이 작품은 베이징 농업 대학 (YQ201603)의 특별 한 과학 연구 프로젝트와 베이징 교육 위원회 (KM201610020005)의 과학 프로젝트에 의해 지원 되었다.

1. 스프레이 접종 M. grisea Conidia의 서 스 펜 션
<ol><li>
M.grisea 에 대 한 곰 팡이 문화
	<ol>
<li>곰 팡이 긴장에 대 한 오트밀 토마토 agar (OTA) 문화 매체를 준비 합니다.</li>
		<li>오트밀의 30-50 g를 무게, 소 주/이온된 수 (ddH2O)의 800 mL에 이것을 추가 하 고 전기 냄비에 30 분을 위한 혼합물을 비등.</li>
		<li>거 즈 조각을 통해 비 커에 끓인된 오트밀 주스를 필터링 합니다.</li>
		<li>토마토 주스와 한 천 20g 150 mL 비 커에 여과 액을 더하고 ddH2O 최대 1000 mL.</li>
	</ol>
</li>
	<li>
실험 자료의 준비
	<ol>
<li>3 d ddH2O에서에서 벼 (Oryza sativa) 품종 Lijiangxintuan-heigu (LTH)의 약 50 씨앗을 담가 또는 보 리의 약 50 씨앗을 담가 (Hordeum vulgare 이력서 골든 약속) ddH2오 약 1 d.</li>
		<li>촉촉한 거 즈에 쌀 이나 보 리의 씨앗을 포장 하 고 직경 10 cm x 10 cm에서 28 ° c.의 약 30 접시에 축축한 필터 종이에 그들을 발 아 쌀 종자 발 아에 약 2-3 d, 보 리 씨앗 발 아 시간이 약 2 d. 상대 습도 온실에서 대략 70% 이다.</li>
		<li>압력가 화분 용 흙 및 급수를 사용 하 여 냄비에 쌀 이나 보 리의 묘를 설치 하 고 질 석의 계층으로 그들을 커버.</li>
		<li>1 약 25 ° C에서 캐비닛 적합 한 유리 공장 또는 성장에 식물을 배치 ~ 2 주.</li>
		<li>원형에서 필터 종이의 3 층 (각 동그라미는 8 cm 직경 있어야 함) 무 균 수술가 위로 잘라 100mm 살 균 플라스틱 접시에 그들을 배치.</li>
		<li>필터 종이를 각 접시 ddH2O를 추가 합니다.</li>
		<li>필터 종이 완전히 젖은 것을 확인 하지만 여분의 물 추가.</li>
		<li>진공 펌프를 가진 어떤 과잉의 물을 제거 합니다.</li>
		<li>쌀/보 리를 지원 하기 위해 문화 접시에 장소 2 살 균 이쑤시개 잎; 이쑤시개는 공간 ~ 2-3 cm 떨어져.</li>
		<li>씨앗을 파 종 후 2 주 쌀의 나뭇잎을 수집 하거나 보 리 7 d의 잎 씨앗을 파 종 후.</li>
		<li>쌀/보 리의 묘 목 4-6 잎을 사용 하 여, 정상에서 ~ 5 센티미터에 줄기의 아래 부분을 잘라 하 고 나뭇잎을 수집 합니다.</li>
	</ol>
</li>
	<li>
스프레이 접종 프로토콜
	<ol>
<li>문화 ~ 4 d 서 모스 탯 인큐베이터 (25 ° C)에서 OTA 접시에 곰 팡이 부담.</li>
		<li>DdH2O 0.5-5 mL 피 펫 각 4-하루-오래 된 접시를 사용 하 여 ~ 2 mL를 추가 합니다.</li>
		<li>접종 루프와 사체 잔해에 M. grisea 야생-타입 긴장과 돌연변이 스트레인의 사체를 다쳤어요.</li>
