122份栽培桃品种（系）黄白肉性状的分子标记辅助鉴定

【目的】 类胡萝卜素裂解双脱氧酶基因（Carotenoid Cleavage Dioxygenases 4,CCD4）控制桃果肉颜色（白/黄）,CCD4存在3种等位基因。本研究利用Indel、SSR荧光标记毛细管电泳及SNP鉴定等基因分型技术分析我国主要桃黄白肉品种（系）中CCD4等位基因的差异,为主要黄/白肉品种（系）的基因型鉴定、亲本选配和选择相应的分子标记对不同来源子代的果肉颜色进行鉴定奠定基础。【方法】利用已经报道的桃不同果肉颜色中CCD4等位基因3种突变类型,合成不同引物进行PCR扩增,LTR反转录转座子插入突变经1%的琼脂糖凝胶电泳检测,CT单元重复的PCR产物在ABI3730XL测序仪上进行SSR荧光标记毛细管电泳检测,SNP标记经Sanger测序后利用ContigExpress软件分析CCD4等位基因的碱基替换（A→T）。综合以上结果,统计每份材料中CCD4等位基因的突变类型与果肉颜色的一致性。【结果】通过对不同来源的122份桃品种（系）材料进行基因型分析,发现CCD4发生LTR反转录转座子插入突变材料的基因型共有31份,占总材料的25.4%,其中纯合插入突变材料的片段扩增长度为729 bp,共有8份,占总突变的25.8%;CCD4发生微卫星重复序列突变材料存在2 bp的插入,扩增片段长度为179 bp,该类型共有68份,占总材料的55.7%,其中纯合插入材料25份,占总突变的36.8%;CCD4发生A→T碱基替换突变的材料较少,仅有1份,占总材料的0.82%,实际应用中可以不考虑该种类型。CT和LTR插入的两种突变类型的黄肉品种（系）有7份,占总材料的5.7%。研究结果表明,LTR反转录转座子插入突变和微卫星序列重复突变是黄肉桃中CCD4等位基因的主要突变类型。其中CCD4发生一种纯合突变或两种杂合突变桃品种（系）为黄肉类型,分子标记鉴定结果与调查的122份桃品种（系）黄白肉表型性状完全一致,准确率为100%。【结论】采用分子标记明确了122个桃品种（系）黄/白肉性状的基因型,为不同亲本组合子代表型鉴定的标记类型选择提供了技术支撑,为建立桃种质材料黄/白性状的分子辅助育种体系和黄肉桃的选育奠定了基础。     

Characterization of Genomic Integration and Transgene Organization in Six Transgenic Rapeseed Events

To characterize the DNA rearrangement of both the T-DNA region and the genomic insertion site during T-DNA insertion, the Genomewalker strategy was used to isolate the junctions between the inserted DNA and the plant genomic DNA in six rapeseed events as well as the genomic DNA at the sites before integration. During transformation in each of the six events, portions of both the right border（RB） and left border（LB） regions of the T-DNA were deleted, ranging from a 7 nucleotide deletion of the LB repeats in event RF1 to a 207 bp deletion of the LB region in event RF2. For the six events, T-DNA integration resulted in a deletion at the target site spanning less than 100 bp. Sequence analysis indicated that the T-DNA was integrated into the coding region of various native rapeseed genes in events RF1 and RF2. Duplications of the genomic DNA target site were observed in events RF2, RF3 and Topas 19/2. And multimerization of transgenes was found in event Topas 19/2, in which, the T-DNA was integrated as a head-to-head（RB-to-RB） concatemer into the recipient genome. In event MS1, chromosomal translocation or a large target-site deletion may have occurred during T-DNA integration, which was identified due to a failure to amplify the presumptive insertion site based on the flanking rapeseed DNA sequences. Our results provide comprehensive data concerning transgene organization and the genomic context of the T-DNA in six rapeseed events, which can aid in the developing of insert fingerprinting and the monitoring of long-term genetic stability and potential unintended effects of transgenic events.     

