海洋生物技术在海水养殖动植物品种培育和病害防治中的应用

海水养殖是我国沿海地区发展经济的重要支柱产业。目前影响我国海水养殖业持续、稳定、健康发展的主要制约因素是苗种和病害问题。本文就海洋生物技术在海水养殖物植物品种培育和病害防治中的应用及有关问题进行了探讨,分别就应用海洋生物技术在群体和个体水平、细胞和染色体水平以及基因和分子水平培育新品种的工作,包括ＳＰＦ和ＳＰＲ苗种的培育、性别控制和杂交育种、多倍体育种、雌核发育、藻类细胞工程育种、转基因技术等进行了讨论。在重要水产养殖疾病病原的快速诊断和检测技术方面,重卢、讨论了核酸探针技术,ＰＣＲ技术和单克隆抗体技术。海洋生物技术在水产养殖疾病防治中的应用,重点讨论了疫苗和免疫促进剂的开发以及基因工程疫苗。本文还对我国海洋生物技术的发展战略及有关问题进行了探讨。     

Restriction site insertion-PCR (RSI-PCR) for rapid discrimination and typing of closely related microbial strains

Taking advantage of point mutations between DNA sequences of closely related microbial strains, PCR primers modified with respect to the target sequence at positions 2-5 near the 3' end were designed to obtain a fragment harbouring an artificial restriction site specific for a given strain. The modified forward primer coupled with a specific reverse primer allows for the amplification of DNA fragments which can be digested with the specific endonuclease only in those strains where the restriction site is inserted by the DNA polymerase. The effectiveness of the method, named restriction site insertion-PCR (RSI-PCR), was tested on isolates of the 'Bacillus cereus group' for the rapid typing and discrimination of these closely related strains.     

Tests of Bacterial Strain Dominance within a Mouse Based on Sampling Faecal Colonies

This paper deals with making inferences about the dominance of a bacterial strain in a mouse gut based on strain typing of a random sample of colonies derived from plated faecal material.     

Cationic (tripyrrinato)palladium(II) complexes

The formation of cationic palladium(II)complexes [TrpyPd^II]⁺X⁻ by salt metathesis of the respective trifluoroacetates with different salts of weakly coordinating anions X⁻ was investigated. With non-hydrolizable counterions, cationic mono- and dinuclear complexes are observed depending on the nature of the anion X⁻ and the solvent. The mononuclear cations, which are only formed with X = BAr^F, most probably carry a weakly bound molecule of dichloromethane at the fourth coordination site of Pd^II. When treated with diazoalkanes, only these are sufficiently reactive to form carbene complexes. Four- and five coordinate Lewis base adducts [TrpyPd^IIL]⁺ with L = CH₃NC, tBuNH₂, PMe₃, PEt₃ and PiPr₃ and [TrpyPd^IIL₂]⁺ with L = PMe₃ were prepared from the mononuclear cations [TrpyPd^II]⁺BAr^F−. From structural studies it becomes apparent, that the formation of stable five coordinate Pd^II species is restricted to medium size ligands and depends on the delicate balance between the steric influence of L and the strain, which is induced on the TrpyPd^II unit.     

Characterization of a novel South American population of the astaxanthin producing yeast <Emphasis Type="Italic">Xanthophyllomyces dendrorhous</Emphasis> (<Emphasis Type="Italic">Phaffia rhodozyma</Emphasis>)

A novel population of the biotechnologically important yeast Xanthophyllomyces dendrorhous, the sexual stage of Phaffia rhodozyma, has been recently isolated for the first time in the southern Hemisphere (Patagonia, Argentina). The aim of the present work was to phenotypically and genotypically characterize two representative strains of this new population, and assess such strains as a potential biotechnological source of astaxanthin, fatty acids and extracellular enzymes. Minor variations were found in physiological tests. PCR fingerprinting studies (MSP-PCR) showed the main differences between X. dendrorhous Patagonian and Type strains. Patagonian strains accumulated a xanthophyll-like pigment, which was identified as astaxanthin. These strains showed low fatty acids content (mainly polyunsaturated fatty acids) and, of a total of six extracellular enzymes tested, only produced amylase. Genetic differences between Patagonian and collection X. dendrorhous strains could be explained by geographic isolation and habitat specificity.     

