细菌表达 dsRNA 介导桔小实蝇 flightin基因的 RNA 干扰

【目的】探索饲喂细菌表达目标基因dsRNA介导桔小实蝇flightin基因RNAi的可行性。【方法】将桔小实蝇flightin基因的相应干扰片段构建至L4440干扰载体,并转化大肠杆菌HT115(DE3)菌株,利用IPTG诱导表达flightin基因对应的dsRNA,命名为flightin-dsRNA。【结果】通过饲喂表达flightin-dsRNA的大肠杆菌10倍浓缩菌液,桔小实蝇flightin基因的表达普遍出现了不同程度的上调,其中,雌虫饲喂后5、10、20 d,雄虫饲喂后5 d与对照差异显著,雄虫饲喂后15 d诱发约43%的下调;该方法对桔小实蝇飞行能力及胸部肌肉发育未产生明显影响。【结论】通过饲喂表达flightin-dsRNA的大肠杆菌对桔小实蝇flightin基因进行RNA干扰不可行或干扰效果不明显。     

甘蓝型油菜木质素合成关键基因F5H、4CL和COMT的定量表达

以甘蓝型油菜(Brassica napus L )抗倒伏品种浙平1号和易倒伏品种高芥1号为材料,采用荧光定量PCR方法对木质素合成关键基因F5H（阿魏酸-5-羟基化酶）、4CL（4-香豆酸CoA连接酶）和COMT（咖啡酸/5-羟基阿魏酸-O-甲基转移酶）在薹期和角果期根颈部、茎部的相对表达量进行了分析。结果显示,F5H在薹期和角果期根颈部的相对表达量,易倒伏材料高于抗倒伏材料;茎部的相对表达量,抗倒伏材料高于易倒伏材料。4CL和COMT基因在薹期根颈部、茎部和角果期根颈部的相对表达量,易倒伏材料高于抗倒伏材料;角果期茎部的相对表达量,抗倒伏材料显著（P＜0.05）高于易倒伏材料。结果表明,油菜角果期茎部木质素合成关键基因F5H、4CL和COMT的相对高量表达可能有助于增强植株的抗倒伏能力。     

小麦籽粒淀粉含量的QTL定位及效应分析

为深入了解小麦籽粒淀粉含量的遗传特性和QTL效应,选择淀粉含量差异较大的小麦品种M344和武春3号杂交,创制F2∶3群体,并利用973对SSR引物对小麦籽粒总淀粉、直链淀粉和抗性淀粉含量进行基于混合线性模型的QTL定位及效应分析。构建了包含有163个标记的遗传连锁图谱,分布于18条染色体,连锁图谱总长度为1 622.5cM,标记间平均距离为9.95cM。QTL分析表明,共检测到4个主效QTL和12对上位性QTL,涉及1B、2B、2D、4A、5D、6A和7A等16条染色体。其中,抗性淀粉含量上位性QTL3对,总淀粉含量上位性QTL 4对,直链淀粉含量上位性QTL 5对,总贡献率分别为8.34%、17.50%和8.13%。总淀粉和直链淀粉含量各检测到1个加性QTL,抗性淀粉含量检测到2个加性QTL,贡献率分别为5.34%、8.49%、11.47%、12.53%。研究发现,2B染色体Xwmc453.3~Xcfd44.2标记区间的QTL位点同时影响抗性淀粉和总淀粉含量,2D染色体Xgwm296~Xgwm455.2标记区间的QTL位点同时影响抗性淀粉和直链淀粉含量,7A染色体Xwmc488.1~Xwmc488.2标记区间的QTL位点同时影响直链淀粉和总淀粉含量。     

Screening,Inheritance and Linkage Marker Analyses of Salt Tolerance in Mutated Scented Japonica Rice (Oryza sativa L.)

Abstract

We grew 1005 mutated scented japonica rice lines to the seventh steady genetic generation, and treated the seedlings with six true leaves with 300 mM NaCl for three days. Only the salt-tolerant line, SM61, survived. We obtained F₁, F₂ and F₃ populations from the cross between SM61 and a salt-susceptible indica variety, TCS17. After culture with 200 mM NaCl for five days, SM61 and F₁ (SM61×TCS17; TCS17×SM61) plants survived (R) while TCS17 plants did not (S). The R to S ratio in 513 F₂ plants showed a good fit to the Mendelian 3 : 1 segregation ratio by a Chi-square test indicating that the salt-tolerance of SM61 was governed by a single dominant gene. The mutated salt-tolerance gene explained close to 100% of the total phenotypic variation, and was tightly linked to RM223 (marker) located on chromosome 8, which was different from the results of previous studies investigating the relationship of QTLs with salt tolerance. This is the first report of mapping tightly linked markers of a single dominant mutated salt-tolerance gene.     

Survival of bacterial DNA and culturable bacteria in archived soils from the Rothamsted Broadbalk experiment

Dried soil samples from many sources have been stored in archives world-wide over the years, but there has been little research on their value for studying microbial populations. Samples collected since 1843 from the Broadbalk field experiment on crop nutrition at Rothamsted have been used to document changes in the structure and composition of soils as agricultural practices evolve, also offering an invaluable record of environmental changes from the pre- to post-industrial era in the UK. To date, the microbial communities of these soils have not been studied, in part due to the well-documented drop in bacterial culturability in dried soils. However, modern molecular methods based on PCR amplification of DNA extracted directly from soil do not require bacterial cells to be viable or intact and may allow investigations into the legacy of bacteria that were present at the time of sample collection.

