Name: the plant infection test: spray and wound-mediated inoculation with the plant pathogen magnaporthe grisea
Uploaded: 2018-08-04T13:30:00
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

この作品は、北京農業大学 (YQ201603) の特別な科学研究プロジェクトと北京教育委員会 (KM201610020005) の科学的なプロジェクトによって支持されました。

1 イネいもち病菌の分生胞子懸濁液の噴霧接種法
<ol><li>
M.griseaの真菌培養
	<ol>
<li>菌のオートミール トマト寒天 (OTA) 培養液を準備します。</li>
		<li>オートミールの 30-50 グラムの重量を量る 800 mL の蒸留水または脱イオンされた水 (ddH2O) にこれを追加し、電気ポットで 30 分間混合物を沸騰させます。</li>
		<li>ガーゼの部分をビーカーにゆでオートミール ジ?...

<ol>
	<li>Li, W. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+natural+allele+of+a+transcription+factor+in+rice+confers+broad-spectrum+blast+resistance.">A natural allele of a transcription factor in rice confers broad-spectrum blast resistance.</a> Cell. 170 (1), 114-126 (2017).</li><li>Chi, M. H., Park, S. Y., Kim, S., Lee, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+pathogenicity+gene+is+required+in+the+rice+blast+fungus+to+suppress+the+basal+defenses+of+the+host.">A novel pathogenicity gene is required in the rice blast fungus to suppress the basal defenses of the host.</a> PLoS Pathogens. 5 (4), 1000401 (2009).</li><li>Jia, Y., Valent, B., Lee, F. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+host+responses+to+Magnaporthe+grisea.on+detached+rice+leaves+using+a+spot+inoculation+method.">Determination of host responses to Magnaporthe grisea.on detached rice leaves using a spot inoculation method.</a> Plant Disease. 87 (2), 129-133 (2003).</li><li>Ebbole, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnaporthe+as+a+model+for+understanding+host-pathogen+interactions.">Magnaporthe as a model for understanding host-pathogen interactions.</a> Annual Review of Phytopathology. 45, 437-456 (2007).</li><li>Hamer, J. E., Talbot, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infection-related+development+in+the+rice+blast+fungus+Magnaporthe+grisea.">Infection-related development in the rice blast fungus Magnaporthe grisea.</a> Current Opinion in Microbiology. 1 (6), 693-697 (1998).</li><li>Howard, R. J., Valent, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breaking+and+entering:+host+penetration+by+the+fungal+rice+blast+pathogen+Magnaporthe+grisea.">Breaking and entering: host penetration by the fungal rice blast pathogen Magnaporthe grisea.</a> Annual Review of Microbiology. 50, 491-512 (1996).</li><li>Chen, X. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=N-Glycosylation+of+Effector+Proteins+by+an+&#945;-1,3-+Mannosyltransferase+Is+Required+for+the+Rice+Blast+Fungus+to+Evade+Host+Innate+Immunity.">N-Glycosylation of Effector Proteins by an &#945;-1,3- Mannosyltransferase Is Required for the Rice Blast Fungus to Evade Host Innate Immunity.</a> The Plant Cell. 26 (3), 1360-1376 (2014).</li><li>Zhang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=M.ARG1,+MoARG5,6+and+MoARG7+involved+in+arginine+biosynthesis+are+essential+for+growth,+conidiogenesis,+sexual+reproduction,+and+pathogenicity+in+Magnaporthe+oryzae.">M.ARG1, MoARG5,6 and MoARG7 involved in arginine biosynthesis are essential for growth, conidiogenesis, sexual reproduction, and pathogenicity in Magnaporthe oryzae.</a> Microbiological Research. 180, 11-22 (2015).</li><li>Du, Y. X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+serine/threonine-protein+phosphatase+PP2A+catalytic+subunit+is+essential+for+asexual+development+and+plant+infection+in+Magnaporthe+oryzae.">A serine/threonine-protein phosphatase PP2A catalytic subunit is essential for asexual development and plant infection in Magnaporthe oryzae.</a> Current Genetics. 59 (1-2), 33-41 (2013).</li><li>Yang, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+protein+com1+is+required+for+normal+conidium+morphology+and+full+virulence+in+Magnaporthe+oryzae.">A novel protein com1 is required for normal conidium morphology and full virulence in Magnaporthe oryzae.</a> Molecular Plant-Microbe Interactions. 23 (1), 112-123 (2010).</li><li>Cao, Z. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+ash1-like+protein+MoKMT2H+null+mutant+is+delayed+for+conidium+germination+and+pathogenesis+in+Magnaporthe+oryzae.">An ash1-like protein MoKMT2H null mutant is delayed for conidium germination and pathogenesis in Magnaporthe oryzae.</a> BioMed Research International. 2016, 1575430 (2016).</li><li>Bryan, G. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+single+amino+acid+difference+distinguishes+resistant+and+susceptible+alleles+of+the+rice+blast+resistance+gene+Pi-ta.">A single amino acid difference distinguishes resistant and susceptible alleles of the rice blast resistance gene Pi-ta.</a> The Plant Cell. 12 (11), 2033-2045 (2000).</li><li>Zhou, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plant+pathology:+a+life+and+death+struggle+in+rice+blast+disease.">Plant pathology: a life and death struggle in rice blast disease.</a> Current Biology. 26 (18), 843-845 (2016).</li><li>Guo, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MoGrr1,+a+novel+F-box+protein,+is+involved+in+conidiogenesis+and+cell+wall+integrity+and+is+critical+for+the+full+virulence+of+Magnaporthe+oryzae.">MoGrr1, a novel F-box protein, is involved in conidiogenesis and cell wall integrity and is critical for the full virulence of Magnaporthe oryzae.</a> Applied Microbiology and Biotechnology. 99 (19), 8075-8088 (2015).</li><li>Talbot, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+trail+of+a+cereal+killer:+Exploring+the+biology+of+Magnaporthe+grisea.">On the trail of a cereal killer: Exploring the biology of Magnaporthe grisea.</a> Annual Review of Microbiology. 57, 177-202 (2009).</li><li>Wilson, R. A., Talbot, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Under+pressure:+investigating+the+biology+of+plant+infection+by+Magnaporthe+oryzae.">Under pressure: investigating the biology of plant infection by Magnaporthe oryzae.</a> Nature Reviews Microbiology. 7, 185-195 (2009).</li><li>Jia, Y. L., Lee, F. N., McClung, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+Resistance+Spectra+of+the+Pi-ta+and+Pi-k+Genes+to+U.S.+Races+of+Magnaporthe+oryzae+Causing+Rice+Blast+in+a+Recombinant+Inbred+Line+Population.">Determination of Resistance Spectra of the Pi-ta and Pi-k Genes to U.S. Races of Magnaporthe oryzae Causing Rice Blast in a Recombinant Inbred Line Population.</a> Plant Disease. 93, 639-644 (2009).</li><li>Peng, Y. L., Shishiyama, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporal+sequence+of+cytological+events+in+rice+leaves+infected+with+Pyricularia+oryzae.">Temporal sequence of cytological events in rice leaves infected with Pyricularia oryzae.</a> Canadian Journal of Botany. 66 (4), 730-735 (1988).</li></ol>

