Name: field experiments of pollination ecology: the case of lycoris sanguinea var. sanguinea
Uploaded: 2016-11-25T15:00:00
Description: Watch this Scientific Journal Video about Field Experiments of Pollination Ecology: The Case of Lycoris sanguinea var. sanguinea at JoVE.com

The authors thank the three anonymous reviewers for their helpful comments on the manuscript. This work was partly supported by Grant-in-Aid for JSPS Fellows (26.11613).

BEMÆRK: Denne artikel er baseret på vores tidligere arbejde 30. Nogle dele er genoptrykt med tilladelse fra Botanisk Society of Japan og Springer Japan. 1. Observation af Floral Besøgende <ol><li> Udvælgelse af Observation Felter <ol><li> Søg de områder, hvor plantematerialer distribueres, og vælg kandidat undersøgelse sites ved hjælp af pålidelige ressourcer, såsom billedbøger, akademiske tidsskrifter osv Juster antallet af undersøgelsessteder at passe forskni...

Flower observationer og sække eksperimenter blev ansat i denne undersøgelse for at afsløre visitation frekvenser og den kvindelige reproduktive succes af planter, hhv. I Dafni (1992) 38, videobåndet metoden var effektiv, fordi det kunne optage timingen og varigheden af de besøgende til analyse og forhindre observatør bias. Men på det tidspunkt, denne metode krævede dyrt udstyr, og observation gange blev begrænset af batteriets levetid. For nylig er omkostningerne til udstyr til fremstilling videoopta...

Plant-bestøver interaktioner er gode eksempler til studiet af evolutionær biologi og økologi. De mutualistic relationer mellem blomster og bestøvere menes at have fremmet diversificering af angiosperm 1,2 som følge af naturlig udvælgelse, selv om andre biotiske og abiotiske faktorer også har udøvet indflydelse 3,4,5. Det menes også, at blomster træk har ændret til at tilpasse sig de mest effektive bestøvere og til at producere mere frugt og frø 6. Disse overbevisninger er blevet bygget om store undersøgelser baseret på forskellige indekser, såsom bestøvning effektivitet, der involverer forskellige fortolkninger 7. Blomstrende planter, der har generelle bestøvning systemer er besøgt af forskellige typer af bestøvere 8. Heri blev en blomst besøgende defineres som en dyreart, der besøgte for at få en blomstret belønning, og bestøvere blev defineret som blomster besøgende, der bestøver.Nogle af disse besøgende bære conspecifikt pollenkorn til stigmata af de besøgte blomster og kan klassificeres som bestøvere. Andre besøgende kan også have nogle intraspecifikke pollen; de kunne foretage mindre bestøvning på grund af adfærdsmæssige eller morfologiske misforhold mellem de bestøvere og blomsterne. Disse sammenlignelige forskelle i bidrag til anlægget reproduktion kunne producere varierende grader af selektivt pres på blomster træk 9 og kan forårsage adaptive divergens af blomstrende planter. Skønt sammensætningen af bestøver samfund og den relative arter overflod er vigtige 10, nøjagtig evaluering af hver besøgendes effektivitet er også kritisk at bestemme det adaptive og / eller evolutionære processer af planterne. I denne undersøgelse blev der kvantitative skøn over bestøver effektivitetsgevinster, defineret som frugt og frøproduktion pr visitation frekvens, bestemt 11. Den specerne og hyppigheden af ​​de enkelte blomster besøgende blev observeret, og reproduktive effekter på de besøgte blomsterne blev estimeret. Optagelsen af ​​blomster besøg gennem menneskelige observationer er en klassisk metode i bestøvning økologi. Men denne metode pålægger en stor byrde for observatører, som skal forblive foran planterne og tage omhyggelige, langsigtede målinger. For nylig har de teknologier filme og optage hurtigt udviklede, og billige digitale videokameraer har gjort det muligt at indføre videooptagelse til bestøver observationer 12,13. Disse metoder kan lette indsamlingen af ​​basisoplysninger om blomster besøgende, og kunne bidrage til at udvikle en forståelse af den bestøvning økologi af et mål plantearter. Imidlertid er visitation frekvenser af bestøvere ikke nødvendigvis korreleret til deres bestøvning effektivitet 7,14, og det er vigtigt at evaluere de kvalitative virkninger af hver bestøverpå blomst fitness. Bestøvning effektivitet er blevet anslået gennem antallet af pollenkorn på stigmata 15,16, pollen rør vækst 17,18 og frugt og / eller frøproduktion 19,20. Sække eksperimenter, udført ved hjælp af besøgende-eksklusiv poser, er de typiske metoder til at teste selv-kompatibilitet, autogamy 21,22, og tilstedeværelsen af apomixes 23. Derudover evaluering af bestøvning effektivitet for en bestemt bestøver i den besøgendes samling er blevet hyppigt udført i miljøer, hvor andre blomster besøgende er blevet begrænset (dvs. en wire bur, net eller pose med en maskestørrelse lille nok til at udelukke større bestøvere, der er indstillet på blomstrende planter). For eksempel blev udført sække eksperimenter med små maske poser til at afsløre bestøvning evne myrer eller thrips 24,25. Desuden har fugl udelukkelse eksperimenter ved hjælp af en wire bur eller netto vist de effektive bestøvere af Aloe taxa26-28. Formålet med denne undersøgelse var: 1) at indføre de metoder, der anvendes i et tidligere papir og 2) at forbedre disse metoder til almindelig brug i andre undersøgelser af blomster besøgende, deres besøg frekvenser, og deres virkninger på planter fitness Lycoris sanguinea var.. sanguinea er en af de arter, der indgår i slægten Lycoris, som distribueres bredt i hele Japan og snævert i Korea 29 og har tragtformet rødlig-orange blomster (figur 1a). En tidligere undersøgelse viste, at L. sanguinea var. sanguinea blev besøgt af flere insektarter, herunder en uidentificerede små bi arter og de større arter Amegilla Florea 29. Imidlertid blev den visitation frekvenser og bestøvning effektiviteten af ​​disse besøgende ikke identificeret. Bestøver observationer til identifikation af blomster besøgende blev udført først. Visitation af små bier blev observed på blomster, der ikke helt havde åbnet endnu (rev knopper fig 1b, c). Små bier flyttede hurtigt rundt om de undehisced støvknapper i bryde knopper og indsamlet pollen ved hjælp af deres kæber. Hypotesen var, at de små bier kunne være bestøvere på breaking-opløbet fase, fordi hullerne mellem støvknapper og stigmata i blomsterne var mindre end kroppen længde bierne. Derfor blev udført sække eksperimenter for at teste bestøvning evne små bier på breaking-opløbet fase, og derudover at undersøge de reproduktive strategier L. sanguinea var. sanguinea. Disse knopper blev sække efter de små bier besøgt, som tillod en vurdering af bestøvning evne bierne. De personer blev bur med uåbnede knopper. En lille-mesh bur blev anvendt, hvorigennem kun små bier kunne passere, så en vurdering af de bestøvning effektiviteten af ​​små bier hele blomsring fase.

