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 border="0" cellpadding="0" cellspacing="0" style="width:797px;" width="796">
	<tbody>
		<tr height="77">
			<td height="77" style="height:77px;width:216px;">&#160;Agar</td>
			<td style="width:177px;">AOBOX Biotechnology&#65288;China&#65289;</td>
			<td style="width:207px;">01-023</td>
			<td style="width:197px;"></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;">Filter paper</td>
			<td style="width:177px;">GE Healthcare brand(Sweden)&#160;&#160;</td>
			<td>10311387</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">50-mL tube</td>
			<td style="width:177px;">CORNING(Amercia)</td>
			<td style="width:207px;">430290</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">Centrifuge</td>
			<td style="width:177px;">Eppendorf(Amercia)</td>
			<td style="width:207px;">5804R</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;">Tween-20</td>
			<td style="width:177px;">Coolaber&#65288;China&#65289;</td>
			<td style="width:207px;">CT11551-100ml</td>
			<td></td>
		</tr>
		<tr height="50">
			<td height="50" style="height:50px;width:216px;">Culture dish</td>
			<td style="width:177px;">Thermofisher(Amercia)</td>
			<td>150326</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">0.5-5 mL pipette</td>
			<td>Eppendorf</td>
			<td style="width:207px;">&#160;4920000105</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">100-1000uL pipette</td>
			<td>Eppendorf</td>
			<td>4920000083</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Vacuum pump</td>
			<td style="width:177px;">Leybold</td>
			<td style="width:207px;">D25B</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Dissection needle</td>
			<td style="width:177px;">FST</td>
			<td style="width:207px;">26000-35</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Incubator</td>
			<td style="width:177px;">MEMMERT</td>
			<td style="width:207px;">PYX313</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Inoculation ring</td>
			<td style="width:177px;">Greiner Bio One</td>
			<td style="width:207px;">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?...

Watch this Scientific Journal Video about The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea at JoVE.com

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 ...

the-plant-infection-test-spray-wound-mediated-inoculation-with-plant

Research

JoVE Journal

Environment

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

12.4K 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

Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses

Setaria viridis is an emerging model system for C4 grasses. It is closely related to the bioenergy feed stock switchgrass and the grain crop foxtail millet. Recently, the 510 Mb genome of foxtail millet, S. italica, has been sequenced 1,2 and a 25x coverage genome sequence of the weedy relative S. viridis is in progress. S. viridis has a number of characteristics that make it a potentially excellent model genetic system including a rapid generation time, small stature, simple growth requirements, prolific seed production 3 and developed systems for both transient and stable transformation 4. However, the genetics of S. viridis is largely unexplored, in part, due to the lack of detailed methods for performing crosses. To date, no standard protocol has been adopted that will permit rapid production of seeds from controlled crosses.
The protocol presented here is optimized for performing genetic crosses in S. viridis, accession A10.1. We have employed a simple heat treatment with warm water for emasculation after pruning the panicle to retain 20-30 florets and labeling of flowers to eliminate seeds resulting from newly developed flowers after emasculation. After testing a series of heat treatments at permissive temperatures and varying the duration of dipping, we have established an optimum temperature and time range of 48 &deg;C for 3-6 min. By using this method, a minimum of 15 crosses can be performed by a single worker per day and an average of 3-5 outcross progeny per panicle can be recovered. Therefore, an average of 45-75 outcross progeny can be produced by one person in a single day. Broad implementation of this technique will facilitate the development of recombinant inbred line populations of S. viridis X S. viridis or S. viridis X S. italica, mapping mutations through bulk segregant analysis and creating higher order mutants for genetic analysis.

Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses

We have developed a methodology for performing crosses in Setaria viridis (S. viridis). The method involves pruning the panicle prior to a hot water treatment to kill viable pollen. Crosses are performed following a well-controlled growth regime and typically result in the recovery of 1 to 7 cross-pollinated seed/s per panicle.

方法中进行杂交<em&gt;狗尾草</em&gt;，新模型体系的草

Setaria viridis is an emerging model system for C4 grasses. It is closely related to the bioenergy feed stock switchgrass and the grain crop foxtail ...

科研

环境科学

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 ...

