水稻落粒性基因qSH1调控区关键SNP的HRM检测体系优化

高分辨率熔解曲线(high resolution melting,HRM)分析是一种新发展起的DNA多态性快速筛查分析技术,具有灵敏、准确和高效等突出优势。q SH1是控制水稻落粒性的主效基因,在该基因的上游12 kb的调节区存在一个SNP位点(命名为q SH1(G/T))调控其表达,从而引起不同品种间落粒性差异。为针对q SH1(G/T)位点建立一种HRM快速分型检测体系,本研究以难落粒的粳稻品种"日本晴"和易落粒的"R1128"为材料,研究了不同的PCR产物长度、DNA浓度、Mg2+浓度、退火温度及扩增模式对HRM分型效果的影响。结果表明:使用q SH1-1引物(产物长度68 bp)能获得平稳的熔解曲线和单一熔解峰;当DNA浓度在5 ng/μL以上、退火温度为60℃、Mg2+浓度为1.5 mmol/L时,HRM分型效果最佳;此外,选择降落PCR模式能取得较好的分型效果。研究结果可为后续该位点的基因分型及水稻落粒性分子遗传改良的研究奠定基础。     

苹果不同HRM反应体系分析效果评价

高分辨率熔解曲线(high resolution melting,HRM)分析是一种新型的DNA多态性快速筛查技术。本研究以苹果品种"富士"和"舞姿"为试材,用来自苹果基因组的SSR标记CH03d11对不同反应体积、不同DNA浓度以及不同PCR退火程序下的HRM分型效果进行了测试与评估。结果表明:采用5μL反应体积可以获得与10μL或20μL反应体积相同的检测效果。在5μL反应体积中,DNA浓度小到0.25ng/μL时,PCR扩增及随后的HRM分析效果依然良好。另外,研究表明,采用降落PCR扩增模式也能取得良好的HRM多态性检测结果。     

高分辨率熔解曲线技术在大豆基因分型中的应用

高分辨熔解曲线(high resolution melting, HRM)分析技术是一种高效,快速,便捷的核苷酸检测方法。本实验从染料用量、模板浓度、产物片段长度3个方面探索了适宜HRM分析的最佳条件,并利用HRM技术对单核苷酸(single nucleotide polymorphism, SNP)及插入/缺失(insertion-deletion, InDel)检测及基因分型等方面进行了分析。结果表明:10μL反应体系中,20×Eva Green饱和染料加入0.1~0.3μL,模板浓度加入10~200 ng不影响HRM分辨率;扩增产物长度在70~190 bp范围内,插入/缺失片段长度3~13 bp范围内,高分辨熔解曲线均有较好的分辨效果。利用HRM分析技术检测分析F4个体基因型,结果与PAGE电泳分析完全一致。本实验结果说明,HRM技术与传统凝胶电泳基因分型方法相比,具有操作简单、分辨率高、准确快速、成本低等优点,是基因型鉴定的最佳选择。     

植物化感作用研究进展

植物的化感作用是当今植物保护领域研究的热点.植物化感物质的研究与应用,建立农作物合理的轮、间、套和连作制度,培育有化感作用的农作物新品种,开发新型安全的除草剂和杀菌剂等,减少人工化学品的投入和对环境的危害,实现有害生物绿色防控.本文概述了近年来植物化感物质的种类、释放途径、作用机理、植物化感研究方法以及在农业生态系统中的应用等方面的研究进展.     

入侵植物含羞草对7种植物的化感作用

以7种植物为受体,采用室内生物测定法研究含羞草(Mimosa pudica Linn.)茎叶水浸提液对7种植物的化感作用.结果表明:含羞草茎叶水浸提液对7种受体植物的种子萌发和幼苗生长均有一定的化感效应,对7种受体植物的化感综合效应均为负值,化感作用综合效应的平均值大小顺序为:皱果苋＞柱花草＞白菜＞豇豆＞萝卜＞绿豆＞苦瓜,佐证了化感作用是含羞草成功入侵的机制之一.     

PAM在土壤重金属污染植物修复中的作用及机理研究进展

聚丙烯酰胺(PAM)作为一种性能优良的高分子聚合物,近年来广泛应用于农业水土保持及土壤改良。通过文献调研,在综述PAM对土壤中重金属作用及机理基础上,分析了其对土壤重金属污染植物修复的应用潜力：（1）PAM及其改性产物施用于土壤后可吸附特定的重金属离子,降低重金属对植物的毒害作用,但效果受pH值、温度等条件影响;（2）PAM在改善土壤理化性质的同时,可通过吸附、螯合及络合等作用抑制重金属的生物有效性,从而改善修复植物的生长条件。尽管PAM降解产物之一的丙烯酰胺单体(AMD)具有神经毒性,但是通过调节施加方式及引入降解微生物等方法可以避免PAM降解对土壤产生不良影响,进而将健康风险控制在安全范围内。因此,PAM是环境友好的重金属污染土壤植物修复促进剂,且其环境风险可控。     

基于高分辨率熔解曲线技术快速筛选黄瓜SNP1

为筛选用以黄瓜纯度检验和品种鉴定的SNP位点,以17个黄瓜杂交品种及其34份亲本为SNP筛选群体,利用高分辨率熔解曲线(high resolution melting,HRM)技术筛选SNP位点.本研究通过HRM分析和焦磷酸测序确认,获得CLA0(TT/--)、CLA1(C/T)和CLA6(A/G)三个SNP位点,群体...     