植物病抵抗性遺伝子は、真菌病原体1,12を含む病原体による感染症の予防に不可欠な役割を果たします。いもち病は、病原体の人口構造の性質を理解して植物抵抗性遺伝子4を識別するためにモデルとして使用されています。したがって、病抵抗性遺伝子型と連続的に栽培することができます病気に強い植物を識別するために大規模...

イネいもち病菌によるイネいもち病は、水稲品種世界中1,2のための最も深刻な病気のひとつです。イネいもち病菌感染宿主植物プロセス分生胞子生産と付着、分生胞子の発芽および付着器形成、形成穿と感染菌糸の分化は、病気を広める3。 これらすべての段階が多く他植物病原糸状菌、一般的で、確かに、任意の 1 つのステージの封鎖ホスト植物の感染が防止します。経済的重要性、遺伝的少ないのためイネいもち病菌は、植物病原菌の相互作用1,4のメカニズムを研究するためのモデル生物として浮上しています。したがって、スクリーニングとデザイン小説候補遺伝子の同定と真菌病原性の分子機構の解明を助けるこれら発達段階のイネいもち病菌の分子基盤を勉強殺菌剤5。
イネいもち病菌感染に関する最近の報告は前侵入の段階、特に、制御、付着器形成、浸透ペグ、伝染の成長3,の分子機構に焦点を当てています。6。 したがって、イネいもち病菌感染をテストする詳細なプロトコルを開発するが不可欠です。本明細書で詳細な胞子懸濁液とイネいもち病菌菌糸のプラグに傷接種スプレーを介した感染アッセイを利用して感染テスト方法を提案する.本報告では、プロトコルは、噴霧のため制御する溶液の調製と植物イネいもち病菌の菌糸のプラグを介した接種菌株のうち、文化に焦点を当てください。詳細は以下で、メソッドの全体のワークフローを示した概略図でこれらの手順を説明し、典型的な病変は、それぞれ図 1 2に示すように。

<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

植物病原性菌による潜在的な脅威から身を守るための強力なシステムを所有しています。農学上重要な植物は、ただし、このような病原体と戦う対策の現状はあまりにも保守的な証明している、したがって、十分に効果的なと潜在的環境は危険が伴います。したがって、それは非常に制御する病害抵抗性遺伝資源の識別、分離抵抗性遺伝子の解析と分子育種を通じて自然を支援する宿主抵抗性要因を識別するために必要です抵抗力がある品種。この点で、繁殖植物抵抗性遺伝子を開発し正確・迅速かつ大規模な接種方法を確立する必要があります。米は、真菌病原体イネいもち病菌原因重症症状を爆破し、損失をもたらします。最近では、イネいもち病菌は、植物病原菌の相互作用のメカニズムを研究するためのモデル生物として浮上しています。したがって、私たちはイネいもち病菌の特定は、植物病原性試験法の開発を報告します。このメソッドは、胞子懸濁液を噴霧接種法と負傷接種キューブ菌糸または胞子懸濁液の液滴の両方を提供します。戸建イネ葉の負傷の接種法の重要なステップは、ホスト貫入抵抗による任意の干渉を回避する植物の葉で傷をすることです。このプロトコルがスプレー/負傷したイネいもち病菌菌株の生じの迅速、正確、かつ大規模なスクリーニングに貢献します。これは統合し、体系的な植物感染症法は植物病理学の問題の広い視野を得るための優れた出発点となります。

技術全体のワークフローは、図 1に示すです。14 日齢の影響を受けやすいイネ幼植物の植物感染アッセイを行った (イネcv CO 39) 7 日齢の影響を受けやすいオオムギ葉または (H. オカダンゴムシcv 黄金の約束)7,8,9。イネ葉の感染症のためにテストするため Com1 削除変...

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

植物感染試験: スプレーと傷を介した植物病原体イネいもち病菌接種

ここでは、提案する植物病原菌と植物病原性をテストするためのプロトコルいもち病。このレポートは菌分離株の pathotype の大規模スクリーニングに貢献し、分子育種中植物の耐性メカニズムを理解するための優れた出発点として役立ちます。

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

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