In a preliminary study, to establish if dried soils can provide a historical record of bacterial communities, samples from the Broadbalk soil archive dating back to 1868 were investigated and plots treated with either farmyard manure (FYM) or inorganic fertilizer (NPK) were compared. As anticipated, the processes of air-drying and milling greatly reduced bacterial viability whilst DNA yields declined less and may be preserved by desiccation. A higher proportion of culturable bacteria survived the archiving process in the FYM soil, possibly protected by the increased soil organic matter. The majority of surviving bacteria were firmicutes, whether collected in 2003 or in 1914, but a wide range of genera was detected in DNA extracted from the samples using PCR and DGGE of 16S rRNA genes. Analysis of DGGE band profiles indicated that the two plots maintained divergent populations. Sequence analysis of bands excised from DGGE gels, from a sample collected in 1914, revealed DNA from - and β-proteobacteria as well as firmicutes. PCR using primers specific for ammonia oxidizing bacteria showed similar band profiles across the two treatments in recently collected samples, however older samples from the NPK plot showed greater divergence. Primers specific for the genus Pseudomonas were designed and used in real-time quantitative PCR to indicate that archived soil collected in 1868 contained 10-fold less pseudomonad DNA than fresh soil, representing around 10⁵ genomes g⁻¹ soil. Prior to milling, dramatically less pseudomonad DNA was extracted from recently collected air-dried soil from the NPK compared to the FYM plot; otherwise, the two plots followed similar trends. Overall bacterial abundance, diversity and survival during the archiving process differed in the two soils, possibly due to differences in clay and soil organic matter content. Nevertheless, the results demonstrate that air-dried soils can protect microbial DNA for more than 150 years and offer an invaluable resource for future research.     

深水抗风浪网箱监测系统研制方案的探讨

探讨一种声学监测方案,用高频、窄脉冲和分裂波束技术,实时监测养殖鱼的进食、生长情况,随时评估网箱内总量的增长情况,并且从水产行业的成本可承受性的实际出发,将成本控制在用户可接受的水平上。     

鳙目标强度和行为特征的水声学研究

使用EY60回声探测仪于2010年5月10-15日、9月28-29日分别对5种不同体长规格的鳙进行了目标强度及昼夜行为的现场测定.结果显示:鳙的全长范围为30.05～80.00cm,体长范围为25.10～68.80 cm,体重范围为0.3～6.0kg,对应的有效目标强度范围为-51.84～-42.06dB,最大目标强度...     

Analysis on Target Genes with Mutation in microRNA of Blue-shelled Chicken and White Leghorn

The study was conducted to explore the potential different characters between Blue-shelled chicken and White leghorn.Global genome microRNA was combined the identified microRNA with complementary lab-predicted microRNA.Then the two breed chicken's SNP data got by GGRS were mapped to the microRNA and focused on SNP that deliberately located in mature-microRNA.Bioinformatics method was adopted for target prediction on microRNA which had SNPs.By further gene enrichment analysis,the study found these genes enriched in 22 GO terms,10 KEGG pathways,and 3 IPA important networks.And they enriched in traits which associated with growth,such as mTOR signaling pathways,Wnt signaling pathways,growth hormone receptor networks and insulin-like growth factor Ⅰ receptor networks.And they also enriched in some laying traits,such as oocyte meiotic signaling pathways and progesterone mature oocytes signaling pathways.The methods and the results might provide references for further studies.     

斑点叉尾鮰暴发性出血溃疡症的病理学研究

【目的】探究两起斑点叉尾鮰Ietalurus punetaus暴发性体表出血溃疡症的发病病因和病理特征。【方法】利用湿片法对鳃组织和体表黏液进行压片,观察寄生虫寄生情况。从发病鱼的肝脏、脾脏和肾脏中分离病原菌,检测细菌感染情况;利用PCR法检测患病斑点叉尾鮰的肝、脾、肾混合组织匀浆中斑点叉尾鮰病毒(Channel catfish virus,CCV)的携带情况。运用组织病理学方法观察病鱼的病理损伤特点,综合分析并推测该病的致病原因。【结果】该出血溃疡症的靶器官主要为脾脏、肾脏、肝胰脏、肠道以及皮肤肌肉。主要表现为严重的血管反应,实质细胞可见肿胀、变性、坏死;而胃、脑和心脏病变较轻,仅有轻微的炎症;鳃丝、眼球和鳔等未见明显病变。所有的患病个体均出现中度至重度出血性坏死性脾炎、中度肾炎、轻度至中度坏死性肝胰腺炎和肠炎。患病斑点叉尾鮰体内未检测到寄生虫、细菌以及CCV。【结论】综合病理学、细菌学和病毒学检测结果,推测此次斑点叉尾鮰暴发性疾病由某种非CCV的病毒感染所致,温度应激可能是引发该病的条件诱因。     

基于文献计量分析的马铃薯晚疫病研究现状

以《中国期刊全文数据库》的数据为检索对象,对1980~2008年的马铃薯晚疫病研究文献采用计量学的方法,从年度载文、核心作者群、合作度、主要产出单位及区域、研究领域等方面进行了统计分析,从而揭示了我国对马铃薯晚疫病的研究现状、研究热点及发展趋势。