水稻内生菌K12G2菌株的鉴定及其促生特性研究

为了开发新的可替代化肥、农药的促植物生长微生物肥料,对从水稻种子中分离得到的1株具有抗氧化活性的内生细菌K12G2进行分类鉴定,检测其促生特性,并通过平板培养和水培试验测定其对水稻幼苗的促生长作用。结合菌株K12G2的形态学、生理生化特征检测及分子生物学鉴定,K12G2为柠檬色短小杆菌(Curtobacterium citreum);该菌株表现为产IAA、溶磷、固氮及淀粉酶活性的促生特性;在限菌平板培养条件下,菌株K12G2培养液浸种1 h处理比4 h处理后对水稻种子的萌发和生长效果明显,培养液处理的水稻幼苗根长和芽长显著增加;在水培条件下,接种K12G2的水稻幼苗根长显著提高47.08%,生长至三叶期的幼苗比例是对照组的4.5倍。结果表明,菌株K12G2促生作用明显,具有一定的应用前景。     

罗勒浸提液对小麦种子萌发及幼苗根系生长的影响

本试验探究了芳香族植物对小麦种子萌发及幼苗根系生长的影响。以白麦207为试验材料，采取室内培养法，分别用浓度为0.5%(5 g/L)、1.0%(10 g/L)、1.5%(15 g/L)、2.0%(20 g/L)、2.5%(25 g/L)的罗勒浸提液对小麦种子进行培养，测定不同浓度罗勒浸提液处理下小麦种子的发芽率、总根长、根鲜重、根干重、根系形态参数等指标。试验表明，低浓度的罗勒浸提液可以更好地促进小麦种子萌发以及幼苗根系生长，高浓度的罗勒浸提液会在一定程度上抑制小麦种子萌发及幼苗根系生长。     

玉米生物诱导孤雌生殖单倍体影响因素研究进展

玉米单倍体育种技术能显著缩短育种周期,提高育种效率,在现代商业化育种中占有重要的地位。规模化高效单倍体的产生是单倍体育种技术应用的前提条件。总结了父本诱导系、母本的遗传背景、诱导的环境、授粉方式等影响玉米生物诱导孤雌生殖单倍体产生的因素,通过文献综合分析得出了获得大量的孤雌生殖单倍体除了选择合适的高频率孤雌生殖诱导系外,同时要考虑其他诱导因素的影响。并就高效生产孤雌生殖单倍体的未来研究及发展方向进行了讨论,以期促进单倍育种技术的规模化应用。     

Validation of two models for self‐incompatibility in autotetraploid perennial ryegrass using high resolution melting‐based markers

Perennial ryegrass (Lolium perenne L.) displays a two‐locus gametophytic self‐incompatibility (SI) system that remains intact at the tetraploid level. Two models are plausible for SI in autotetraploids. In Model I: both alleles at the S locus and both at the Z locus in diploid pollen matching the female genotype results in incompatibility. In Model II: only one allele at S and one at Z locus in diploid pollen matching the female results in incompatibility. The goals were to determine which of the models best explains SI in our autotetraploid ryegrass population and to evaluate the efficiency of high‐resolution melting (HRM) genotyping for discriminating different iso‐allelic genotypes. The progeny of a cross between two autotetraploids was characterized with three HRM‐based markers co‐segregating with Z. Segregation ratios were used to make inferences about the mode of action of the SI system. The observed segregation differed significantly (P < 0.001) from the expected under Model I, but not from the expected under Model II (P = 0.463). Thus, Model II explains SI in this population, and HRM is an efficient tool to distinguish different iso‐allelic genotypic classes.     

采用基于高分辨率熔解曲线技术的SNP标记定位徐州142无絮的短绒控制基因

【目的】定位徐州142无絮(XZ142w)突变体的短绒控制基因n2。【方法】以陆地棉(Gossypium hirsutum L.)徐州142(XZ142)×XZ142w的F2群体为研究对象,利用108个简单重复序列(Simple sequence repeat,SSR)标记对n2进行初步定位,再根据2个亲本材料中有单核苷酸多态性(Single nucleotide polymorphic,SNP)的差异基因设计50对SNP引物,用高分辨率熔解曲线(High resolution melting,HRM)技术从中筛选在亲本间有多态性的SNP引物,并用于后代的基因分型。【结果】利用108个SSR标记将n2初步定位在26号染色体的20.2c M的遗传区间内;用HRM技术筛选到9对亲本间有多态性的SNP引物,成功实现基因分型;并结合以SSR构建的连锁图谱,将n2的遗传区间缩小为19.5 c M,n2与最近的SNP标记Cricaas20158遗传距离为5.5 c M,且遗传图谱上的标记与四倍体陆地棉测序物理图谱基本一致。【结论】HRM技术可用于棉花中的SNP检测和n2基因的定位。     

