Name: the use of high-resolution infrared thermography (hrit) for the study of ice nucleation and ice propagation in plants
Uploaded: 2015-05-08T14:00:00
Description: Watch this Scientific Journal Video about The Use of High-resolution Infrared Thermography HRIT for the Study of Ice Nucleation and Ice Propagation in Plants at JoVE.com

Denne forskning blev finansieret af den østrigske Science Fund (FWF): P23681-B16.

1. Forberedelse af plantematerialer <ol><li> Bruge enten blade eller hele planter af emne plantemateriale (Hosta spp. Eller Phaseolus vulgaris). </li></ol> 2. Udarbejdelse af Water Solutions indeholdende isnukleering Aktive (INA) Bakterier <ol><li> Kultur INA bakterien Pseudomonas syringae (Stamme Cit-7) i petriskåle ved 25 ° C på Pseudomonas Agar F fremstillet med 10 g / l af 100% glycerol pr fabrikantens retning. </li><li> Efter kulturer er vokset tilstrækkeli...

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Vand har evnen til at blive underafkølet til temperaturer et stykke under 0 ° C og den temperatur, hvor vand vil fryse kan være ganske variabel. 36. Grænsen temperatur i underafkøling af rent vand er ca. -40 ° C, og er defineret som det homogene nukleationspunkts. Når vand fryser ved temperaturer varmere end -40 ° C er det som følge af tilstedeværelsen af heterogene kimdannelsesmidler at sætte små is embryoner til at danne, der udgør en katalysator for isdannelse og vækst. 37 Der er e...