		<li>사체 잔해를 수집 하 고 새로운 OTA 접시에 그것을 전송. 받아 사체 파편 타격 건조 깨끗 한 벤치에.</li>
		<li>24-48 시간 (10 h) 밤에 25 ° C (14 h) 하루에 23 ° C에서 온실에서 conidia의 성장에 필요한 습도 되도록 거 즈의 3 레이어 플레이트 커버.</li>
		<li>각 접시 ddH2O의 2 개 mL를 추가 하 고 뒤에 렌즈 종이의 2 층을 통해 여과 멸 균 면봉으로 부드럽게는 conidia를 다쳤어요. 문화 매체의 표면을 긁을 조심 해야.</li>
		<li>100-1000 µ L 피펫으로 새로운 50 mL 튜브에 conidia 현 탁 액을 전송 합니다.</li>
		<li>25 ° c.에 x g 5000의 최소한 5 분 동안 원심 분리기</li>
		<li>제거는 상쾌한 고 mL 당 2 × 104 conidia 0.025% (v/v) 트윈 20 솔루션에에서 주려고 펠 릿 resuspend. 트윈-20 솔루션은 일반적으로 약 10-20 mL.</li>
		<li>휴대용 분무기에 포자 현 탁 액을 붓는 다.</li>
		<li>Spay 2 주 된 쌀 묘 또는 보 리 쌀 잎에 conidial 서 스 펜 션의 약 10 mL 보 리 7-하루-오래 된 묘 목의 잎 하 고 0.025% (v/v) 트윈 20 솔루션 컨트롤 식물 25 ° c ~ 24 h. 스프레이 대 한 어두운, 습 한 약 실에서 그들을 품 어.</li>
		<li>12 h photoperiod 잎 25 ° C에서 형광등 불빛 아래에서 또 다른 습 챔버로 전송.</li>
		<li>접종 후 5 d에 질병 증상을 기록 합니다. 길이 ~ 6 cm의 병 쌀/보 리 잎을 검사 합니다. 표준 평가 국제 쌀 연구 연구소 (IRRI)의 폭발 저항에 대 한 채 점 시스템에 따라 이다. 폭발 저항에 대 한 채 점 시스템의 세부 정보는 표 1에 나와 있습니다.</li>
		<li>사진 테스트 긴장의 감염을 평가 하는 나뭇잎. 감염 병 변 3.6 c m2당 수로 평가 했다.</li>
	</ol>
</li>
</ol>2. 접종 균 큐브 또는 방울 Conidial 정지의 M. grisea 부상
<ol><li>분리 된 쌀/보 리 잎;의 주요 혈관에 3 2-3 cm 긴 상처를 긁어 해부학 바늘을 사용 하 여, 알아서 하지 잎에 침투.</li>
	<li>이쑤시개에 긁힌 것된 잎을 넣고 0.02% (v/v) 트윈 20 솔루션 방울의 층을 형성 하는 잎에 스프레이.</li>
	<li>OTA 접시에서 각 M. grisea 스트레인 (야생-타입, 돌연변이, 보완 긴장, 또는 다른 테스트 긴장)에 대 한 0.5 c m x 0.5 c m 바로 플러그를 잘라.</li>
	<li>바로 플러그를 넣어 또는 부상에 conidial 서 스 펜 션의 25 µ L 방울 단풍 잎 3-8 d의 습기 챔버에 25 ° c를 품 어.</li>
	<li>5-7 d 후 접종에 장애를 검사 합니다. 시험 방법은 같은 스프레이 접종 방법으로 (단계 1.3.13 참조).</li>
	<li>길이 ~ 6 cm의 병 쌀/보 리 잎을 검토 하 고 테스트 긴장의 감염을 평가 하기 위해 사진을. 감염 병 변 3.6 c m2당 수로 평가 했다.</li>
</ol>

