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.     

云南特有野生果树资源及其分布特点

 云南有各类野生果树资源约500 余种, 其中云南特有的164种, 分属35科54属, 多为热带、亚热带果树和浆果类果树, 形成滇南—滇东南、滇西—滇西北和滇中—滇东北3个特有野生果树集中分布中心。这些果树资源用途广泛, 优异性状明显, 有待深入研究和综合开发利用。     

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.     

Microscopic detection of reactive oxygen species generation in the compatible and incompatible interactions of Alternaria alternata Japanese pear pathotype and host plants

 Reactive oxygen species (ROS) generation was examined in the interaction of Alternaria alternata Japanese pear pathotype and host plants using three methods: nitro blue tetrazolium (NBT) method for microscopic detection of O₂ ⁻, diaminobenzidine (DAB) methods for microscopic detection of H₂O₂, and cerium chloride methods for ultrastructural detection of H₂O₂. ROS generation was detected by NBT and DAB methods at appressoria on leaves of susceptible cultivars and heat-shocked leaves of resistant cultivars but not in leaves of resistant cultivars. Ultrastructural detection by the cerium chloride method identified ROS generation at cell walls of appressoria and penetration pegs in susceptible, resistant leaves and heat-shocked leaves. These differences in the ultrastructural and microscopic data in resistant areas were due to the restriction of ROS generation in limited areas, the side facing the plant surface, of appressoria and penetration pegs. Therefore, ROS generation was apparently induced regardless of the resistance or susceptibility of the cultivar with the difference being in the volumes generated. After evaluating the pathological role of ROS generation in fungal structures, such generation was found to be associated with early penetration of cell walls in pear plants. Additionally, ROS generation in plants was also found in degrading pectin layers near infected hyphae and in plasma membrane modification sites in susceptible leaves but not in resistant leaves. ROS generation in susceptible leaves might be accompanied with plasma membrane damage, although the role of ROS generation in the pectin layers is not clear. ROS generation in both fungal and plant cells during their interaction was likely associated with the expression of susceptibility. Received: June 3, 2002 / Accepted: July 31, 2002     

Rapid detection of Phytophthora cinnamomi using PCR with primers derived from the Lpv putative storage protein genes

Phytophthora cinnamomi is an ecologically and economically important pathogen. In this study, PCR assays were developed with primer pair LPV2 or LPV3 for rapid detection and identification of this organism. Both primer pairs were selected from putative storage protein genes. The specificity of these primer pairs was evaluated against 49 isolates of P. cinnamomi , 102 isolates from 30 other Phytophthora spp., 17 isolates from nine Pythium spp. and 43 isolates of other water moulds, bacteria and true fungi. PCR with both primer pairs amplified the DNA from all isolates of P. cinnamomi regardless of origin. The LPV3 primers showed adequate specificity among all other species tested. The LPV2 primers cross-reacted with some species of Pythium and true fungi, but not with any other Phytophthora species. PCR with the LPV3 primers detected the pathogen at levels of a single chlamydospore or 10 zoospores in repeated tests. The PCR assay was at least 10 times more sensitive than the plating method for detection of the pathogen from artificially infested soilless medium, and, to a lesser extent, from naturally infected plants. PCR with LPV3 primers can be a useful tool for detecting P. cinnamomi from soilless media and plant tissues at ornamental nurseries, whereas the LPV2 primers can be an effective alternative for identification of this species from pure culture. Applications of these assays for detection of P. cinnamomi in other environments were also discussed.     

