이것은 생산 및 실험 작업은 여기에 친절 리나 아귀레, 길버토 코르도바, 윌리엄 메라, Ximena Bonilla, 그리고 다리오 Viveros 지원 레이 날도 Pareja의 헌신적이고 열정적인 도움을 반영 제시. 우리는 또한 우리가 비디오와 원고를 향상 도움이 덧글에 대한 존 마일즈 세 익명 리뷰어 감사합니다.

1. 식물 <ol><li> 성숙한 식물의 단일 줄기 잘라는 실험 단위로 사용됩니다. 잘라는 심는 재료의 균일을 위해 10cm로 손질합니다. </li><li> 식물 병원균에 의해 오염을 방지하기 위해, 잘라 2 분 3 % 나트륨 차아 염소 산염의 용액에 씻은 후 수돗물로 씻어서 thoughroughly 있습니다. </li><li> 각각의 절단은 CA에 심어져 있습니다. 원통형 PVC 튜브에 무균 토양 36g는 (6.5 cm 높이의 직경 외부 5.3 ㎝) 스티로폼 컵에있는 낮은 끝에 봉인 및 PVC 부싱로 덮인. 절단은 부시의 중심 오프닝에 배치 거품 링에 의해 장소에서 개최됩니다. </li><li> 적절한 토양 수분을 유지하기 위해 필요에 따라 식물은 수정된하고 물결 무늬가있는 있습니다. </li><li> 식물은 뿌리 - 목부 수유의 spittlebug의 nymphs을위한 사이트를 먹이 역할을 외면 뿌리의 개발을 허용, 곤충에 의해 도전되기 전에 한 달 동안 성장을합니다. </li><li> 뿌리 잘라와 PVC 튜브는 부분적으로 루트 아키텍처 고유의 차이에 대한 조정, 더 이상 외면 뿌리의 성장을 촉진하기 전에 침입 8 D에 대한 반전됩니다. 이 단계는 우리가 24헥타르 일을 제공 빛의 인공​​ 소스를 필요로합니다. 이 역전의 치료 실험에서 제거 후 herbivore 저항과 혼란함을 주죠 herbivory에서 탈출을 피하기 위해 각각의 공장은 거의 또는 전혀 외면 뿌리로 복제합니다. 이 제거는 그들의 유전자형의 철폐로 이어지는 결코 드문 있어야합니다. </li></ol> 2. 곤충 <ol><li> 어른 spittlebugs는 현장에서 수집한 종을 발견하고, 그들에 대한 피드에 감염될 brachiariagrasses의 단풍을 제공 산란 연습장에 소개합니다. 여자들은 토양에 알을은 새장의 바닥에 놓인 얕은 이동식 트레이에 포함되어 있다고 믿는다. </li><li> 며칠 후, 흙은 물에 중단되고 계란을 수집 sieves 일련의 (42, 60 및 150 메쉬) 통과. 30 % 생리 식염수에 sieved 자료를 중지하면 미숙 계란 및 잔류 유기물 뒤에두고, 성숙한 난자가 부동하게됩니다. </li><li> 곤충 병원체의 오염을 방지하기 위해, 계란은 5 분 0.5 % 나트륨 차아 염소 산염의 용액에 소독 후 증류수로 씻어서 있습니다. </li><li> 각각의 시험 공장 (실험 장치) 6 성숙 난자로 토양 표면에 감염됩니다. 부화에 가까운만이 성숙한 난자가 infestations 위해 선택됩니다. 곤충의 복부에 대응하는 자신의 사후 부분에 핑크색 표시,,, 그리고 완벽하게 확장 operculum 그들은 두 개의 빨간 곤충의 눈을에 대응하는 자신의 앞쪽에 부분 점의 존재에 의해 확인할 수 있습니다. </li><li> 성공적인 부화은 어떤 안깐 계란은 우리의 식민지에서 neonate spittlebug으로 대체되는 시점에서 4 D 나중에 확인됩니다. </li><li> 테스트 식물은 유전자형 최대 6 복제와 함께 무작위로 전체 블록 설계에 정렬됩니다. </li></ol> 3. 평가 <ol><li> 실험은 CA를 평가합니다. 침입 후 30 D (정확한 시간은 공부 벌레 종류에 따라 다릅니다.) </li><li> 피해 1 눈에 띄는 손상과 죽은 식물 (그림 1) 5에 해당에 해당하는 1-5 시각적 척도에 점수입니다. 식물의 피해를 채점하는 것은 내성을 예측할 수 있습니다. </li></ol><img src="/files/ftp_upload/3047/3047fig1.jpg" alt="그림 1" /> Brachiaria 종의 spittlebug herbivory에 내성을 평가하기 위해 사용 그림 1. 피해 규모. 1 = 없음 감지 손상, 2 = 0~25% 괴사성 단풍, 3 = 25-50% 괴사성 단풍, 4 = 50~75% 괴사성 단풍, 5 = 75-100% 괴사성 단풍. <ol start="3"><li> 곤충 생존들은 최종 instar 또는 성인 무대 (그림 2) 중 하나를 도달하는 곤충을 계산하여 평가합니다. 곤충 생존을위한 채점하는 것은 항생의 예측을 제공합니다. </li></ol><img src="/files/ftp_upload/3047/3047fig2.jpg" alt="그림 2" /> 첫 instar (왼쪽)에서 성인 (오른쪽)에 그림 2. Spittlebug 개발. 평가 날짜 적어도 최종 instar을 (화살표)에 도달만이 벌레가로 계산됩니다 &quot;생존자.&quot; 4. 대표 결과 spittlebug Aeneolamia reducta에 대한 저항을위한 대표적인 분석 심사 Brachiaria의 하이브리드의 결과는 그림 3에 제공됩니다. <img src="/files/ftp_upload/3047/3047fig3.jpg" alt="그림 3" /> 그림 3. spittlebug Aeneolamia reducta에 저항 Brachiaria의 하이브리드를 검사의 결과. herbivory 자신의 응답이 음모의 절반 아랫부분에 일인 경우 우리는 식물이 저항하는 것이 좋습니다. 오른쪽에서 왼쪽 및 관용 위에서 아래로 증가 항생 증가. 화살표 우리 내성 검사에서 가리 킵니다.

