Adaptive diversification of bitter taste receptor genes in mammalian evolution

The diversity and evolution of bitter taste perception in mammals is not well understood. Recent discoveries of bitter taste receptor (T2R) genes provide an opportunity for a genetic approach to this question. We here report the identification of 10 and 30 putative T2R genes from the draft human and mouse genome sequences, respectively, in addition to the 23 and 6 previously known T2R genes from the two species. A phylogenetic analysis of the T2R genes suggests that they can be classified into three main groups, which are designated A, B, and C. Interestingly, while the one-to-one gene orthology between the human and mouse is common to group B and C genes, group A genes show a pattern of species- or lineage-specific duplication. It is possible that group B and C genes are necessary for detecting bitter tastants common to both humans and mice, whereas group A genes are used for species-specific bitter tastants. The analysis also reveals that phylogenetically closely related T2R genes are close in their chromosomal locations, demonstrating tandem gene duplication as the primary source of new T2Rs. For closely related paralogous genes, a rate of nonsynonymous nucleotide substitution significantly higher than the rate of synonymous substitution was observed in the extracellular regions of T2Rs, which are presumably involved in tastant-binding. This suggests the role of positive selection in the diversification of newly duplicated T2R genes. Because many natural poisonous substances are bitter, we conjecture that the mammalian T2R genes are under diversifying selection for the ability to recognize a diverse array of poisons that the organisms may encounter in exploring new habitats and diets.

Direct characterization of bitter acids in a crude hop extract by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry

The applicability of on-line coupling of reversed-phase high-performance liquid chromatography to atmospheric pressure ionization tandem mass spectrometry for the separation and characterization of hop acids mixture from the crude extract of Humulus lupulus was investigated. The solvent system consisting of acetonitrile-aqueous formic acid was used to give proper separation of the six main hop bitter acids within 30 min. Further structural information about the components was acquired by collision-induced dissociation (CID). On the basis of analyses of the fragmentation patterns of the major alpha- and beta-bitter acids respectively, identification of the minor ones was performed using selected reaction monitoring (SRM) with a group of qualitatively relevant selected precursor-product ion transitions for each bitter acid in a single high performance liquid chromatography (HPLC) run. Using this technique, six minor hop acids, including "adprelupulone" observed for the first time in natural resources, were detected along with the six major acids. This hyphenated techniques provides potency for rapid qualitative determination of analogs and homologs in mixtures. (C) 2004 American Society for Mass Spectrometry.

