P23681-B16：この研究は、オーストリア科学基金（FWF）によって資金を供給されました。

植物材料の調製<ol><li>対象植物材料（ ギボウシ属またはインゲンマメ ）のいずれかの葉や植物全体を使用してください。 </li></ol>氷核活性（INA）細菌を含有する水溶液の調製<ol><li>メーカーの方向に100％グリセロールを10g / Lを用いて調製シュードモナス寒天Fに25℃でペトリ皿で培養INA菌、 シュードモナスシリン （株CIT-7）。 </li><li>必要になるまで培養した後、十分に4℃の場所に成長しているが、氷核活性の高いレベルを確保する前に2日間4℃で維持。 </li><li>使い捨て、プラスチックで寒天の表面から一枚板からの細菌を掻き取りまたは再利用可能な金属へらを使用して場所の時に脱イオン水を10〜15ミリリットルに25ミリリットルの使い捨てキュベットに。濃度が1×10 9〜1×10 7の範囲であるべきです。· </sup&gt;ミリリットル-1。溶液が濁って表示されます。濃度はおおよそのことのみ必要があるとして、血球計または分光光度計を用いて濃度を確認する必要はありません。 </li><li>細菌を配布する10秒の最低キュベットを渦。 注：結果INA混合物の特定の濃度は重要ではなく、記載されているプロトコルは、氷核活性の適切なレベル以上のものを提供します。 INA菌とこの水の混合物を、核実験の後半で使用されます。 </li></ol> 3.凍結実験のセットアップ<ol><li>保護アクリルボックス内の高解像度赤外線カメラ（SC-660）を配置しているので、箱の前面の開口部を介してレンズのプロジェクトやボックスの後部開口部を介してノートパソコンや記録装置出口にカメラを接続する配線。箱の蓋を固定して、そのすべての意志の場所に環境室や冷凍庫の内部にボックスを配置OW主題植物材料が見られます。 <ol><li>反射された赤外線エネルギーからの干渉を防止するために、黒画用紙にチャンバの壁の内側を覆うことにより、植物材料の周りに暗い背景を提供します。 </li><li>可視波長での記録画像が必要とされる場合、光源からの発熱を最小限に抑えるためにLED照明でチャンバーを取り付けます。植物がカメラで見えるようにするために、このようなバッテリ駆動クローゼット光または他の小型LED素子としての照明の唯一の最小値は、必要とされます。 <ol><li>対象植物材料の可視画像が撮影されると、LED照明をオフにします。ポートまたはチャンバ内の他の開口を介して、カメラへのすべての外部有線接続（コンピュータにFireWire接続、電源コードなど）を配布します。 </li></ol></li><li>チャンバ内の温度勾配を回避または低減するために発泡絶縁材料を使用してポートまたは開口部に余分なスペースを埋めます。 1℃のチャンバーの初期温度を設定します。 </li></ol></li><li>植物材料は、カメラの視野内にあり、植物材料は、リモート表示画面または選択されたソフトウェアの中に表示されるように、植物または植物の部分を合わせ。 </li><li>植物は、従来の制御凍結実験を開始する、植物材料の大きさに応じて、1時間に30分1℃で平衡化することを可能にします。これは、凍結実験が開始されると、植物の温度は何度により空気温度に遅れないことを保証します。植物材料の温度が空気温度の0.5℃以内である場合に平衡が達成されます。 <ol><li>鉢植えの植物を使用する場合には鉢植えの土の上に発泡スチロール断熱材の層を配置します。植物が平衡化した後は、室内の冷却開始。 注：ポットの土壌表面上に絶縁層が植物の周囲の空気をポットから継続熱損失の量を低減し、FREから根を防ぎezing、このように一般的に起因する土壌中の残留熱本の巨大なタンクに自然の中で霜イベント中に発生しません。 </li></ol></li><li> 3.4.1-3.4.4で説明したように、所望のカメラパラメータ（カラー·パレット、温度範囲、関心の特定の領域など ） を設定します。 <ol><li>ライブ画像を見ながら温度変化を表示するには、虹のパレットを選択します。 </li><li>ソフトウェアだけで画像の下に配置された温度·バーを調整することにより、5℃に温度スパンを設定します。 </li><li>選択されたパレット（レインボー）で定義されているように擬似カラー画像に赤外線データを変換するためのリニアスケール（アルゴリズム）を選択し、5℃に温度の範囲を設定し、画像に基づいて自動的に追跡します。実験を行いながら別の方法として、手動で設定された範囲を調整します。 <ol><li> SOFTWが提供する目的の定義された領域内の特定の点の温度または平均温度を使用してくださいです。 。記録されたビデオシーケンスから、または画像ファイルに埋め込 ​​まれた情報から、全ての画素の温度データを取得する3 ResearchIRソフトウェアの中から代表的なスクリーンショットを示します。 </li></ol></li><li>興味のある特定のポイントを表す植物組織上の位置にカーソルを置きます。ポイント（サイズは1 -3画素）、ボックス、線、楕円、または円として関心領域を定義します。ポイントまたは形状の複数の組み合わせは、画像の上に配置することができます。 </li></ol></li><li>ビデオシーケンスを記録します<ol><li> 60 Hzで、記録を手動で停止されるのを記録するためにカメラを設定します。 </li><li>録画したビデオファイルが配置されるコンピュータまたは外部ドライブ上の場所を示しています。 </li><li>録音開始。 注：大規模なビデオファイルが生成されますので、外付けハードドライブへの録音を強くお勧めします。録画したビデオファイルは、後にNEを含む部分のみを含むように編集することができますcessary情報。これは、大幅にファイルサイズを小さくします。 </li><li> 0.5 -1.0°Cで増分室の温度を下げます。植物の温度が空気温度と平衡になるまで待ってから、0.5〜1.0°Cで再び温度を下げます。観察される植物組織の質量とその形態に応じて、平衡化は10〜15分かかることがあります。従って、約4℃/ hrの冷却速度を与えます。 </li><li>植物のフリーズとの観測が完了するまで、このように続けます。凍結プロセスが完了したときに記録を終了します。 注意：それらは同じ温度であるため、植物材料と背景が同じ色である場合に、植物組織は、空気温度で平衡化しました。背景温度や植物組織の温度が同じであるため、再度温度を下げるまで、植物材料を可視化することが困難であり、植物組織とAとの間の温度差がありますIR温度。 </li></ol></li></ol>

