转基因植物非预期效应检测评价技术的发展

检测和评价转基因植物非预期效应是转基因植物安全评价不可或缺的重要内容之一。介绍了转基因植物非预期效应的定义、成因及其检测评价技术的发展,指出了转基因植物非预期效应检测评价技术的难点和存在的问题。阐述了基因芯片分析技术在检测和评价转基因植物非预期效应中的应用前景,特别介绍了转基因植物在不同发育阶段和生长条件下差异表达基因的分布模型,并提出了利用基因芯片分析技术,从差异基因到代谢途径,再到非预期效应的转基因植物安全评价模式。这一评价模式可为今后创建高效、全面、客观、公正的转基因植物非预期效应检测和评价技术提供可能的实验模型和理论依据。     

基于阵列芯片现场高灵敏同步快速检测粮油中氨基甲酸酯类农药残留混合污染

The quality and safety of agricultural products is the technique focus of the strategy of rural vitaliza⁃tion. Carbamate pesticide residues are seriously threatening the safety of agricultural product and human life and health. Nowadays, the common detection methods are instrument-based for carbamate pesticide residue, which dis⁃ satisfy the request of an on-site rapid, sensitive and simultaneous detection for multiple carbamate pesticide resi⁃ dues. Based on the home-made monoclonal antibodies against carbofuran and carbaryl respectivity, a microarray for rapid and simultaneous detection of multiple carbamate pesticide residues was developed within 30 min. Under the optimized conditions, the limits of detection（LODs）were found to be 0.11 and 0.09 ng/g, while the linear ranges were 0.33-3000 and 0.18-1500 ng/g for carbofuran and carbaryl respectively. By using spiked peanut samples, it recorded recoveries of 88.4%-98.3%, 90.1%-118.8% for carbofuran and carbaryl respectively. The RSD of intraarray were found at 9.0%, 3.5%, while the RSD of inter-array were recorded at 12.3%, 16.3% respectively. The de⁃ tection results of the microarray were highly consistent with that of HPLC-MS/MS. This proposed microarray provid⁃ ed an on-site high-sensitivity, simultaneous and rapid detection of carbofuran and carbaryl, and could be widely used in the fields of agricultural product quality safety, food safety and environmental monitoring.     

全基因组选择育种策略及在水产动物育种中的应用前景

全基因组选择的概念自2001年由Meuwissen等提出后便引起了动物育种工作者的广泛关注.目前,澳大利亚、新西兰、荷兰、美国的研究小组已经应用该方法进行了优质种牛的选择育种,并取得了很好的效果.此外在鸡和猪的选择育种中也有该方法的应用,但在水产动物选育中尚未见该方法使用的报道.本文对“全基因组选择育种”的概念和提出背景进行了归纳,对全基因组选择育种的优势进行了阐述,并详细介绍了其具体的策略,总结了目前全基因组育种所广泛采用的方法以及取得的成果,旨在为该方法在水产动物育种方面的应用研究提供科学参考.     

水稻稻瘟病菌胁迫应答cDNA片段的表达及定位

以抗稻瘟病品系G205为材料,应用cDNA微阵列分别获得了一个受稻瘟病菌诱导的含NBS LRR的cDNA克隆（暂命名为RIM1, rice induced by Magnaporthe grisea）和一个受稻瘟病菌抑制的编码腈水解酶（Nitrilase）的cDNA克隆（暂命名为NIT）,并通过Northern得到证实。RFLP分析将RIM1和NIT分别定位于水稻第2和第3染色体上,它们均位于控制水稻稻瘟病部分抗性QTL区间。     

基于不完全双列杂交设计的水稻农艺性状配合力基因组预测

[目的]在亲本一般配合力的基础上优选特殊配合力高的杂交种,是水稻杂种育种的关键。基因组选择基于覆盖全基因组的分子标记和样本的表型数据建立预测模型,实现对品种更加可靠的选择。[方法]本研究利用一组基于不完全双列杂交(NCII 设计)的水稻数据集,考查其多个农艺性状配合力的基因组预测能力。并比较了不同训练群体构建方法对杂交种表型预测能力的影响。[结果]8个农艺性状一般配合力的预测能力由其遗传率主导,从0.3888 到0.7367。杂交种特殊配合力的预测能力较低,但是直接预测杂交种的表型可以获得较高的预测能力。[结论]基因组预测水稻亲本一般配合力是有效的,能够帮助育种家实现对亲本的科学选择。如果要选配杂交种,直接预测杂交种表型是最有效的手段。此时让更多的亲本均衡地参与杂交种训练集的组配,有利于获得更高的预测能力。     