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관심 없음 충돌 선언하지 않습니다.

herbivore 저항 메커니즘 간의 차별하는 것은 작물 번식 노력 3 밝게하실 수 있습니다. 강력한 항생에 따라 저항이 어떤 상황에서보다 적극적인 해충 biotypes에 대한 선택 압력을 만들 수 있습니다. 한편, 관용에 따라 저항은 오차가 결국 압도 때까지 해충 인구가 증가 허용할 수 있습니다. 튼튼한 저항 번식, 따라서 어떤 저항 메커니즘은 특정 작물 / 해충 시스템에 대한 대부분의 안정성을 부여 것입니다 [케네디 외 볼의 신중하게 고려해야합니다. (1987) 주제] 3 종합 토론. 이 비디오 데모는 brachiariagrasses 6-9에서 spittlebug 저항하는 항생과 관용의 상대적인 기여도를 평가하기위한 몇 가지 이전의 연구에 빌드. 선택 시험 또는 현장 실험과 대조적으로, 노 - 선택 시험 따라서 herbivore 행동 (예, 호스트 환경 설정, 집합)에서 발생하는 차이에 대한 제어를 제공하는 모든 식물이 같은 herbivore 압력을 받게 보장합니다. 이러한 이유로, 그러나, - 선택 시험은 antixenosis에 의해 저항을 평가하기위한 적합하지 않습니다. 인위적으로 곤충 식민지를 양육하는 것은 저항 심사 assays 10 테스트 곤충을 얻기 위해 선호하는 방법입니다. 식민지은 연중 11 시간을 실시하는 실험 알려진 시대의 spittlebugs의 신뢰할 수와 유니폼 소스를 제공합니다. 그것은, 그러나, 정기적으로 식민지 관련 현장 인구 10 유전 이탈하지 않도록하기 위해 식민지에 야생 개인 달이다해야 할 수도 있습니다. 이러한 인위적 - reared herbivores의 적합성을 평가하고, 시간이 지남에 따라 자신의 공격성에 가능한 변화를 모니터링하기 위해서는 모든 검사는 알려진 저항 수준의 적절한 검사를 포함하는 것이 중요합니다. 여러 개의 매개 변수는 신중하게 적절한 저항 심사 분석을 설계 정의해야합니다. 우리가 테스트를 설계하는 생각이 몇 가지 주요 매개 변수는 호스트 식물의 나이, 종 및 spittlebugs의 발달 단계, 침입 수준 및 식물 곤충 담당자 8,9,12 기간을 포함합니다. 실험 및 프로토콜 세련의 여러 반올림은 적절하게 현장에서 herbivore 저항을 예측 신속하고 비용 효율, 신뢰성, 분석에 도달하는 데 필요한 수 있습니다.