Molecular markers for improving control of soil-borne pathogen <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in sugar beet

Fusarium spp. cause severe damage in many agricultural crops, including sugar beet, with Fusarium oxysporum historically being considered as the most damaging of all species. Sugar beet needs to be protected from this class of soil-borne pathogens in order to ensure an optimal sugar yield in the field. Genetic control of the disease is crucial in managing these pathogens. Identification of single nucleotide polymorphism (SNP) markers linked to resistance can be a powerful tool for the introgression of valuable genes needed to develop Fusarium-resistant varieties. A candidate gene approach was carried out to identify SNP markers linked to putative Fusarium resistance sources in sugar beet. Five resistant analogue genes (RGAs) were screened by means of high resolution melting (HRM) analysis in a set of sugar beet lines, considered as resistant and susceptible to Fusarium oxysporum. HRM polymorphisms were observed in 80% of amplicons. Two HRM polymorphisms were significantly associated with Fusarium resistance (P < 0.05). The amplicons that showed association were sequenced and two SNPs were identified. The association was further validated on 96 susceptible and 96 resistant plants using competitive allele-specific PCR (KASPar) technology. The selected SNPs could be used for marker-assisted breeding of Fusarium resistance in sugar beet.     

DNA fingerprinting — A useful tool in fruit breeding

Summary Identification of cultivars and selections, pedigree analysis, estimation of genetic relatedness and genome mapping — all of these objects are achieved with various methods of DNA fingerprinting which can distinguish even closely related genotypes. Initial results were obtained by a variation of the well-known RFLP-technique (Restriction Fragment Length Polymorphism): hybridization of restriction enzyme-digested DNA samples with hypervariable minisatellite DNA probes. Technical improvements, including a number of additional DNA probes and various DNA fragment detection methods, have since been published. With the recent advent of the PCR technique (Polymerase Chain Reaction) and PCR-based methods like RAPD (Random Amplified Polymorphic DNA), DNA fingerprinting has widened its applicability and attracted a growing number of scientists.     

5-氮杂胞苷调节植物基因表达研究进展与应用展望

5-氮杂胞苷是一种DNA甲基化抑制剂,可以通过降低DNA的甲基化水平来调节基因的表达,从而对生物的生长发育进行表观遗传调控,其在动物与医学领域已有广泛的研究与应用,近年来在植物基因表达调节方面的研究与应用报道也逐渐增多。本研究对植物DNA甲基化的类型、引发、影响因素和抑制剂的作用进行了概述,重点对5-氮杂胞苷对植物基因表达水平的调节和作用效果进行了阐述。并对其在水稻、小麦、棉花等重要作物生长发育、逆境适应、育性调节及次生代谢的影响与作用进行归纳,可为植物基因表达表观调节研究与应用提出新的关注点。     

Research on the Human Resource Management Information System in Colleges and Universities

This paper illustrates the existent of universities. Discussing the necessity for colleges issues in the traditional personnel management system in colleges and and universities to open out HRMIS, the paper puts forward the aims of soft development concerning HRM. According to the practice of the human resource management and exploitation in colleges and universities, the frame and functions and system implement of HRMIS founded on date-base based Web and E2Q business flat roof in universities are brought forward.     

Effect of Mischmetal Additions on the Ignition Temperature ofMolten ZM5 Magnesium Alloy

Magnesium alloy is prone to burning during its melting and casting in air, which is a major factor in obstructing its application. Fluxes and cover gases are currently used for the melting and production processes, and semi solid thixocasting is also used to lower the operation temperature, but there still remain many problems. Alloying is a promising method of preventing Mg from burning. The effect of mischmetal additions on the ignition temperature of ZM5 Mg alloy was investigated and compared with that of pure rare earths. It is shown that an addition of 0.12 % mischmetal can greatly heighten the ignition point and such an alloy can be widely applied.     

基因组原位杂交技术在植物研究中的应用

基因组原位杂交是以亲本之一的总基因组DNA做探针,另一亲本的基因组DNA做封阻,在荧光原位杂交技术的基础上发展起来的一种染色体/染色质检测技术。在其发展的十几年里,已在植物的基因组研究中发挥了重要的作用。应用这一技术可对多倍体中基因组之间的亲缘关系、基因组组成及起源进行研究;对杂交种中染色体组的组成进行分析;对代换系、附加系和易位系进行有效的鉴定,并对其中的外源染色体或染色体片段的来源、大小、数目及发生位点进行检测和定位。此外,利用基因组原位杂交技术还有助于确定物种间的同源性;研究杂交种中来源不同的染色质在核中的分布;探索B染色体的起源、染色体间的配对、重组、交换等现象。随着基因组荧光原位杂交技术体系的不断发展、完善和改进,其应用范围不断拓展,在植物基因组研究领域中发挥了越来越重要的作用。