Name: the plant infection test: spray and wound-mediated inoculation with the plant pathogen magnaporthe grisea
Uploaded: 2018-08-04T13:30:00.000+00:00
Duration: 7 min 14 s
Description: Watch this Scientific Journal Video about The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea at JoVE.com

这项工作得到了北京农业大学 (YQ201603) 的专项科研项目和北京教育委员会 (KM201610020005) 科学项目的支持。

1. 用悬浮菌孢子进行喷雾接种
<ol><li>
菌的真菌培养
	<ol>
<li>为真菌菌株制备燕麦番茄琼脂 (OTA) 培养基。</li>
		<li>重 30-50 克燕麦片, 加入到800毫升的蒸馏/去离子水 (ddH2O) 和煮沸的混合物为30分钟在电热锅。</li>
		<li>用一块纱布把煮过的燕麦汁过滤到烧杯中。</li>
		<li>在烧杯中加入150毫升番茄汁和20克琼脂, 加入 ddH2O 至1000毫升。</li>
	</ol>
</li>
	<li>
实验材料的制备
	<ol>
<li>在大麦2o 中浸泡约50 种水稻 (稻) 品种 Lijiangxintuan-heigu (LTH) 在 ddH2o 3 d 或浸泡约50粒大麦 (ddH的 cv 金承诺), 约 1 d。</li>
		<li>将大米或大麦的种子裹在潮湿的纱布中, 在湿润的滤纸上发芽, 在大约30培养皿中, 直径为10厘米 x 10 厘米, 28 摄氏度。水稻种子萌发时间约为 2-3 维, 大麦种子萌发时间约为2。温室相对湿度约为70%。</li>
		<li>用蒸压灌封土和浇水, 然后用一层蛭石将大米或大麦的幼苗种植在盆中。</li>
		<li>将植物放置在合适的温室或生长柜中, 温度在25摄氏度左右, 约 1 ~ 2 周。</li>
		<li>用无菌手术剪刀切开3层滤纸, 用不育的外科剪 (每个圆应该有一个8厘米直径) 并且安置他们到100毫米无菌的塑料板材。</li>
		<li>添加 ddH2O 到每道菜浸泡过滤纸。</li>
		<li>确保过滤纸是完全湿的, 但添加没有额外的水。</li>
		<li>用真空泵除去多余的水。</li>
		<li>将2不育牙签放入培养皿中, 以支持大米/大麦叶;间隔 2-3 厘米的牙签。</li>
		<li>在播种种子后, 在播种后的2周内收集大米的叶子, 或取去大麦的叶子 7 d。</li>
		<li>用4到6叶的水稻/大麦幼苗, 从顶部到5厘米处切下茎下部, 收集叶子。</li>
	</ol>
</li>
	<li>
喷雾接种协议
	<ol>
<li>在恒温孵化器 (25 °c) 中, 对 OTA 板上的真菌菌株进行培养, 4 d。</li>
		<li>添加〜2毫升的 ddH2O 使用 0.5-5 毫升吸管, 每4天老板。</li>
		<li>用接种循环, 将m . 型菌的菌丝和突变株刮成菌丝碎屑。</li>
		<li>收集菌丝碎片, 并将其转移到一个新的 OTA 板块。把菌丝碎片和吹干在干净的长凳上。</li>
		<li>盖板与3层纱布, 以确保在温室中的孢子生长所需的湿度在25摄氏度在白天 (14 小时) 和23°c 在夜间 (10 h) 24-48 h。</li>
		<li>在每道菜上加入2毫升的 ddH2O, 用不育的棉签轻轻地刮出孢子, 然后通过2层透镜纸过滤。注意不要刮伤培养基的表面。</li>
		<li>将孢子悬浮液转移到一个新的50毫升管与 100-1000 µL 吸管。</li>
		<li>离心机为5分钟, 最小为 5000 x 克25摄氏度。</li>
		<li>除去上清和并用重悬颗粒, 在 0.025% (v/v) Tween-20 溶液中每毫升 2 x 104的孢子。Tween-20 溶液通常约为 10-20 毫升。</li>
		<li>将孢子悬浮液倒入手持式喷雾器中。</li>
		<li>将约10毫升的孢子悬浮液喷洒在2周大的水稻幼苗上, 或在7天大的大麦幼苗上的大麦叶上, 在黑暗中孵化25摄氏度, 湿润室为 ~ 24 h. 用 0.025% (v/v) Tween-20 溶液喷洒控制装置。</li>
		<li>在25摄氏度的光照下, 将叶子转移到另一个潮湿的房间中, 周期为12小时。</li>
		<li>在接种后记录 5 d 的疾病症状。检查长度为6厘米的病米/大麦叶片。评价标准是根据国际水稻研究所 (IRRI) 抗稻瘟病评分系统进行的。表 1列出了抗爆性评分系统的详细情况。</li>
		<li>拍摄树叶以评估受试菌株的感染情况。感染是由每3.6 厘米2的病灶数量来评估的。</li>
	</ol>
</li>
</ol>2. 用菌丝体或孢子悬浮液滴进行的伤杆菌接种
<ol><li>使用解剖针, 在分离的大米/大麦叶片的主脉上刮三 2-3 厘米长的伤口;注意不要穿透树叶。</li>
	<li>把刮过的叶子放在牙签上, 在叶子上喷洒 0.02% (v/v) Tween-20 溶液, 形成一层水滴。</li>
	<li>从 OTA 板上切下0.5 厘米 x 0.5 厘米的菌丝塞, 为每株m菌菌株 (野生型、突变体、补体菌株或其他试验菌株)。</li>
	<li>将菌丝塞或25µL 滴孢子悬浮在受伤的叶子上, 在潮湿的房间里孵化25摄氏度的叶子, 3-8 d。</li>
	<li>检查 5-7 d 接种后的病变。检查方法与喷雾接种方法相同 (见步骤 1.3.13)。</li>
	<li>检查长度为6厘米的病米/大麦叶片, 并拍摄它们以评估受试菌株的感染情况。感染是由每3.6 厘米2的病灶数量来评估的。</li>
</ol>