characterizing herbivore resistance mechanisms: spittlebugs on brachiaria spp. as an example

antixenosis, 항생 및 관용 1 : 식물 세 확장 메커니즘을 통해 herbivore 손상에 저항하실 수 있습니다. Antixenosis은 herbivore 다른 식물에게 2를 선택할 수있을 때 발전소를 피할 수있다 정도입니다. 항생은 식물이 1 먹이 herbivore의 적합성에 영향을 미치는 정도입니다. 공차가 공장이 herbivore의 성장과 재생산 1 손상시키지 않고 견딜 또는 herbivore으로 인한 손상을 복구할 수있는 정도입니다. 농업 환경에서 herbivore 저항의 내구성은 농작물 번식 노력 셋 중에 선호하는 저항 메커니즘에 큰 정도에 따라 달라집니다. 우리는 Brachiaria 종에 spittlebug 저항하는 항생과 관용의 상대적인 기여도를 추정하기위한 노 선택 실험을 보여줍니다. 속 Brachiaria의 아프리카 목초의 여러 종의 Neotropics에 귀중한 마초와 파스투레 식물, 그러나 그들이 심각 spittlebugs 여러 토착 종 (Hemiptera : Cercopidae)에 의해 도전을하실 수 있습니다. spittlebugs들의 저항을 평가 4, 식물 vegetatively - 전달됩니다 줄기 잘라 의해 및 spittlebugs는 피드 수있는 피상적인 뿌리의 성장을 허용, 약 한 달 동안 성장을 수있었습니다. 그 시점에서, 각 시험 공장은 개별적으로 부화 근처 여섯 spittlebug 달걀과 도전이다. Infestations는 공장 손상과 곤충의 생존을 평가하기 전에 한 달 동안 진행하기 위해 사용할 수 있습니다. 곤충의 생존을 득점하는 것은 항생의 추정치를 제공하면서 설비의 손상을 득점하는 것은 내성을 예측할 수 있습니다. 이 프로토콜은 상업 brachiariagrases 5 spittlebug 저항을 강화하기 위해 식물 번식 목표를 촉진하고 있습니다.

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Characterizing Herbivore Resistance Mechanisms: Spittlebugs on Brachiaria spp. as an Example

이 동영상은 herbivory에 호스트 식물 저항의 메커니즘을 설명 및 저항을 spittlebug하기 위해 항생과 관용의 상대적인 기여도를 추정 노 선택 테스트를 보여줍니다<em&gt; Brachiaria</em&gt; 종.

특성화 Herbivore 저항 메커니즘 : Spittlebugs에 Brachiaria 종. 예로

Plants can resist herbivore damage through three broad mechanisms: antixenosis, antibiosis and tolerance1. Antixenosis is the degree to which the plant is ...

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Characterizing Herbivore Resistance Mechanisms: Spittlebugs on Brachiaria spp. as an Example

13.7K 조회수.  CIAT. 이 동영상은 herbivory에 호스트 식물 저항의 메커니즘을 설명 및 저항을 spittlebug하기 위해 항생과 관용의 상대적인 기여도를 추정 노 선택 테스트를 보여줍니다 Brachiaria 종.

비디오: Characterizing Herbivore Resistance Mechanisms: Spittlebugs on Brachiaria spp. as an Example

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 ...

연구

환경과학

Choice and No-Choice Assays for Testing the Resistance of A. thaliana to Chewing Insects

Larvae of the small white cabbage butterfly are a pest in agricultural settings. This caterpillar species feeds from plants in the cabbage family, which include many crops such as cabbage, broccoli, Brussel sprouts etc. Rearing of the insects takes place on cabbage plants in the greenhouse. At least two cages are needed for the rearing of Pieris rapae. One for the larvae and the other to contain the adults, the butterflies. In order to investigate the role of plant hormones and toxic plant chemicals in resistance to this insect pest, we demonstrate two experiments. First, determination of the role of jasmonic acid (JA - a plant hormone often indicated in resistance to insects) in resistance to the chewing insect Pieris rapae. Caterpillar growth can be compared on wild-type and mutant plants impaired in production of JA. This experiment is considered "No Choice", because larvae are forced to subsist on a single plant which synthesizes or is deficient in JA. Second, we demonstrate an experiment that investigates the role of glucosinolates, which are used as oviposition (egg-laying) signals. Here, we use WT and mutant Arabidopsis impaired in glucosinolate production in a "Choice" experiment in which female butterflies are allowed to choose to lay their eggs on plants of either genotype. This video demonstrates the experimental setup for both assays as well as representative results.