<ol>
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저자는 공개 없다.

식물 질병 저항 유전자 곰 팡이 병원 균1,12를 포함 하 여, 병원 균에 의해 감염 방지에 필수적인 역할을 재생 합니다. 쌀 폭발 병원 체 인구 구조의 특성을 이해 하 고 식물 저항 유전자4를 식별 하는 모델로 사용 되었습니다. 따라서, 그것은 질병 저항 유전자를 지속적으로 재배 될 수 있는 질병 저항 식물을 식별 하는 농업 식물의 주요 ...

쌀 폭발, M. grisea를 기인한 쌀 품종 전세계1,2에 대 한 가장 심각한 질병 중 하나입니다. 프로세스는 M. grisea 감염 호스트 식물 conidia 생산 및 표면 부착, conidia 발 아 및 appressorium 형성, 침투 못 감염 hypha 차별화의 형성을 포함 하 고 질병 확산 3. 이러한 모든 단계는 많은 다른 식물 병원 성 곰 팡이에서 일반적이 고, 실제로, 모든 단일 단계 봉쇄 호스트 식물의 감염을 방지. 때문에 그것의 경제 중요성 및 유전 추적성, M. grisea 식물 곰 팡이 병원 체 상호 작용1,4의 메커니즘을 공부에 대 한 모델 생물으로 떠오르고 있다. 따라서, 기본 균 pathogenicity 및 심사 및 디자인 소설 후보 대상 유전자의 식별 분자 메커니즘을 명료 하 도움이 될 것입니다 이러한 발달 단계 M. grisea 에 분자 기초 공부 살 균 제5.
최근 보고서 M. grisea 감염에 관한 사전 침투 단계, 특히에서 conidiation, appressorium 형성, 침투 못 및 전염 성 성장3, 의 분자 메커니즘에 초점을 맞춘합니다 6. 따라서, M. grisea 감염 테스트 상세한 프로토콜을 개발 하는 것이 필수적입니다. 여기, 우리 conidial 서 스 펜 션와 M. grisea의 사체 플러그와 상처의 접종 감염 스프레이 중재 분석을 활용 한 감염 테스트에 대 한 자세한 방법 제시. 이 보고서는 프로토콜, conidiation 솔루션의 준비와 바로 플러그 중재 접종 M. grisea와 식물의 종자, 문화에 중점을 둡니다. 다음이 단계는 아래의 세부 사항 그리고 방법의 전체 워크플로 보여주는 회로도 보기에 설명 하 고 전형적인 병 변은 각각 그림 1 과 2에 표시 됩니다.

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			<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>
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		<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>
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		<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

식물 병원 성 곰 팡이 의해 잠재적인 위협 으로부터 방어 하는 강력한 시스템을가지고 있습니다. 그러나 농업으로 중요 한 식물에 대 한, 현재 조치 같은 병원 체에 대처 하기 위해 너무 보수적인 증명 그리고, 따라서, 충분 하지 효과적인, 그리고 그들은 수 있습니다 잠재적으로 포즈 환경 위험. 따라서, 그것은 매우 내성 germplasm의 식별, 격리 및 저항 유전자의 특성 및 분자 사육을 통해 자연스럽 게 제어 식물 질병을 호스트 저항 요소를 식별 하는 데 필요한 저항 하는 재배 품 정. 이와 관련, 번 식 하 고 식물 저항 유전자를 개발, 신속, 정확 하 고 대규모 접종 메서드를 설정할 필요가 있다. 쌀 버섯 모양 병원 체 Magnaporthe grisea 원인 심각한 질병 증상을 폭발 하 고 손실을 항복. 최근에, M. grisea 식물 곰 팡이 병원 체 상호 작용의 메커니즘을 공부에 대 한 모델 생물으로 떠오르고 있다. 따라서, 우리 보고 M. grisea위해 특정 식물 독성 시험 방법의 개발. 이 메서드는 conidial 현 탁 액을 분무 접종 및 균 사체 큐브 또는 conidial 서 스 펜 션의 방울 부상 접종에 대 한 제공합니다. 분리 된 쌀 잎 부상 접종 방법의 주요 단계는 호스트 침투 저항으로 인 한 간섭 방지 식물 잎에 상처를 만드는 것입니다. 이 M. grisea 격리의 pathotypes의, 정확, 신속 하 고 대규모 심사에 기여 스프레이/부상 프로토콜. 이 통합 하 고 체계적인 식물 감염 방법 식물 병리학에서 문제의 광범위 한 관점을 확보에 대 한 훌륭한 출발점 될 것입니다.

