典型旱作农田土壤氧化亚氮排放的氨氧化微生物相对贡献

氨氧化过程对氧化亚氮（N2O）排放具有重要贡献。在不同土壤类型和农田管理下,氨氧化微生物类群对N2O排放的相对贡献组成规律还缺乏系统的研究。本研究选取典型农田耕层土壤（潮土、黑土、砖红壤）,以及有机肥改良的砖红壤剖面土壤,采用选择性抑制法（乙炔和辛炔）研究氨氧化细菌（AOB）、氨氧化古菌和全程硝化菌（AOA+comammox）以及异养硝化菌对土壤硝化潜势、净硝化速率及N2O排放的相对贡献。结果表明,在耕层土壤中,潮土、黑土和砖红壤的pH分别是8.0、6.7和5.7,硝化潜势分别是N 32.5、6.6和4.8 mg?kg-1?d-1,净硝化速率分别是N 7.1、3.0和0.5 mg?kg-1?d-1,7天N2O累积排放量分别是N 38.0、35.4和8.7 μg?kg-1。AOB主导耕层土壤的硝化潜势,对硝化潜势的贡献分别是82%、58%和100%。对于净硝化速率,在潮土和砖红壤中,AOB和AOA+comammox贡献相当（均在30%~40%）,而黑土中由AOB主导（72%）。AOB主导耕层土壤的N2O排放,对N2O排放的贡献分别是72%、92%和58%。在改良的砖红壤剖面土壤中,在0~20 cm、20~40 cm和40~60 cm,pH分别是7.0、5.5和4.9,硝化潜势分别是N 6.6、2.0和1.1 mg?kg-1?d-1,净硝化速率分别是N 4.1、0.9和0.2 mg?kg-1?d-1,N2O排放分别是N 16.3、6.5和2.8 mg?kg-1?d-1。随土壤由深层至表层,硝化潜势、净硝化速率及N2O排放显著提高。AOA+comammox主导表层硝化潜势及净硝化速率的提高（分别贡献63%和54%）,AOB主导N2O排放的增加（贡献54%）。本研究为制定与土壤氨氧化特性及土壤性质相匹配的N2O减排措施提供了新的科学依据。     

GM1 gangliosidosis in a Japanese domestic cat: a new variant identified in Hokkaido,Japan

A male Japanese domestic cat with retarded growth in Hokkaido, Japan, showed progressive motor dysfunction, such as ataxia starting at 3 months of age and tremors, visual disorder and seizure after 4 months of age. Finally, the cat died of neurological deterioration at 9 months of age. Approximately half of the peripheral blood lymphocytes had multiple abnormal vacuoles. Magnetic resonance imaging showed bisymmetrical hyperintensity in the white matter of the parietal and occipital lobes in the forebrain on T2-weighted and fluid-attenuated inversion recovery images, and mild encephalatrophy of the olfactory bulbs and temporal lobes. The activity of lysosomal acid β-galactosidase in leukocytes was negligible, resulting in the biochemical diagnosis of GM1 gangliosidosis. Histologically, swollen neurons characterized by accumulation of pale, slightly granular cytoplasmic materials were observed throughout the central nervous system. Dysmyelination or demyelination and gemistocytic astrocytosis were observed in the white matter. Ultrastructually, membranous cytoplasmic bodies were detected in the lysosomes of neurons. However, genetic analysis did not identify the c.1448G>C mutation, which is the single known mutation of feline GM1 gangliosidosis, suggesting that the cat was affected with a new variant of the feline disease.     

ABA， H2O2与NO对木枣果实成熟生理的影响

在木枣白熟期分别用ABA、H2O2与NO供体硝普纳（SNP）喷洒果实,对其调控木枣果实成熟的生理基础进行了研究。结果显示用不同浓度的ABA,H2O2与SNP处理,枣果实中丙二醛（MDA）的含量明显升高,其中100 mg?L-1浓度时MDA含量为最高;Ca、Vc和β-胡萝卜素含量呈现先降低后升高再下降的趋势,其中75 mg?L-1浓度时Vc和β-胡萝卜素含量最高,50 mg?L-1时Ca含量最高;CAT和APX活性的变化呈现先升高后降低的趋势,75 mg?L-1时CAT和APX活性均最大;DNA含量的变化呈现先缓慢降低后升高再下降的趋势,ABA比H2O2、SNP处理对DNA含量影响更明显,100 mg?L-1时DNA含量最高。结论：ABA、H2O2与SNP对木枣果实发育成熟的影响效果较为明显,其生理效果在多方面表现基本一致,有加快木枣果实成熟衰老的作用。     