<ol>
	<li>Taschler, D., Beikircher, B., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Frost+resistance+and+ice+nucleation+in+leaves+of+five+woody+timberline+species+measured+in+situ+during+shoot+expansion.">Frost resistance and ice nucleation in leaves of five woody timberline species measured in situ during shoot expansion.</a> Tree Physiol. 24, 331-337 (2004).</li><li>Neuner, G., Hacker, J., L&#252;tz, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ice+formation+and+propagation+in+alpine+plants.">Ice formation and propagation in alpine plants.</a> Plants in alpine regions: Cell Physiology of adaptation and survival strategies. , 163-174 (2012).</li><li>Ladinig, U., Hacker, J., Neuner, G., Wagner, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+endangered+is+sexual+reproduction+of+high-mountain+plants+by+summer+frosts?+-+Frost+resistance,+frequency+of+frost+events+and+risk+assessment.">How endangered is sexual reproduction of high-mountain plants by summer frosts? - Frost resistance, frequency of frost events and risk assessment.</a> Oecologia. 171, 743-760 (2013).</li><li>Wisniewski, M. E., Gusta, L. V., Fuller, M. P., Karlson, D., Gusta, L. V., Wisniewski, M. E., Tanino, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ice+nucleation,+propagation+and+deep+supercooling:+the+lost+tribes+of+freezing+studies.">Ice nucleation, propagation and deep supercooling: the lost tribes of freezing studies.</a> Plant Cold Hardiness: from the laboratory to the field. , 1-11 (2009).</li><li>Bokhurst, S., Bjerke, J. W., Davey, M. P., Taulavuori, K., Taulavuori, E., Laine, K., Callaghan, T. V., Phoenix, G. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impacts+of+extreme+winter+warming+events+on+plant+physiology+in+a+sub-Arctic+heath+community.">Impacts of extreme winter warming events on plant physiology in a sub-Arctic heath community.</a> Physiol. Plant. 140, 128-140 (2010).</li><li>Taulavuori, K., Laine, K., Taulavuori, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+studies+on+Vaccinium+myrtillus.and+Vaccinium+vits-idea.in+relation+to+air+pollution+and+global+change+at+northern+high+latitudes:+A+review.">Experimental studies on Vaccinium myrtillus.and Vaccinium vits-idea.in relation to air pollution and global change at northern high latitudes: A review.</a> Env. Exp. Bot. 87, 191-196 (2013).</li><li>Wisniewski, M., Glenn, D. M., Gusta, L., Fuller, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+infrared+thermography+to+study+freezing+in+plants.">Using infrared thermography to study freezing in plants.</a> HortScience. 43, 1648-1651 (2008).</li><li>Gu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+2007+eastern+US+spring+freeze:+increased+cold+damage+in+a+warming+world.">The 2007 eastern US spring freeze: increased cold damage in a warming world.</a> BioScience. 58, 253-262 (2008).</li><li>Augspurger, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spring+warmth+and+frost:+phenology,+damage,+and+refoliation+in+a+temperate+deciduous+forest.">Spring warmth and frost: phenology, damage, and refoliation in a temperate deciduous forest.</a> Func. Ecol. 23, 1031-1039 (2007).</li><li>Neuner, G., Erler, A., Hacker, J., Ladinig, U., Wagner, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Frost+resistance+of+reproductive+tissues+in+various+reproductive+stages+of+high+alpine+plant+species.">Frost resistance of reproductive tissues in various reproductive stages of high alpine plant species.</a> Physiol. Plant. 147, 88-100 (2013).</li><li>Skre, O., Taulavuori, K., Taulavuori, E., Nilsne, J., Igeland, B., Laine, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+importance+of+hardening+and+winter+temperature+for+growth+in+mountain+birch+populations.">The importance of hardening and winter temperature for growth in mountain birch populations.</a> Env. Exp. Bot. 62, 254-266 (2008).</li><li>H&#228;nninen, H., Tanino, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tree+seasonality+in+a+warming+climate.">Tree seasonality in a warming climate.</a> Trends in Plant Science. 16, 412-416 (2011).