Modelling Plant Disease Epidemics

An epidemic is the progress of disease in time and space. Each epidemic has a structure whose temporal dynamics and spatial patterns are jointly determined by the pathosystem characteristics and environmental conditions. One of the important objectives in epidemiology is to understand such spatio-temporal dynamics via mathematical and statistical modelling. In this paper, we outline common methodologies that are used to quantify and model spatio-temporal dynamics of plant diseases, with emphasis on developing temporal forecast models and on quantifying spatial patterns. Several examples of epidemiological models in cereal crops are described, including one for Fusarium head blight.     

Evidence that a Leaf-Disk Test Allows Assessment of Isolate-Specific Resistance in Brassica oleracea Crops Against Downy Mildew (Peronospora parasitica)

A rapid resistance/susceptibility test for Peronospora parasitica (downy mildew) was established by inoculating leaf-disks of four Brassica oleracea accessions. Several conditions were tested: disk disinfection or not, agar medium with or without nutrients and with 50 or 100 ppm of benzimidazole. Using disinfected disks placed on agar (no nutrient and benzimidazole at 50 or 100 ppm), the responses of leaf-disks to four isolates were similar to those obtained using the classical cotyledon test, whereas undesired contaminations occurred in all other conditions. The possible effect of the particular leaf used for obtaining the disks was also studied. In each incompatible interaction tested, disks were resistant whatever the leaf used. In compatible interactions, susceptible phenotypes were observed on disks derived from the six lowest leaves, but disks from upper leaves were resistant. The genetic basis of resistance in a F1 hybrid broccoli was assessed, by testing six isolates on an F2 population derived from this hybrid. The cotyledon test only allows inoculation of two isolates per seedling, whereas many isolates can be tested on each plant by using leaf-disks. The segregation of the resistance to each of the six isolates was analysed: two dominant genes (tightly linked) control resistance to all isolates (one to five isolates; the other to only one isolate).     

利用RAPD标记评价桃种间杂交一代群体的分离方式

以毛桃2-7(Prunus persica)、山桃白山碧桃(Prunus davidiana)以及两者的F_1代为试材,对RAPD标记的分离方式进行分析。结果表明,RAPD标记在F_1代呈3种分离方式:孟德尔分离、偏离孟德尔分离规律、异常分离。3种分离位点出现的频率和数量分别为:80%、240,14%、42,6%、18。呈孟德尔分离的情况有:不分离、1:1分离和3:1分离,其中不分离的频率为64％。并对偏离孟德尔分离比例和异常分离的RAPD标记进行分析。其研究结果可为该群体是否适合构建遗传连锁图谱进行评价及进一步利用该群体奠定良好基础。     

对鳞翅目害虫有活性的cry1C基因的克隆和表达

在鉴定苏云金芽孢杆菌(Bacillus thuringiensis，简称Bt)Btc001菌株cry基因型的基础上，构建了Btc001菌株质粒DNA HindⅢ片段的文库，并利用聚合酶链式反应-限制性酶切片段多态性(PCIR-RFIP)方法筛选出含有crylC6全长基因的13．5kb大片段，酶切分析得到该片段的物理图谱，BamHI和EcoRI切完成了6．5kb含全长基因的亚克隆，并对这条6．5kb片段亚克隆、测序，序列在国际核酸序列数据库(GenBank)登记(AY007686)，并由Bt杀虫晶体蛋白基因国际命名委员会命名为cry1Cb2基因。根据序列设计了一对用于扩增全长基因的引物S581CB和S381CB，扩增产物插入表达载体pET-21b中，诱导后在大肠杆菌BL21(DE3)中获得高效表达。表达产物对小菜蛾(Plutella xylostella)表现出较高活性，LC50达到7．9μg/ml。     