Single-plant, Sterile Microcosms for Nodulation and Growth of the Legume Plant Medicago truncatula with the Rhizobial Symbiont Sinorhizobium meliloti

Rhizobial bacteria form symbiotic, nitrogen-fixing nodules on the roots of compatible host legume plants. One of the most well-developed model systems for studying these interactions is the plant Medicago truncatula cv. Jemalong A17 and the rhizobial bacterium Sinorhizobium meliloti 1021. Repeated imaging of plant roots and scoring of symbiotic phenotypes requires methods that are non-destructive to either plants or bacteria. The symbiotic phenotypes of some plant and bacterial mutants become apparent after relatively short periods of growth, and do not require long-term observation of the host/symbiont interaction. However, subtle differences in symbiotic efficiency and nodule senescence phenotypes that are not apparent in the early stages of the nodulation process require relatively long growth periods before they can be scored. Several methods have been developed for long-term growth and observation of this host/symbiont pair. However, many of these methods require repeated watering, which increases the possibility of contamination by other microbes. Other methods require a relatively large space for growth of large numbers of plants. The method described here, symbiotic growth of M. truncatula/S. meliloti in sterile, single-plant microcosms, has several advantages. Plants in these microcosms have sufficient moisture and nutrients to ensure that watering is not required for up to 9 weeks, preventing cross-contamination during watering. This allows phenotypes to be quantified that might be missed in short-term growth systems, such as subtle delays in nodule development and early nodule senescence. Also, the roots and nodules in the microcosm are easily viewed through the plate lid, so up-rooting of the plants for observation is not required.

Single-plant, Sterile Microcosms for Nodulation and Growth of the Legume Plant Medicago truncatula with the Rhizobial Symbiont Sinorhizobium meliloti

Growth of Medicago truncatula plants in symbiosis with the nitrogen-fixing bacteria Sinorhizobium meliloti in individual, sterile microcosms made from standard laboratory plates permits frequent examination of root systems and nodules without compromising sterility. Plants can be maintained in these growth chambers for up to 9 weeks.

单厂，无菌的缩影和结瘤的豆科植物生长<em&gt;苜蓿</em&gt;与根瘤菌共生体<em&gt;根瘤菌</em

Rhizobial  bacteria form symbiotic, nitrogen-fixing nodules on the roots of compatible host  legume plants.  One  of the most well-developed model systems ...

Transcript and Metabolite Profiling for the Evaluation of Tobacco Tree and Poplar as Feedstock for the Bio-based Industry

The global demand for food, feed, energy, and water poses extraordinary challenges for future generations. It is evident that robust platforms for the exploration of renewable resources are necessary to overcome these challenges. Within the multinational framework MultiBioPro we are developing biorefinery pipelines to maximize the use of plant biomass. More specifically, we use poplar and tobacco tree (Nicotiana glauca) as target crop species for improving saccharification, isoprenoid, long chain hydrocarbon contents, fiber quality, and suberin and lignin contents. The methods used to obtain these outputs include GC-MS, LC-MS and RNA sequencing platforms. The metabolite pipelines are well established tools to generate these types of data, but also have the limitations in that only well characterized metabolites can be used. The deep sequencing will allow us to include all transcripts present during the developmental stages of the tobacco tree leaf, but has to be mapped back to the sequence of Nicotiana tabacum. With these set-ups, we aim at a basic understanding for underlying processes and at establishing an industrial framework to exploit the outcomes. In a more long term perspective, we believe that data generated here will provide means for a sustainable biorefinery process using poplar and tobacco tree as raw material. To date the basal level of metabolites in the samples have been analyzed and the protocols utilized are provided in this article.

Plant biomass offers a renewable resource for multiple products, including fuel, feed, food, and a variety of materials. In this paper we investigate the properties of tobacco tree (Nicotiana glauca) and poplar as suitable sources for a biorefinery pipeline.

成绩单及代谢物谱烟草树和杨树的评价作为原料的生物基工业

The global demand for food, feed, energy, and water poses extraordinary challenges for future generations. It is evident that robust platforms for the ...

Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies

The two-spotted spider mite, Tetranychus urticae, is a ubiquitous polyphagous arthropod herbivore that feeds on a remarkably broad array of species, with more than 150 of economic value. It is a major pest of greenhouse crops, especially in Solanaceae and Cucurbitaceae (e.g., tomatoes, eggplants, peppers, cucumbers, zucchini) and greenhouse ornamentals (e.g., roses, chrysanthemum, carnations), annual field crops (such as maize, cotton, soybean, and sugar beet), and in perennial cultures (alfalfa, strawberries, grapes, citruses, and plums)1,2. In addition to the extreme polyphagy that makes it an important agricultural pest, T. urticae has a tendency to develop resistance to a wide array of insecticides and acaricides that are used for its control3-7.
T. urticae is an excellent experimental organism, as it has a rapid life cycle (7 days at 27 &#176;C) and can be easily maintained at high density in the laboratory. Methods to assay gene expression (including in situ hybridization and antibody staining) and to inactivate expression of spider mite endogenous genes using RNA interference have been developed8-10. Recently, the whole genome sequence of T. urticae has been reported, creating an opportunity to develop this pest herbivore as a model organism with equivalent genomic resources that already exist in some of its host plants (Arabidopsis thaliana and the&#160;tomato Solanum lycopersicum)11. Together, these&#160;model organisms could provide insights into molecular bases of plant-pest interactions.
Here, an efficient method for quick and easy collection of a large number of adult female mites, their application on an experimental plant host, and the assessment of the plant damage due to spider mite feeding are described. The presented protocol enables fast and efficient collection of hundreds of individuals at any developmental stage (eggs, larvae, nymphs, adult males, and females) that can be used for subsequent experimental application.

Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies

Protocols for efficient preparation of homogenous samples of spider mites, infestation of experimental plants, and assessment of plant damage, as required for studies of plant-pest interaction were developed.

二斑叶螨的应用<em&gt;二斑叶螨</em&gt;植物病虫害的相互作用研究

The two-spotted spider mite, Tetranychus urticae, is a ubiquitous polyphagous arthropod herbivore that feeds on a remarkably broad array of species, with ...

The Use of High-resolution Infrared Thermography (HRIT) for the Study of Ice Nucleation and Ice Propagation in Plants

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events often have devastating effects on agricultural production and can also play an important role in shaping community structure in natural populations of plants, especially in alpine, sub-arctic, and arctic ecosystems. Therefore, a better understanding of the freezing process in plants can play an important role in the development of methods of frost protection and understanding mechanisms of freeze avoidance. Here, we describe a protocol to visualize the freezing process in plants using high-resolution infrared thermography (HRIT). The use of this technology allows one to determine the primary sites of ice formation in plants, how ice propagates, and the presence of ice barriers. Furthermore, it allows one to examine the role of extrinsic and intrinsic nucleators in determining the temperature at which plants freeze and evaluate the ability of various compounds to either affect the freezing process or increase freezing tolerance. The use of HRIT allows one to visualize the many adaptations that have evolved in plants, which directly or indirectly impact the freezing process and ultimately enables plants to survive frost events.

The Use of High-resolution Infrared Thermography HRIT for the Study of Ice Nucleation and Ice Propagation in Plants

Here we present a protocol that allows one to visualize sites of ice formation and avenues of ice propagation in plants utilizing high resolution infrared thermography (HRIT).

高分辨率红外热成像（HRIT）的冰核和冰传播的植物研究中的应用

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events ...

A New Application of the Electrical Penetration Graph (EPG) for Acquiring and Measuring Electrical Signals in Phloem Sieve Elements

Electrophysiological properties of cells are often studied in vitro, after dissociating them from their native environments. However, the study of electrical transmission between distant cells in an organism requires in vivo, artifact-free recordings of cells embedded within their native environment. The transmission of electrical signals from wounded to unwounded areas in a plant has since long piqued the interest of botanists. The phloem, the living part of the plant vasculature that is spread throughout the plant, has been postulated as a major tissue in electrical transmission in plants. The lack of suitable electrophysiological methods poses many challenges for the study of the electrical properties of the phloem cells in vivo. Here we present a novel approach for intracellular electrophysiology of sieve elements (SEs) that uses living aphids, or other phloem-feeding hemipteran insects, integrated in the electrical penetration graph (EPG) circuit. The versatility, robustness, and accuracy of this method made it possible to record and study in detail the wound-induced electrical signals in SEs of central veins of the model plant Arabidopsis thaliana1. Here we show that EPG-electrodes can be easily implemented for intracellular electrophysiological recordings of SEs in marginal veins, as well as to study the capacity of SEs to respond with electrical signals to several external stimuli. The EPG approach applied to intracellular electrophysiology of SEs can be implemented to a wide variety of plant species, in a large number of plant/insect combinations, and for many research aims.

A New Application of the Electrical Penetration Graph EPG for Acquiring and Measuring Electrical Signals in Phloem Sieve Elements

Electrical Penetration Graph (EPG) is a well-established technique for studying the feeding behavior of stylet-bearing insects. Here we show a new application of EPG as a non-invasive tool for the acquisition of intracellular electrophysiology recordings of sieve elements (SEs), the cells that form the phloem vasculature in plants.

在电气贯穿图（EPG）的韧皮部筛分子获取和测量电信号中的新应用

Electrophysiological properties of cells are often studied in vitro, after dissociating them from their native environments. However, the study of ...