</li><li>Katz, R. W., Brown, B. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extreme+events+in+a+changing+climate:+variability+is+more+important+than+averages.">Extreme events in a changing climate: variability is more important than averages.</a> Climate Change. 21, 289-302 (1992).</li><li>Wisniewski, M., Gusta, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+plant+cold+hardiness:+an+opinion.">Understanding plant cold hardiness: an opinion.</a> Physiol. Plant. 147, 4-14 (2013).</li><li>Wisniewski, M., Gusta, L., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptive+mechanisms+of+freeze+avoidance+in+plants.+A+brief+update.">Adaptive mechanisms of freeze avoidance in plants. A brief update.</a> Env. Exp. Bot. 99, 133-140 (2014).</li><li>Burke, M. J., Gusta, L. V., Quamme, H. A., Weiser, C. J., Li, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Freezing+and+injury+in+plants.">Freezing and injury in plants.</a> Annu. Rev. Plant Physiol. 27, 507-528 (1976).</li><li>Ishikawa, M., Price, W. S., Ide, H., Arata, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualization+of+freezing+behaviors+in+leaf+and+flower+buds+of+full-moon+maple+by+nuclear+magnetic+resonance+microscopy.">Visualization of freezing behaviors in leaf and flower buds of full-moon maple by nuclear magnetic resonance microscopy.</a> Plant Physiol. 115 (4), 1515-1524 (1997).</li><li>Ishikawa, M., Sakai, A., Li, P. H., Sakai, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+of+freezing+avoidance+in+comparison+with+freezing+tolerance:+a+demonstration+of+extra-organ+freezing.">Characteristics of freezing avoidance in comparison with freezing tolerance: a demonstration of extra-organ freezing.</a> Plant cold hardiness and freezing stress. , 325-340 (1982).</li><li>Buchner, O., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Freezing+cytorrhysis+and+critical+temperature+thresholds+for+photosystem+II+in+the+peat+moss+Sphagnum+capillifolium.">Freezing cytorrhysis and critical temperature thresholds for photosystem II in the peat moss Sphagnum capillifolium.</a> Protoplasma. 243 (1), 63-71 (2010).</li><li>Pearce, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+ice+and+cell+shape+in+frost-stressed+cereal+leaves:+A+low+temperature+scanning+electron+microscopy+study.">Extracellular ice and cell shape in frost-stressed cereal leaves: A low temperature scanning electron microscopy study.</a> Planta. 175, 313-324 (1988).</li><li>Ashworth, E. N., Anderson, J. A., Davis, G. A., Lightner, G. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ice+formation+in+Prunus+persica.+under+field+conditions.">Ice formation in Prunus persica. under field conditions.</a> J. Am. Soc. Hort. Sci. 110 (3), 322-324 (1985).</li><li>Ashworth, E. N., Davis, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ice+formation+in+woody+plants+under+field+conditions.">Ice formation in woody plants under field conditions.</a> HortSci. 21, 1233-1234 (1986).</li><li>Pramsohler, M., Hacker, J., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Freezing+pattern+and+frost+killing+temperature+of+apple+(Malus+domestica.)+wood+under+controlled+conditions+and+in+nature.">Freezing pattern and frost killing temperature of apple (Malus domestica.) wood under controlled conditions and in nature.</a> Tree Physiol. 32 (7), 819-828 (2012).</li><li>Wisniewski, M., Lindow, S. E., Ashworth, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Observations+of+ice+nucleation+and+propagation+in+plants+using+infrared+video+thermography.">Observations of ice nucleation and propagation in plants using infrared video thermography.</a> Plant Physiol. 113 (2), 327-334 (1997).</li><li>Lutze, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elevated+atmospheric+[CO2]+promotes+frost+damage+in+evergreen+tree+seedlings.">Elevated atmospheric [CO2] promotes frost damage in evergreen tree seedlings.</a> Plant Cell Environ. (6), 631-635 (1998).</li><li>Ball, M. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Space+and+time+dependence+of+temperature+and+freezing+in+evergreen+leaves).">Space and time dependence of temperature and freezing in evergreen leaves).