Internal transcribed spacer (ITS)

内部转录间隔区 (ITS)
  • 文章类型: Journal Article
    水泡枯病(BB)是由专性生物营养真菌病原体Exobasidiumvexexansmassee引起的,严重影响了山茶的产量和品质。在茶叶上使用化学农药大大增加了茶叶消费的毒性风险。植物性杀真菌剂异叶阿瓦卡酮(IBC)具有控制许多作物的真菌病害的潜力,但尚未在茶树上使用。在这项研究中,通过与天然激发子壳寡糖(COSs)和化学农药吡唑醚酯(Py)的比较和组合,评价了IBC的田间防治效果,并对IBC的初步作用模式进行了研究。IBC或其与COSs的组合的生物测定结果显示出对BB的显着控制效果(61.72%和70.46%)。IBC,像COSs一样,可以通过增强茶树相关防御酶的活性来提高茶树的抗病性,包括多酚氧化酶(PPO),过氧化氢酶(CAT),苯丙氨酸氨基解酶(PAL),过氧化物酶(POD),超氧化物歧化酶(SOD),β-1,3-葡聚糖酶(Glu),和几丁质酶。使用核糖体rDNA基因的内部转录间隔区(ITS)区域的IlluminaMiSeq测序检查了患病茶叶的真菌群落结构和多样性。很明显,IBC可以显着改变受影响植物地点的物种丰富度和真菌群落的多样性。本研究拓宽了IBC的应用范围,为BB病的防治提供了重要的策略。
    Blister blight (BB) disease is caused by the obligate biotrophic fungal pathogen Exobasidium vexans Massee and seriously affects the yield and quality of Camellia sinensis. The use of chemical pesticides on tea leaves substantially increases the toxic risks of tea consumption. Botanic fungicide isobavachalcone (IBC) has the potential to control fungal diseases on many crops but has not been used on tea plants. In this study, the field control effects of IBC were evaluated by comparison and in combination with natural elicitor chitosan oligosaccharides (COSs) and the chemical pesticide pyraclostrobin (Py), and the preliminary action mode of IBC was also investigated. The bioassay results for IBC or its combination with COSs showed a remarkable control effect against BB (61.72% and 70.46%). IBC, like COSs, could improve the disease resistance of tea plants by enhancing the activity of tea-plant-related defense enzymes, including polyphenol oxidase (PPO), catalase (CAT), phenylalanine aminolase (PAL), peroxidase (POD), superoxide dismutase (SOD), β-1,3-glucanase (Glu), and chitinase enzymes. The fungal community structure and diversity of the diseased tea leaves were examined using Illumina MiSeq sequencing of the internal transcribed spacer (ITS) region of the ribosomal rDNA genes. It was obvious that IBC could significantly alter the species\' richness and the diversity of the fungal community in affected plant sites. This study broadens the application range of IBC and provides an important strategy for the control of BB disease.
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  • 文章类型: Journal Article
    几乎没有证据表明已经描述和接受的asc虫类群(Ascarislumbricoides,A.suum,和A.Ovis)感染分类学上遥远群体的个体(原始人,猪,绵羊,山羊,和狗)可以在遗传或形态上区分。然而,尽管描述了形态差异,例如,由于种内变异,这些不足以确定物种,并且可能表明由于交叉感染而导致的a虫之间的差异,混合生产,以及对宿主的特定适应。在这里,寄生在苏门答腊猩猩身上的蛔虫的分子和形态分析结果(PongoabeliiLesson,1827)在本地人口中呈现。这项研究发生在武吉拉昂地区,印度尼西亚,2009年。全年,定期从24只猩猩中收集新鲜粪便样本,并检查了所有线虫成虫的存在。在定期收集过程中,仅从两只猩猩雌性中发现了五只成虫。使用综合分类学方法,发现的线虫被鉴定为A.lumbricoides。这一发现的重要性及其罕见性通过以下事实得到了证明:这是130多年来(包括过去20年的长期研究,重点是猩猩寄生虫和天然抗寄生虫药物)首次确认发现来自原始猩猩遗址(不是来自动物园)的成虫。建立了更准确的形态计量学参数和遗传差异来鉴定a虫。这些参数将有助于大猿的其他发现,也适用于进一步和精确地确定这种寄生虫。男性和女性标本之间的区别细节也有陈述和明确定义。对寄生猩猩的A虫物种的状况进行了综合评估,包括与先前描述的猩猩寄生虫的比较(即,A.satyri-speciesinquirenda),正在讨论。
    There is little evidence that the already described and accepted taxa of ascarids (Ascaris lumbricoides, A. suum, and A. ovis) infecting individuals of taxonomically distant groups (hominids, pigs, sheep, goats, and dogs) can be genetically or morphologically distinguished. However, despite described morphological differences, e.g., due to intraspecific variation, these are insufficient for species determination and may indicate differences amongst ascarids because of cross infections, hybrid production, and specific adaptations to hosts. Herein, the results of a molecular and morphological analysis of ascarids parasitising Sumatran orangutans (Pongo abelii Lesson, 1827) in native populations are presented. The research took place in the Bukit Lawang area, Indonesia, in 2009. Throughout the year, fresh faecal samples were collected regularly from 24 orangutans, and all were examined for the presence of nematode adults. Only five adult worms from two orangutan females were found during regular collection. Using the integrative taxonomic approach, the nematodes found were identified as A. lumbricoides. The significance of the find and its rarity is documented by the fact that this is the first confirmed finding of adult ascarids from an original orangutan site (not from a zoo) in more than 130 years (including the long-term study spanning the last 20 years focusing on orangutan parasites and natural antiparasitic drugs). More accurate morphometric parameters and genetic differences for the identification of ascarids were established. These parameters will be helpful for other findings in great apes and will also be suitable for further and precise determination of this parasite. The details distinguishing between male and female specimens are also stated and well defined. A comprehensive evaluation of the situation of Ascaris species parasitising orangutans, including a comparison with previously described orangutan parasite (i.e., A. satyri-species inquirenda), is discussed.
