小反刍兽疫病毒及山羊痘病毒N-ITR基因二联实时荧光定量RT-PCR检测方法的建立

Peste des petits ruminants virus (PPRV) and goat pox virus (GTPV) are the causative agents of two kinds of goats’ diseases-peste des petits ruminants and goat pox which can cause disaster economic losses. In order to detect the two viruses simultaneously and quickly, two sets of primers and relative probes were designed based on the nucleoprotein (N) gene of PPRV and the inverted terminal repeat (ITR) segment of GTPV， respectively. In order to work together in the same reaction, the probes were labeled with different fluorescent materials 5′FAM-TAMRA3′and 5′JOE-Eclipse3′，respectively. Results showed that the duplex Real-time RT-PCR assay was identified to be specific for PPRV and GTPV only and specific fluorescent signal could be detected, but the related viruses including fowl pox virus（FPV）and canine distemper virus (CDV) had no specific fluorescent signal. Positive recombinant plasmids (PPRV pMD18-T-N and GTPV pMD18-T-ITR) were built and used for positive quantitative templates to establish duplex standard curves. The developed assay based on the probe N-ITR was found to be highly specific and sensitive with a detection limit of 10² copies/μL cDNA and 10³ copies/μL DNA for PPRV and GTPV， respectively. Finally, the duplex Real-time RT-PCR assay for simultaneous detection of PPRV and GTPV was established preliminarily in the study.     

Identification and verification of quantitative trait loci for eating and cooking quality of rice (Oryza sativa)

This study was conducted to identify new quantitative trait loci (QTLs) that have stable effects for eating and cooking quality (ECQ) of rice. Three recombinant inbred line populations of indica rice were each planted in two years. Three traits for ECQ, amylose content (AC), gel consistency (GC) and alkali spreading value (ASV), were measured for QTL analysis. A total of 13 QTLs were detected, including four for AC, six for ASV and three for GC. Two QTLs, qGC4 in the interval RM16252–RM335 on the short arm of chromosome 4 and qGC6.2 in the Alk region, were validated in a population derived from a residual heterozygote that was homozygous at the major locus Wx. In the absence of segregation at the Wx locus, qGC4 and qGC6.2 had additive effects of 2.46 and 8.18 mm, respectively, offering a potential for improving GC property of rice varieties. Comparison between qGC4 and previous results suggests that qGC4 is likely a new QTL for GC, providing a candidate for gene cloning and functional characterization.     

基于气象要素影响的柑橘定量滴灌模型研究

为了准确估算柑橘水肥一体化的滴灌量，实现手机APP远程操控滴灌起止时间。基于广西桂林1994—2015年柑橘物候期与同期气象条件的分析，构建了桂林柑橘不同生育期土壤需水量模型及降水量等气象要素与土壤重量含水率变化的转化算法模式，给出了柑橘滴灌量技术流程。应用模型估算广西农垦国有明阳农场（南宁）柑橘种植示范园区1997年3月干旱时的滴灌量。结果表明：应用模型估算滴灌量比传统模型节水40%以上。此结果对“气象监测预报信息+规模化生产管理设施+生产信息反馈与互动”创新服务模式有积极参考意义。     

茶多酚高通量检测技术研究进展

茶多酚是茶叶的主要活性成分,具有抗氧化、抗肿瘤、抗衰老、降血糖、防辐射等多种生物学活性。然而,茶多酚的不稳定性极大限制其进一步开发应用。茶多酚易受光线、温度、pH等因素影响而发生氧化、聚合和缩合,从而使其活性改变。因此,茶多酚组分的高通量快速检测在生物学及临床上具有重要意义。高效液相色谱法、毛细管电泳法、近红外光谱法、核磁共振等均是茶多酚组分高通量检测的主要方法,但这些方法各有优劣。对茶多酚高通量快速检测技术及其相关应用进行了归纳和总结,并对其优势及存在的问题进行分析,以期为茶多酚组分的高通量检测提供参考。     

hrpZ_(Psta)在转基因大豆中定量表达与疫霉根腐病和灰斑病抗性相关研究

以T5转hrpZPsta基因大豆JN29-705-15和JL30-187为材料,利用实时荧光定量PCR技术(Quantitative Real Time PCR,qRT-PCR)检测了目标基因在转基因大豆不同组织中的表达量,分别利用下胚轴侵染法和叶面喷施法鉴定疫霉根腐病抗性和灰斑病抗性,并分析目的基因表达量与疫霉根腐病和灰斑病抗性的相关性。结果表明:hrpZPsta基因在大豆的叶、茎、根、籽粒中均有表达,二个株系平均相对表达量分别为8.2/6.1、0.9/0.7、6.5/4.6和0.8/0.7;T5转hrpZPsta基因大豆抗疫霉根腐病和灰斑病能力与野生型相比均有所提高,JN29-705-15对疫霉根腐病抗性从感病提高到中抗,而JL30-187从中抗提高到抗病;hrpZPsta基因在叶中的表达量也与抗灰斑病能力呈正相关,与病情级别呈极显著负相关。试验结果初步证明了外源基因hrp ZPsta在大豆植株中的表达量与受体植株对疫霉根腐病和灰斑病抗性存在一定的相关性。     