</a> Func. Plant Biol. 29 (11), 1259-1272 (2002).</li><li>Sekozawa, Y., Sugaya, S., Gemma, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Observations+of+ice+nucleation+and+propagation+in+flowers+of+Japanese+Pear+(Pyrus+Pyrifolia).">Observations of ice nucleation and propagation in flowers of Japanese Pear (Pyrus Pyrifolia).</a> Nakai) using infrared video. 73 (1), 1-6 (2004).</li><li>Hacker, J., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ice+propagation+in+plants+visualized+at+the+tissue+level+by+IDTA+(infrared+differential+thermal+analysis).">Ice propagation in plants visualized at the tissue level by IDTA (infrared differential thermal analysis).</a> Tree Physiol. 27, 1661-1670 (2007).</li><li>Hacker, J., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ice+propagation+in+dehardened+alpine+plant+species+studied+by+infrared+differential+thermal+analysis+(IDTA).">Ice propagation in dehardened alpine plant species studied by infrared differential thermal analysis (IDTA).</a> Arc. Antarc. Alp. Res. 40 (4), 660-670 (2008).</li><li>Hacker, J., Spindelb&#246;ck, J., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesophyll+freezing+and+effects+of+freeze+dehydration+visualized+by+simultaneous+measurement+of+IDTA+and+differential+imaging+chlorophyll+fluorescence.">Mesophyll freezing and effects of freeze dehydration visualized by simultaneous measurement of IDTA and differential imaging chlorophyll fluorescence.</a> Plant Cell Environ. 31, 1725-1733 (2008).</li><li>Neuner, G., XU, B., Hacker, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Velocity+and+pattern+of+ice+propagation+and+deep+supercooling+in+woody+stems+of+Castanea+sativa.,+Morus+nigra.+and+Quercus+robur.+measured+by+IDTA.">Velocity and pattern of ice propagation and deep supercooling in woody stems of Castanea sativa., Morus nigra. and Quercus robur. measured by IDTA.</a> Tree Physiol. 30, 1037-1045 (2010).</li><li>Hacker, J., Ladinig, U., Wagner, J., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflorescences+of+alpine+cushion+plants+freeze+autonomously+and+may+survive+subzero+temperatures+by+supercooling.">Inflorescences of alpine cushion plants freeze autonomously and may survive subzero temperatures by supercooling.</a> Plant Sci. 180, 149-156 (2011).</li><li>Kuprian, E., Briceno, V., Wagner, J., Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ice+barriers+promote+supercooling+and+prevent+frost+injury+in+reproductive+buds,+flowers+and+fruits+of+alpine+dwarf+shrubs+throughout+the+summer.">Ice barriers promote supercooling and prevent frost injury in reproductive buds, flowers and fruits of alpine dwarf shrubs throughout the summer.</a> Env. Exp. Bot. 106, 4-12 (2014).</li><li>Wisniewski, M., Glenn, D. M., Fuller, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+hydrophobic+particle+film+as+a+barrier+to+extrinsic+ice+nucleation+in+tomato+plants.">Use of a hydrophobic particle film as a barrier to extrinsic ice nucleation in tomato plants.</a> HortScience. 127, 358-364 (2002).</li><li>Aryal, B., &amp;Neuner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Leaf+wettability+decreases+along+an+extreme+altitudinal+gradient.">Leaf wettability decreases along an extreme altitudinal gradient.</a> Oecologia. 162, 1-9 (2010).</li><li>Wisniewski, M., Fuller, M. P., Margesin, R., Schinner, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ice nucleation and deep supercooling in plants: new insights using infrared thermography. In: Cold adapted organisms. Ecology, physiology, enzymologyandmolecularbiology. , 105-118 (1999).</li><li>Franks, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biophysics and biochemistry at low temperatures. , (1985).</li><li>Neuner, G., Kuprian, E., Hincha, D., Zuther, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infrared+thermal+analysis+of+plant+freezing+processes.">Infrared thermal analysis of plant freezing processes.</a> Methods in Molecular Biology: Plant Cold Acclimation. , 91-98 (2014).</li></ol>