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作者没有什么可透露的。

植物抗病基因在预防病原体感染方面起着至关重要的作用, 包括真菌病原体1、12。稻瘟病已被用来作为一个模型, 以了解病原体种群结构的性质, 并确定植物抗病基因4。因此, 有必要对主要品种的抗病基因型和无毒基因型进行大规模的检测, 以确定可持续栽培的抗病植物。此外, 无毒基因型在田间病原体中的优势, 使得寄主品种<sup class="xr...

水稻稻瘟病是1、2世界水稻品种中最严重的病害之一. 附着胞菌感染寄主植物的过程包括孢子的产生和表面附着、孢子萌发和的形成、渗透 peg 的形成和传染性菌丝分化以及疾病的传播。3. 所有这些阶段在许多其他植物病原真菌中很常见, 事实上, 任何单一阶段的封锁都可以防止寄主植物的感染。由于其经济重要性和遗传驯良, m . 菌已成为研究植物-真菌病原体相互作用机制1,4的模型有机体。因此, 研究m . 菌的这些发育阶段的分子基础将有助于阐明真菌致病性的分子机制和候选靶基因的鉴定, 以筛选和设计新的杀菌剂5。
最近有关m . 菌感染的报告着重于前侵染阶段的分子机制, 特别是 conidiation、附着胞形成、穿透钉和传染性生长3,6. 因此, 必须制定一项详细的议定书, 以测试m . 菌感染。在此, 我们提出了一个感染测试的详细方法, 利用喷雾介导的感染化验与孢子悬浮和接种的伤口, 菌丝插头的m。在本报告中, 该议定书的重点是菌株的培养, conidiation 溶液的制备, 以及菌丝栓介导的植物与m. 菌的接种。下面详细介绍了这些步骤, 并分别显示了图 1和表2中显示方法的整个工作流和典型病变的示意图视图。

<table><tbody><tr><td>Filter paper</td><td>GE Healthcare brand(Sweden)&amp;nbsp;&amp;nbsp;</td><td>10311387</td><td></td></tr><tr><td>50-mL tube</td><td>CORNING(Amercia)</td><td>430290</td><td></td></tr><tr><td>Centrifuge</td><td>Eppendorf(Amercia)</td><td>5804R</td><td></td></tr><tr><td>Culture dish</td><td>Thermofisher(Amercia)</td><td>150326</td><td></td></tr><tr><td>0.5-5 mL pipette</td><td>Eppendorf</td><td>&amp;nbsp;4920000105</td><td></td></tr><tr><td>100-1000uL pipette</td><td>Eppendorf</td><td>4920000083</td><td></td></tr><tr><td>Vacuum pump</td><td>Leybold</td><td>D25B</td><td></td></tr><tr><td>Dissection needle</td><td>FST</td><td>26000-35</td><td></td></tr><tr><td>Incubator</td><td>MEMMERT</td><td>PYX313</td><td></td></tr><tr><td>Inoculation ring</td><td>Greiner Bio One</td><td>731175</td><td></td></tr></tbody></table>

the plant infection test: spray and wound-mediated inoculation with the plant pathogen magnaporthe grisea

