AtSOS基因在紫花苜蓿中的表达及其耐盐性研究

本研究采用基因工程技术改良紫花苜蓿耐盐性,通过种植转基因耐盐紫花苜蓿达到改良土壤的目的。以紫花苜蓿子叶节为外植体,通过农杆菌介导法将来源于拟南芥的AtSOS1-AtSOS2-AtSOS3多基因表达载体导入阿尔冈金紫花苜蓿中,经PCR检测、抗除草剂筛选和RT-PCR鉴定,获得了能稳定表达的转基因株系。以转基因的紫花苜蓿和野生型紫花苜蓿为材料进行盐处理,每个处理重复3次,测定其生理生化指标、株高、Na⁺和K⁺含量、细胞膜透性、叶绿素含量。结果显示,在不同盐浓度处理下,所有植株的株高均有所增长,但在100和200 mmol·L^-1的NaCl处理下,转基因植株的长势显著高于野生型植株;随着处理时间的增加,所有植株的叶绿素含量均呈先上升后下降的趋势,且野生型植株叶绿素含量均低于转基因植株;在100和200 mmol·L^-1的NaCl处理下,转基因植株的细胞膜透性、超氧化物歧化酶活性和脯氨酸含量的增加量均小于野生型植株,而过氧化物酶、过氧化氢酶活性和可溶性糖含量的增加量均大于野生型;各植株中丙二醛含量均下降,且野生型植株下降的更为明显;盐处理后,转基因植株根系中Na⁺的积累比野生型植株少,而K⁺的吸收多于野生型植株。转AtSOS基因的紫花苜蓿通过发挥AtSOS途径的作用,促进了植物体将细胞内的Na⁺外排,从而减轻盐胁迫对植物体的伤害,提高了转基因植株的耐盐性。     

Research Progress on Bacterial Resistance in Environment

Antibiotic resistance genes have become a recognized environmental pollutant, threatening the ecological safety and human health. The application of antibiotics in the clinical and animals breeding, making the environmental microorganisms living with the impact of the residue of antibiotics and elements of resistance genetic and leading to antibiotic resistant bacteria to gain a competitive advantage and destroyed the stability of ecosystem. In this paper, we expounded the concept of resistance gene transmission by the view of macro environment. Through analyzing the mechanism of resistance development,environmental pollution caused by drug resistance and the impact of environmental microorganism for drug resistance, we clarified the key role of the environment in the development of the characteristics of bacterial resistance and analysis environmental resistance.Such as the ecological diversity of flora, the types and the spread of resistant bacteria, the residues of antibiotics and the transmission of the resistance genes.     

养殖废弃物堆肥中抗生素和抗性基因的降解研究

抗生素的滥用及排放会造成细菌产生耐药性以及抗生素抗性基因(Antibiotic resistance genes, ARGs)的传播和扩散。畜禽粪便是导致环境中抗生素污染的主要来源之一。本文综述了四环素类、大环内酯类、喹诺酮类、β-内酰胺类、磺胺类和氨基糖苷类等在水土环境中广泛存在的抗生素及其环境残留水平和对动植物、微生物的影响,分析了当前利用堆肥技术降解畜禽粪便中抗生素和ARGs效果及机制的研究情况。总结得出,猪粪中抗生素残留量最高,其中四环素类残留量为1390~354 000 mg·kg^-1,磺胺类170.6~89 000 mg·kg^-1,氟喹诺酮类411.3~1 516.2 mg·kg^-1,硝基呋喃类85.1~158.1 mg·kg^-1,大环内酯类1.4~4.8 mg·kg^-1。堆肥对大部分抗生素具有好的降解效果,其中四环素类抗生素降解率为62.7%~99%,磺胺类为0~99.99%,对大环内酯类几乎可以完全降解,但是,堆肥无法降解喹诺酮类抗生素。养殖废弃物堆肥过程中,ARGs的降解情况同样因抗生素种类和堆肥方式而不同。已有的研究表明,除大环内酯类ARGs外,堆肥对其他ARGs均具有有效的降解效果,降解率为50.03%~100%。堆肥初期的优势菌门是厚壁菌门、放线菌门、变形菌门和拟杆菌门;堆肥结束后放线菌门成为最优势菌门。初始抗生素的浓度不影响堆肥结束时微生物的群落组成。温度和pH是影响抗生素降解的最主要因素,而ARGs的降解效果主要受温度影响。     