著者らは、全く競合する金融利害や利害の衝突を持っていません。

水は十分に0℃以下の温度に過冷却する能力を有しており、水が凍結する温度は非常に可変であることができる。36純水の過冷却温度の上限は、約-40℃であり、均一な核形成点として定義されます。暖かい温度で水が凍結がより-40℃、それは次いで、氷の形成と成長のための触媒として機能する形成するために、小さな氷の胚を可能にする核不均一の存在によってもたらされた場合...

植物が活発に増殖している場合に発生する凍結温度は、植物がほとんど、あるいは全く凍結耐性を持っている場合は特に、致死することができます。このような霜のイベントは、多くの場合、農業生産に壊滅的な影響を持っていると、特に高山、サブ北極および北極の生態系1-6に、植物の自然集団におけるコミュニティ構造を形成する上で重要な役割を果たすことができます。深刻な春の霜のエピソードは、近年7-9でアメリカと南アメリカの果実生産に大きな影響を持っていた、より一般的な平均低温が続く暖かい天候の早期発症によって悪化されています。初期の暖かい天気がない霜耐性1,3,10-12にはほとんど持っているすべてのそれらの新芽、葉、花の成長を活性化し、破壊するために芽を誘導します。このような不安定な天候パターンは、進行中の気候変動の直接的反映であることが報告されており、foresための一般的な天候パターンであることが予想されますeeable今後13。増加霜耐性を提供することができ、経済的で効果的、かつ環境に配慮した経営手法や農薬を提供する努力は、理由のホストのための限られた成功を収めているが、これは、部分的に凍結耐性植物に回避メカニズムを凍結の複雑な性質に起因することができます。 14 植物における霜の生存に関連する適応メカニズムは、伝統的に凍結耐性と回避を凍結、2つのカテゴリに分類されています。前者は、植物が存在し、その組織中の氷の脱水効果に関連する応力を許容することができ、遺伝子の特定のセットで規制生化学的機構に関連しています。後者は、典型的であるが、単独で、プラント14であれば、氷の形を決定し、植物の構造的側面 ​​と関連しないが。広告としての凍結回避の有病率にもかかわらず、aptive機構は、少し研究は、基礎となるメカニズム、凍結回避の調節を理解する最近で専念してきました。読者は、この問題に関するより詳細のための最近の総説15と呼ばれています。 低温での氷の形成は簡単なプロセスのように見えるかもしれないが、多くの要因が植物組織中での氷の核形成温度を決定することに寄与し、それが工場内に広がりますか。このような外因性および内因性の核の氷の存在などのパラメータ、不均一な対均一核イベント、熱ヒステリシス（不凍液）タンパク質、特定の糖および他のオスモライトの存在、および植物の構造的側面のホストがすべての重要なを再生することができます植物における凍結過程における役割。まとめると、これらのパラメータは、氷が開始され、それがどのように成長される植物は、凍結する温度に影響を与えます。また、得られた氷結晶の形態に影響を与えることができます。種々の方法は、磁気共鳴イメージング（MRI）17、低温顕微鏡18-19、低温走査型電子顕微鏡（LTSEM、核磁気共鳴分光法（NMR）16を含む実験室条件下で植物における凍結過程を研究するために使用されています）。実験室とフィールドの設定で、植物全体の20凍結は、しかし、主に熱電対を用いて監視されています。凍結を研究するための熱電対の使用は、水が液体から固体への相転移を受ける熱の放出（融解エンタルピー）に基づいています。凍結次に、発熱事象として記録される。21-23を熱電対は、植物中で凍結研究において選択される典型的な方法であるにもかかわらず、その使用は、凍結事象の間に得 ​​られた情報の量を制限する多くの制限を有します。例えば、熱電対とそれがどのように伝播するか、氷が植物で開始されている場所を特定することはほぼ不可能に困難であり、それも速度で伝播している場合、一部の組織は、氷のない残っている場合。 高解像度の赤外線サーモグラフィ（HRIT）24-27の進歩は、しかし、差動撮影モードで使用する場合は特に、植物全体で凍結プロセスに関する情報を取得する能力を大幅に増加している。28-33本報告書では、凍結プロセスの様々な態様およびどこで、どのような温度で氷植物において開始される様々なパラメータに影響を研究するためのこの技術の使用を記載しています。プロトコルは高い、零下の温度での草本植物での凍結開始外因性核剤として作用する氷核活性（INA）細菌、 シュードモナスシリン （CIT-7）の能力を実証することが発表されます。 高解像度赤外線カメラこのレポートに記載されたプロトコルおよび実施例は、高解像度の赤外線を利用しますビデオ放射計。放射計（ 図1）は、赤外線および可視スペクトル画像と温度データの組合せを供給する。カメラのスペクトル応答は、7.5から13.5ミクロンの範囲であり、640×480ピクセルの解像度を提供します。内蔵カメラで生成された可視スペクトルの画像が複雑な、熱画像の解釈を容易にする、リアルタイムでのIR画像と融合させることができます。カメラ用レンズの範囲は、クローズアップ、顕微鏡観察を行うために使用することができます。カメラは、スタンドアロンモードで使用、またはpropietaryソフトウェアを使用してラップトップコンピュータとインターフェースし、制御することができます。ソフトウェアが記録されたビデオに埋め込まれた温度データの様々な得るために使用することができます。これは、赤外線放射計の多種多様な商業的に入手可能であることに注意することが重要です。したがって、研究者が知識豊富な製品エンジニアとの意図された用途を議論していることと、研究者は、任意の仕の能力をテストすることが不可欠ですC放射計は、必要な情報を提供します。記載されているプロトコルで使用される撮像放射計は、アクリルボックスに発泡スチロールI N温暖化と冷却のプロトコルの間に結露への暴露を抑止するためで絶縁（ 図2）が配置されています。この保護は、すべてのカメラやアプリケーションのために必要とされません。