植物拥有强大的系统来抵御致病真菌的潜在威胁。然而, 对于农业上重要的植物来说, 目前防治这种病原体的措施过于保守, 因而没有足够的效率, 而且可能造成环境风险。因此, 通过对抗病种质的鉴定、抗性基因的分离和鉴定以及分子育种, 确定寄主抗性因子, 以帮助自然控制植物病害是非常必要的。抗病品种。在这方面, 需要建立一个准确、快速、大规模的接种方法来培育和发展植物抗病基因。稻瘟病真菌病原菌引起严重的疾病症状和产量损失。近年来, m . 菌已成为研究植物真菌病原体相互作用机制的模型有机体。因此, 我们报告了植物毒性试验方法的发展, 这是特定的m. 菌。这种方法提供两种喷雾接种与孢子悬浮和伤人接种与菌丝立方体或孢子悬浮液滴。分离水稻叶片伤后接种方法的关键步骤是在植物叶片上制造伤口, 避免因寄主侵彻阻力而引起的干扰。这种喷雾/伤后的协议有助于快速, 准确, 大规模筛选致病型菌的分离株. 这种综合和系统的植物感染方法将作为一个良好的出发点, 以获得广泛的观点, 植物病理学的问题。

该技术的整个工作流如图 1所示。对14天的易感水稻幼苗 (CO-39 cv) 或7天大的大麦叶片进行了植物感染检测,7、8、9。为了测试水稻叶片的感染情况, 制备了 P131 菌野生型菌株的孢子悬浮 (1.0 x 105孢子 /毫升) 和 Com1 删除突变株, 然后喷洒到14天大易感性的叶?...

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The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

植物感染试验: 以植物病原菌为载体的喷雾和伤口介导接种

在这里, 我们提出了一个测试植物致病力的植物病原菌稻瘟病的协议.这份报告将有助于大规模筛选真菌分离株的致病型, 作为了解植物在分子育种过程中耐药性机制的良好出发点。

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current ...

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The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

12.5K 次观看.  Beijing University of Agriculture. 在这里, 我们提出了一个测试植物致病力的植物病原菌稻瘟病的协议.这份报告将有助于大规模筛选真菌分离株的致病型, 作为了解植物在分子育种过程中耐药性机制的良好出发点。

视频: The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is a biotrophic fungal pathogen and the causal agent of corn smut on maize. This disease is responsible for significant yield losses of approximately $1.0 billion annually in the U.S.1 Several methods including crop rotation, fungicide application and seed treatments are currently used to control corn smut2. However, host resistance is the only practical method for managing corn smut. Identification of crop plants including maize, wheat, and rice that are resistant to various biotrophic pathogens has significantly decreased yield losses annually3-5. Therefore, the use of a pathogen inoculation method that efficiently and reproducibly delivers the pathogen in between the plant leaves, would facilitate the rapid identification of maize lines that are resistant to U. maydis. As, a first step toward indentifying maize lines that are resistant to U. maydis, a needle injection inoculation method and a resistance reaction screening method was utilized to inoculate maize, teosinte, and maize x teosinte introgression lines with a U. maydis strain and to select resistant plants.
Maize, teosinte and maize x teosinte introgression lines, consisting of about 700 plants, were planted, inoculated with a strain of U. maydis, and screened for resistance. The inoculation and screening methods successfully identified three teosinte lines resistant to U. maydis. Here a detailed needle injection inoculation and resistance reaction screening protocol for maize, teosinte, and maize x teosinte introgression lines is presented. This study demonstrates that needle injection inoculation is an invaluable tool in agriculture that can efficiently deliver U. maydis in between the plant leaves and has provided plant lines that are resistant to U. maydis that can now be combined and tested in breeding programs for improved disease resistance.

