Progress 01/01/24 to 12/31/24
Outputs
Target Audience:The target audience is researchers and plant breeders working in the area of agricultural pest control. By identifying the mechanisms by which aphids influence plant defenses, it will be possible to identify new protein targets for controlling aphid infestations. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Dr. Leila Feiz, a postdoc working on this project, received further experience in research on plant-insect interactions. Dr. Feiz also received guidance with her job applications and interviews. A graduate student, Syed Najeebullah, received training in plant molecular biology and the preparation of reports about his research. How have the results been disseminated to communities of interest?Two journal publications describing plant-aphid interactions include research that was conducted with funding from this award. Dr. Feiz presented the research on AAF1 in three invited talks entitled 'Investigating the Interplay of Factors in Plant Cell Physiology and Development' at the Institute of Biological Chemistry, Washington State University, Pullman; the Department of Biology, Purdue University, Fort Wayne (both in person); and for scientists at the Cotton Fiber Bioscience & Utilization Research Unit, USDA-ARS, Southern Regional Research Center (via Zoom). What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period we will continue research on autophagy induction by aphid salivary proteins. Research results will be published. An undergraduate student will receive research training.
Impacts
What was accomplished under these goals?
By generating and analyzing Arabidopsis plants transformed with autophagy-related factor 1 (AAF1, formerly SP11), it was shown that the expression of AAF1 in Arabidopsis significantly increases rosette size. This increase is autophagy-related, as it does not occur in the Arabidopsis autophagy mutant atg5, which was also transformed with the AAF1 effector. Optimized methods and demonstrated that AAF1 induces autophagy flux in the vacuoles of Nicotiana benthamiana leaves by showing an increase in mCherry to mCherry-ATG8d, which is a standard method for measuring autophagy flux. Performed a large-scale lipidomics analysis on Nicotiana benthamiana leaves transiently expressing different levels of AAF1 and or GFP control protein and demonstrated that AAF1 increases the abundance of sphingolipids, sterols, and triglycerides in leaves expressing AAF1 compared to control leaves overexpressing GFP. The observed increase in triglycerides and esterified sterols supports the hypothesis that AAF1 plays a role in the recruitment of lipid droplets to starvation-induced autophagosomes. It is well established that the induction of autophagy requires extensive lipid remobilization and recruitment and that triglycerides and esterified sterols constitute the major components of lipid droplets. Reciprocal interaction assays demonstrated that a plant phosphatidylinositol transfer protein binds to AAF1. The binding of AAF1 to this lipid transfer protein, along with its interaction with RabD1/D2a--two major vesicle trafficking proteins involved in autophagosome formation--further confirmed that AAF1 confers its function by facilitating lipid and vesicle remobilization. This process provides the essential components for the formation of aphid-mediated starvation-induced autophagosomes.
Publications
- Type:
Other Journal Articles
Status:
Published
Year Published:
2025
Citation:
Negin, B, Wang, F., Fischer, H.D., and Jander, G. (2025) Acylsugars, nicotine, and a protease inhibitor provide variable protection for Nicotiana benthamiana in a natural setting, Plant Cell and Environment, 48:1073-1087
- Type:
Other Journal Articles
Status:
Published
Year Published:
2025
Citation:
Najeebullah S, Umer N, Zahra Naqvi R, Arshad M, Jander G, Asad S, Mukhtar Z, Asif M. (2025) Transgenic cotton expressing Allium sativum leaf agglutinin exhibits resistance to whiteflies and aphids without negative effects on ladybugs
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Progress 01/01/23 to 12/31/23
Outputs
Target Audience:The target audience is researchers working in the area of agricultural pest control. By identifying the mechanisms by which aphids suppress plant defenses, it will be possible to identify new protein targets for controlling aphid infestations. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A postdoc, Dr. Leila Feiz, received futher experience in plant-aphid interactions and protein biochemistry research. An undergraduate student received training from Dr. Feiz through this project. How have the results been disseminated to communities of interest?Two reseach articles related to aphid-plant interactions were published in the past year. Both include research funded by this award. A postdoc, Dr. Leila Feiz, presented a talk at an Autophagy Gordon Conference. Georg Jander presented a talk describing results from this research at Cornell University. What do you plan to do during the next reporting period to accomplish the goals?We will finish analysis of autophagy suppression by aphid salivary proteins. Results of this research will be submitted for publication. An undergraduate student will receive research research training.
Impacts
What was accomplished under these goals?
Research in the past year was focused mainly on verifying in vivo interactions between an aphid salivary protein and plant proteins involved in triggering autophagy. This included pull-down experiments, fluorescence complementation, and analysis of plant gene expression in response to expression of the salivary protein. These results conclusively showed an induction of autophagy, a function that has not been previously confirmed as a function during aphid feeding. Additionally, new evidence shows that the protein actually is injected into the plant during aphid feeding, rather than just being collected from artificial diet on which aphids are feeding.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2024
Citation:
Serine proteinase inhibitors from Nicotiana benthamiana, a nonpreferred host plant, inhibit the growth of Myzus persicae (green peach aphid).