Silicon-mediated genotoxic alterations in Brassica juncea under arsenic stress: A comparative study of biochemical and molecular markers

Arsenic (As), one of the most harmful toxicant at the global level, severely affects plant metabolism when taken up. Interestingly, the presence of silicon (Si) as a fertilizer in As-contaminated soil is an effective strategy to decrease As accumulation in plants. Brassica juncea (var. Varuna) were grown hydroponically to investigate the role of Si at biochemical and molecular levels under arsenite (As³⁺) stress. Seedlings of B. juncea were exposed to As³⁺, Si, and a combination of both elements. Our data demonstrated that seedlings exposed to As³⁺ showed an inhibition in shoot length, chlorophyll, carotenoid, and protein, while co-application of Si improved these growth parameters. Silicon supplementation reduced As accumulation in shoot. Increase/decrease was observed in stress-related parameters (cysteine and proline), antioxidant enzymes (superoxide dismutase, ascorbate peroxidase, and catalase), and oxidative stress markers (malondialdehyde and H₂O₂), which were improved upon co-application of Si as compared to As³⁺ alone treatment. Random amplified polymorphic DNA (RAPD) is a suitable biomarker assay for plants for assessing the genotoxicity. Seven RAPD primers produced a total of 39 and 48 bands in the leaves of the untreated and treated seedlings, respectively. The RAPD band-profiles and genomic template stability were consistent with other growth and physiological parameters. In conclusion, the genotoxic alterations along with the biochemical parameters indicate that the exposure to Si mitigates As³⁺-induced oxidative stress by improving the stress-related parameters and antioxidant system in B. juncea.     

基于系谱和基因组信息估计荷斯坦青年母牛体重性状遗传参数

旨在设计利用不同信息来源的模型估计荷斯坦后备牛不同月龄体重性状的遗传参数。本研究于2014—2020年测定并收集了7 122头荷斯坦牛32 338条0～12月龄体重数据,分别利用系谱信息(linear mixed model with pedigree relationship matrix, LM＿A)和系谱-基因组信息构建亲缘关系矩阵(linear mixed model with genotype-pedigree joint relationship matrix, LM＿H),基于母体效应动物模型估计初生重,基于是否考虑初生重作为协变量的单性状动物模型估计2～12月龄各月龄体重遗传力,并利用双性状动物模型估计初生重与其它月龄体重的遗传相关。结果显示,对于初生重,根据赤池信息量准则(Akaike information criterion, AIC),LM＿H方法的拟合程度显著优于LM＿A方法,但两种方法估计的遗传参数相差不大：直接遗传力分别为0.30和0.32,母体遗传力分别为0.08和0.09,个体直接遗传效应和母体遗传效应遗传相关系数分别为-0.65和-0.64;对于2～...     

应用cDNA微阵列芯片技术筛选猪蛔虫性别差异表达基因的研究

采用cDNA微阵列芯片技术，从所构建的猪蛔虫雌、雄成虫cDNA消减文库分别挑取1044和1119个克隆，PCR扩增其插入片段，经纯化后点样于预先处理好的基片上（双点杂交），制备成cDNA微阵列芯片。将分别标记荧光素Cy3-dUTP和Cy5-dUTP的雌虫和雄虫cDNA探针，与制备好的cDNA芯片杂交（平行进行反标杂交试验）。根据每个点杂交后的Ratio值，筛选出双点杂交和正反标中都同时具有表达差异的基因克隆共1559个。将表达差异最明显的前831个克隆进行测序，获得720个有效序列，经生物信息学分析发现，雄虫特异表达的主要精于蛋白和雌虫特异表达的卵巢信息蛋白的基因序列多数与新杆属线虫存在同源性，有31个可能是新的ESTs。性别差异表达基因及其相关生物信息的获得为下一步研究基因功能奠定了基础。     

用改进的酚-氯仿法提取鱼类基因组DNA效果的分析

提取鱼类基因组DNA的研究报道并不多见,效果也各有差异。本文以泰山螭霖鱼、东平湖鲤鱼、东平湖鲫鱼为材料,采用改进的酚-氯仿法提取基因组DNA,并进行了紫外分光光度、琼脂糖凝胶电泳、微卫星PCR扩增等方法的鉴定。结果表明,本方法提取的基因组DNA浓度在0.9μg/μL~3.25μg/μL之间,A260/A280比值在1.79~1.87之间,电泳条带整齐明亮,适合微卫星PCR扩增,且整个操作过程使用试剂较少,简单易行。