牧草产业是我国家兔产业可持续发展的重要支撑

针对我国家兔规模化养殖的快速发展,牧草作为家兔生产主要饲料原料来源,供需矛盾日益突出。本文重点从家兔消化生理特点、兔产品安全生产、农民增收、需求现状与问题等方面进行了论述,并提出了相应对策。     

三种融合标签对大熊猫轮状病毒结构蛋白VP6-VP7可溶性表达的影响

大熊猫轮状病毒(giant panda rotavirus,GPRV)是引起幼龄大熊猫腹泻的主要病原,对圈养大熊猫产生了较大的危害。轮状病毒结构蛋白VP6是一种载体蛋白,可介导黏膜免疫反应。VP7是轮状病毒结构蛋白中主要的中和抗原。因此,VP6-VP7的融合表达作为候选抗原对该病的防治具有重要的意义。传统大肠埃希菌原核表达存在表达量低、可溶性差以及纯度低等弊端。本研究使用醛缩酶(EDA)、谷胱甘肽S-转移酶(GST)、麦芽糖结合蛋白(MBP)3种融合标签,以实现获得表达量高和纯度高的GPRV-VP6-VP7重组表达蛋白。将扩增的VP6、VP7基因片段利用同源重组酶构建到含3种融合标签的表达载体pET21b上,将重组质粒转化至大肠埃希菌Rosetta(DE3)感受态细胞中进行低温诱导表达。用Ni-柱亲和层析法纯化目的蛋白,SDS-PAGE和Image J分析蛋白表达量和可溶性,Western blot分析得到表达的重组表达蛋白正确且具有蛋白活性。实验结果证明,EDA标签能显著促进VP6-VP7蛋白的原核可溶性表达,提高VP6-VP7蛋白表达量。     

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.     

非会计专业《会计学》课程教学方法刍议

目前,许多高校非会计专业都将《会计学》课程作为一门专业必修的基础课程。结合多年的非会计专业会计教学经验,笔者认为根据专业特点,采用灵活多样的教学方法是实现非会计专业《会计学》课程教学目标,提高《会计学》教学效果的重要手段。     

美国农产品分布对我国农产品区域发展的影响研究

美国农业一直处于领先地位,不仅得益于其丰富的自然资源和高科技的农业技术,其合理的农产品分布也不可少的影响因素。中国是一个农业大国,农业的发展离不开农产品,农产品的种植种类、农产品的区域分布、农产品的质量发展都直接影响农产品的发展。美国虽然面积和我国差不多,但因为有合理的布局,农产品贸易一直发展良好。要借鉴美国农产品的发展,研究和分析其农产品分布现状很有必要。     

基于S基因序列分析新型猪流行性腹泻病毒的遗传演化

最近，笔者实验室在青藏高原地区发现两种新亚型藏猪源猪流行性腹泻病毒（PEDV），为进一步调查新型PEDV是否在四川腹泻猪群中存在或流行，对实验室2018-2019年保存的116份猪腹泻粪便或肠组织样本进行PEDV的检测及其纤突蛋白基因（spike）分子特征研究。结果表明：腹泻样本的PEDV检出率为42.2%（49/116，95% CI=33.1%~51.8%），并获得了13条完整的S基因序列，全长为4 149~4 170 bp，序列相似性为94.2%~99.9%，其中SWUN-H3-CH-SCYA-2019的S基因与藏猪源新G1亚群PEDV的序列相似性高达97.0%~98.6%。遗传演化研究结果表明13株PEDV S基因划分为G1和G2大群，其中SWUN-H3-CH-SCYA-2019位于藏猪源新G1亚群；SWUN-19-CH-SCZY-2018、SWUN-4-CH-SCXC-2018、SWUN-1-CH-SCNJ-2019和SWUN-3CH-CH-SCZG-2019位于G2亚群中一个独立的分支，且与藏猪源新G2亚群毒株有着较近的亲缘关系。为了进一步研究13株PEDV的演化过程，以贝叶斯进化分析软件包（BEAST）进行分歧时间估算，结果表明SWUN-H3-CH-SCYA-2019的分歧时间约为2012.3年，早于藏猪源新G1亚群其余毒株的最早分歧时间（2015.7年）；SWUN-4-CH-SCXC-2018、SWUN-19-CH-SCZY-2018和SWUN-3CH-CH-SCZG-2019的分歧时间约为2014.2年，早于G2亚群的藏猪源毒株2014.7年，所有藏猪源PEDV的分歧时间均晚于四川毒株。本研究在四川地区首次发现了藏猪源PEDV，并且从毒株的分歧时间推断青藏高原的藏猪源PEDV来源于四川，为新型PEDV分子遗传进化的监测提供了依据。     

灰霉菌侵染对番茄幼苗活性氧积累及抗氧化酶活性的影响

为进一步分析活性氧和抗氧化防御系统在番茄抗病信号调控机制中的作用,为提高产量和品质提供科学依据,以4叶1心的番茄幼苗为试验材料,比较番茄幼苗受灰霉菌侵染后叶片O_2~-·以及抗氧化酶、内源褪黑素等的变化。结果表明:番茄幼苗接种灰霉菌后,随着时间的延长,幼苗叶片上病斑斑点逐渐增多,灰霉菌含量显著增多,叶色逐渐变浅,植株表现明显的发病症状;灰霉病菌侵染后番茄体内O_2~-·活性爆发,超氧化物歧化酶(SOD)和过氧化物酶(POD)活性下降,过氧化氢酶(CAT)和谷胱甘肽还原酶(GR)活性上升,褪黑素含量增加;上述变化都是番茄幼苗为了抵御灰霉菌的生物胁迫而发生的应激反应。综上,植物在遭受病原菌等生物胁迫时不能简单地推断SOD、CAT和POD等抗氧化酶活性一定升高,植物抵御病原菌侵染更多地依赖于抗性信号路径中信号物质的激活和积累。     

Whistle signal variations among three Indo-Pacific humpback dolphin populations in the South China Sea: a combined effect of the Qiongzhou Strait's geographical barrier function and local ambient noise?

Geographic variations in the dolphin whistles could be useful in assessing association and isolation among populations. Whistle of free-ranging Indo-Pacific humpback dolphins (Sousa chinensis) among the Pearl River Estuary (PRE), Leizhou Bei (LZB) and Sanniang Bay (SNB) populations were investigated. A total of 2850 whistles with legible fundamental contour were extracted and 15 acoustic parameters were measured. Contrary to SNB, PRE and LZB had the same relative proportion of tonal type compositions with flat and sine representing the most frequent types. The generalized linear model analysis showed significant acoustic difference among populations and tonal types. All frequency parameters in SNB were significantly higher than those in PRE and LZB, where no significant variation was observed in most of the parameters either at the population level or within each tonal type. Canonical discriminant functions analysis showed a smaller difference between PRE and LZB than between PRE and SNB and between LZB and SNB. Compared with previous recordings, recent recordings demonstrated a consistent pattern of becoming higher in whistle frequency parameters in both LZB and SNB populations, suggesting that noise pollution in LZB and SNB increasing with time according to the acoustic niche hypothesis. Dolphin whistle's geographic variations could be shaped by the combined function of the geographical barrier function of the Qiongzhou strait and local ambient noise. Considering the isolated condition and the relatively smaller population size of the humpback dolphin in the SNB, more effective and proactive conservation actions should be taken to prevent the extinction of small populations.