Frysning temperaturer, der forekommer, når planterne er aktivt voksende kan være dødbringende, især hvis planten har ringe eller ingen frysning tolerance. Sådanne frost begivenheder ofte ødelæggende virkninger på landbrugsproduktionen og kan også spille en vigtig rolle i udformningen af samfundet struktur i naturlige populationer af planter, især i alpin, sub-arktiske og arktiske økosystemer 1-6. Episoder med svær forårsfrost har haft store konsekvenser for frugtproduktion i USA og Sydamerika i de seneste år 7-9 og er blevet forværret af den tidlige debut af varmt vejr, efterfulgt af mere typiske gennemsnitlige lave temperaturer. Den tidlige varmt vejr inducerer knopper til at bryde, aktivering af væksten af nye skud, blade og blomster som alle har meget lidt at ingen frost tolerance 1,3,10-12. Sådanne uberegnelige vejrmønstre er blevet rapporteret til at være en direkte afspejling af igangværende klimaændringer og forventes at være en fælles vejr mønster for foreseeable fremtid 13. Bestræbelser på at give økonomiske, effektive og miljøvenlige management teknikker eller landbrugskemikalier, som kan give øget frost tolerance har haft begrænset succes for et væld af grunde, men dette kan til dels tilskrives den komplekse karakter af frysning tolerance og frysning unddragelse mekanismer i planter. 14 De adaptive mekanismer forbundet med frost overlevelse i planter er traditionelt blevet opdelt i to kategorier, frysning tolerance og frysning unddragelse. Førstnævnte kategori er forbundet med biokemiske mekanismer reguleres af et bestemt sæt af gener, der giver planter til at tolerere de belastninger, der er forbundet med tilstedeværelsen og den dehydratiserende virkning af is i sine væv. Mens sidstnævnte kategori er typisk, men ikke udelukkende, i forbindelse med de strukturelle aspekter af en plante, der bestemmer, om, hvornår og hvor is former i et anlæg 14. Til trods for forekomsten af ​​fryse undgåelse som en adaptive mekanisme, er lidt forskning er blevet afsat i den seneste tid på at forstå de underliggende mekanismer og regulering af fryse unddragelse. Læseren henvises til en nylig 15 for nærmere detaljer om dette emne. Mens isdannelse ved lave temperaturer kan virke som en enkel proces, mange faktorer bidrager til at bestemme den temperatur, hvor is nukleerer i plantevæv, og hvordan den spreder inden planten. Parametre, såsom tilstedeværelsen af ​​extrinsiske og iboende is kimdannelsesmidler, heterogene vs. homogene nukleeringsbegivenheder, termisk-hysterese (frostvæske) proteiner, tilstedeværelsen af ​​specifikke sukkere og andre osmolytter, og et væld af strukturelle aspekter af planten kan alle spille en væsentlig rolle i fryseprocessen i planter. Kollektivt, disse parametre påvirker temperatur, ved hvilken en plante fryser, hvor isen er indledt, og hvordan det vokser. De kan også påvirke morfologien af ​​de resulterende iskrystaller.Forskellige metoder er blevet anvendt til at undersøge fryseprocessen i planter under laboratoriebetingelser, herunder kernemagnetisk resonans spektroskopi (NMR) 16, magnetisk resonans (MRI) 17, cryo-mikroskopi 18-19, og lav temperatur scanningselektronmikroskopi (LTSEM ). 20 Indefrysning af hele planter i laboratorie- og field indstillinger har dog primært blevet overvåget med termoelementer. Anvendelsen af ​​termoelementer for at studere frysning er baseret på frigivelse af varme (smeltevarme), når vandet undergår en faseovergang fra en væske til et faststof. Frysning derpå registreres som en exoterm begivenhed. 21-23 Selvom termoelementer er de typiske foretrukne metode i at studere frysning i planter, deres anvendelse har mange begrænsninger, der begrænser mængden af information indsamlet under selve frysningen. For eksempel med termoelementer er det vanskeligt at næsten umuligt at afgøre, hvor isen initieres i planter, hvordan det udbreder,hvis det udbreder med en ensartet hastighed, og hvis nogle væv forbliver fri for is. Fremskridt i høj opløsning infrarød termografi (HRIT) 24-27, men har øget muligheden for at indhente oplysninger om indefrysning processen i hele planter, især når de anvendes i en differential billedbehandlingstype. 28-33 I nærværende rapport, vi beskriver anvendelsen af ​​denne teknologi til at studere forskellige aspekter af fryseprocessen og forskellige parametre, som påvirker, hvor og og til hvilken temperatur is initieres i planter. En protokol vil blive præsenteret, der vil demonstrere evne til is-nukleation-aktiv (INA) bakterie, Pseudomonas syringae (Cit-7) til at fungere som en ydre kimdannelsesmiddel indlede frosset for urteagtig plante med en høj, frost temperatur. Høj opløsning Infrarød kamera Protokollen og eksempler dokumenteret i denne rapport anvender en høj opløsning infrarødvideo radiometer. Den radiometer (figur 1) leverer en kombination af infrarøde og synlige spektrum billeder og temperaturdata. Det spektrale respons på kameraet er i området fra 7,5 til 13,5 um og giver 640 x 480 pixel opløsning. Synlige spektrum billeder genereret af den indbyggede kamera kan fusioneres med IR-billeder i realtid, hvilket letter fortolkningen af ​​komplekse, termiske billeder. En række linser til kameraet kan bruges til at lave close-up og mikroskopiske observationer. Kameraet kan bruges i en stand-alone mode eller grænseflader og styres med en bærbar computer ved hjælp propietary software. Kan bruges softwaren til at få en bred vifte af termiske data indlejret i de optagede videoer. Det er vigtigt at bemærke, at en lang række af infrarøde radiometre er kommercielt tilgængelige. Derfor er det vigtigt, at forskeren diskutere deres påtænkte anvendelse med en kyndig produkt ingeniør og at forskeren teste evnen af ​​enhver specific radiometer til at give de nødvendige oplysninger. Det billeddannende radiometer anvendes i den beskrevne protokol anbringes i en akryl boks (figur 2) isoleret med Styrofoam i n for at afskrække eksponering for kondensation under opvarmning og køling protokoller. Er ikke nødvendigt for alle kameraer eller programmer denne beskyttelse.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Infrared Camera</td>
			<td>FLIR</td>
			<td>SC-660</td>
			<td>Many models available depending on application</td>
		</tr>
		<tr>
			<td>Infrared Analytical Software</td>
			<td>FLIR</td>
			<td>ResearchIR 4.10.2.5</td>
			<td align="right">$3,500</td>
		</tr>
		<tr>
			<td rowspan="2">Pseudomonas syringae (strain Cit-7)</td>
			<td rowspan="2"></td>
			<td rowspan="2"></td>
			<td rowspan="2"><a href="mailto:icelab@berkeley.edu">Kindly provided by Dr. Steven Lindow, University of California&#160; Berkeley icelab@berkeley.edu</a></td>
		</tr>
		<tr>
		</tr>
		<tr>
			<td>Pseudomonas Agar F</td>
			<td>Fisher Scientific</td>
			<td>DF0448-17-1</td>
			<td></td>
		</tr>
	</tbody>
</table>

the use of high-resolution infrared thermography (hrit) for the study of ice nucleation and ice propagation in plants

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events often have devastating effects on agricultural production and can also play an important role in shaping community structure in natural populations of plants, especially in alpine, sub-arctic, and arctic ecosystems. Therefore, a better understanding of the freezing process in plants can play an important role in the development of methods of frost protection and understanding mechanisms of freeze avoidance. Here, we describe a protocol to visualize the freezing process in plants using high-resolution infrared thermography (HRIT). The use of this technology allows one to determine the primary sites of ice formation in plants, how ice propagates, and the presence of ice barriers. Furthermore, it allows one to examine the role of extrinsic and intrinsic nucleators in determining the temperature at which plants freeze and evaluate the ability of various compounds to either affect the freezing process or increase freezing tolerance. The use of HRIT allows one to visualize the many adaptations that have evolved in plants, which directly or indirectly impact the freezing process and ultimately enables plants to survive frost events.