草原生态奖补政策实施效应——基于政治社会学视角的实证分析

草原生态奖补政策有助于保护草原生态,促进畜牧业的发展转型,有利于民族地区的和谐稳定。生态奖补政策的实施对牧民生计、基层社会治理等方面也产生了一些负面影响。未来的政策实践过程中,需要建立政策评估反馈机制,不断提高生态奖补标准,完善奖补资金发放办法,着力调动地方政府积极性,力求实现奖补手段的多样化,不断提高政策的法制化、系统化水平。     

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells

Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.     

Effects of Different Nitrogen Treatments on N use Efficiency and Grain Yield of Mechanically-transplanted Hybrid Rice

In order to investigate effects of topdressing on grain yield and N use efficiency of mechanically-transplanted hybrid rice,this experiment was conducted with 2-line hybrid rice cultivars,Quanliangyou 1 and Quanliangyou 681 with 3 application levels of N fertilizer,No,N1,N2,respectively.The results showed that there were no significant differences in grain yields of the 2 varieties between treatments N1 and N2.It was indicated that N is not the factor which could limit grain yield within limited application levels of N fertilizer.High N fertilizer treatment could contribute to the formation of effective panicles and spikelets per panicle,but it could also lead to the decline in grain filling rate.It was suggested that lowering the amount of Nfertilizer could facilitate improvement of N-use efficiency.     

Cloning and Sequence Analysis of Buffalo Keap1 Gene and Investigation of Its Expression Pattern in Different Tissues

In this study,the CDS sequence of buffalo Keap1 gene was cloned and analyzed,then its expression pattern in different tissues was also investigated.A pair of primers of buffalo Keap1 gene was designed based on the nucleotide sequence of Bos taurus Keap1 gene from GenBank,and then the buffalo Keap1 gene was amplified.Using the bioinformation techniques,the gene sequence and the protein structure were analyzed.The expression level of Keap1 gene in different tissues were detected with Real-time quantitative PCR.The results showed that the length of buffalo Keap1 gene coding sequence was 1 875 bp and encoded 624 amino acids.The multiple sequence alignment results showed that buffalo Keap1 gene shared 99%,96%,92% and 90% of similar nucleotide sequence with that of Bos taurus,Ovis aries,Sus scrofa and Homo sapiens,respectively.And the phyogenetic tree also showed the conservatism between several different species.The second structure of buffalo Keap1 protein was predicted as 24 alpha regions,40 beta regions,38 turn regions and 27 coil regions.In addition,the results of Real-time quantitative PCR showed that Keap1 mRNA exists in all seven tissues,but the most abundant expression was in heart and the minimal expression was in liver and spleen.The results provided an foundation for further study of Keap1-Nrf2-ARE signal pathway,for enhancing the ability of antioxidant of buffalo embryo in vitro culture.     

江西省新型农业社会化服务体系的特点与运行模式

实地调研江西省新型农业社会化服务体系建设现状，对江西省目前的新型农业社会化服务体系的特点与运行模式进行分析，针对当地在服务体系构建过程中出现的问题，提出新时期的发展建议。     

不同药剂组合对马铃薯晚疫病的防治效果

马铃薯晚疫病是马铃薯生产中的重要病害,是影响马铃薯产量和品质的重要因素之一,在云南春作马铃薯上普遍发生,且危害较重,给当地马铃薯产业带来了巨大的经济损失。为减轻马铃薯晚疫病对马铃薯生产造成的损失,试验研究了11种药剂7种药剂组合(包衣剂+保护剂+治疗剂)通过播种期种薯包衣,现蕾期叶面喷施一次保护剂,发病初期和发病中期叶面各喷一次治疗剂的方法防治马铃薯晚疫病。结果表明,药剂组合种薯包衣+甲霜·锰锌+氟吡菌胺·霜霉威的防治效果最好,防治效果达到60.99%,增产率176.28%,增效率152.27%;其次是种薯包衣+霜脲·锰锌+氟吡菌胺·霜霉威组合,防治效果达到60.15%,增产率173.20%,增效率150.55%。这2个药剂组合对马铃薯晚疫病均有极显著的防治效果,增产增效明显。