鸭亲本间遗传距离与杂交后代杂种优势关系的研究

运用随机扩增多态DNA方法,对樱桃谷鸭、麻鸭及其杂交一代和用F1作为母本再与父本番鸭属间杂交培育的三元杂交半番鸭(F2)基因组DNA进行了遗传分析。结果表明,RAPD可用于有效标记鸭品种之间遗传亲缘关系和杂种优势预测;在所使用的20种随机引物中,5个鸭种共产生760条带,其中多态性带568条,占总条带数的 74．7％,说明鸭群体具有较为丰富的遗传多样性;杂交一代多态性位点比例为67．9％,遗传相似度为0．822 7,三元杂交二代多态性位点比例为83．1％,遗传相似度为0．826 4。说明三元杂交子二代的杂种优势可以用杂种优势理论中的显形假说来解释。     

贮梨冷库生产金针菇时的CO2浓度变化及调控研究

针对闲置贮梨冷库进行金针菇生产易造成库内CO2过度积累而影响金针菇商品性能问题,采用观测和模拟试验,对贮梨冷库生产金针菇时的CO2浓度变化及排除方法进行了研究.结果表明:金针菇菌丝体和子实体发育过程中不断产生CO2,且所产生的CO2浓度随时间延长而增高;在冷库近底层增加新风口,与库门构成平抽气流的改建设计,能有效提高CO2的排除效果,机械通风并辅以化学吸收剂Ca(OH)2,可有效保障库内CO2浓度与金针菇子实体生长发育的不同阶段相适应.     

减蛋综合征病毒六邻体蛋白基因的克隆与真核表达载体的构建

应用降落PCR技术扩增出减蛋综合征病毒六邻体蛋白基因并将其克隆至pMDT-18载体上,酶切、PCR及测序结果表明插入的片段为目的基因.切下该目的基因定向克隆至pcDNA 3质粒构建六邻体蛋白基因真核表达载体pcDNA 3-hexon,经各种酶切、PCR鉴定及进一步测序鉴定,证明六邻体蛋白基因片段所插入的位置、大小、核苷酸序列和阅读框架正确无误,从而为下一步的转染、表达及进一步阐明EDSV六邻体基因结构与功能的关系和减蛋综合征基因工程苗的研究奠定了良好的基础.     

编码杜果丝氨酸/苏氨酸蛋白激酶类蛋白抗病基因同源序列的克隆与分析

根据丝氨酸／苏氨酸蛋白激酶类催化结构域Ⅰ和Ⅸ的氨基酸保守序列设计简并引物,PCR扩增‘紫花芒’杧果嫩绿色叶片基因组DNA。在NCBI中用Blast X比对发现,有6个阳性克隆是抗病基因同源序列,并命名为PK1～PK6,GenBank注册序列号为AY693369-AY693371和AY776277-AY776279。在PK3、PK4和PK6中发现了丝氨酸／苏氨酸蛋白激酶的9个催化结构域Ⅰ～Ⅸ,而其它3个克隆具有部分结构域。氨基酸序列同源性分析还发现,6个克隆编码的氨基酸序列都与受体样蛋白激酶有较高的氨基酸序列同源性。系统进化树分析还表明它们都是可能的抗病基因同源序列。总之,6个克隆不仅是抗病基因同源序列而且是可能的受体样蛋白激酶基因同源序列。     