A Rapid and Efficient Method for Assessing Pathogenicity of Ustilago maydis on Maize and Teosinte Lines

The use of a needle injection method to inoculate maize and teosinte plants with the biotrophic pathogen Ustilago maydis is described. The needle injection inoculation method facilitates the controlled delivery of the fungal pathogen in between the plant leaves where the pathogen enters the plant through the formation of appresoria. This method is highly efficient, enabling reproducible inoculations with U. maydis.

评估 乌西拉戈 病原性的快速高效方法，可在玉米和特奥辛特线上使用

Maize is a major cereal crop worldwide. However, susceptibility to biotrophic pathogens is the primary constraint to increasing productivity. U. maydis is ...

科研

环境科学

Establishing Fungal Entomopathogens as Endophytes: Towards Endophytic Biological Control

Beauveria bassiana is a fungal entomopathogen with the ability to colonize plants endophytically. As an endophyte, B. bassiana may play a role in protecting plants from herbivory and disease. This protocol demonstrates two inoculation methods to establish B. bassiana endophytically in the common bean (Phaseolus vulgaris), in preparation for subsequent evaluations of endophytic biological control. Plants are grown from surface-sterilized seeds for two weeks before receiving a B. bassiana treatment of 108 conidia/ml (or water) applied either as a foliar spray or a soil drench. Two weeks later, the plants are harvested and their leaves, stems and roots are sampled to evaluate endophytic fungal colonization. For this, samples are individually surface sterilized, cut into multiple sections, and incubated in potato dextrose agar media for 20 days. The media is inspected every 2-3 days to observe fungal growth associated with plant sections and record the occurrence of B. bassiana to estimate the extent of its endophytic colonization. Analyses of inoculation success compare the occurrence of B. bassiana within a given plant part (i.e. leaves, stems or roots) across treatments and controls. In addition to the inoculation method, the specific outcome of the experiment may depend on the target crop species or variety, the fungal entomopathogen species strain or isolate used, and the plant's growing conditions.

This protocol demonstrates two inoculation methods to introduce the fungal entomopathogen Beauveria bassiana as an endophyte in the common bean (Phaseolus vulgaris), in preparation for subsequent evaluations of endophytic biological control.

建立真菌Entomopathogens作为内生菌：建立内生生物控制

 Beauveria bassiana is a fungal entomopathogen with the ability to colonize plants endophytically. As an endophyte, B. bassiana may play a role in ...

生物工程

An Effective Inoculation Method for Phytophthora capsici on Black Pepper Plants

Piper nigrum L. (black pepper) is a typical woody vine that is an economically important spice crop across the world. Black pepper production is significantly impacted by root rot disease caused by Phytophthora capsici, which has seriously influenced the industry development as a "choke point" problem. However, the molecular genetic mechanism of resistance in black pepper is unclear, leading to slow progress in the development of new black pepper varieties. An effective inoculation and precise sampling system for Phytophthora capsici on black pepper plants is essential for studying this plant-pathogen interaction. The main aim of this study is to demonstrate a detailed methodology where the basal head of black pepper is inoculated with Phytophthora capsici, while also providing a reference for the inoculation of woody vine plants. The basal head of the black pepper plant was pinpricked to damage it, and mycelial pellets covered the three holes to retain the moisture so the pathogen could infect the plant well. This method provides a better way of solving the instability that is caused by traditional inoculation methods including soil drench or root dipping. It also provides a promising means for studying the mode of action between plants and other soil-borne plant pathogens in agricultural precision breeding.

An Effective Inoculation Method for Phytophthora capsici on Black Pepper Plants

Pinpricking the basal head of the black pepper plant is a brief and time-saving method to damage it. Here, we provided detailed steps with a video for infecting black pepper plants.

一种有效的黑胡椒植 酸菌 接种方法

Piper nigrum L. (black pepper) is a typical woody vine that is an economically important spice crop across the world. Black pepper production is ...

