大叶藤黄叶片角质层的酶分离技术

角质层运输特征的研究在生态学、环境和农用化学等方面都具有非常重要的意义 ,完好地分离角质层是进行这方面研究的前提。本文以生长在西双版纳热带雨林中的大叶藤黄为材料 ,探讨了角质层的酶分离技术。结果表明 ,30℃时用 2 % (w/v) ,pH 3 0的果胶酶和纤维素酶溶液浸泡大叶藤黄叶圆片 9h ,叶圆片边缘角质层分离 ,浸泡 5d可分离到完整的角质层 ,角质层在硼酸盐缓冲液 (0 0 1mol·L-1,pH 9 0 )中充分浸泡数日去除石炭酸 ,空气中干燥 ,室温下于聚乙烯的盒子保存备用     

一株鸭源传染性支气管炎病毒的分离鉴定及结构蛋白基因和血清型分析

【目的】对从广西某鸭场发生呼吸道感染的11天龄樱桃谷肉鸭分离到的病毒株进行鉴定,并探索此鸭源病毒分离株的遗传变异情况。【方法】通过血凝试验、鸡胚接种实验、3?端非编码区(3'UTR)基因扩增与序列测定对分离株进行鉴定,并对该分离株的结构基因S1、E、M和N分别进行序列测定以及相似性、系统进化树分析和血清型鉴定。【结果】血凝试验为阴性,接种鸡胚盲传5代后出现侏儒胚,3?UTR基因测序结果表明为传染性支气管炎病毒(IBV)序列。该分离株S蛋白的裂解位点为RRSRR,S1、E、M和N基因与IBV毒株H120、4/91、LTD3核苷酸相似性分别为:78.6%–99.7%、85.4%–100.0%、91.6%–93.2%、86.7%–91.7%。除N基因存在点突变外,S1、E和M基因均存在氨基酸的突变、插入和(或)缺失。系统进化树分析显示,其S1基因属于4/91型,E、M和N基因均为LDT3型。血清型分析表明,该分离株的血清型不同于疫苗株H120和4/91。【结论】此鸭源病毒分离株为IBV,且该分离株的基因型与血清型均发生了变异。本研究结果暗示禽类传染性支气管炎的防控面临着更严峻的挑战。     

广西岩溶区烟草黑胫病拮抗细菌的筛选鉴定及其抗病机理

从广西岩溶区靖西县优质烟叶生产区分离烟草黑胫病病原及土壤细菌,通过拮抗试验、离体及田间抗病能力测试等方法筛选拮抗细菌;利用形态观察、生理生化测试和16SrDNA序列分析三种方法相结合,对抗病效果良好的菌株进行分类鉴定;并对拮抗菌抗黑胫病机理进行初步研究。结果表明:从8份土壤样品中,共分离出土壤细菌340株,获得抗黑胫病效果良好的拮抗细菌3株,编号为8-23、6-70和13,它们分别属芽孢杆菌属、溶菌杆属和假单胞杆菌属细菌;三个拮抗细菌的抗黑胫病机理是通过胞外分泌一些可溶解病原菌细胞壁的酶或其它化学物质,破坏菌丝的细胞壁和细胞膜等生理结构,使细胞质渗漏、凝集,从而导致病原死亡;其中,菌株8-23和13的抗病活性物质主要为蛋白质类化合物;而菌株6-70除蛋白质外,还有其它一些非蛋白因子起作用。     

FEMALE HETEROGAMETY AND SPECIATION: REDUCED INTROGRESSION OF THE Z CHROMOSOME BETWEEN TWO SPECIES OF NIGHTINGALES

Several lines of evidence suggest that the X chromosome plays a large role in intrinsic postzygotic isolation. The role of the Z chromosome in speciation is much less understood. To explore the role of the Z chromosome in reproductive isolation, we studied nucleotide variation in two closely related bird species, the Thrush Nightingale ( Luscinia luscinia ) and the Common Nightingale ( L. megarhynchos ). These species are isolated by incomplete prezygotic isolation and female hybrid sterility. We sequenced introns of four Z-linked and eight autosomal loci and analyzed patterns of polymorphism and divergence using a divergence-with-gene flow framework. Our results suggest that the nightingale species diverged approximately 1.8 Mya. We found strong evidence of gene flow after divergence in both directions, although more introgression occurred from L. megarhynchos into L. luscinia . Gene flow was significantly higher on the autosomes than on the Z chromosome. Our results support the idea that the Z chromosome plays an important role in intrinsic postzygotic isolation in birds, although it may also contribute to the evolution of prezygotic isolation through sexual selection. This highlights the similarities in the genetic basis of reproductive isolation between organisms with heterogametic males and organisms with heterogametic females during the early stages of speciation.     