Ice-kernedannende aktivitet Ice + bakterie, Pseudomonas syringae (stamme Cit-7) En 10 pi dråbe vand og 10 pi vand indeholdende P. syringae (Cit-7) blev placeret på abaxial overflade af en Hosta blad (Hosta spp.) (figur 4). Som vist dråbe vand indeholdende INA bakterier frøs første og var ansvarlig for at inducere bladet til at fryse, medens dråbe vand på bladoverfladen forblev frosne. Frysning og Ice formering i ...

Watch this Scientific Journal Video about The Use of High-resolution Infrared Thermography HRIT for the Study of Ice Nucleation and Ice Propagation in Plants at JoVE.com

The Use of High-resolution Infrared Thermography HRIT for the Study of Ice Nucleation and Ice Propagation in Plants

Here we present a protocol that allows one to visualize sites of ice formation and avenues of ice propagation in plants utilizing high resolution infrared thermography (HRIT).

Anvendelsen af ​​høj opløsning Infrarød termografi (HRIT) for Studiet af isnukleering og Ice Formering i Plants

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events ...

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Environment

The Use of High-resolution Infrared Thermography (HRIT) for the Study of Ice Nucleation and Ice Propagation in Plants

Expression of Recombinant Cellulase Cel5A from Trichoderma reesei in Tobacco Plants

Expression of Recombinant Cellulase Cel5A from Trichoderma reesei in Tobacco Plants

Ekspression af rekombinant Cellulase Cel5A fra<em&gt; Trichoderma reesei</em&gt; I tobaksplanter

Cellulose degrading enzymes, cellulases, are targets of both research and industrial interests. The preponderance of these enzymes in difficult-to-culture ...

Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields

Forbedring af Initial Growth Rate Jordbrugsvidenskabelige Planter efter hjælp statiske magnetfelter

Electronic devices and high-voltage wires induce magnetic fields. A magnetic field of 1,300-2,500 Gauss (0.2 Tesla) was applied to Petri dishes containing ...

Use of a Battery of Chemical and Ecotoxicological Methods for the Assessment of the Efficacy of Wastewater Treatment Processes to Remove Estrogenic Potency

Anvendelse af et batteri af kemiske og økotoksikologiske metoder til vurdering af effekten af ​​spildevandsrensningen kan fjerne østrogene styrke

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in ...

Empirical, Metagenomic, and Computational Techniques Illuminate the Mechanisms by which Fungicides Compromise Bee Health

Empiriske, Metagenomic og beregningsmæssige teknikker belyse mekanismerne af som fungicider kompromis biers sundhed

Growers often use fungicide sprays during bloom to protect crops against disease, which exposes bees to fungicide residues. Although considered ...

Automated, High-resolution Mobile Collection System for the Nitrogen Isotopic Analysis of NOx

Automated, High-resolution Mobile Collection System for the Nitrogen Isotopic Analysis of NOx

Automatiseret, høj opløsning Mobile Collection System for Nitrogen isotopanalysen af ​​NO<sub&gt; x</sub

Nitrogen oxides (NOx = NO + NO2) are a family of atmospheric trace gases that have great impact on the environment. NOx concentrations directly influence ...

The Calibration and Use of Capacitance Sensors to Monitor Stem Water Content in Trees

Kalibrering og brug af kapacitans sensorer til at overvåge stilk vandindholdet i træer

Water transport and storage through the soil-plant-atmosphere continuum is critical to the terrestrial water cycle, and has become a major research focus ...

Stress Distribution During Cold Compression of Rocks and Mineral Aggregates Using Synchrotron-based X-Ray Diffraction

Stress Distribution under kolde komprimering af klipper og mineralske aggregater ved hjælp af synkrotron-baserede røntgen diffraktion

We report detailed procedures for performing compression experiments on rocks and mineral aggregates within a multi-anvil deformation apparatus (D-DIA) ...

Protocol for Producing Three-Dimensional Infrared Video of Freezing in Plants

Protokol for at producere tredimensionale infrarød Video af frysning i planter

Freezing in plants can be monitored using infrared (IR) thermography, because when water freezes, it gives off heat. However, problems with color contrast ...

Generating Homo- and Heterografts Between Watermelon and Bottle Gourd for the Study of Cold-responsive MicroRNAs

Generering af Homo- og Heterografts mellem vandmelon og flasken centnergræskar for studiet af kolde-responderende MicroRNA

MicroRNAs (miRNAs) are endogenous small non-coding RNAs of about 20 - 24 nt, known to play important roles in plant development and adaptation. There is ...

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

Anvendelsen af ​​høj opløsning Infrarød termografi (HRIT) for Studiet af isnukleering og Ice Formering i Plants

Anvendelsen af høj opløsning Infrarød termografi (HRIT) for Studiet af isnukleering og Ice Formering i Plants