京张生态关系与张家口市贫困问题关联分析

张家口是河北省最为贫困的地区,也是全国范围内贫困县较为集中的地区之一,而且,贫困势态愈演愈烈。在众多的贫困成因中,京张生态关系对张家口贫困影响最为深远。从京张生态关系、张家口市工农业生产发展及资源开发受限、与周边地区的政策反差、京张生态补偿机制的缺失等角度,分析了京张生态关系与张家口贫困的关联关系。     

Identification of a novel miRNA from the ovine ovary by a combinatorial approach of bioinformatics and experiments

MicroRNAs (miRNAs) are a class of short endogenous, single-stranded, non-coding small RNA molecules, about 19–25 nucleotides in length that regulate gene expression at the translation level and influence many physiological process, such apoptosis, metabolism, signal transduction, and occurrence and development of diseases. In this study, we constructed a library from the ovine luteal phase ovary by using next-generation sequencing technology (Solexa high-throughput sequencing technique) and identified 267 novel miRNAs by bioinformatics. One of the novel miRNAs (ovis_aries_ovary-m0033_3p), which expressed in the sheep ovary and testis, was confirmed by real time PCR and northern blot. Ovis_aries_ovary-m0033_3p was 21 nucleotides in length and located on chromosome 12, and it had 100% similarity to hsa-miR-214-3p, mmu-miR-214-3p, dre-miR-214and ssc-miR-214. Meanwhile, the pre-miRNA was 82 nucleotides in length and had a standard hairpin stem-loop structure. From the consistency of the sequence and structure, we speculated that ovis_aries_ovary-m0033_3p had a function similar to hsa-miR-214-3p, which is involved in the fine regulation of cell survival, embryonic development, breeding activities and resistance to ovarian cancer, so we defined it as oar-miR-214-3p. These experimental results will enrich the miRNA database for ovis aries and provide the basis for researching the regulation mechanism of miRNA in relation to breeding activities of seasonal breeding animals.     

河北省农业龙头企业技术推广现状与存在问题及对策

概述了农业龙头企业技术推广的内涵和意义,结合河北省农业龙头企业技术推广现状,深入剖析其在推广过程中所面临的不足和问题,并在此基础上探索进一步完善和改进的对策.     

Fusing multi-source data to map spatio-temporal dynamics of winter rape on the Jianghan Plain and Dongting Lake Plain,China

Mapping crop distribution with remote sensing data is of great importance for agricultural production, food security and agricultural sustainability. Winter rape is an important oil crop, which plays an important role in the cooking oil market of China. The Jianghan Plain and Dongting Lake Plain(JPDLP) are major agricultural production areas in China. Essential changes in winter rape distribution have taken place in this area during the 21 st century. However, the pattern of these changes remains unknown. In this study, the spatial and temporal dynamics of winter rape from 2000 to 2017 on the JPDLP were analyzed. An artificial neural network(ANN)-based classification method was proposed to map fractional winter rape distribution by fusing moderate resolution imaging spectrometer(MODIS) data and high-resolution imagery. The results are as follows:(1) The total winter rape acreages on the JPDLP dropped significantly, especially on the Jianghan Plain with a decline of about 45% during 2000 and 2017.(2) The winter rape abundance keeps changing with about 20–30% croplands changing their abundance drastically in every two consecutive observation years.(3) The winter rape has obvious regional differentiation for the trend of its change at the county level, and the decreasing trend was observed more strongly in the traditionally dominant agricultural counties.     