Ancestral and neo-sex chromosomes contribute to population divergence in a dioecious plant

Empirical evidence from several animal groups suggests sex chromosomes disproportionately contribute to reproductive isolation. This effect may be enhanced when sex chromosomes are associated with turnover of sex determination systems resulting from structural rearrangements to the chromosomes. We investigated these predictions in the dioecious plant Rumex hastatulus, which is composed of populations of two different sex chromosome cytotypes caused by an X-autosome fusion. Using population genomic analyses, we investigated the demographic history of R. hastatulus and explored the contributions of ancestral and neo-sex chromosomes to population genetic divergence. Our study revealed that the cytotypes represent genetically divergent populations with evidence for historical but not contemporary gene flow between them. In agreement with classical predictions, we found that the ancestral X chromosome was disproportionately divergent compared with the rest of the genome. Excess differentiation was also observed on the Y chromosome, even when we used measures of differentiation that control for differences in effective population size. Our estimates of the timing of the origin of neo-sex chromosomes in R. hastatulus are coincident with cessation of gene flow, suggesting that the chromosomal fusion event that gave rise to the origin of the XYY cytotype may have also contributed to reproductive isolation.     

WIDESPREAD HOST‐DEPENDENT HYBRID UNFITNESS IN THE PEA APHID SPECIES COMPLEX

Linking adaptive divergence to hybrid unfitness is necessary to understand the ecological factors contributing to reproductive isolation and speciation. To date, this link has been demonstrated in few model systems, most of which encompass ecotypes that occupy relatively early stages in the speciation process. Here we extend these studies by assessing how host‐plant adaptation conditions hybrid fitness in the pea aphid, Acyrthosiphon pisum. We made crosses between and within five pea aphid biotypes adapted to different host plants and representing various stages of divergence within the complex. Performance of F1 hybrids and nonhybrids was assessed on a “universal” host that is favorable to all pea aphid biotypes in laboratory conditions. Although hybrids performed equally well as nonhybrids on the universal host, their performance was much lower than nonhybrids on the natural hosts of their parental populations. Hence, hybrids, rather than being intrinsically deficient, are maladapted to their parents’ hosts. Interestingly, the impact of this maladaptation was stronger in certain hybrids from crosses involving the most divergent biotype, suggesting that host‐dependent postzygotic isolation has continued to evolve late in divergence. Even though host‐independent deficiencies are not excluded, hybrid maladaptation to parental hosts supports the hypothesis of ecological speciation in this complex.     

Isolation of DNA for RAPD analysis from dry leaf material of some<Emphasis Type="Italic">Hesperis</Emphasis> L. specimens

An improved protocol for the isolation of DNA from dry material of someHesperis specimens is described. The isolated DNA is suitable for random amplification of polymorphic DNA (RAPD) analysis. Different DNA extraction protocols were examined to determine which might yield DNA from dry leaf tissue ofHesperis specimens. The methods examined include the protocols with hexadecyltrimethylammonium bromide (CTAB) described by Doyle and Doyle (1987); sodium dodecyl sulfate (SDS) by Dellaporta et al. (1983); and CTAB and SDS, the modified minipreparation, by Dellaporta et al (1983). None of these procedures yielded DNA of suitable purity for RAPD assay. We established an improved procedure involving CTAB and enzymatic digestion of proteins and RNA. The recovery of DNA with an average yield of 25 mg/g of leaf material was possible with this procedure. RAPD bands, which could be used to distinguish amongHesperis specimens, were generated.     

1株高产多糖红曲霉菌株的分离鉴定及生物学特性研究

从市售红曲米和红腐乳中分离纯化得到56株红曲霉菌株,通过摇瓶发酵试验筛选出1株胞外多糖产量为4.73 g/L的红曲霉菌株M-6;依据形态特征、生理生化特征和ITS序列将菌株M-6鉴定为红色红曲霉（Monascus rubber）。并对其部分生物学特性进行了研究。     

类芒齿黄芪地下部分化学成分研究

为了解类芒齿黄芪( Astragalus camptodontoides)主要化学成分,从其地下部分甲醇提取物的乙酸乙酯部位分离出19个单体化合物,通过现代波谱分析及理化性质等手段分别鉴定为异补骨脂黄酮(1),4′-hydroxy-isolonchocarpin (2),5-去氧山豆根黄酮(3),shinflavanone (4),khonklonginols H (5),4′-O-methylpreglabridin (6),3′-hydroxy-4′-O-methylglabridin (7),4′-O-methylglabridin (8),8-prenyl-phaseollinisoflavan (9),xambioona (10),光甘草酚(11),粗毛甘草素H (12),methylnissolin (13),邻苯二甲酸异丁酯(14),邻苯二甲酸丁酯异丁酯(15),β-谷甾醇(16),胡萝卜苷(17),齐墩果酸(18),(2S,3S,4R,9E)-1,3,4-trihydroxy-2-[(2′R)-2′- hydr-oxytetraco-sanoylamino]-9-octadecene (19)。化合物1~19均为首次从该植物中获得,化合物1~7为首次从黄芪属(Astraga-lus)植物中分离得到。