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  • 文章类型: Journal Article
    In the present study, Neorhadinorhynchus nudus (Harada, 1938) is reported from the frigate tuna Auxis thazard (Lacepéde) (Perciformes: Scombridae), in the South China Sea for the first time. The detailed morphology of N. nudus was studied using light and scanning electron microscopy based on the newly collected material. The results showed some morphometric variability between our specimens and previous studies, including the number of hooks per longitudinal row and the size of copulatory bursa and eggs. Our SEM observations also revealed all proboscis hooks emerged from elevated round rims on proboscis surface. In addition, N. nudus was firstly characterised using molecular methods by sequencing and analysing the ribosomal ITS and mitochondrial cox1 regions. There is no nucleotide divergence found in the ITS sequences, but a low level of nucleotide variability detected in the cox1 regions (the level of intraspecific nucleotide variability being 0.75% to 2.54%). The DNA sequence data obtained herein will indeed be a useful reference for rapid and accurate species identification of Neorhadinorhynchus.
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  • 文章类型: Evaluation Study
    背景:环境DNA(eDNA)监测在水生系统中越来越受欢迎,作为常规监测的一种有价值的补充方法。然而,这些工具尚未广泛用于后生动物鱼类寄生虫监测。鱼外寄生虫,1975年引入挪威,对大西洋鲑鱼种群和渔业造成了严重破坏。在挪威的几个河流系统中成功根除了寄生虫,大西洋鲑鱼只在7条河流中被感染,包括三个在Drammen地区.在这个特殊的感染区域,治疗的先决条件是确定鲑鱼迁徙屏障上游的虹鳟鱼上是否也存在G.salaris。这里,我们开发并测试了eDNA方法来补充常规监测方法.
    方法:通过Drammen水道9个地点的玻璃纤维过滤器现场过滤水样(2×5升),用CTAB方案提取DNA。我们开发了针对针对核核糖体ITS1区域的G.salaris的qPCR检测方法,我们实施了针对大西洋鲑鱼和虹鳟鱼的线粒体细胞色素b和NADH区域的公开试验,分别。将所有测定成功转移至液滴数字PCR(ddPCR)。
    结果:所有qPCR/ddPCR检测在组织样本和现场样本中均表现良好,证明eDNA检测对S.salaris的适用性,虹鳟鱼和大西洋鲑鱼在自然的水系统。使用ddPCR,我们消除了使用qPCR观察到的Gyrodactylusderjavinoides的低交叉扩增,从而大大提高了特异性和敏感性。已成功实施了针对G.salaris和大西洋鲑鱼的DuplexddPCR,可在未来的监测计划中用作方法。记录了受感染的Lierelva河中存在G.salariseDNA,而不是其他地方。虹鳟鱼eDNA仅在某些地方被检测到,这些地方的阳性可归因于上游陆基虹鳟鱼养殖场的eDNA释放。电捕捞支持所有地区都没有虹鳟鱼。
    结论:我们提供了可靠的现场和实验室方案,用于G.salaris的eDNA检测,大西洋鲑鱼和虹鳟鱼,这可以补充传统的监测计划,并大大减少活鱼的牺牲。我们还表明,ddPCR优于qPCR的特异性检测。
    BACKGROUND: Environmental DNA (eDNA) monitoring is growing increasingly popular in aquatic systems as a valuable complementary method to conventional monitoring. However, such tools have not yet been extensively applied for metazoan fish parasite monitoring. The fish ectoparasite Gyrodactylus salaris, introduced into Norway in 1975, has caused severe damage to Atlantic salmon populations and fisheries. Successful eradication of the parasite has been carried out in several river systems in Norway, and Atlantic salmon remain infected in only seven rivers, including three in the Drammen region. In this particular infection region, a prerequisite for treatment is to establish whether G. salaris is also present on rainbow trout upstream of the salmon migration barrier. Here, we developed and tested eDNA approaches to complement conventional surveillance methods.
    METHODS: Water samples (2 × 5 l) were filtered on-site through glass fibre filters from nine locations in the Drammen watercourse, and DNA was extracted with a CTAB protocol. We developed a qPCR assay for G. salaris targeting the nuclear ribosomal ITS1 region, and we implemented published assays targeting the mitochondrial cytochrome-b and NADH-regions for Atlantic salmon and rainbow trout, respectively. All assays were transferred successfully to droplet digital PCR (ddPCR).
    RESULTS: All qPCR/ddPCR assays performed well both on tissue samples and on field samples, demonstrating the applicability of eDNA detection for G. salaris, rainbow trout and Atlantic salmon in natural water systems. With ddPCR we eliminated a low cross-amplification of Gyrodactylus derjavinoides observed using qPCR, thus increasing specificity and sensitivity substantially. Duplex ddPCR for G. salaris and Atlantic salmon was successfully implemented and can be used as a method in future surveillance programs. The presence of G. salaris eDNA in the infected River Lierelva was documented, while not elsewhere. Rainbow trout eDNA was only detected at localities where the positives could be attributed to eDNA release from upstream land-based rainbow trout farms. Electrofishing supported the absence of rainbow trout in all of the localities.
    CONCLUSIONS: We provide a reliable field and laboratory protocol for eDNA detection of G. salaris, Atlantic salmon and rainbow trout, that can complement conventional surveillance programs and substantially reduce the sacrifice of live fish. We also show that ddPCR outperforms qPCR with respect to the specific detection of G. salaris.
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