遗传学

Inoculation Strategies to Infect Plant Roots with Soil-Borne Microorganisms

The rhizosphere harbors a highly complex microbial community in which plant roots are constantly challenged. Roots are in close contact with a wide variety of microorganisms, but studies on soil-borne interactions are still behind those performed on aboveground organs. Although some inoculation strategies for infecting model plants with model root pathogens are described in the literature, it remains difficult to get a comprehensive methodological overview. To address this problem, three different root inoculation systems are precisely described that can be applied to gain insights into the biology of root-microbe interactions. For illustration, Verticillium species (namely, V. longisporum and V. dahliae) were employed as root invading model pathogens. However, the methods can be easily adapted to other root colonizing microbes - both pathogenic and beneficial. By colonizing the plant xylem, vascular soil-borne fungi such as Verticillium spp. exhibit a unique lifestyle. After root invasion, they spread via the xylem vessels acropetally, reach the shoot, and elicit disease symptoms. Three representative plant species were chosen as model hosts: Arabidopsis thaliana, economically important oilseed rape (Brassica napus), and tomato (Solanum lycopersicum). Step-by-step protocols are given. Representative results of pathogenicity assays, transcriptional analyses of marker genes, and independent confirmations by reporter constructs are shown. Furthermore, the advantages and disadvantages of each inoculation system are thoroughly discussed. These proven protocols can assist in providing approaches for research questions on root-microbe interactions. Knowing how plants cope with microbes in the soil is crucial for developing new strategies to improve agriculture.

This protocol presents a detailed summary of strategies to inoculate plant roots with soil-borne microbes. Exemplified for the fungi Verticillium longisporum and Verticillium dahliae, three different root infection systems are described. Potential applications and possible downstream analyses are highlighted, and advantages or disadvantages are discussed for each system.

用土壤微生物感染植物根系的接种策略

The rhizosphere harbors a highly complex microbial community in which plant roots are constantly challenged. Roots are in close contact with a wide ...

Visualizing Early Infection Sites of Rice Blast Disease (Magnaporthe oryzae) on Barley (Hordeum vulgare) Using a Basic Microscope and a Smartphone

Understanding how plants and pathogens interact, and whether that interaction culminates in defense or disease, is required to develop stronger and more sustainable strategies for plant health. Advances in methods that more effectively image plant-pathogen samples during infection and colonization have yielded tools such as the rice leaf sheath assay, which has been useful in monitoring infection and early colonization events between rice and the fungal pathogen, Magnaporthe oryzae. This hemi-biotrophic pathogen causes severe disease loss in rice and related monocots, including millet, rye, barley, and more recently, wheat. The leaf sheath assay, when performed correctly, yields an optically clear plant section, several layers thick, which allows researchers to perform live-cell imaging during pathogen attack or generate fixed samples stained for specific features. Detailed cellular investigations into the barley-M. oryzae interaction have lagged behind those of the rice host, in spite of the growing importance of this grain as a food source for animals and humans and as fermented beverages. Reported here is the development of a barley leaf sheath assay for intricate studies of M. oryzae interactions during the first 48 h post-inoculation. The leaf sheath assay, regardless of which species is being studied, is delicate; provided is a protocol that covers everything, from barley growth conditions and obtaining a leaf sheath, to inoculation, incubation, and imaging of the pathogen on plant leaves. This protocol can be optimized for high-throughput screening using something as simple as a smartphone for imaging purposes.

Visualizing Early Infection Sites of Rice Blast Disease Magnaporthe oryzae on Barley Hordeum vulgare Using a Basic Microscope and a Smartphone

This is a straightforward protocol of a barley leaf sheath assay using minimal reagents and common laboratory equipment (including a basic smartphone). The purpose is to visualize the early infection process of blast disease in labs without access to advanced microscopy equipment.

使用基本显微镜和智能手机可视化大麦（大麦）稻瘟病（稻瘟病）的早期感染部位

Understanding how plants and pathogens interact, and whether that interaction culminates in defense or disease, is required to develop stronger and more ...

发育生物学

An Easy and Flexible Inoculation Method for Accurately Assessing Powdery Mildew-Infection Phenotypes of Arabidopsis and Other Plants

Reducing crop losses due to fungal diseases requires improved understanding of the mechanisms governing plant immunity and fungal pathogenesis, which in turn requires accurate determination of disease phenotypes of plants upon infection with a particular fungal pathogen. However, accurate disease phenotyping with unculturable biotrophic fungal pathogens such as powdery mildew is not easy to achieve and can be a rate-limiting step of a research project. Here, we have developed a safe, efficient, and easy-to-operate disease phenotyping system using the Arabidopsis-powdery mildew interaction as an example. This system mainly consists of three components: (i) a wooden inoculation box fitted with a removable lid mounted with a stainless steel or nylon mesh of ~50 &#181;m pores for inoculating a flat of plants with fungal spores, (ii) a transparent plastic chamber with a small front opening for minimizing spore escape while conducting inoculation inside, and (iii) a spore-dislodging and distribution method for even and effective inoculation. The protocols described here include the steps and parameters for making the inoculation box and the plastic chamber at a low cost, and a video demonstration of how to use the system to enable even inoculation with powdery mildew spores, thereby improving accuracy and reproducibility of disease phenotyping.

