正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

Genetic diversity of herbicide-resistant and -susceptible Avena fatua populations in North Dakota and Minnesota

Genetic diversity within and among 20 herbicide-resistant (HR) and 16 herbicide-susceptible (HS) Avena fatua multi-field populations was determined using 82 polymorphic loci resulting from two intersimple sequence repeat (ISSR) primers and one long-primer random amplified polymorphic DNA (LP-RAPD) primer. Collections from the Red River Valley of North Dakota and Minnesota, sampled in 1964 and 2000, represented A. fatua populations before and after intensive exposure to herbicides. A 1995 collection from south-west North Dakota represented A. fatua exposed to low herbicide selection. Despite differences in years of herbicide exposure among collections, both HR and HS populations from every collection maintained nearly similar levels of ISSR and RAPD diversity. Genetic differentiation among populations (G_ST) varied from 11% to 13% among HR populations and from 9% to 16% among HS populations, indicating that 84–91% of total variation remained within HS or within HR populations. Minimal difference in gene diversity between HR and HS is consistent with multiple origins of resistance, where HR A. fatua most likely evolved from diverse founding individuals.     

狗舌草提取物诱导淋巴细胞性白血病L1210细胞分化的研究

以单猪屎豆碱（MO）、槲皮素（QU）和环磷酰胺（CY）为参照，观察了狗舌草600mL／L乙醇提取物（EX）对淋巴细胞性白血病L1210细胞体外试验的形态变化；利用流式细胞术，从DNA分子水平上检查了EX对L1210细胞各周期相的影响，探讨EX对L1210细胞的分化机理。结果发现，EX能够使L1210细胞向淋巴细胞方向发展；经EX作用24h后，L1210细胞G0＋G1期的百分比较对照组明显升高。提示EX对L1210细胞增殖的抑制作用可能是由于G1期的阻滞所致。     

分子标记用于赤眼蜂分子监测的研究

通过对松毛虫赤眼蜂（Trichogramma dedrolimi Matsumura）以及玉米螟赤眼蜂（Trichogramma ostriniae Pang et Chen）rDNA-ITS2基因的克隆测序，并联机检索GenBank核酸序列库中其它赤眼蜂的相关序列，利用核酸分析软件找到松毛虫赤眼蜂和玉米螟赤眼蜂ITS2序列中一段有鉴别意义的标志，并据此设计出蜂种的特异PCR引物，实现了松毛虫赤眼蜂和玉米螟赤眼蜂rDNA特异区带的PCR扩增。研究表明该特异区带具有种的特异性。我们认为该分子标记技术可用于目前我国生防中常用的松毛虫赤眼蜂和玉米螟蒌眼蜂的分子监测，如田间种群动态监控和寄生效果评估。     

柑橘抗CTV转基因与分子标记研究进展

综述了柑橘抗衰退病基因工程中两方面的研究进展。介绍多种来源于柑橘衰退病毒(Citrustriztezavirus,CTV)核酸序列的转基因柑橘和抗性种质资源中抗性基因的分子标记,以及所涉及的方法和遇到的问题。目前研究表明,虽然已成功实现对病毒衣壳蛋白(CP)等基因的转化和Ctv等抗性基因的标记,但尚未获得对CTV有高度抗性的转基因柑橘,而抗性基因亦不能实现定点克隆和转化。因此上述两方面研究还有待深入。     

Modelling spatio-temporal dynamics of herbicide resistance

A mathematical model has been developed for the risk assessment of the spread of genes conferring herbicide resistance in plant populations. The model combines an age-and-stage-structured population dynamic model, a population genetic model and a model of spatial spread. This is achieved by embedding a local matrix population model into a cellular automaton model with raster cells as spatial units. The dynamics of each cell is determined by both its local dynamics and the interaction with neighbouring cells. The model is applied to the evaluation of management strategies to delay or even to prevent long-term evolution of resistance in an annual grass weed. The results show that the appearance and spread of resistant genes is a highly non-linear process exhibiting threshold phenomena, which occur for a wide range of parameters. The properties of the seed survival curve constitute the `genetic memory' of the system and thus determine its long-term dynamics. It is possible to delay the evolution of resistance by suspension of treatment, reduction in herbicide application rate and introducing fallow periods. Spatial spread from an infested plot is inhibited by leaving untreated strips between adjacent fields.     

Orobanche species and population discrimination using intersimple sequence repeat (ISSR)

Summary Orobanche species are commonly identified using morphological characteristics. In many cases, the distinction of closely related species is difficult, and a molecular tool is more suitable to differentiate them. In this study, genomic polymorphism between morphologically distinct species was investigated through amplification by polymerase chain reaction (PCR) of intersimple sequence repeat (ISSR) regions. Five primers were used to study genetic variation in the morphologically distinct species O. hederae and O. amethystea, as well as the closely related species O. cernua and O. cumana. For the first two species, all the primers detected genetic polymorphism. Anchored primers allowed the identification of more specific molecular markers than non‐anchored tri‐ and tetranucleotide primers. Genetic polymorphism was investigated among three O. hederae populations using the two types of primer. One non‐anchored and two anchored primers detected intraspecific variation, which was not correlated with the geographical location of those populations. The primer (GATA)₄ detected polymorphism between five specimens each of O. cernua and O. cumana species collected from different countries, permitting these two closely related species to be clearly differentiated. This study demonstrated that ISSR markers can be highly reliable for precise identification of Orobanche species.     

Variation in the response of melon genotypes to Fusarium oxysporum f.sp. melonis race 1 determined by inoculation tests and molecular markers

Screening of genotypes of melon ( Cucumis melo ) for resistance to wilt caused by Fusarium oxysporum f.sp. melonis is often characterized by wide variability in their responses to inoculation, even under carefully controlled conditions. The variability at the seedling stage of 17 genotypes susceptible to race 1 was examined in growth-chamber experiments. Disease incidence varied from 0 to 100% in a genotype-dependent manner. Using four combinations of light (60 and 90  µ E m⁻² s⁻¹) and temperatures of (27 and 31°C), only light intensity showed a statistically significant effect. Marker-assisted selection for fusarium resistance breeding using cleaved amplified polymorphic sequence (CAPS) and sequence-characterized amplified region (SCAR) markers were compared using a single set of genotypes that included 24 melon accessions and breeding lines whose genotype regarding the Fom-2 gene was well characterized. The practical value of the markers for discriminating a range of genotypes and clarifying the scoring of phenotypes was also tested using a segregating breeding population which showed codominant SCAR markers to be useful in marker-assisted selection.     

苹果基因组分子生物学研究进展

借助分子标记手段苹果基因组研究取得了巨大成就,涉及苹果的抗性、生长发育、果实品质等各方面基因,综合有关文献,将其研究的主要成果,包括已发现的主要苹果遗传标记、已构建的苹果遗传连锁图谱及苹果的转基因研究进行综述,并对目前苹果基因组研究存在的问题和对策进行了分析,以期为苹果遗传育种提供参考。