Operation of a 25 KWth Calcium Looping Pilot-plant with High Oxygen Concentrations in the Calciner

Calcium looping (CaL) is a post-combustion CO2 capture technology that is suitable for retrofitting existing power plants. The CaL process uses limestone as a cheap and readily available CO2 sorbent. While the technology has been widely studied, there are a few available options that could be applied to make it more economically viable. One of these is to increase the oxygen concentration in the calciner to reduce or eliminate the amount of recycled gas (CO2, H2O and impurities); therefore, decreasing or removing the energy necessary to heat the recycled gas stream. Moreover, there is a resulting increase in the energy input due to the change in the combustion intensity; this energy is used to enable the endothermic calcination reaction to occur in the absence of recycled flue gases. This paper presents the operation and first results of a CaL pilot plant with 100% oxygen combustion of natural gas in the calciner. The gas coming into the carbonator was a simulated flue gas from a coal-fired power plant or cement industry. Several limestone particle size distributions are also tested to further explore the effect of this parameter on the overall performance of this operating mode. The configuration of the reactor system, the operating procedures, and the results are described in detail in this paper. The reactor showed good hydrodynamic stability and stable CO2 capture, with capture efficiencies of up to 70% with a gas mixture simulating the flue gas of a coal-fired power plant.

Operation of a 25 KWth Calcium Looping Pilot-plant with High Oxygen Concentrations in the Calciner

This manuscript describes a procedure for operating a calcium looping pilot-plant for post-combustion carbon capture with high oxygen concentrations in the calciner in order to reduce or eliminate the flue gas recycle.

在煅烧炉中使用高氧浓度的 25 KWth钙循环试验装置的操作

Calcium looping (CaL) is a post-combustion CO2 capture technology that is suitable for retrofitting existing power plants. The CaL process uses limestone ...

Adherence of Bacteria to Plant Surfaces Measured in the Laboratory

This manuscript describes a method to measure bacterial binding to axenic plant surfaces in the light microscope and through the use of viable cell counts. Plant materials used include roots, sprouts, leaves, and cut fruits. The methods described are inexpensive, easy, and suitable for small sample sizes. Binding is measured in the laboratory and a variety of incubation media and conditions can be used. The effect of inhibitors can be determined. Situations that promote and inhibit binding can also be assessed. In some cases it is possible to distinguish whether various conditions alter binding primarily due to their effects on the plant or on the bacteria.

An easy method for measuring and characterizing bacterial adhesion to plants, particularly roots and sprouts, is described in this article.

细菌附着在实验室测量的植物表面

This manuscript describes a method to measure bacterial binding to axenic plant surfaces in the light microscope and through the use of viable cell ...

Specific and Accurate Detection of the Citrus Greening Pathogen Candidatus liberibacter spp. Using Conventional PCR on Citrus Leaf Tissue Samples

Citrus greening, also known as huanglongbing, is a destructive citrus disease ravaging citrus farms globally. This disease causes asymmetrical yellow leaf mottling, vein yellowing, defoliation, root decay, and ultimately, the death of the citrus plant. When infected, the citrus plants have stunted growth and produce flowers out of season. These flowers rarely yield fruit, and those that do yield small, bitter, irregularly shaped citrus fruit that are not desirable. This disease is spread by the Asian citrus psyllid, Diaphorina citri, and by the grafting of infected citrus tissue. The pathogen has a long and variable incubation period within the citrus plant&#8212;sometimes years, before symptoms appear. Attempts to culture this pathogen in vitro have been unsuccessful, possibly due to the low and uneven concentration of the pathogen within infected citrus tissue, or because it is difficult to replicate the environmental conditions conducive to growth of the pathogen. It is very difficult to identify the disease before it has spread, due to its long incubation period and researchers&#39; inability to culture the pathogen. As a result, the disease only becomes apparent after suddenly destroying a citrus farmer&#39;s entire yield. Presented here is a method for the accurate and specific detection of the citrus greening pathogen, Candidatus liberibacter spp. using a genomic DNA extraction kit and PCR. This method is simple, efficient, cost effective, and adaptable for quantitative analysis. This method can be adapted for use on any citrus tissue; however, it is potentially limited by the amount of pathogen present in the tissue. Nevertheless, this method will allow citrus farmers to identify infected citrus plants earlier, and curb the spread of this destructive disease before it can further spread.

Specific and Accurate Detection of the Citrus Greening Pathogen Candidatus liberibacter  spp. Using Conventional PCR on Citrus Leaf Tissue Samples

Citrus Greening is a particularly destructive disease affecting citrus crops globally. Presented here is a simple method using PCR and genomic DNA extraction of citrus leaf tissue for the accurate and precise identification of the citrus greening pathogen, Candidatus liberibacter spp.

利用常规 PCR 技术对柑桔叶片组织样品Candidatus liberibacter的特异和准确检测

Citrus greening, also known as huanglongbing, is a destructive citrus disease ravaging citrus farms globally. This disease causes asymmetrical yellow leaf ...