Sequence Analysis and Prokaryotic Expression of NSP1 Gene of Bovine Rotavirus UK Strain

Rotavirus is a major cause of acute diarrhea in both many kinds of young animals and children under 5 years old.Rotavirus NSP1, a 55 ku RNA binding protein, is the product of gene 5, which can subvert innate immune responses and be one of virulent determinant factors.According to the sequence in GenBank, specific primers targeting to NSP1 gene were designed and the gene was amplified by RT-PCR, following by being cloned into the pET-28a(+) vector.It showed that the full length of NSP1 gene was 1 473 bp, encoding 491 amino acids.The NSP1 shared the highest identity with WC3 strain.The recombinant protein was induced in E.coli Rosetta(DE3) by IPTG and was analyzed by SDS-PAGE and Western blotting.The results revealed that NSP1 recombinant protein existed in the form of inclusion body with the molecular weight of 55 ku.The purified recombinant protein could be recognized by His-tag antibody.This study laid the foundation for further research on the relationship between the intracytoplasmic location of NSP1 protein and its activity.     

增密减氮对弱筋小麦‘宁麦13’产量和品质的影响

为实现弱筋小麦优质稳产,解决当前弱筋小麦存在品质稳定性差的问题。本试验以弱筋小麦‘宁麦13’为试材,结合方差分析等方法研究增密减氮对弱筋小麦的产量、群体质量指标以及籽粒品质的影响。结果表明,在240 kg/hm²施氮水平条件下,随着密度的增加,小麦LAI、干物质积累量均呈先增加后下降的趋势,密度超过240×10⁴/hm²会导致LAI、干物质积累量、产量下降。在240×10⁴/hm²密度条件下,施氮量超过240 kg/hm²会导致小麦叶面积指数、SPAD值、花后干物质积累量和产量下降。适当的增密减氮有利于提高弱筋小麦的优质稳产,而过量增密减氮则会导致小麦产量下降,品质不稳定。为实现产量和品质的最优化,生产上推荐采用种植密度为240×10⁴/hm²,施氮量为180 kg/hm²,氮肥运筹为7:1:2:0的栽培模式。     

草果AtNCED基因克隆、表达和植物表达载体构建

9-顺式环氧类胡萝卜素双加氧酶(9-cis-epoxycarotenoid dioxygenase, NCED)是ABA生物合成的限速酶。诸多研究表明NCED基因表达量的变化与ABA含量显著相关,在种子休眠过程中发挥重要作用。为探究草果AtNCED基因的序列特征和功能,本研究以草果(Amomum tsaoko Crevost et Lemarie)种子转录组为基础,采用RT-PCR技术克隆了一个AtNCED基因。该基因全长2 372 bp,包含一个1 830 bp的开放阅读框(open reading frame, ORF),编码610个氨基酸。生物信息学分析显示,AtNCED基因编码的蛋白在N-末端包含典型的叶绿体转运肽序列,在催化结构域含有4个高度保守的组氨酸残基。AtNCED蛋白与芭蕉科马来西亚蕉同源性较高,与其他单子叶植物处在同一进化分支。实时荧光定量PCR分析结果表明,AtNCED基因表达量随着层积时间的增加逐渐下降,该基因可能正向调控种子休眠,在草果种子休眠诱导与维持中发挥重要作用。进一步地,利用同源重组方法,成功构建了pBI121-AtNCED过表达载体,并转化至农杆菌EHA105。该研究结果为弄清ABA激素信号在种子休眠中的调控作用提供了帮助。     

甘薯耐旱性品种苗期筛选及耐旱性指标研究

为实现甘薯耐旱性品种的快速筛选,本试验采用室内PEG模拟连续干旱法对22个甘薯品种(系)进行了研究。结果表明:济薯21、12057、11001和零9281等品种(系)的根系持水力、根系活力和幼苗成活率较高,具有较强的苗期耐旱性;通过统计分析发现,根系持水力、薯苗质量和含水量、根系活力等可作为重要的苗期耐旱性筛选指标;干旱胁迫下根系的持水能力和采苗后薯苗状况是影响甘薯苗期耐旱性的重要因素。在生产上,选用耐旱性品种和采用壮苗是抵御干旱胁迫的重要措施。     

果菜类蔬菜早熟性育种研究进展

早熟性是蔬菜作物育种的重要指标,对农业增产和农民增收意义重大。本文对果菜类蔬菜早熟性评价指标,早熟性相关农艺性状和早熟性育种方法三个方面的研究进展进行综述,并对基因工程辅助早熟育种进行了展望。     

Genome-wide Association Study of Teats Number Traits in Kele Pigs

Teats number is one of the most important reproductive traits,and closely related to the economic benefit in pig industry.In order to reveal the underlying genetics of left teats number,right teats number and total teats number traits,a genome-wide association study（GWAS）was performed.Samples of DNA were collected to genotyping for 22 Kele pigs using the Illumina Porcine SNP 60K Chip.The GWAS was performed using a mixed-effects model and linear regression approach.When a genome-wide threshold was determined using the Bonferroni method（P<2.06E-5),4 single nucleotide polymorphism（SNP）markers were potentially associated with left teats number,right teats number and total teat number.However,3 SNPs were significant associated and 18 SNPs were potentially associated in chromosomes level.304 Ensembl genes were retrieved around 1 cM of the associated SNPs.The candidate genes in Wnt and Fgf signaling pathway（BTRC,FGF5,FGF8,BMP3,RASGEF1B and HMGB3）might have effect on target traits.These results provided valuable information about the selective breeding for Kele pigs.     

YWS-1型烟田挖坑施肥机的研究设计与应用

为了更好地解决“烟草三农”问题，针对我国烟草种植落后的现状和发展趋势，研究和应用具有稳定挖坑和精量施肥等功能YWS-1型烟田挖坑施肥机。通过性能和生产试验测定和效益分析证明，该机具各项技术、性能指标符合设计要求，解决了烟草种植的问题，具有一定的创新特点和较好的推广和应用价值。     

益生菌大肠杆菌Nissle 1917抗逆性能、猪肠上皮细胞黏附率及抑菌效果研究

本试验旨在研究益生菌大肠杆菌Nissle 1917(Ec N)抗逆性能、猪肠上皮细胞黏附率及抑菌效果。采用体外法对Ec N进行生长曲线绘制和耐酸、耐胆盐、耐热性能的测定;以猪肠上皮细胞IPEC-J2细胞为体外细胞模型,考察了Ec N对该细胞的黏附率以及对致病菌大肠杆菌K88的黏附抑制率;同时通过蛋白质印迹法检测了Ec N对IPEC-J2细胞β-防御素-2和Toll样受体4的水平的影响。结果表明:1)Ec N对高酸、高胆盐和高温环境具有一定耐受能力。2)Ec N对IPEC-J2细胞的黏附作用以对数期最佳,黏附率达33.96%,显著高于迟缓期、稳定期和衰亡期(P0.05)。3)Ec N对致病菌大肠杆菌K88具有良好的抑制效果,黏附抑制率达87.84%。4)Ec N还能上调IPEC-J2细胞β-防御素-2和Toll样受体4水平。结果提示,益生菌Ec N具有较好的抗逆性能,能够良好地黏附猪肠上皮细胞,对致病菌大肠杆菌K88具有良好的抑制作用。