大功率微波与臭氧等复合因子对大棚蔬菜根结线虫及土壤的作

虫（nematodes）是世界农业生产的一大障碍，每年给世界农业生产造成约1000亿美元的巨额损失，是世界各国一直关注的重点病害之一。线虫可以危害各种大田作物和各种温室植物，几乎所有的栽培植物都有线虫危害发生。随着人们环保意识的增强以及健康意识的觉醒，传统防治根结线虫的药物如溴甲烷等因对环境的破坏作用，或者由于高毒、使用不便、抗药性等原因而迅速退出历史舞台，研究环保高效防治线虫成为世界各国竞相发展的技术之一。本研究尝试通过物理方式防治蔬菜根结线虫，并调查了对土壤质量的影响，主要内容包括： 1. 设计制造了功率20 Kw，频率为915 MHz的大功率土壤微波处理机，并通过了田间试验; 2. 设计制造了每小时120 g臭氧产量的便携式臭氧土壤处理机，并通过了田间试验; 3. 采用4因素3水平正交法L9(34)在山东寿光蔬菜大棚进行了蔬菜根结线虫的防治试验，综合考察微波、臭氧、微波吸收剂、EM菌等不同处理水平对根结线虫和对土壤质量的影响，结果表明： 1) 大棚土壤经微波60 s照射能够显著降低甜瓜根结线虫数和根结指数（p < 0.05），盆栽试验只需处理10 s即可显著防治甜瓜根结线虫（p < 0.01）;以120 g/h臭氧浓度处理5 s也能显著降低甜瓜根结线虫的数量（p < 0.05）;田间施入5 g/m2的EM菌，能够显著降低甜瓜根结线虫的数量（p < 0.05）;盆栽试验中微波吸收剂具有降低根结线虫数量的趋势，但是不同处理水平之间差异不显著，田间试验则具有增加根结线虫的趋势，不同处理水平之间也没有显著差异。 2) 微波不同处理水平之间对甜瓜单瓜重的影响没有显著差异;以120 g/h臭氧浓度处理甜瓜能够显著增加甜瓜单瓜重（p < 0.05）;200 g/m2微波吸收剂能够显著提高甜瓜单瓜重（p < 0.05）;以5 g/m2EM菌处理土壤则显著降低甜瓜单瓜重（p < 0.05）。 3) 微波、臭氧、微波吸收剂不同处理水平对甜瓜糖度的影响没有显著差异;以3 g/m2的EM菌处理甜瓜能够显著降低甜瓜的糖度。 4) 不同因素对甜瓜硬度的影响没有显著差异。 5) 微波处理60 s能够显著提高甜瓜的果型指数（p < 0.1），臭氧、微波吸收剂、EM菌等因素不同处理水平之间对果型指数影响差异不显著。 6) EM菌5 g/m2处理浓度能够显著降低甜瓜株高（p < 0.1），微波、臭氧、微波吸收剂不同处理水平之间对株高的影响没有显著差异。 7) EM菌5 g/m2处理浓度能够显著降低甜瓜根径（p < 0.1），微波、臭氧、微波吸收剂不同处理水平之间对根径的影响没有显著差异。 8) 微波处理60 s能够显著降低表层土壤（0-5 cm）的有机质含量（p < 0.01）;以120 g/h的臭氧浓度处理，则可以显著提高10-15 cm土壤的有机质含量（p < 0.05），其它处理因素的不同水平对有机质的影响差异不显著。 9) 微波不同处理水平对土壤全氮的影响没有显著差异（p < 0.05）;120 g/h臭氧处理浓度能够显著降低表层土壤（0-5 cm）的土壤全氮量（p < 0.1）;100 g/m2微波吸收剂处理水平能够显著提高10-15 cm土壤全氮量（p < 0.05）;EM菌3 g/m2的处理水平则显著降低5-10 cm土壤全氮量（p < 0.05）。 10) 微波、微波吸收剂、EM菌不同处理水平之间对土壤全磷没有显著影响;240 g/h臭氧浓度处理5 s能够显著降低10-15 cm土层的土壤全磷含量(p < 0.05)。 11) 微波、微波吸收剂对土壤全磷没有显著影响;臭氧处理具有降低土壤有效磷的趋势，EM菌处理则有提高土壤有效磷的趋势，但是 不同处理水平之间差异不显著（p > 0.05）。 12) 微波处理对土壤EC值没有显著影响;臭氧、微波吸收剂、EM菌处理具有降低土壤EC值的趋势，但是不同处理水平之间差异不显著（p > 0.05）。 13) 微波处理30 s能够显著降低表层土壤的pH值（p < 0.05），臭氧、微波吸收剂、EM菌处理具有提高土壤pH值的趋势，但是不同处理水平之间差异不显著。

Effects of Prenatal Exposure to a 50-Hz Magnetic Field on One-Trial Passive Avoidance Learning in 1-Day-Old Chicks

We investigated memory impairment in newly hatched chicks following in ovo exposure to a 50-Hz magnetic field (MF) of 2 mT (60 min/day) on embryonic days 12-18. Isolated and paired chicks were used to test the effect of stress during training, and memory retention was tested at 10, 30, and 120 min, following exposure to a bitter-tasting bead (100% methylanthranilate). Results showed that memory was intact at 10 min in both isolated and paired chicks with or without MF exposure. However, while isolated chicks had good memory retention levels at 30 and 120 min, those exposed to MF did not. The results suggest a potential disruption of memory formation following in ovo exposure to MF, with this effect only evident in the more stressed, isolated chicks. Bioelectromagnetics 31:150-155, 2010. (C) 2009 Wiley-Liss. Inc.

