惯于利用川桂作为辛辣类佐料 ,烹调制作食品 ,或当作桂皮类中药材用以治疗疾病。因此 ,关于川桂挥发油 (精油 )的化学组成颇为引人关注。我们曾经研究报道过 ,湖北长阳和咸丰两县产川桂叶精油的主成分均为柠檬醛 ,其含量分别高达77.99%和 86.47% [1]。另据报道 ,四川产川桂叶精油的主要成分为芳樟醇 ( 2 4 .1 4% )和 1 ,8-桉叶…     

Crystal structure of a pro-inflammatory lectin from the seeds of Dioclea wilsonii Standl

The crystal structure and pro-inflammatory property of a lectin from the seeds of Dioclea wilsonii (DwL) were analyzed to gain a better understanding of structure/function relationships of Diocleinae lectins. Following crystallization and structural determination by standard molecular replacement techniques, DwL was found to be a tetramer based on PISA analysis, and composed by two metal-binding sites per monomer and loops which are involved in molecular oligomerization. DwL presents 96% and 99% identity with two other previously described lectins of Dioclea rostrata (DRL) and Dioclea grandiflora (DGL). DwL differs structurally from DVL and DRL with regard to the conformation of the carbohydrate recognition domain and related biological activities. The structural analysis of DwL in comparison to other Diocleinae lectins can be related to the differences in the dose-dependent pro-inflammatory effect elicited in Wistar rats, probably via specific interactions with mast cells complex carbohydrate, resulting in significant paw edema. DwL appears to be involved in positive modulation of mast cell degranulation via recognition of surface carbohydrates. Since this recognition is dependent on site volume and CRD configuration, edematogenesis mediated by resident cells varies in potency and efficacy among different Diocleinae lectins.