Feng H, Jander G. Pest Manag Sci. 2024 Sep;80(9):4470-4481. doi: 10.1002/ps.8148
- Type:
Journal Articles
Status:
Accepted
Year Published:
2024
Citation:
Negin, B, Wang, F., Fischer, H.D., and Jander, G. (2024) Acylsugars, nicotine, and a protease inhibitor provide variable protection for Nicotiana benthamiana in a natural setting, Plant Cell and Environment, in press.
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Progress 01/01/22 to 12/31/22
Outputs
Target Audience:The target audience is researchers working in the area of agricultural pest control. By identifying the mechanisms by which aphids suppress plant defenses, it will be possible to identify new protein targets for controlling aphid infestations. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Leila Feiz, a postdoc working on this project, received training in research on plant-insect interactions. An undergraduate student, Miguel Barrera, received training in plant molecular biology and the preparation of reports about his research. How have the results been disseminated to communities of interest?A paper has been published describing aphid RNA interference methods that were developed with funding from this project. Research related to this project was presented in the form of a poster in a symposium at Cornell University. What do you plan to do during the next reporting period to accomplish the goals? In the next reporting period, we will continue our functional analysis of two aphid salivary proteins. We will publish a paper about SP11. In a new direction, we will be looking for protein-small molecule interactions in aphid saliva that could be an indication of enzymatic activity.
Impacts
What was accomplished under these goals?
We have focused research primarily on studying the function of the SP11 aphid salivary protein in the past year. Several lines of evidence suggest that it may be a specific trigger of autophagy in plant cells. These include Inhibitors of autophagy reduce the effects of SP11 Co-immunoprecipitation with known autophagy-related proteins Biomolecular fluorescence complementation assays Subcellular localization of fusion proteins Increased ubiquitination of plant proteins RNA interference of aphid SP11 gene expression when feeding on plants In separate experiments, we have been developing assays and methods for RNA interference in aphids. Horizontally transferred genes, but also SP11, were targets in these experiments. There are potential practical applications both for aphid research and in pest control.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Feng H, Chen W, Hussain S, Shakir S, Tzin V, Adegbayi F, Ugine T, Fei Z, Jander G. (2023) Horizontally transferred genes as RNA interference targets for aphid and whitefly control. Plant Biotechnol J. doi: 10.1111/pbi.13992. Online ahead of print
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Progress 01/01/21 to 12/31/21
Outputs
Target Audience:The target audience is researchers working in the area of agricultural pest control. By identifying the mechanisms by which aphids suppress plant defenses, it will be possible to identify new protein targets for controlling aphid infestations. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Leila Feiz, a postdoc working on this project, received training in research on plant-insect interactions. An undergraduate student, Miguel Barrera, received training in plant molecular biology and the preparation of reports about his research. How have the results been disseminated to communities of interest?Leila Feiz presented a talk about this research at the American Society of Plant Biologists annual meeting in 2021. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, we will continue our functional analysis of two aphid salivary proteins: SP11, which elicits autophagy in plant cells, and SP15, which moves to the nucleus and binds to plant transcription factors.
Impacts
What was accomplished under these goals?
From a pool of 48 aphid salivary proteins that we transiently expressed in Nicotiana benthamiana, we chose two proteins, SP11 and SP15, for further analysis as part of Objectives 1 and 2. Objective 1: Analyze salivary proteins that induce hypersensitive responses An aphid salivary protein, SP11, was found to induce a hypersensitive response in Nicotiana benthamiana. Further experiments with N. benthamiana plants transiently expressing SP11 showed that aphids reproduce better on plants expressing SP11 than on empty vector control plants. This suggested that the cell death is selectively induced by aphids, rather than being a plant defense response. Antibodies raised against SP11 showed that the protein is injected into plants when aphids are feeding. Co-immunoprecipitation and bimolecular fluorescence complementation assays showed that SP11 interacts with proteins that are localized in the plant peroxisomes. One of the co-localize proteins was catalase, which appeared to be degraded in response to SP11 overexpression in N. benthamiana. Concomitant over expression of catalase with SP11 reduced cell death in N. benthamiana. Objective 2: Determine regulatory functions of salivary proteins in plant nuclei Based on GFP fusions, the SP15 protein was found to be localized to the cell nuclei in N. benthamiana plants on which the aphids were feeding. Pull-down experiments and proteomic analysis of plant proteins interacting with SP15 indicated that this protein is interacting with at least two different N. benthamiana transcription factors. Thus, SP15 is likely to alter plant gene expression. Objective 3: Investigate salivary protein targeting to plastids and mitochondria In the current project year, no research was done on proteins that are targeted to plastids or mitochondria.
Publications
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