<table border="0" cellpadding="0" cellspacing="0" width="535">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:200px;">recording sheet</td>
			<td style="width:159px;">any</td>
			<td style="width:112px;">NA</td>
			<td style="width:64px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">insect net</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">pooter</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">ethyl acetate</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">100% Ethanol</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">plastic tube</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">plastic case</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">soft bag</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">digital video camera(s)</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">tripod(s)</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">bags</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">wire or plastic mesh boards</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">iron wires</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">labeling tape</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">stick supporters</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">soft strings or wire</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">pincette(s)</td>
			<td>any</td>
			<td>NA</td>
			<td></td>
		</tr>
	</tbody>
</table>

field experiments of pollination ecology: the case of lycoris sanguinea var. sanguinea

Plant-pollinator interactions have been studied for approximately one hundred years. During that time, many field methods have been developed to clarify the pollination effectiveness of each pollinator for visited flowers. Pollinator observations have been one of the most common methods to identify pollinators, and bagging and cage experiments have been conducted to show the effectiveness of specific pollinators. In a previous study of Lycoris sanguinea var. sanguinea, its effective pollinators, the visitation frequencies of each floral visitor, and its reproductive strategies were not identified. This study reports the observation that small bees visited flowers that were partially opened (breaking buds). To the best of our knowledge, this phenomenon has not been reported previously. Further, this study investigates the hypothesis that small bees can pollinate at that flowering stage. This study demonstrates the basic methods of field experiments in pollination ecology using L. sanguinea var. sanguinea. Pollinator observations and digital video showed the visitation frequencies of each floral visitor. Bagging and cage experiments revealed that these flowers could be pollinated fully and that breaking-bud pollination could be important for the pollination of this plant species. The advantages and disadvantages of each method are discussed, and recent developments, including laboratory experiments, are described.