기술에 대 한 전체 워크플로 그림 1에 표시 됩니다. 14 일 된 취약 밥 묘 종에 식물 감염 분석 수행 (O. sativa 이력서 공동-39) 취약 7-하루-오래 된 보 리 잎 또는 (H. vulgare 이력서 골든 약속)7,,89. 쌀 잎에 감염에 대 한 테스트, M. grisea 야생-타입 변형과 P131 Com1 삭제 ?...

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

식물 감염 테스트: 스프레이 식물 병원 체 Magnaporthe Grisea 와 상처-중재 접종

여기, 우리 식물 병원 체와 식물 독성 테스트 프로토콜을 제시 Magnaporthe grisea. 이 보고서 균 분리의 pathotypes의 대규모 검열을 하 고 분자 번 식 하는 동안 식물의 저항 메커니즘을 이해 하기 위한 훌륭한 시작 지점으로 제공.

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 조회수.  Beijing University of Agriculture. 여기, 우리 식물 병원 체와 식물 독성 테스트 프로토콜을 제시 Magnaporthe grisea. 이 보고서 균 분리의 pathotypes의 대규모 검열을 하 고 분자 번 식 하는 동안 식물의 저항 메커니즘을 이해 하기 위한 훌륭한 시작 지점으로 제공.

비디오: 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.

에 십자가를 수행하는 방법<em&gt; 강아지풀</em목초에 대한&gt;, 새로운 모델 시스템

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 ...

연구

환경과학

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.

옥수수와 테오신테 라인에 우스틸라고 성 실증의 병원성을 평가하기위한 신속하고 효율적인 방법

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.

유괴와 콩과 식물의 성장을위한 단일 공장, 무균 소우주<em&gt; 개자리 truncatula</em&gt; Rhizobi​​al 공생과<em&gt; Sinorhizobi​​um meliloti</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.

바이오 기반 산업을위한 원료로 담배 나무와 포플러의 평가에 대한 성적 증명서 및 대사 산물 프로파일

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.

두 발견 거미 진드기의 적용<em&gt; 점박이 응애</em&gt; 식물 해충의 상호 작용 연구

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).

식물의 얼음 핵 얼음 전파의 연구에 대한 고해상도 적외선 체열 (HRIT)의 사용

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.

체관부 체 요소에 전기 신호를 획득 및 측정을위한 전기 침투 그래프 (EPG)의 새로운 응용 프로그램

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.

25 KW회 칼슘 반복 파일럿-식물은 Calciner에서 높은 산소 농도와 운영

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.

공장 실험실에서 측정 하는 표면에 박테리아의 부착

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.

특정 및 감귤 류 녹화 병원 체 Candidatus liberibacter 종 감귤 잎 조직 샘플에서 사용 하 여 기존의 PCR의 정확한 검출

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

식물 감염 테스트: 스프레이 식물 병원 체 Magnaporthe Grisea 와 상처-중재 접종

요약

더 많은 비디오 탐색

이 비디오의 챕터

에 십자가를 수행하는 방법

옥수수와 테오신테 라인에 우스틸라고 성 실증의 병원성을 평가하기위한 신속하고 효율적인 방법

유괴와 콩과 식물의 성장을위한 단일 공장, 무균 소우주

바이오 기반 산업을위한 원료로 담배 나무와 포플러의 평가에 대한 성적 증명서 및 대사 산물 프로파일

두 발견 거미 진드기의 적용

식물의 얼음 핵 얼음 전파의 연구에 대한 고해상도 적외선 체열 (HRIT)의 사용

체관부 체 요소에 전기 신호를 획득 및 측정을위한 전기 침투 그래프 (EPG)의 새로운 응용 프로그램

25 KW_회 칼슘 반복 파일럿-식물은 Calciner에서 높은 산소 농도와 운영

공장 실험실에서 측정 하는 표면에 박테리아의 부착

특정 및 감귤 류 녹화 병원 체 Candidatus liberibacter 종 감귤 잎 조직 샘플에서 사용 하 여 기존의 PCR의 정확한 검출