Plant resistance to chewing insect herbivores can be tested in several ways. Here, we demonstrate how to set-up a choice and a no-choice experiment with the model plant Arabidopsis thaliana to identify resistance against the pest species Pieris rapae.

씹는 곤충에 A.의 thaliana의 저항을 테스트하기 위해 선택과 노 - 선택 Assays

Larvae of the small white cabbage butterfly are a pest in agricultural settings. This caterpillar species feeds from plants in the cabbage family, which ...

생물학

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.

Endophytes로 곰팡이 Entomopathogens 구축 : Endophytic 생물학적 방제 향해

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

생체공학

Harvesting Venom Toxins from Assassin Bugs and Other Heteropteran Insects

Heteropteran insects such as assassin bugs (Reduviidae) and giant water bugs (Belostomatidae) descended from a common predaceous and venomous ancestor, and the majority of extant heteropterans retain this trophic strategy. Some heteropterans have transitioned to feeding on vertebrate blood (such as the kissing bugs, Triatominae; and bed bugs, Cimicidae) while others have reverted to feeding on plants (most Pentatomomorpha). However, with the exception of saliva used by kissing bugs to facilitate blood-feeding, little is known about heteropteran venoms compared to the venoms of spiders, scorpions and snakes.
One obstacle to the characterization of heteropteran venom toxins is the structure and function of the venom/labial glands, which are both morphologically complex and perform multiple biological roles (defense, prey capture, and extra-oral digestion). In this article, we describe three methods we have successfully used to collect heteropteran venoms. First, we present electrostimulation as a convenient way to collect venom that is often lethal when injected into prey animals, and which obviates contamination by glandular tissue. Second, we show that gentle harassment of animals is sufficient to produce venom extrusion from the proboscis and/or venom spitting in some groups of heteropterans. Third, we describe methods to harvest venom toxins by dissection of anaesthetized animals to obtain the venom glands. This method is complementary to other methods, as it may allow harvesting of toxins from taxa in which electrostimulation and harassment are ineffective. These protocols will enable researchers to harvest toxins from heteropteran insects for structure-function characterization and possible applications in medicine and agriculture.

Although many insects in the suborder Heteroptera (Insecta: Hemiptera) are venomous, their venom composition and the functions of their venom toxins are mostly unknown. This protocol describes methods to harvest heteropteran venoms for further characterization, using electrostimulation, harassment, and gland dissection.

독 독 소 암살자 버그와 다른 Heteropteran 곤충에서 수확

Heteropteran insects such as assassin bugs (Reduviidae) and giant water bugs (Belostomatidae) descended from a common predaceous and venomous ancestor, ...

Detached Leaf Assays to Simplify Gene Expression Studies in Potato During Infestation by Chewing Insect Manduca sexta

The multitrophic nature of gene expression studies of insect herbivory demands large numbers of biological replicates, creating the need for simpler, more streamlined herbivory protocols. Perturbations of chewing insects are usually studied in whole plant systems. While this whole organism strategy is popular, it is not necessary if similar observations can be replicated in a single detached leaf. The assumption is that basic elements required for signal transduction are present within the leaf itself. In the case of early events in signal transduction, cells need only to receive the signal from the perturbation and transmit that signal to neighboring cells which are assayed for gene expression.
The proposed method simply changes the timing of the detachment. In whole plant experiments, larvae are confined to a single leaf which is eventually detached from the plant and assayed for gene expression. If the order of excision is reversed, from last in whole plant studies, to first in the detached study, the feeding experiment is simplified.
Solanum tuberosum var. Kennebec is propagated by nodal transfer in a simple tissue culture medium and transferred to soil for further growth if desired. Leaves are excised from the parent plant and relocated to Petri dishes where the feeding assay is conducted with the larval stages of M. sexta. Damaged leaf tissue is assayed for the expression of relatively early events in signal transduction. Gene expression analysis identified infestation-specific Cys2-His2 (C2H2) transcription factors, confirming the success of using detached leaves in early response studies. The method is easier to perform than whole plant infestations and uses less space.

The presented method creates natural herbivore damaged plant tissue through the application of Manduca sexta larvae to detached leaves of potato. The plant tissue is assayed for expression of six transcription factor homologs involved in early responses to insect herbivory.