低温胁迫下弓葵幼苗膜脂过氧化及保护酶活性的变化

 低温胁迫下弓葵( Butia capitata Becc) 幼苗叶片的MDA 含量逐渐增加, 膜脂过氧化作用增强。- 8 ℃条件下的膜脂过氧化作用明显强于2 ℃。细胞膜透性在2 ℃条件下变化不大, - 8 ℃时则随低温胁迫时间延长而急剧上升, 细胞膜受到伤害。- 8 ℃胁迫下细胞保护酶SOD、POD 和CAT 活性短期(6 h) 内升高,然后下降, 24 h 以后3 种保护酶受到低温胁迫的严重抑制。在2 ℃胁迫下, SOD 活性在6 h 内变化不大, 随后下降; CAT活性变化趋势与- 8 ℃时相似, 但变化幅度较小; POD 虽也呈现先升后降的趋势, 但降幅明显小于升幅, 至48 h 时POD 活性仍维持较高水平。2 ℃低温胁迫不是抑制而是促进POD 活性的提高。     

外来入侵种的危害与防治对策

外来入侵种对当地的自然生态系统结构和功能以及生物多样性都有着深远的影响,而且可能对地方的生态环境和经济造成严重危害。四川已经成为遭受外来入侵种威胁的省份之一,全省现有20多个外来入侵种。为了防止外来入侵种的泛滥,应当建立外来物种的引入评价机制,慎重进行有意引种;加强口岸管理,防止外来入侵种通过贸易途径无意引入,同时采取措施控制已经产生危害的外来入侵种。     

棉铃虫核型多角体病毒分子生物学和基因工程研究进展

棉铃虫核型多角体病毒(HaSNPV)是棉铃虫专一性病原物,隶属于杆状病毒科核型多角体病毒属.对其分子生物学和基因工程的研究主要包括以下几个方面:测定了HaSNPV G4株和C1株基因组核苷酸全序列,并与其它病毒进行了同源性比较;研究了HaSNPV部分基因的结构、转录、表达及其功能.构建了HaSNPV Bac to Bac杆状病毒表达系统;重组病毒杀虫剂的研究为HaSNPV大面积防治棉铃虫展示了广阔的前景.随着棉铃虫核型多角体病毒分子生物学和基因工程研究的不断深入,重组病毒杀虫剂将在棉铃虫综合防治中发挥更为重要的作用.     

Mariner转座子与小菜蛾抗性关系的研究

以抗溴氰菊酯、杀虫双、杀螟丹小菜蛾种群及其敏感品系为研究对象,借助聚合酶链式反应(PCR)、载体筛选、琼脂糖凝胶电泳等实验技术,检测了小菜蛾4个品系的Mariner转座子的存在及其与抗性的关系。结果表明,在抗溴氰菊酯、杀虫双、杀螟丹以及敏感品系的小菜蛾中都存在两种大约500 bp的Mariner转座子片断,初次证明在小菜蛾4个品系中均含有两种Mariner转座子,并发现一种新的转座子基因片断,但是没有发现Mariner转座子与抗性之间存在直接的联系。研究结果将为利用Mariner转座子标签法在小菜蛾中分离定位基因、转化外源基因、转基因昆虫防治害虫等研究提供理论基础。     

半自然条件下几种赤眼蜂及品系对亚洲玉米螟卵寄生能力比较

在半自然条件(田间网罩)下,将人工饲养的玉米螟卵按4、8、16块/株3个处理分别固定在网罩内的玉米植株中上部叶片上,然后每网罩分别引入供试赤眼蜂20头,24 h后更换玉米螟卵块,连续3 d。7 d后调查供寄生的玉米螟卵被寄生率及羽化率。结果表明:在半自然条件下广赤眼蜂伊朗1-1品系虽寄生能力高于其他供试品系,但到第2、3天明显降低;玉米螟赤眼蜂北京6-2-2品系每天均维持一个较平稳的寄生能力,产卵量在时间上的分配比较分散,并在第2、3天显示出比其他品系较高的寄生潜能。广赤眼蜂伊朗1-1品系的卵块寄生数量可随卵块密度的增加而增加,而其他品系则在各处理密度下,寄生数量基本没有变化;从卵粒寄生率看,广赤眼蜂伊朗1-1和吉林1-2两个品系的寄生数量随卵块密度的增加而大幅度提高,其他品系增加幅度较小。