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Anthocyanin Production in a Carnivorous Plant, Pinguicula Eastern Illinois University The Keep Masters Theses Student Theses & Publications 2016 Seeing Red in a Sea of Green: Anthocyanin Production in a Carnivorous Plant, Pinguicula planifolia Jenna Annis Eastern Illinois University This research is a product of the graduate program in Biological Sciences at Eastern Illinois University. Find out more about the program. Recommended Citation Annis, Jenna, "Seeing Red in a Sea of Green: Anthocyanin Production in a Carnivorous Plant, Pinguicula planifolia" (2016). Masters Theses. 2478. https://thekeep.eiu.edu/theses/2478 This is brought to you for free and open access by the Student Theses & Publications at The Keep. It has been accepted for inclusion in Masters Theses by an authorized administrator of The Keep. For more information, please contact [email protected]. The Graduate School � MTERNILLINOIS UNIVERSITY­ Thesis Maintenance and Reproduction Certificate FOR: Graduate Candidates Completing Theses in Partial Fulfillment of the Degree Graduate Faculty Advisors Directing the Theses RE: Preservation, Reproduction, and Distribution of Thesis Research Preserving, reproducing, and distributing thesis research is an important part of Booth Library's responsibility to provide access to scholarship. In order to further this goal, Booth Library makes all graduate theses completed as part of a degree program at Eastern Illinois University available for personal study, research, and other not-for-profit educational purposes. Under 17 U.S.C. § 108, the library may reproduce and distribute a copy without infringing on copyright; however, professional courtesy dictates that permission be requested from the author before doing so. Your signatures affirm the following: D The graduate candidate is the author of this thesis. D The graduate candidate retains the copyright and intellectual property rights associated with the original research, creative activity, and intellectual or artistic content of the thesis. D The graduate candidate certifies her/his compliance with federal copyright law (Title 17 of the U. S. Code) and her/his right to authorize reproduction and distribution of all copyrighted materials included in this thesis. D The graduate candidate in consultation with the faculty advisor grants Booth Library the non­ exclusive, perpetual right to make copies of the thesis freely and publicly available without restriction, by means of any current or successive technology, including by not limited to photocopying, microfilm, digitization, or internet. D The graduate candidate acknowledges that by depositing her/his thesis with Booth Library, her/his work is available for viewing by the public and may be borrowed through the library's circulation and interlibrary loan departments, or accessed electronically. D The graduate candidate waives the confidentiality provisions of the Family Educational Rights and Privacy Act (FERPA) (20 U. S. C. § 1232g; 34 CFR Part 99) with respect to the contents of the thesis and with respect to information concerning authorship of the thesis, including name and status as a student at EasternIllinois University. I have conferred with my graduate faculty advisor. My signature below indicates that I have read and agree with the above statements, and hereby give my permission to allow Booth Library to reproduce and distribute my thesis. My adviser's signature indicates concurrence to reproduce and distribute the thesis. liracf].(ate Candidate Signature Facultv A\ldviser Siiroature Printed Name Printed Name radjl��(� G Date Please submiJ in duplicate. Seeing Red in a Sea of Green: Anthocyanin Production in a Carnivorous Plant, Pinguicula plani{Olia (TITLE) BY Jenna Annis THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in BIOLOGICAL SCIENCES IN THE GRADUATE SCHOOL, EASTERN ILLINOIS UNIVERSITY CHARLESTON, ILLINOIS 2016 YEAR I HEREBY RECOMMEND THAT THIS THESIS BE ACCEPTED AS FULFILLING THIS PART OF THE GRADUATE DEGREE CITED ABOVE ;1/0/:;>-o/v 144 •' THESIS COMMITTEE CHAIR DATE DEPARTMjz�'r/SCHOOL CHAIR DATE OR CHAIR'S DESIGNEE __±J/2,1( l }u.\\� THESIS COMMITTEE MEMBER DATE THESIS COMMITTEE MEMBER DATE l. I . 