분리 된 잎 어설아는 곤충 만두카 sexta를 츄잉에 의해 감염 하는 동안 감자에 유전자 발현 연구를 단순화 하기 위해

The multitrophic nature of gene expression studies of insect herbivory demands large numbers of biological replicates, creating the need for simpler, more ...

Preference Assessment of White-backed Planthopper Feeding on Rice

Exploiting insect-resistant rice germplasm resources and related genes is the primary need for breeding insect-resistant varieties, but the accuracy of the identification of insect-resistant phenotypes of rice is a major difficulty. It is urgent to develop a new method or improve existing methods to screen rice for insect resistance. This article describes a simple and feasible method to assess nonpreference-type resistance of rice to the white-backed planthopper (WBPH), Sogatella furcifera, in the laboratory. The preference of adult WBPHs feeding or inhabiting on maturing rice plants is continuously analyzed by pairwise comparison. The dynamic changes of WBPHs on rice plants are recorded and compared as an index of resistance identification. The current method is simply operable and easily observable and has a short cycle. The use of this method could be extended to investigate the feeding and oviposition preference of similar hemipterans, such as the brown planthopper (BPH), Nilaparvata lugens(St&#229;l).

This study presents a simple and feasible method to assess the nonpreference resistance to white-backed planthoppers who are feeding on rice under laboratory conditions. Improvement of the strategies and makeup of the current method of identification of the resistance to white-backed and brown planthoppers are discussed.

Exploiting insect-resistant rice germplasm resources and related genes is the primary need for breeding insect-resistant varieties, but the accuracy of ...

A Visual Guide for Studying Behavioral Defenses to Pathogen Attacks in Leaf-Cutting Ants

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies have described the behaviors connected with disease defense, and the associated use of chemicals and antimicrobials, no common visual reference has been made. The main aim of this study was to record short clips of behaviors involved in disease defense, both prophylactically and directly targeted towards an antagonist of the colony following infection. To do so we used an infection experiment, with sub-colonies of the leaf-cutting ant species Acromyrmex echinatior, and the most significant known pathogenic threat to the ants&#39; fungal crop (Leucoagaricus gongylophorus), a specialized pathogenic fungus in the genus Escovopsis. We filmed and compared infected and uninfected colonies, at both early and more advanced stages of infection. We quantified key defensive behaviors across treatments and show that the behavioral response to pathogen attack likely varies between different castes of worker ants, and between early and late detection of a threat. Based on these recordings we have made a library of behavioral clips, accompanied by definitions of the main individual defensive behaviors. We anticipate that such a guide can provide a common frame of reference for other researchers working in this field, to recognize and study these behaviors, and also provide greater scope for comparing different studies to ultimately help better understand the role these behaviors play in disease defense.

We present a visual guide to disease defense behaviors in leaf-cutting ants, with individual clips and accompanying definitions, illustrated in an experimental infection scenario. Our main aim is to help other researchers recognize key defensive behaviors and to provide a common understanding for future research in this field.

공부 하는 병원 체에 행동 방어 위한 비주얼 가이드 잎-절단 개미에 공격

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies ...

행동분석학

Bacterial Leaf Infiltration Assay for Fine Characterization of Plant Defense Responses using the Arabidopsis thaliana-Pseudomonas syringae Pathosystem

In the absence of specialized mobile immune cells, plants utilize their localized programmed cell death and Systemic Acquired Resistance to defend themselves against pathogen attack. The contribution of a specific Arabidopsis gene to the overall plant immune response can be specifically and quantitatively assessed by assaying the pathogen growth within the infected tissue. For over three decades, the hemibiotrophic bacterium Pseudomonas syringae pv. maculicola ES4326 (Psm ES4326) has been widely applied as the model pathogen to investigate the molecular mechanisms underlying the Arabidopsis immune response. To deliver pathogens into the leaf tissue, multiple inoculation methods have been established, e.g., syringe infiltration, dip inoculation, spray, vacuum infiltration, and flood inoculation. The following protocol describes an optimized syringe infiltration method to deliver virulent Psm ES4326 into leaves of adult soil-grown Arabidopsis plants and accurately screen for enhanced disease susceptibility (EDS) towards this pathogen. In addition, this protocol can be supplemented with multiple pre-treatments to further dissect specific immune defects within different layers of plant defense, including Salicylic Acid (SA)-Triggered Immunity (STI) and MAMP-Triggered Immunity (MTI).