Genomewide association study reveals novel quantitative trait loci associated with resistance towards Septoria tritici blotch in North European winter wheat

Fungal diseases are a major constraint for wheat production. Effective disease resistance is essential for ensuring a high production quality and yield. One of the most severe fungal diseases of wheat is Septoria tritici blotch (STB), which influences wheat production across the world. In this study, genomewide association mapping was used to identify new chromosomal regions on the wheat genome conferring effective resistance towards STB. A winter wheat population of 164 North European varieties and breeding lines was genotyped with 15K single nucleotide polymorphism (SNP) wheat array. The varieties were evaluated for STB in field trials at three locations in Denmark and across 3 years. The association analysis revealed four quantitative trait loci, on chromosomes 1B, 2A, 5D and 7A, highly associated with STB resistance. By comparing varieties containing several quantitative trait loci (QTL) with varieties containing none of the found QTL, a significant difference was found in the mean disease score. This indicates that an effective resistance can be obtained by pyramiding several QTL.     

Mapping and validation of the quantitative trait loci for leaf stay‐green‐associated parameters in maize

Functional stay‐green is generally regarded as a desirable trait of varieties in major crops including maize. In this study, we used an F_3:4 recombinant inbred line population with 165 lines from a cross between a stay‐green inbred line (Zheng58) and a model inbred line (B73) using 211 polymorphic simple sequence repeat markers to map quantitative trait loci for three stay‐green‐associated parameters, chlorophyll content, photosystem II photochemical efficiency and stay‐green area, at maturity stage, detected a total of 23 quantitative trait loci (QTL) on nine chromosomes. Single QTL explained 3.7–13.5% of the phenotypic variance. Additionally, we validated some important stay‐green QTL using a heterogeneous inbred family approach and found that the stay‐green‐associated parameters were significantly correlated with the plant yield. This study may contribute to a better insight into the regulatory mechanism behind leaf stay‐green in maize and a novel development of elite maize varieties with delayed leaf senescence through molecular marker‐assisted selection.     

茶树小分子量热激蛋白基因CsHSP17.2的克隆与表达分析

[目的]进一步了解茶树小分子量热激蛋白基因CsHSP17.2在逆境胁迫条件下的分子生物学功能.[方法]利用RT-PCR技术从茶树‘迎霜’中克隆得到CsHSP1 7.2基因,运用生物信息学软件分析其核苷酸和编码蛋白,通过Real-timePCR分析其表达模式.[结果]该基因开放阅读框长度为453 bp,编码150个氨基酸,蛋白质相对分子质量为17.2×10a,理论等电点5.56;无信号肽位点,属于非分泌型蛋白;被定位于细胞质中.系统发育树分析表明,茶树CsHSP1 7.2与水稻(GenBank登录号:P27777)和花生(ABC41131)的进化关系较近,属于小分子量热激蛋白基因家族第Ⅰ亚族.qRT-PCR分析发现,茶树CsHSP17.2属于组成型基因;高温(38℃)处理1h能显著提高CsHSP17.2 mRNA的相对表达量(P＜0.05);干旱(100 g·L-1 PEG 6000)、高盐(200 mmol· L-1 NaCl)和外源脱落酸(200 mg· L-1 ABA)处理条件下,该基因的转录水平均出现不同程度的上调.[结论]克隆得到茶树‘迎霜’小分子量热激蛋白基因CsHSP17.2,其在花中表达量最高,且响应高温、干旱、高盐和外源脱落酸胁迫.     

苹果属无融合生殖SERKs基因家族的克隆和表达分析

本研究利用同源克隆法以苹果属平邑甜茶和杂种后代叶片为试验材料,克隆得到4个SERK基因家族片段,并分析了SERKs家族基因在平邑甜茶和杂种后代不同器官和组织中的表达情况。研究结果表明:分离获得的4个SERK基因家族片段的氨基酸序列与模式植物拟南芥、烟草等植物的同源性均在84%以上,同时与其他物种的氨基酸序列进行Blast比对都具有较高的同源性;RT-PCR定量分析结果表明SERK1、SERK2、SERK3、SERK4基因主要在生殖器官中表达。因此,推测SERKs基因在平邑甜茶和杂种后代生殖发育过程中起着重要的调控作用。本研究结果为进一步揭示无融合生殖的发生机理奠定了基础。