We present a protocol for constructing a simple spore-distribution system consisting of an inoculation box with a ~50 &#181;m mesh and a transparent plastic chamber. This can be used to evenly inoculate plants with powdery mildew spores, thereby enabling accurate and reproducible assessment of disease phenotypes of plants under study.

一种简单灵活的接种方法，用于准确评估拟南芥和其他植物的白粉病感染表型

Reducing crop losses due to fungal diseases requires improved understanding of the mechanisms governing plant immunity and fungal pathogenesis, which in ...

生物学

Isolation and Selection of Entomopathogenic Fungi from Soil Samples and Evaluation of Fungal Virulence against Insect Pests

Entomopathogenic fungi (EPF) are one of the microbial control agents for integrated pest management. To control local or invasive pests, it is important to isolate and select indigenous EPF. Therefore, the soil bait method combined with the insect bait (mealworm, Tenebrio molitor) system was used in this study with some modifications. The isolated EPF were then subjected to the virulence test against the agricultural pest Spodoptera litura. Furthermore, the potential EPF strains were subjected to morphological and molecular identifications. In addition, the conidia production and thermotolerance assay were performed for the promising EPF strains and compared; these data were further substituted into the formula of effective conidia number (ECN) for laboratory ranking. The soil bait-mealworm system and the ECN formula can be improved by replacing insect species and integrating more stress factors for the evaluation of commercialization and field application. This protocol provides a quick and efficient approach for EPF selection and will improve the research on biological control agents.

Here we present a protocol based on the&#160;mealworm (Tenebrio molitor)-bait system that was used for isolating and selecting entomopathogenic fungi (EPF) from soil samples. An effective conidia number (ECN) formula is used to select high stress tolerant EPF based on physiological characteristics for pest microbial control in the field.

土壤样本中昆虫病原真菌的分离与选择及真菌对害虫的毒力评价

Entomopathogenic fungi (EPF) are one of the microbial control agents for integrated pest management. To control local or invasive pests, it is important ...

A Contrast of Three Inoculation Techniques used to Determine the Race of Unknown Fusarium oxysporum f.sp. niveum Isolates

Fusarium wilt of watermelon (Citrullus lanatus), caused by Fusarium oxysporum f. sp. niveum (Fon), has reemerged as a major production constraint in the southeastern USA, especially in Florida. Deployment of integrated pest management strategies, such as race-specific resistant cultivars, requires information on the diversity and population density of the pathogen in growers' fields. Despite some progress in developing molecular diagnostic tools to identify pathogen isolates, race determination often requires bioassay approaches.
Race typing was conducted by root-dip inoculation, infested kernel seeding method, and the modified tray-dip method with each of the four watermelon differentials (Black Diamond, Charleston Grey, Calhoun Grey, Plant Introduction 296341-FR). Isolates are assigned a race designation by calculation of disease incidence five weeks after inoculation. If less than 33% of the plants for a particular cultivar were symptomatic, they were categorized as resistant. Those cultivars with incidence greater than 33% were regarded as susceptible. This paper describes three different methods of inoculation to ascertain race, root-dip, infested kernel, and modified tray-dip inoculation, whose applications vary according to the experimental design.

A Contrast of Three Inoculation Techniques used to Determine the Race of Unknown Fusarium oxysporum f.sp. niveum Isolates

Managing Fusarium wilt of watermelon requires knowledge of the pathogen races present. Here, we describe the root-dip, infested kernel seeding, and modified tray-dip inoculation methods to demonstrate their efficacy in race-typing of the pathogenic fungus Fusarium oxysporum f. sp niveum (Fon).

用于确定未知 尖孢镰刀菌 f.sp的种族的三种接种技术的对比。 尼维姆 分离

Fusarium wilt of watermelon (Citrullus lanatus), caused by Fusarium oxysporum f. sp. niveum (Fon), has reemerged as a major production constraint in the ...