Bacterial Leaf Infiltration Assay for Fine Characterization of Plant Defense Responses using the Arabidopsis thaliana-Pseudomonas syringae Pathosystem

Quantification of pathogen growth is a powerful tool to characterize various Arabidopsis thaliana (hereafter: Arabidopsis) immune responses. The method described here presents an optimized syringe infiltration assay to quantify the Pseudomonas syringae pv. maculicola ES4326 growth in adult Arabidopsis leaves.

을 사용해서 식물 방어 응답의 미세 특성에 대한 세균성 잎 침투 분석<em&gt; 애기 장대-슈도모나스 시린 개 (Pseudomonas syringae)</em&gt; Pathosystem

In the absence of specialized mobile immune cells, plants utilize their localized programmed cell death and Systemic Acquired Resistance to defend ...

면역학 및 감염병학

Herbivore-induced Blueberry Volatiles and Intra-plant Signaling

Herbivore-induced plant volatiles (HIPVs) are commonly emitted from plants after herbivore attack1,2. These HIPVs are mainly regulated by the defensive plant hormone jasmonic acid (JA) and its volatile derivative methyl jasmonate (MeJA)3,4,5. Over the past 3 decades researchers have documented that HIPVs can repel or attract herbivores, attract the natural enemies of herbivores, and in some cases they can induce or prime plant defenses prior to herbivore attack. In a recent paper6, I reported that feeding by gypsy moth caterpillars, exogenous MeJA application, and mechanical damage induce the emissions of volatiles from blueberry plants, albeit differently. In addition, blueberry branches respond to HIPVs emitted from neighboring branches of the same plant by increasing the levels of JA and resistance to herbivores (i.e., direct plant defenses), and by priming volatile emissions (i.e., indirect plant defenses). Similar findings have been reported recently for sagebrush7, poplar8, and lima beans9..
Here, I describe a push-pull method for collecting blueberry volatiles induced by herbivore (gypsy moth) feeding, exogenous MeJA application, and mechanical damage. The volatile collection unit consists of a 4 L volatile collection chamber, a 2-piece guillotine, an air delivery system that purifies incoming air, and a vacuum system connected to a trap filled with Super-Q adsorbent to collect volatiles5,6,10. Volatiles collected in Super-Q traps are eluted with dichloromethane and then separated and quantified using Gas Chromatography (GC). This volatile collection method was used n my study6 to investigate the volatile response of undamaged branches to exposure to volatiles from herbivore-damaged branches within blueberry plants. These methods are described here. Briefly, undamaged blueberry branches are exposed to HIPVs from neighboring branches within the same plant. Using the same techniques described above, volatiles emitted from branches after exposure to HIPVs are collected and analyzed.

A push-pull method for collecting plant volatiles is described. The method allows for a comparison of volatiles induced by herbivore feeding, exogenous methyl jasmonate, and mechanical damage. This technique is also used to investigate the volatile response of undamaged branches to exposure to volatiles from herbivore-damaged branches within blueberry plants.

Herbivore 유도 블루베리 휘발성 물질과 내부 식물 신호

Herbivore-induced plant volatiles (HIPVs) are commonly emitted from plants after herbivore attack1,2.  These HIPVs are mainly regulated by the defensive ...

Two-layered Membrane Sandwich Method for Laodelphax striatellus Saliva Collection

Rice stripe virus (RSV), which causes significant economic loss of agriculture in East Asia, entirely depends on insect vectors for its effective transmission among host rice. Laodelphax striatellus (small brown planthopper, SBPH) is the primary insect vector that horizontally transmits RSV while sucking sap from the phloem. Saliva plays a significant role in insects' feeding behavior. A convenient method that will be useful for research on insects' saliva with piercing-sucking feeding behavior is described here. In this method, insects were allowed to feed on an artificial diet sandwiched between two stretched paraffin film layers. The diet containing the saliva was collected each day, filtered, and concentrated for further analysis. Finally, the quality of collected saliva was examined by protein staining and immunoblotting. This method was exemplified by detecting the presence of RSV and a mucin-like protein in the saliva of SBPH. These artificial feeding and saliva collection method will lay a foundation for further research on factors in insect saliva related to feeding behavior and virus transmission.

Two-layered Membrane Sandwich Method for Laodelphax striatellus Saliva Collection

The present protocol describes a method to collect sufficient saliva from piercing-sucking insects using an artificial medium. This is a convenient method for collecting insect saliva and studying salivary function on insect feeding behavior and vector-borne virus transmission.

특성화 Herbivore 저항 메커니즘 : Spittlebugs에 Brachiaria 종. 예로

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