Program Announcement/Closing Date Global and Regional Environmental Change BSF-24-1 Investigators Title of Proposed Project: Heat Effect on Herbivory Damage on growth and leaf quality of Helianthus Annuus Requested Starting Date:

2/12/2024 1.Melinna Cabral 2.Nora Abouelsaad 3.Alexa Szymborski Biology 321, Laboratory Section C14 Teaching Assistant: Emel Kangi Triad No: 16 Date to Resubmit Date Approved Authorization For Consideration by BSF Organization Unit Authorization Ecology and Evolution Program Proposed Duration February 12, 2024 - March 26, 2024 (7 weeks) Signature Melinna Cabral Nora Abouelsaad Alexa Szymborski 1 Table of Contents: I. Project Summary… Project Description... A. Introduction.. B. Background..... II. III. C. Objectives, Hypotheses, or Questions…... D. Research Plan… 1. Experimental design.. 2. Methods.. 3. Safety precautions…... 4. Statistical analysis... 5. Research schedule... 1. Helianthus Annuus.. 2. Response of Helianthus Annuus to temperature….... 3. Response of Helianthus Annuus to wounding stress.. 4. Response of other species to temperature and wounding stress.......9 E. Expected Significance. F. Literature Cited... G. Biographical Sketches.. Budget. IV. Budget Explanation………. A. Equipment and Supplies. B. Computing Services.. C. Facilities... D. Special Equipment... .22-23 .22 .22 .23 23 .11-12 .3-16 11-14 12-13 13 13 .3-5 14 5-9 .6-8 .8 .8-9 .10-11 .14-16 .17-19 .20 .21 2 I. Project Summary Climate change is becoming a major threat to many ecosystems. The abrupt climate changes are forcing plants to adapt or go extinct. In this increasingly problematic time, insect herbivores can migrate and adapt to a greater capacity than plant species can keep up with. The proposed study will attempt to determine if rising temperature influences the productivity of Helianthus Annuus. We aim to observe how Helianthus Annuus survives under heat while experiencing wounding stress. Helianthus Annuus will be grown in a growth chamber while exposed to the optimal temperatures of 25°C. Some of the species will then be exposed to an elevated temperature of 42°C. Half of the Helianthus Annuus will receive wounding stress by hole punches to account for the effect of herbivory damage. The species root-to-shoot ratio, greenness, and overall growth will be studied. Under these treatments, we will be able to observe the effect the association of temperature and herbivory have on Helianthus Annuus, and how it reacts under these abiotic and biotic stresses. II. Project Description A. Introduction In today's day and age climate change is becoming a pressing issue. Escalating temperatures are increasing the frequency of droughts, wildfires, and invasive pest outbreaks. Plant function is intricately linked to the weather conditions. Climate affects a plant's environment and therefore its physiological processes (Morison and Morecroft 2006). Climate change is shifting plant-herbivore interactions, which could change the ecological and evolutionary states of entire ecosystems (Hamann et al. 2021). Traits of insect herbivores such as high reproduction and mobility rates can enable them to adapt to changing climate conditions better than plant species (Hamann et al. 2021). With this reasoning under continued climate change, plants could experience surging herbivory levels. 3 A primary factor affecting plant development is temperature. The Intergovernmental Panel Climate on Change expects temperature change over the next couple of years, heat waves are proposed to become more intense and frequent (Hatfield and Prueger 2015). With these expectations, understanding the growth and impacts on plants will help us develop strategies to overcome these impacts. Environmental stresses observed in studies have different effects depending on the tissues and organs of a plant (Brinkman and Sombroek 1996). In previous research, elevated temperature is causing developmental issues such as altered size and number of leaves. These changes alter function and thus hurt fitness in natural populations (Brinkman and Sombroek 1996). Herbivore damage can be harmful to a plant's fitness. Plant populations and their evolutionary response to this damage can involve either resistance or tolerance (Pilson and Decker 2002). Studies have shown the fitness of undamaged compared to the fitness of damaged plant species, but provide no evidence as to what the factors of tolerance are (Pilson and Decker 2002). We are aiming to see the relation between high temperature and ambient temperatures on plant growth while observing the wounding damage tolerance. Climate change stress is altering the biological processes of plants causing changes in development, productivity, and growth. In the face of these environmental conditions, it is essential to study plants that have great adaptability (Agüera and Purificación 2021). Of particular interest is the climate change effects on Helianthus Annuus, wild sunflowers. The species is typically located in disrupted areas and has a greater adaptive capacity to stress conditions concerning climate change (Agüera and Purificación 2021). A large number of herbivores are found to feed on them (Pilson 2000). Helianthus Annuus' fast early growth and ability to adapt to stress make it a great choice for the study of herbivory and climate change. 4 Little research has been done about the effects of temperature and wounding stress on Helianthus Annuus simultaneously. Therefore, our experiment will focus on the effects of these two factors on root-to-shoot ratio, greenness, and wounding stress. B. Background Information The increasing amount of heat due to global warming throughout the United States has been an issue in the cultivation of H. annuus. Plant biomass and growth can be decreased by heat due to increasing the production of reactive oxygen species while the plant is growing under these stressful conditions (Parveen et al. 2023). Heat stress at temperatures of 45°C also causes a decrease in shoot length, root length, and overall weight of the plant. The optimal temperature for H. annuus is around 21°C. It is also known that certain strains of H. annuus tolerate heat stress better than others and are considered to be more resistant. These strains are Hysin-39, FH-825, and Hysin-33 since they developed well in heat-stress environments (Parveen et al. 2023). However, H. annuus, like other plants, tends to grow better in warmer conditions so there must be a line between too much heat where the plant becomes overstressed and a possible temperature above optimal where if the plant was growing or healing they would benefit more from the extra heat. Higher heat units accumulated by planting H. annuus crops early showed that the optimum sowing time of a particular crop is early planting to have good output since the plant is exposed to heat for a longer period. This is because the fatty acid composition of H. annuus, as an oil seed crop, is influenced by temperature which regulates the ratio of oleic and linoleic acid that are essential for plant biosynthesis (Qadir et al. 2007). H. annuus typically undergoes stress to their leaves due to grazing and herbivory of insects like moths and sunflower beetles. This leaves holes in their leaves and the plant 5