Fem populationer blev udvalgt til bestøver observationer. I den præ-observation fase blev visitations af forskellige insektarter til åbning blomster og små bier til at bryde knopper bekræftet. Blomster besøgende observationer viste, at de fleste af de besøgende på alle fem studier steder var individer af de små biarter Lasioglossum japonicum. Den samlede visitation rekord viste, at visitation nøgletal af denne art var mere end 90% ved tre lokaliteter (Figur 2)....

Watch this Scientific Journal Video about Field Experiments of Pollination Ecology: The Case of Lycoris sanguinea var. sanguinea at JoVE.com

Field Experiments of Pollination Ecology: The Case of Lycoris sanguinea var. sanguinea

For at afsløre bestøver effektiviteten af ​​en given planteart, er der blevet udviklet flere metoder til markforsøg. Denne undersøgelse viser de grundlæggende metoder til markforsøg til bestøvning økologi ved hjælp af casestudie af Lycoris sanguinea var. Sanguinea og romanen bestøvning mekanisme, bryde-opløbet bestøvning.

Field Eksperimenter af Bestøvning Økologi: Sagen om<em&gt; Lycoris sanguinea</em&gt; Var.<em&gt; sanguinea</em

Plant-pollinator interactions have been studied for approximately one hundred years. During that time, many field methods have been developed to clarify ...

The overall goal of this experiment is to investigate the pollinators of Lycoris sanguinea, variety sanguinea, and their pollination abilities. This method can help researchers in the pollination ecology field. For example, in the decreasing of observers around the time of the will focus on specific pollinators'effect per plant.The main advantage of this technique is that it facilitates the investigation of floral visitor species and frequencies for analysis of specific visitors'flower pollination efficiency. For floral visitor pre-observation first decide which target individuals in each area will be observed during the same time interval. Next, select 5 to 10 freshly opened flowers who are breaking buds as target flowers, and begin the visual observation for the period of an entire day, as the flowers can be visited by various pollinators in different time ranges.On the recording sheets, record the species name of each floral visitor, and the time of each visitation per hour, per flower. Closely observe whether floral visitors touch the anthers and/or stigmata. If they do, record the visitors as pollinators.To capture the floral visitors for their identification and preservation, use a handmade aspirator. Then kill the trapped visitors quickly with ethyl acetate and preserve them in a plastic tube with 100%ethanol. At the end of the observation period, use the morphological features of the visitors to identify the specific names of the specimens.Select the target flowers as just demonstrated. Then fix the video cameras to tripods and set the tripods approximately 50 centimeters away from the individual targets. Confirm the appropriate focus and light quantity on the screen displays of each target.Then begin the visual observation and video filming for the appropriate time zones as estimated by the pre-observations. Identify the species names of the visitors and pollinators. At the end of the observation period, check the video clips and note the species'names and their visit times in the same way as for the pre-observation.For a bagging experiment, select 30 individuals per treatment that have no damage from herbivores or from a severe environment. Label the target flowers in the field. Then, taking care not to touch the anthers or stigmata, bag the flowers with a bag with 0.5 to 1 millimeter mesh holes.Use supports to keep the individuals upright as necessary, and use a wire to carefully fix the bags to the flowers. Then, adjust the positions of the flowers to prevent contact and pollen depositions between the different stigmata to reject the influences from before the cage establishment. Afterwards, cover the target plant individuals with prepared cages.Using iron poles, attach the cages firmly to the ground to prevent entry by visitors between the cage bases and the ground. At the end of the flowering season, collect all of the caged and labeled flowers by cutting and separating them from the maternal individuals. Preserve each labeled sample separately to prevent contamination.Then check for the presence or absence of a fruit set on each labeled flower, recording the seed numbers of each fruit in the case of setting fruit. In this representative experiment, floral visitor observations revealed that most of the visitors to all five study sites were individuals of the small bee species Lasioglossum Japonicum, with a visitation ratio of more than 90%at three sites. In contrast, the ratios of the second most frequent visitor, Amegilla Florea, were lower than 10%in these fields.Both bee species also touched the stigmata of visited flowers and were recorded as pollinators. At one site, the visitation frequencies of small bees recorded by visual observations were significantly higher than those recorded by video. The breaking bud treatment of bagged or caged flowers demonstrates the pollination ability of small bees visiting breaking buds with a higher fruit set than the auto self-pollination observed in this experiment, indicating the presence of pollen deposition by small bees to stigmata at the breaking bud stage.Comparing the fruit and seed sets between the treatments reveals the reproductive traits of Lycoris sanguinea, variety sanguinea, which is predominantly animal-pollinated, with partially pollen-limited conditions. Further, these comparisons also suggest that the plant in this experiment has a self-compatibility consistent with previous reports. Moreover, comparisons of the other treatments confirm the pollination abilities of breaking bud pollination in Lycoris sanguinea, variety sanguinea.Once mastered, the plant cage technique can be performed on most growing plants for a variety of visitors. For the pollinator with patience, pre-research of the floral visitors, applying suitable recording techniques and selection of optimal are essential for accurate data collection. The introduction of recording equipment reduces incidence of possible human recording errors, such as over-observation of floral visitors.Following these procedures as a method, like molecular analysis using polymorphic micro saccharide markers can be performed to answer additional questions about pollinator-mediated gene flow between individuals. After watching this video, you should have a good understanding of how to record the interactions of target plants and pollinators, through observation of floral visitors and estimations of their pollination efficiencies.