不同高精料耐受时间对泌乳期山羊肝脏糖脂代谢的影响

为了研究饲喂高精料日粮对泌乳期山羊肝脏糖脂代谢的影响,试验选用16只健康经产泌乳中期关中奶山羊,随机分为3组,低精料组(LC, n=5)、长期高精料组(HL, n=7)和短期高精料组(HS, n=4)。实验结束后采集血液和肝脏组织。结果显示,与LC组相比,HL和HS组奶山羊血糖和肝糖原水平显著升高(P<0.05),血液中甘油三酯(TG)水平显著降低(P<0.05),胆固醇(Tch)含量没有显著变化(P>0.05);HS组山羊肝脏TG和Tch水平显著增加(P<0.05),HL组无显著变化 (P>0.05)。HL组肝脏中丙酮酸羧化酶(PC)和葡萄糖转运蛋白2(GLUT2)mRNA表达水平显著上调;HS组肝脏PC、固醇调节元件结合蛋白1(SREBP1)、硬脂酰辅酶A去饱和酶(SCD)、二酰基甘油酰基转移酶(DGAT)和3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR) mRNA表达水平显著升高(P<0.05),SREBP-2有升高的趋势(P=0.76),肉碱棕榈酰转移酶-1(CPT-1)和胆固醇7α-羟化酶(CYP7α1)mRNA表达显著下降(P<0.05);线粒体型磷酸烯醇式丙酮酸羧激酶(PCK2)蛋白表达水平显著升高(P<0.05)。以上结果表明,短期饲喂高精料日粮能增强肝脏糖异生能力,促进肝脏胆固醇和脂肪合成;而长期饲喂高精料日粮能增强肝脏糖异生作用,但对胆固醇和脂肪代谢相关基因的表达没有影响。     

肠炎沙门菌Ⅰ型菌毛主要亚单位FimA原核表达及纯化

为探讨肠炎沙门菌非编码小RNA(non-coding small RNA)IsrE在转录后水平是否参与对fimA的调控作用,在大肠杆菌中构建并表达FimA重组蛋白,并对其培养条件进行优化,制备高纯度FimA重组蛋白。利用原核表达载体p ET-28a(+)构建出能表达fimA的重组质粒,并分别对IPTG浓度、诱导温度和诱导时间等条件进行优化,以提高目的蛋白的产量和增大其可溶性,最后经Ni-NTA亲和层析分离纯化FimA重组蛋白。通过酶切、测序和Western blot鉴定分析,成功构建并表达肠炎沙门菌I型菌毛主要亚单位FimA,蛋白分子质量约为19.3 kDa,且主要以包涵体的形式表达,经Ni-NTA亲和层析纯化后的重组蛋白在SDS-PAGE中显示单一条带。本研究构建、表达并获得纯度较高的重组蛋白FimA,为进一步研究和验证肠炎沙门菌非编码sRNA IsrE是否在转录后水平参与对fimA的调控奠定了基础。     

不同动物源大肠杆菌gapA看家基因的克隆及序列比较

3-磷酸甘油醛脱氢酶(Glyceraldehyde 3-phosphate dehydrogenase,GAPDH/G3PDH)是糖酵解反应的关键酶,为生命体的各种生命活动提供能量.在大肠杆菌中,GAPDH由看家基因gapA编码.本研究对不同动物源的大肠杆菌菌株(包括禽源致病性大肠杆菌(APEC)分离株O1和CE129、人源肠致病性大肠杆菌(EPEC)菌株738、人源益生菌Nissle1917和猪源表达F18菌毛大肠杆菌标准株F107/86的gapA基因进行PCR扩增、克隆测序,并与GenBank中8株菌株序列比较,发现其核酸序列虽有少数位点的突变,但是氨基酸序列仍保持完全一致,从分子基础上保证了GAPDH的酶活性.本研究验证了原核病原微生物,尤其是大肠杆菌gapA基因转录翻译的GAPDH蛋白的高度保守性,为制备大肠杆菌专用内参蛋白抗体提供可能,并进一步为对致病性大肠杆菌的蛋白代谢通路和合成生物学研究提供了基础材料.     

红麻细胞质雄性不育系与保持系总蛋白提取及双向电泳体系优化

为进一步研究高等植物体内控制蔗糖合成的关键酶基因蔗糖合酶基因Susy在甘蔗中的表达情况，本研究以甘油醛-3-磷酸脱氢酶（GAPDH）基因为内参基因，通过对退火温度、循环数的优化建立了一个稳定、方便的半定量RT-PCR体系。通过该体系检测干旱处理下甘蔗叶的Susy基因表达，并结合叶片酶活性和蔗糖含量进行分析。结果表明：甘蔗Susy基因的半定量RT-PCR扩增，以循环数30个、退火温度为54 ℃、内参基因与目的基因同批异管进行扩增为宜。甘蔗Susy基因在不同叶位中的表达有很大差异，总体上表现为：+1叶、+2叶>+5叶、+6叶>心叶，干旱胁迫可诱导幼嫩叶中Susy基因的表达量上升，基因表达的变化与其酶活性和蔗糖含量变化的情况相一致。