秸秆还田对麦玉系统土壤有机碳稳定性的影响

为揭示不同秸秆还田量对华北小麦-玉米轮作系统土壤有机碳官能团结构及稳定性的影响,研究了秸秆还田5 a后土壤有机碳官能团结构、团聚体组成及有机碳含量、活性有机碳含量、土壤铁离子的变化。田间实验设置4个处理:秸秆不还田作为对照(CK)、秸秆1/3还田(S1)、秸秆2/3还田(S2)、秸秆全部还田(S3)。采用常规方法测定土壤理化性质、粒径、铁离子及土壤微生物量碳含量,13C核磁共振波谱技术(NMR)检测分析土壤有机碳官能团结构。结果表明:秸秆还田5 a后,土壤总有机碳(TOC)、2mm与2.00～0.25 mm团聚体有机碳、可溶性有机碳(DOC)、易氧化态碳(EOC)和微生物量碳(MBC)含量,均随还田量增加而逐渐增加,且不同处理增加量不同,与CK相比,S3处理显著增加了这些有机碳的含量(P0.05)。各处理土壤有机碳以烷基碳与烷氧基碳为主,其次是芳香碳与羰基碳,秸秆还田增加了烷氧基碳、羰基碳(易分解碳组分)含量,降低了烷基碳和芳香碳(难分解碳组分)含量,与CK相比,S3处理显著增加烷氧基碳含量(P0.05)而显著降低了芳香碳含量(P0.05)。与CK相比,S2、S3处理也显著降低了有机碳的芳香度、疏水碳/亲水碳、烷基碳/烷氧基碳比值(P0.05),而对脂族碳/芳香碳影响不明显。与CK相比,S3处理显著增加了2.00 mm团聚体组分,增加了2.00～0.25 mm组分,而降低了0.25～0.053 mm组分和显著降低了0.053 mm组分(P0.05)。秸秆还田对土壤游离铁、活性铁、螯合铁含量的影响不明显。有机碳官能团组成与土壤因子间的冗余分析表明土壤TOC、MBC含量、团聚体组分、铁离子的改变是导致不同处理间有机碳官能团结构存在差异的重要原因。综上所述,由于短期秸秆还田增加了活性有机碳含量、易分解有机碳组分,减少了难分解有机碳组分,降低了微团聚体物理保护作用,改变了微生物活性和铁离子络合作用,在一定程度上降低了土壤有机碳稳定性,可能导致麦玉复种系统土壤碳排放水平的增加。     

Attributes of wheat cultivars for late autumn sowing in genes expression and field estimates

Due to the growing interest in Central and Eastern Europe on cropping of wheat in optional late autumn terms, called facultative, genetic research and field evaluation were taken on four spring cultivars: Tybalt (NL), Monsun (DE), Ostka Smolicka (PL) and Bombona (PL), currently being recommended by breeders. The PPD gene analyze, expression level of dehydrine genes (WCS120 and WDHN13) in cooling test, and qPCR for RNA isolation and analyses of WCS120 and WDHN13 gene expression at the BBCH12 stage of wheat were estimated. Molecular analysis of PPD-D1 gene confirmed the presence of photoperiod sensitive allele ppd-D1b in all tested genotypes. The highest level of NRE WCS120 gene was detected in cultivars Tybalt and Bombona. Two-year field experimental study assessed the growth, development and productivity of facultative and spring crops of studied cultivars. Based on our results from field experiments and result of molecular analysis of alleles of PPD-D1 gene, the tested genotypes can be considered as potentially facultative genotypes.     

Soybean plant height QTL mapping and meta‐analysis for mining candidate genes

Plant height is closely related to seed yield of soybean. The goal of this study was to identify important loci affecting soybean plant height using meta‐analysis based on a reference physical map. Plant height related to QTLs was mapped across eight years with a RIL population by WinQTLCart v2.5. 182 QTLs related to plant height of soybean from database and our research were collected, and each QTL was projected onto the soybean physical map by software BioMercator v2.1. The confidence interval of meta‐QTL ranged from 0.09 to 5.07 Mb, and the mean phenotypic variance ranged from 4.9% to 73.0%. Furthermore, 4,259 candidate genes were located in these consensus QTLs, and 40 of them were involved in the plant growth and stem elongation and annotated as plant hormone signal transduction (pathway ID ko04075) in KEGG pathway. These results would lay a foundation for fine mapping of QTLs/genes related to plant height and marker‐assisted selection for breeding in soybean.     

Bioinformatics Analysis of Bone Morphogenetic Protein 4 (BMP4) of Sheep

The assay was aimed to explore the biological characteristics of bone morphogenetic protein 4 (BMP4) of sheep,NCBI,DNAMAN DNAStar,TMHMM Server v.2.0,PsortⅡ,SignalP various bioinformatical softwares were used to speculate the physical and chemical properties,hydrophobic property,phosphorylation site,conservative structure domain,protein secondary structure of BMP4 protein.Also,the three-dimensional structure was forecasted with the SWISS-MODEL Workspace software.The results indicated that the BMP4 of sheep had high homologies with the BMP4 of various species.The encoded protein was a hydrophilic protein which was unstable.There was no transmembrane regions and it was likely to be located in the nucleus.What was more,there was signal peptide and eighteen phosphorylation sites.Through the forecast of functional domains,the protein had two functional domains,including the transforming growth factor-beta (TGF-beta) superfamily and TGF-beta propeptide superfamily.The result was consistent with the function of BMP4 gene family,it also demonstrated that BMP4 was a growth factor and it had the function of signal transduction.The amino acid homology between the predicted 3D structure of protein and template 3bmp.1.A was 88.29%.The bioinformatics analysis of BMP4 gene could provide reference for the further study in practice.     

低温胁迫下马铃薯的数字基因表达谱分析

以马铃薯品种合作88为材料,利用数字基因表达谱(DGE)技术,对–2℃低温胁迫处理后的马铃薯叶片c DNA文库进行差异基因表达谱分析。结果表明,有28 505个基因受低温胁迫诱导差异表达,其中上调表达基因13 703个,下调表达基因14 802个。GO功能显著性富集分析表明,DEGs主要涉及信号生物代谢过程、氧化还原过程、能量代谢、次生代谢过程以及催化活性。KEGG富集分析表明,上调表达基因主要富集于苯丙烷、光合作用天线蛋白、类胡萝卜素的生物合成、苯丙氨酸代谢及淀粉与蔗糖代谢途径,而下调表达基因主要富集于植物激素信号转导途径。利用实时荧光定量PCR(q RT-PCR)验证4个DEGs在低温胁迫条件下的差异表达,其结果与DGE分析结果基本一致,证实了DGE测序结果的可靠性。