一个与稻瘟病菌无毒基因AVR-Pik~m连锁的SCAR标记的分离

 本研究将以前在稻瘟病菌菌株S1522获得的与决定对水稻品种梅雨明无毒性的基因(AVR-Pik^m)相连锁的1个RAPD标记OPO12₁₀₀₀进行了克隆和鉴定。核苷酸序列测定与分析结果表明:OPO12₁₀₀₀的大小为946个碱基,不含有与已报道的稻瘟病菌Mg-SINE、Fosburry、Magyy、Grasshopper、Pot2以及Pot3等同源的重复序列。根据OPO12₁₀₀₀的核苷酸序列,设计了1对24个核苷酸的特异引物,对无毒表型亲本S1522和毒性表型亲本S159、无毒表型群体基因池、毒性表型基因池以及有性杂交后代108个菌株进行了PCR扩增,所有无毒表型的菌株均能特异性地扩增出1条与OPO12₁₀₀₀大小相同的DNA条带,而毒性表型的菌株除5个重组个体外,均不能扩增出这条特异带。此结果表明,与稻瘟病菌无毒基因AVR-Pik^m连锁的RAPD标记OPO12₁₀₀₀被成功地转化为SCAR标记,为进一步通过染色体步移克隆该无毒基因奠定了基础。     

转基因水稻对稻瘟病的抗性研究

 采用苗期初筛、复筛、抗谱测定和田间自然诱发试验等不同鉴定方法,对经分子检测证明已整合有碱性几丁质酶基因和β-1,3-葡聚精酶基因的22个转化系的转基因水稻植株进行稻瘟病抗性鉴定研究,筛选出对稻瘟病的抗性比原种对照七丝软占有明显提高的一系列转基因水稻品系,其中表现高抗的有来自F4-9转化株系的7个品系。高抗材料的R7代品系,经室内抗谱测定及田间病圃试验结果,仍然表现高抗稻瘟病。本研究通过转基因技术,成功地将优质感病品种改良成高抗品系,研究结果证明了利用基因工程手段培育抗病水稻新品种是一个非常有希望的育种途径。     

植物苯丙氨酸解氨酶基因的表达调控与研究展望

苯丙氨酸解氨酶(phenylalanineammonia-lyase,PAL,EC4.3.1.5)是催化苯丙烷代谢途径第一步反应的酶,也是这个途径的关键酶,对植物有非常重要的生理意义。根据有关文献综述了植物PAL的分布与定位、酶学性质,总结了生长发育、钝化因子与调节因子、末端产物等内部因素及光、温、机械损伤与生长调节剂等外部因素对PAL的调控作用,得出外部因子是在转录水平上对酶活性实施调控的结论,并运用酶学和分子生物学方面的知识阐述了其调控机理。还着重阐述了PAL酶在果树上的研究现状与进展,并对其今后的研究及在果树上的应用进行了展望。     

利用PCR技术同时鉴定番茄抗根结线虫和抗斑萎病毒基因

 利用同一PCR反应体系，对分别与番茄抗根结线虫的 基因和抗斑萎病毒(1w V)的Sw一5基因紧密连锁的SCAR标记进行了同时扩增筛选，扩增的特异性片段与单引物扩增片段完全吻合，其中与基因紧密连锁的SCAR1标记为共显性标记，抗感试材均产生750 bp的特异片段，纯合和杂合抗病基因型试材存在 I酶切位点，酶切后分别产生了570 bp、160 bp和750 bp、570 bp、160 bp的不同特异性片段，而感病基因型试材无 I酶切位点；与Sw一5基因紧密连锁的SCAR2标记为显性标记，只有抗病试材扩增出400 bD的特异性片段。经反复验证，结果稳定准确可靠，可用于在同一PCR反应体系中对两个抗病基因进行同时筛选鉴定。     

大白菜细胞核雄性不育基因向自交系97A407的转育

根据复等位基因遗传假说 ,利用大白菜核不育系 9810 9和已知基因型的甲型两用系可育株 99135 2作为不育源 ,以大白菜自交系 97A40 7为目标亲本 ,经过 5个世代的转育 ,获得了具有 97A40 7遗传基因型的新甲型两用系、临时保持系和雄性不育系。