7/ 6 /Zo/{- I THESIS COMMITTEE MEMBER DATE THESIS COMMITTEE MEMBER DATE Abstract The Southeastern United States is a biological hot-spot for carnivorouspla nts, with over half of the North American species occurring along the Gulf Coast. Pinguicula planifolia Chapman is one of six carnivorous perennial butterwort species found in the Florida Panhandle. Among these species, only P. planifolia expresses a distinct variation of red coloration on its leaves. The functionalrole of foliar anthocyanins may include stress response to drought and nutrient deficiency, herbivory defense, freeradical scavenging, and photoprotection. Carnivorousplant leaves facestrong selection pressures to optimize prey capture andnutrient absorption. Given the frequency of these red pigments in such specialized trapping leaves of carnivorous plants, their presence may be providing some type of physiological adaptation. The objectives of this study were: 1) to determine how environmental factors impact foliar anthocyanin production in P. planifolia, 2) to investigate how light manipulation affects foliar anthocyanin content of green andred plants over time, 3) to determine how foliar anthocyanins affects prey capture, and4) to investigate how leaf gland density differs in red and green leaves. Three fieldstudies ( quadrat, light manipulation, and prey capture) were conducted at 7 populations within the Florida Panhandle, and2 lab studies (chamber and gland density) were conducted at Eastern Illinois University. In the quadrat study, chlorophyll and anthocyanin content indexes in leaves of plants selected in quadrats along a transect were measured as a factor of light intensity, water depth, soil nutrients, water nutrients, and habitat structure. In the light manipulation study, three treatments (control, clipped, and caged) were established for red and green plants. These treatments tested the effects of normal, heightened, and lowered solar radiation on red and green plants over time. The prey capture study employed artificial sticky traps painted to match leaf colors (red or green), which were established next to plants with red or green leaves. Prey was collected from artificial traps and leaves to determine how color affects prey capture. The chamber study tested ex situ responses of P. planifolia anthocyanin production to artificial environmental cues by growing plants in controlled growth chambers under varying conditions including light intensity, nutrient levels, and substrate moisture levels. In the gland density study, leaf clearings were performed to conduct density counts of stalked and sessile glands on red and green leaves. Foliar anthocyanin content is correlated positively with light intensity and negatively with water depth and vegetation height. In soil, calcium and pH levels are positively correlated with foliar anthocyanins. In water, bicarbonates and pH levels are positively correlated with foliar anthocyanins. In water, ammonium is negatively correlated with foliar anthocyanins. In field and laboratory settings, exposing red plants to decreased light intensity significantly reduced foliar anthocyanin content, while exposing green plants to increased light intensity significantly increased foliar anthocyanin content. Rosette diameter and leaf number of plants gr own in artificial high light conditions (1200-1500 µmol/m2/s) were significantly higher than of plants grown in artificial low light conditions (500-800 µmol/m2/s). Growing plants with fertilizer(16 -16-16 diluted to 0.9 g/lL) added to underlying water reduced survival by over 70%. Green traps (both artificial and leaves) captured significantly more prey than red traps. The most abundant taxon captured on leaves, regardless of color, was Collembola (62-67%). Red leaves contained a significantly higher number of stalked glands (12.7) compared to green leaves (9.9), while sessile gland density did not differ between leaf colors. This study provides evidence that light intensity alone can effectively increase or decrease anthocyanin production in P. planifolia. Further work is needed to determine if anthocyanins may serve as a type of protective screening from UV radiation in this species. Increased calcium availability along with higher pH in soils may serve to enhance anthocyanin production. Increased bicarbonates and higher pH in water also may be linked to increased foliar anthocyanins. Plants at the germination to seedling stage thrive in higher light intensities, likely due to increased photosynthetic activity. This species exhibits a strong resiliency to low nutrient/anoxic soil conditions, as plants thrived in completely submerged conditions for over 4 months, producing the largest plants of all chamber study treatments. Anthocyanins do not appear to play a role in prey attraction. The target ii prey (Collembola) may be attracted to olfactory or other cues emitted by these plants. While leaf color affected stalked gland density on P. planifolia leaves, where they had sporadic occurrence compared to sessile glands, which were more evenly spaced. Since stalked glands are the first step to prey consumption, they may be more sensitive to the environmental changes that affect leaf color, compared to sessile glands. While there is much more to learnabout the physiological role of foliar anthocyanins and its importance to the conservation of this species, we can conclude
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