哺乳动物适应性进化的分子机制探讨——若味受体基因和犁鼻器受体1基因的研究

动物的适应进化是生物学研究的最基本的问题之一。虽然，人们早从形态学上研究了动物的适应现象，但是对适应的遗传学机制知之甚少。动物感觉系统对周围环境的适应是动物适应进化中的一个关键问题。因此，我们选取了动物感觉系统中苦味受体基因家族（T2R）和犁鼻器受体1基因家族（VIR）为研究对象，对哺乳动物适应性进化的分子机制进行探讨。通过生物信息学和分子生物学手段相结合，我们在哺乳动物6个目共16个物种中获得了157个的苦味味觉受体基因，并对这些TZR基因的系统发育关系进行分析，结果显示哺乳动物的TZR基因家族经历了"生一和一灭"的进化模式，即频繁的基因重复和假基因化。另外，结果还显示这些基因可以被分为3个主要类群，分别命名为A，B和C。有趣的是，B和C类群的基因在所研究的物种间普遍是一对一的直系同源的，而A类群基因则显示了种属或世系特异性。有可能B和C类群的基因是识别哺乳动物共同的苦味物质所必需的，而A类群基因则是用于识别具有种属特异性的苦味物质。这个分析还揭示了在系统发育关系上近相关的基因在它们的染色体位置上也是靠近的，这证明了串联重复是新的TZR基因产生的主要方式。此外，通过核昔酸的异义替换数和同义替换数的比较显示不同物种新近产生的基因所编码的受体蛋白在膜外区的异义替换数显著地大于同义替换数，提示着这些通过基因重复产生的新基因受到了正选择的作用。在自然界中，许多的天然有毒物质一般都是苦的，因此，我们推测哺乳动物TZR基因的分"化选择是为了使其在探索新的生活环境和寻找新食物时能够辨别出更多不同的有毒物质，更好地适应新环境。此外，我们还研究了苦味受体基因和甜味／鲜味受体基因的进化途径。结果显示苦味受体基因和甜味／鲜味受体基因在进化上具有远相关，并具有不同的进化途径，提示着这可能是导致了这些受体基因具有不同功能，传导不同味觉的原因。犁鼻受体噬因家族（VIR）是哺乳动物的信息素受体。应用生物信息学手，我们从大鼠和小鼠的基因组中分别得到了152和115个VIR基因。大鼠VIR基因家族包含11个亚家族，其中10个是与小鼠共享的，而M家族是大鼠特有的；另夕卜大鼠缺少了H和I亚家族，而这两个亚家族存在于小鼠的基因组中。系统发育关系分析发现，"生一和一灭"进化模式也在V1R基因的进化过程中占了主导地位。所有检测到的亚家族都出现于啮齿目和灵长目分歧之后，这说明V1R基因的多样性反映了这一基因家族在啮齿目内基因重复、丢失，基因漂变及自然选择等作用的动态过程。我们的分析还表明大部分不同亚家族下的基因簇爆发的时间接近于大、小鼠分歧的时间。此外，用最大似然法分析的结果表明在这些基因簇中异义替换和同义替换的比值远大于1，揭示了正选择在这些基因的分化过程的作用。一般认为V1R在动物识别信息素过程中起重要的作用，因此我们推测V1R基因的适应性进化是为了使不同的哺乳动物能够识别不同的、复杂的信息素。