七彩神仙鱼 BTN1A1 基因克隆及其在嗜水气单胞菌胁迫下的表达分析

【目的】克隆七彩神仙鱼（Symphysodon aequifasciatus）嗜乳脂蛋白 1A1（Butyrophilin 1A1, BTN1A1）基因,分析其在病原刺激下的表达模式,为了解七彩神仙鱼 BTN1A1 基因功能提供依据。【方法】利用生物信息学对七彩神仙鱼 2 个 BTN1A1 基因（BTN1A1-1 和 BTN1A1-2）进行结构和进化分析。根据前期获得的七彩神仙鱼皮肤转录组数据,选取 BTN1A1-1 和 BTN1A1-2 基因的 CDS 区设计引物进行克隆。采用 qRT-PCR 分析 BTN1A1 在各组织中的表达及嗜水气单胞菌刺激下的表达模式。【结果】BTN1A1-1 和 BTN1A1-2 的 ORF 序列长度为 894、1 275 bp,分别编码 298、424 个氨基酸。BTN1A1-1 和 BTN1A1-2 蛋白均具有 1 个信号肽、1 个 Ig 结构域、1 个 lg_like 结构域和 1个跨膜结构域的经典结构,BTN1A1-2 还具有 1 个胞质结构域。七彩神仙鱼 BTN1A1-1 与慈鲷科其他鱼类相似性较高,与尼加拉瓜湖始丽鱼（Archocentrus centrarchus）BTNIAI 对应氨基酸序列同源性最高、为 94.14%。但 BTN1A1-2 与其他鱼类发生分离,形成独特分支。BTN1A1-1 和 BTN1A1-2 在七彩神仙鱼各组织中均有表达,但在鳃、肠道、皮肤等与免疫相关的组织中表达量相对较高。七彩神仙鱼 BTN1A1-1 在嗜水气单胞菌胁迫后表达量先显著下降后上升,而 BTN1A1-2 表达量则先显著上升后下降。【结论】七彩神仙鱼 BTN1A1-1 基因相对保守,而 BTN1A1-2 基因在进化上较为特殊。二者在免疫刺激下的差异表达暗示其可能在七彩神仙鱼免疫防御中发生功能分化。     

Impact of Honey Bee Migratory Management on Pathogen Loads and Immune Gene Expression is Affected by Complex Interactions With Environment,Worker Life History,and Season

The effects of honey bee management, such as intensive migratory beekeeping, are part of the ongoing debate concerning causes of colony health problems. Even though comparisons of disease and pathogen loads among differently managed colonies indicate some effects, the direct impact of migratory practices on honey bee pathogens is poorly understood. To test long- and short-term impacts of managed migration on pathogen loads and immunity, experimental honey bee colonies were maintained with or without migratory movement. Individuals that experienced migration as juveniles (e.g., larval and pupal development), as adults, or both were compared to control colonies that remained stationary and therefore did not experience migratory relocation. Samples at different ages and life-history stages (hive bees or foragers), taken at the beginning and end of the active season, were analyzed for pathogen loads and physiological markers of health. Bees exposed to migratory management during adulthood had increased levels of the AKI virus complex (Acute bee paralysis, Kashmir bee, and Israeli acute bee paralysis viruses) and decreased levels of antiviral gene expression (dicer-like). However, those in stationary management as adults had elevated gut parasites (i.e. trypanosomes). Effects of environment during juvenile development were more complex and interacted with life-history stage and season. Age at collection, life-history stage, and season all influenced numerous factors from viral load to immune gene expression. Although the factors that we examined are not independent, the results illuminate potential factors in both migratory and nonmigratory beekeeping that are likely to contribute to colony stress, and also indicate potential mitigation measures.     

菠萝酸性转化酶基因家族初步生物信息学及表达分析

酸性转化酶（acid invertase, AIN）在菠萝采后蔗糖降解过程中起着重要作用,基于菠萝全基因组数据库,预测菠萝AIN家族基因并进行生物信息学分析,解析其在采后菠萝不同贮藏温度下的表达变化情况,为阐明AIN基因在采后菠萝果实贮藏特性中的作用奠定基础。以水稻AIN家族基因为探针,在菠萝全基因组中鉴定到2个菠萝细胞壁酸性转化酶基因（cell wall acid invertase, CWIN）和2个液泡酸性转化酶基因（vacuolar acid invertase, VIN）,分别命名为AcCWIN1、AcCWIN2、AcVIN1、AcVIN2,设计编码区引物进行测序验证,并进行生物信息学分析。进化分析结果表明,AcCWIN1、AcCWIN2和AcVIN1、AcVIN2蛋白分别归于细胞壁酸性转化酶和液泡酸性转化酶2个进化支上,且均属于糖基水解酶家族GH32,基因结构、保守域和保守基序均一致。荧光定量分析结果表明,菠萝果肉中AcVIN1和AcVIN2在果实采后贮藏过程中表达量升高,且AcVIN1在发生黑心病的部位大量表达,而AcCWIN1和AcCWIN2在采后贮藏过程中表达量逐渐降低,且随着贮藏温度的升高其表达量降低,预示AcVIN1、AcVIN2较AcCWIN1、AcCWIN2在菠萝采后蔗糖降解和黑心病的发生方面发挥着更为重要的作用。