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Plant Science

Categories
+ Phycology
+ Plant biochemistry
+ Plant breeding
+ Plant cell biology
+ Plant developmental biology
- Plant immunity
Disease bioassay
Disease symptom
Host-microbe interactions
Perception and signaling
Plant-insect interaction
+ Plant metabolism
+ Plant molecular biology
+ Plant physiology
+ Plant transformation
Protocols in Past Issues

Controlled Transmission of a Fijivirus Under Field Conditions Using Mass-Reared Planthoppers
AD Analía D. Dumón
MB Milene R. Barcenilla
JB Julieta B. Bariles
NP Nicolás A. Pereyra
SR Sandra M. Rodriguez
MM María F. Mattio
Q&A 0 Q&A
136 Views
Mar 20, 2026

Mal de Río Cuarto disease, caused by a Fijivirus, is a major constraint for maize production in Argentina. The traditional evaluation of resistant hybrids is limited by the low efficiency of natural virus transmission and the lack of standardized field inoculation methods. We developed a protocol that combines laboratory mass-rearing of the planthopper vector Delphacodes kuscheli with a controlled field transmission system. The method involves the synchronized production of large insect populations, acquisition of viruliferous vectors under controlled conditions, and their safe transport to the field using specialized containers. Transmission is achieved through individual cages placed on maize seedlings, ensuring high inoculation pressure under field-like conditions. This protocol enables reliable and reproducible virus transmission, facilitating large-scale screening of maize hybrids and other cereals. Its main advantages are the high throughput of vector production, improved transmission efficiency, and adaptability to diverse experimental designs.

Image-Based Lignin Detection in Nematode-Induced Feeding Sites in Arabidopsis Roots
MA Muhammad Amjad Ali
Krzysztof Wieczorek Krzysztof Wieczorek
Q&A 0 Q&A
1895 Views
May 5, 2025

Cyst and root-knot nematodes are sedentary biotrophic parasites that infect a wide range of plant species, causing significant annual yield and economic losses. Cyst nematodes (genera Heterodera and Globodera) induce specialized feeding structures called syncytia in host plant roots, while root-knot nematodes (Meloidogyne spp.) form galls containing feeding cells known as giant cells. This protocol describes the visualization of lignin in Arabidopsis roots infected by beet cyst nematode H. schachtii and root-knot nematode M. incognita using histochemical staining. We present two distinct approaches for lignin detection: direct staining of root segments containing syncytia and galls and histopathological detection in thin longitudinal sections of the feeding sites.

Quantification of Methylglyoxal Levels in Cowpea Leaves in Response to Cowpea Aphid Infestation
JM Jacob R. MacWilliams
MO Monika Ostaszewska-Bugajska
KB Klaudia Borysiuk
BS Bożena Szal
IK Isgouhi Kaloshian
Q&A 0 Q&A
3554 Views
Oct 20, 2020
Aphids are a serious pest of crops across the world. Aphids feed by inserting their flexible hypodermal needlelike mouthparts, or stylets, into their host plant tissues. They navigate their way to the phloem where they feed on its sap causing little mechanical damage to the plant. Additionally, while feeding, aphids secrete proteinaceous effectors in their saliva to alter plant metabolism and disrupt plant defenses to gain an advantage over the plant. Even with these arsenals to overcome plant responses, plants have evolved ways to detect and counter with defense responses to curtail aphid infestation. One of such response of cowpea to cowpea aphid infestation, is accumulation of the metabolite methylglyoxal. Methylglyoxal is an α,β-dicarbonyl ketoaldehyde that is toxic at high concentrations. Methylglyoxal levels increase modestly after exposure to a number of different abiotic and biotic stresses and has been shown to act as an emerging defense signaling molecule at low levels. Here we describe a protocol to measure methylglyoxal in cowpea leaves after cowpea aphid infestation, by utilizing a perchloric acid extraction process. The extracted supernatant was neutralized with potassium carbonate, and methylglyoxal was quantified through its reaction with N-acetyl-L-cysteine to form N-α-acetyl-S-(1-hydroxy-2-oxo-prop-1-yl)cysteine, a product that is quantified spectrophotometrically.

Insect Feeding Assays with Spodoptera exigua on Arabidopsis thaliana
YY Yanrong You
CA Chunpeng An
Chuanyou Li Chuanyou Li
Q&A 0 Q&A
5377 Views
Mar 5, 2020
Plant-insect interaction is an important field for studying plant immunity. The beet armyworm, Spodoptera exigua, is one of the best-known agricultural pest insects and is usually used to study plant interactions with chewing insects. Here, we describe a protocol for insect feeding assays with Spodoptera exigua lavae using model host plant Arabidopsis thaliana, which is simple and easy to conduct, and can be used to evaluate the effect of host genes on insect growth and thus to study plant resistance to chewing insects.

Peptide Feeding and Mechanical Wounding for Tomato Seedlings
LW Lei Wang
YW Yan Wang
GF Georg Felix
Q&A 0 Q&A
6250 Views
Mar 20, 2019
Plants need to respond appropriately to wounding and herbivorous insects. Peptide signals have been implicated in local and systemic induction of appropriate plant defense responses. To study these peptide signals and their perception in host plants, it is important to have reproducible bioassays. Several assays, such as treatment of peptide solution via pressure infiltration, have been developed. Here, we provide detailed protocols for peptide feeding and mechanical wounding for tomato seedlings. To directly introduce peptides into tomato seedlings, peptide solution is fed through the excised stem via the transpiration stream. To mimic the wounding caused by insect feeding, leaflets of tomato seedlings are mechanically damaged with a hemostat; and wounded and systemic unwounded leaves are harvested and analyzed separately. Samples from both assays may be further assessed by examining the transcript level of marker genes by quantitative real-time PCR (qRT-PCR).

Quantification of Serotonin in Rice and Insect Pest and its Functional Analysis in Insects Using Artificial Diet Feeding
LW Long Wang
XG Xiaohao Guo
WH Wenqian Huang
HL Haiping Lu
QS Qing-yao Shu
Q&A 0 Q&A
5676 Views
Feb 20, 2019
Rice is one of the world’s most important crops, but its production suffers from insect pests. Rice brown planthopper (BPH; Nilaparvata lugens Stål) and striped stem borer (SSB, Chilo suppressalis Walker) are the two most serious pests in rice production. We reported that serotonin is an essential mediator in the interaction between rice and insect. Here, we established a method for extraction and determination of serotonin in rice and BPH.

Arabidopsis-Green Peach Aphid Interaction: Rearing the Insect, No-choice and Fecundity Assays, and Electrical Penetration Graph Technique to Study Insect Feeding Behavior
Vamsi Nalam Vamsi Nalam
JL Joe Louis
MP Monika Patel
Jyoti Shah Jyoti Shah
Q&A 1 Q&A
10506 Views
Aug 5, 2018
Aphids constitute a large group of Hemipterans that use their slender stylets to tap into the sieve elements of plants from which they consume copious amounts of phloem sap, thus depriving the plant of photoassimilates. Some aphids also transmit viral diseases of plants. Myzus persicae Sülzer, commonly known as the green peach aphid (GPA), which is a polyphagous insect with a host range that covers 50 plant families, is considered amongst the top 3 insect pest of plants. The interaction between Arabidopsis thaliana and the GPA is utilized as a model pathosystem to study plant-aphid interaction. Here we describe the protocol used in our laboratories for rearing the GPA, and no-choice and fecundity bioassays to study GPA performance on Arabidopsis. In addition, we describe the procedure for the electrical penetration graph (EPG) technique to monitor feeding behavior of the GPA on Arabidopsis.