泸州老窖古酿酒作坊空气微生物分布特征研究

本文主要研究了泸州老窖古酿酒作坊内外环境空气真菌和空气细菌的群落结构和分布特征。结果如下： 作坊内外环境空气微生物浓度差别显著，并随季节变换而变化，春、夏季微生物浓度较高，秋、冬季较低，空气真菌在夏季达到最高，细菌在春季最高。 古作坊内外环境检测到的真菌均为16 属，但优势菌属不同，作坊外的优势菌属为青霉属(Penicillium)、曲霉属(Aspergillus)、无孢菌(non-sporing)、枝孢霉属(Cladosporium)和链格孢属(Alternaria)；而作坊内优势菌属为曲霉属、青霉属、酵母菌(Yeast)、无孢菌，作坊内还含有较高浓度的根霉属（Rhizopus）、毛霉属（Mucor）、短梗霉属（Aureobasidiu），枝孢霉属和链格孢属等，曲霉属、酵母菌、根霉属、毛霉属为古酿酒作坊重要的酿酒真菌，青霉属、链格孢属为酿酒不利菌群。对古作坊内曲霉属进行了初步鉴定，主要是小冠曲霉（A.cristatellus）、米曲霉（A.oryzae）、黑曲霉（A.niger）和白曲霉（A.cadidus）。 空气细菌10 属21 种，作坊内外环境的优势菌属均为芽孢杆菌属（Bacillus）、微球菌属(Micrococcus)、葡萄球菌属(Staphylococcus)、假单胞菌属(Pseudomonad)，其中芽孢杆菌属在作坊内占有绝对的优势，浓度比在40℅以上，是古酿酒作坊重要的酿酒细菌，另外还检测到较高浓度的乳酸杆菌（lactobucillus），这类菌容易使酒味发涩发苦，为酿酒不利菌。 作坊内外环境空气微生物表现出明显的交流现象。作坊内，青霉属、枝孢霉属、链格孢属、葡萄球菌属等杂菌占有一定比例；而在作坊外,芽孢杆菌属、曲霉属、根霉属(Rhizopus)、酵母菌等处于相对较高水平，绿化环境较好的营沟头作坊内的短梗霉属，枝孢霉属和链格孢属等杂菌含量低于什字头和新街子作坊。 The community structure and distribution characteristic of airborne microbes was investigated in ancient brewage workshops of luzhoulaojiao. The results are as follows: The concentration of airborne microbes was different in interior and exterior environment of ancient workshops, and also varied by seasons. microbial concentration was higher in spring and summer, and lower in fall and winner. The highest levels of airborne bacteria was in spring, but the fungal’s in summer. The identified genus of fungi were 16 in interior and exterior environment of the ancient workshops. But the dominant genus were different , The advantage genus in the interior were Aspergillus, Yeasts, Penicillum and Nonsporing and in the exterior were Penicillum, Nonsporing, Cladosporium, Aspergillus and Aureobasidiu. Rhizopus ,mucor, Aureobasidiu, Cladosporium, Alternaria and all also were at a higher level. Among these, Aspergillus, Yeasts, Rhizopus ,mucor are important vintage flora . Penicillum, Alternaria do harm to vintage. Aspergillus of ancient workshops was identified , the preponderant aspergillus species were A.cristatellus, A.oryzae, A.niger and A.cadidus in ancient brewage workshops. 10 genus 21 species bacteria were identified, the advantage genuses among the interior and exterior of the three workshops were bacillus, microccus, Staphylococcus Pseudomonas. Bacillus, which account for beyond 40℅ of the total bacteria concentration in all sampling pots, was the most dominant genus. Lactobacillus was identified at a high level in ancient workshops, it makes spirit taste bitter and astringent. So it is not a kind of good bacterium for vintage. The fungus in the interior and exterior atmosphere characterized intercommunion phenomenon. Obviously, the concentration of profitless fungus such as Penicillum, Cladosporium, Alternaria appeared in the interior, and the fungus such as Bacillus, Aspergillus, Rhizopus and Yeasts in the exterior were at a relatively high level. the harmfull fungus in yinggoutou workshops such as Aureobasidiu, Cladosporium, Alternaria and all were lower than shenzitou and xinjiezi workshops.