Research

JoVE Journal

Environment

Field Experiments of Pollination Ecology: The Case of Lycoris sanguinea var. sanguinea

Experimental Protocol for Manipulating Plant-induced Soil Heterogeneity

Eksperimentel protokol til at manipulere Plant-induceret Soil Heterogenitet

Coexistence theory has often treated environmental heterogeneity as being independent of the community composition; however biotic feedbacks such as ...

Modifying the Bank Erosion Hazard Index (BEHI) Protocol for Rapid Assessment of Streambank Erosion in Northeastern Ohio

Modifying the Bank Erosion Hazard Index BEHI Protocol for Rapid Assessment of Streambank Erosion in Northeastern Ohio

Ændring af brederosion Hazard Index (Behi) Protokol til hurtig vurdering af Streambank Erosion i det nordøstlige Ohio

Understanding the source of pollution in a stream is vital to preserving, restoring, and maintaining the stream&#8217;s function and habitat it provides. ...

Calibrated Passive Sampling - Multi-plot Field Measurements of NH3 Emissions with a Combination of Dynamic Tube Method and Passive Samplers

Calibrated Passive Sampling - Multi-plot Field Measurements of NH3 Emissions with a Combination of Dynamic Tube Method and Passive Samplers

Kalibreret Passiv Prøveudtagning - Multi-plot Field Målinger af NH<sub&gt; 3</sub&gt; Emissioner med en kombination af Dynamic Tube Metode og Passive samplere

Agricultural ammonia (NH3) emissions (90% of total EU emissions) are responsible for about 45% airborne eutrophication, 31% soil acidification and 12% ...

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

VacuSIP, en forbedret Inex Metode til<em&gt; In Situ</em&gt; Måle partikler og opløst Forbindelser Bearbejdet af aktiv affjedring Feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

Modification and Application of a Leaf Blower-vac for Field Sampling of Arthropods

Ændring og anvendelse af en løvblæser-vac for Field Sampling af leddyr

Rice fields host a large diversity of arthropods, but investigating their population dynamics and interactions is challenging. Here we describe the ...

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

Pålidelig metode til at vurdere Seed Spiring, dvaletilstand, og Dødelighed under markforhold

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

Construction of a Compact Low-Cost Radiation Shield for Air-Temperature Sensors in Ecological Field Studies

Opførelse af et kompakt Low-Cost stråling skjold for luft-temperatur sensorer i økologiske feltundersøgelser

Low cost temperature sensors are increasingly used by ecologists to assess climatic variation and change on ecologically relevant scales. Although ...

Visualizing Efficacy of Pesticides Against Disease Vector Mosquitoes in the Field

Visualisere effekten af pesticider mod sygdom vektor myg i feltet

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform ...

Long-term Video Tracking of Cohoused Aquatic Animals: A Case Study of the Daily Locomotor Activity of the Norway Lobster (Nephrops norvegicus)

Long-term Video Tracking of Cohoused Aquatic Animals: A Case Study of the Daily Locomotor Activity of the Norway Lobster Nephrops norvegicus

Langsigtet Video sporing af Cohoused vanddyr: et casestudie af den daglige bevægeapparatet aktivitet af jomfruhummer (Nephrops norvegicus)

We present a protocol related to a video-tracking technique based on the background subtraction and image thresholding that makes it possible to ...

Standard Operating Procedure for Lyssavirus Surveillance of the Bat Population in Taiwan

Standard Operations procedure for lyssavirus overvågning af bat-populationen i Taiwan

Viruses within the genus Lyssavirus are zoonotic pathogens, and at least seven lyssavirus species are associated with human cases. Because bats are ...