Quantifying the instability of core temperature in rodents

Radiotelemetry provides researchers with the ability to sample the core temperature of rodents and other species rapidly. Compared to large mammals, the time-course of core temperature of rodents is variable and their data are often averaged into bins of one or more hours for publication. When viewing averaged data, a stable core temperature over a wide range of ambient temperatures is observed; however, if the time-course of core temperature of individual animals is examined closely with a sampling period of 1–10 min, a continuous waxing and waning of temperature is observed. It is proposed that calculating the change in temperature from one time point to the next (i.e., temperature differential) is an informative measure of the performance of the thermoregulatory system. A quantitative measure of the stability of the thermoregulatory system can be determined by calculating the absolute value of the temperature differential. It is shown that the absolute temperature differential (ATD) is dependent on ambient temperature, genetic strain, type of cage bedding, and body size. The ATD is simple to calculate from telemetry data and may be an important parameter for the study of rodent thermal physiology.     

Biological control of take-all in wheat by endophytic Bacillus subtilis E1R-j and potential mode of action

The bacterial strain E1R-j, isolated as an endophyte from wheat roots, exhibited high antifungal activity to Gaeumannomyces graminis var. tritici (Ggt). Strain E1R-j was identified as Bacillus subtilis based on morphological, physiological and biochemical methods as well as on 16S rDNA analysis. This strain inhibited mycelium growth in vitro of numerous plant pathogenic fungi, especially of Ggt, Coniothyrium diplodiella, Phomopsis sp. and Sclerotinia sclerotiorum. In greenhouse experiments, soil drenches with cell densities of 10⁶, 10⁹ and 10¹² CFU ml⁻¹ E1R-j reduced significantly take-all disease, caused by Ggt, in wheat seedling by 62.6%, 68.6% and 70.7%, respectively, compared to the inoculated control, 4 weeks after sowing. Growth parameters such as lengths and fresh weights of roots and shoots of Ggt-inoculated control plants were significantly lower compared to Ggt-inoculated and E1R-j treated plants. Field experiments in the season 2006/2007, heights of wheat plants in the Ggt inoculated plots were significantly reduced compared to the non inoculated treatments. Yield parameters such as kernels per head and thousand kernel weight (TKW) in inoculated control plants were lower compared to the other treatments. In the experimental year 2007/2008, independent treatments with the bacterial strain E1R-j and the fungicide Triadimefon reduced take-all disease in wheat roots by 55.3% and 61.9%, compared to the inoculated control plants. In this season plant height in inoculated control was significantly lower and also the yield parameters seeds per head and especially TKW were drastically reduced compared to the other treatments. E1R-j treatment alleviated the detrimental effects of take-all on grain yield parameters to a similar extent as Triadimefon application. SEM studies revealed that in the presence of E1R-j, hyphae of Ggt showed leakage, appeared ruptured, swollen and shriveled. Following root drench, strain E1R-j was able to colonize endophytically roots and leaves of wheat seedlings. While the population of the bacterial strain in wheat roots steadily increased from the second to the fourth leaf stage, in the leaf tissue the population of the strain rapidly declined. TEM studies also showed that cells of E1R-j were present in roots of wheat seedlings and effectively retarded infection and colonization of Ggt in root tissue; suppression of Ggt by E1R-j was accompanied by disintegration of hyphal cytoplasm. In addition, in the presence of E1R-j cells in Ggt-infected root tissue morphological defense reactions were triggered such as formation of wall appositions and papillae. The results presented indicate that the endophytic strain E1R-j of B. subtilis meets demands required for biocontrol of take-all.     

三个鲤品种微卫星丰度与遗传多样性分析

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栓孔菌属漆酶高产菌株的初步筛选及其产酶条件的优化

利用显色反应对栓孔菌属(Trametes)进行了漆酶高产菌株的筛选,并对目标菌株的产酶条件进行了优化,在添加愈创木酚的固体培养基中,通过显色反应初步筛选出漆酶高产菌株东方栓孔菌Trametes orientalis Cui 6300;进一步通过单因子分析、正交试验和ABTS法确定了菌株Cui 6300的最适产酶条件:麦芽糖15 g/L,蛋白胨3 g/L,pH 4.8,Cu2+2.0 mmol/L,培养温度28°C,接种饼直径1.5 cm,此时酶活最高可达19.923 U/mL;同时探索了Cu2+浓度及添加时间对其菌丝生物量和漆酶活力的影响。研究表明,Cu2+最适添加浓度为2.0 mmol/L,添加时间为接种后第3天。