玉米鞘和真菌之间细胞相互作用的可视化

Detached Maize Sheaths for Live-Cell Imaging of Infection by Fungal Foliar Maize Pathogens

We have optimized a protocol to inoculate maize leaf sheaths with hemibiotrophic and necrotrophic foliar pathogenic fungi. The method is modified from one originally applied to rice leaf sheaths and allows direct microscopic observation of fungal growth and development in living plant cells. Leaf sheaths collected from maize seedlings with two fully emerged leaf collars are inoculated with 20 &#181;L drops of 5 x 105 spores/mL fungal spore suspensions and incubated in humidity chambers at 23 &#176;C under continuous fluorescent light. After 24-72 h, excess tissue is removed with a razor blade to leave a single layer of epidermal cells, an optically clear sample that can be imaged directly without the necessity for chemical fixation or clearing. Plant and fungal cells remain alive for the duration of the experiment and interactions can be visualized in real-time. Sheaths can be stained or subjected to plasmolysis to study the developmental cytology and viability of host and pathogen cells during infection and colonization. Fungal strains transformed to express fluorescent proteins can be inoculated or co-inoculated on the sheaths for increased resolution and to facilitate the evaluation of competitive or synergistic interactions. Fungal strains expressing fluorescent fusion proteins can be used to track and quantify the production and targeting of these individual proteins in planta. Inoculated sheath tissues can be extracted to characterize nucleic acids, proteins, or metabolites. The use of these sheath assays has greatly advanced the detailed studies of the mechanisms of fungal pathogenicity in maize and also of fungal protein effectors and secondary metabolites contributing to pathogenicity.

作者聚焦:研究玉米中的真菌致病机制 

This manuscript details an optimized inoculation protocol that uses detached maize leaf sheaths for reproducible cytological, physiological, and molecular studies of maize interactions with fungal plant pathogens. The leaf sheaths facilitate real-time observation of cellular interactions between the living plant and fungus in unfixed tissues.

用于真菌叶面玉米病原体感染的活细胞成像的分离玉米鞘

We have optimized a protocol to inoculate maize leaf sheaths with hemibiotrophic and necrotrophic foliar pathogenic fungi. The method is modified from one ...

植物感染试验: 以植物病原菌为载体的喷雾和伤口介导接种

摘要

探索更多视频

此视频中的章节

评估乌西拉戈病原性的快速高效方法，可在玉米和特奥辛特线上使用

建立真菌Entomopathogens作为内生菌：建立内生生物控制

一种有效的黑胡椒植酸菌接种方法

用土壤微生物感染植物根系的接种策略

使用基本显微镜和智能手机可视化大麦（大麦）稻瘟病（稻瘟病）的早期感染部位

一种简单灵活的接种方法，用于准确评估拟南芥和其他植物的白粉病感染表型

土壤样本中昆虫病原真菌的分离与选择及真菌对害虫的毒力评价

用于确定未知尖孢镰刀菌 f.sp的种族的三种接种技术的对比。尼维姆分离

用于真菌叶面玉米病原体感染的活细胞成像的分离玉米鞘

用于真菌叶面玉米病原体感染的活细胞成像的分离玉米鞘

植物感染试验: 以植物病原菌为载体的喷雾和伤口介导接种

摘要

探索更多视频

此视频中的章节

评估 乌西拉戈 病原性的快速高效方法，可在玉米和特奥辛特线上使用

建立真菌Entomopathogens作为内生菌：建立内生生物控制

一种有效的黑胡椒植 酸菌 接种方法

用土壤微生物感染植物根系的接种策略

使用基本显微镜和智能手机可视化大麦（大麦）稻瘟病（稻瘟病）的早期感染部位

一种简单灵活的接种方法，用于准确评估拟南芥和其他植物的白粉病感染表型

土壤样本中昆虫病原真菌的分离与选择及真菌对害虫的毒力评价

用于确定未知 尖孢镰刀菌 f.sp的种族的三种接种技术的对比。 尼维姆 分离

用于真菌叶面玉米病原体感染的活细胞成像的分离玉米鞘

用于真菌叶面玉米病原体感染的活细胞成像的分离玉米鞘

评估乌西拉戈病原性的快速高效方法，可在玉米和特奥辛特线上使用

一种有效的黑胡椒植酸菌接种方法

用于确定未知尖孢镰刀菌 f.sp的种族的三种接种技术的对比。尼维姆分离