<table><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>$3,500</td></tr><tr><td>Pseudomonas syringae (strain Cit-7)</td><td></td><td></td><td>&lt;a href=&quot;mailto:icelab@berkeley.edu&quot;&gt;Kindly provided by Dr. Steven Lindow, University of California&amp;nbsp; Berkeley icelab@berkeley.edu&lt;/a&gt;</td></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.

アイス+細菌の氷核形成活性を、 シュードモナスシリン （株CIT-7） 10μlの水の低下とPを含む水10μl syringaeの （CIT-7）ギボウシの葉の背軸面上に置いた（ ギボウシ属 ）（ 図4）。図示のように、INA菌を含む水の低下は最初凍結し、葉の表面に水滴が凍結されていないままフリーズする葉の誘導を担当していました。 <p class="jove_cont...

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).

植物の氷核と氷伝播の研究のための高解像度赤外線サーモグラフィ（HRIT）の使用

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 ...

the-use-high-resolution-infrared-thermography-hrit-for-study-ice

Universidad de Cartagena UDEC

Research

JoVE Journal

Environment

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

9.5K ビュー.  Agricultural Research Service (USDA-ARS), Kearneysville, WV. 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).

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

A Rapid Laser Probing Method Facilitates the Non-invasive and Contact-free Determination of Leaf Thermal Properties

Plants can produce valuable substances such as secondary metabolites and recombinant proteins. The purification of the latter from plant biomass can be streamlined by heat treatment (blanching). A blanching apparatus can be designed more precisely if the thermal properties of the leaves are known in detail, i.e., the specific heat capacity and thermal conductivity. The measurement of these properties is time consuming and labor intensive, and usually requires invasive methods that contact the sample directly. This can reduce the product yield and may be incompatible with containment requirements, e.g., in the context of good manufacturing practice. To address these issues, a non-invasive, contact-free method was developed that determines the specific heat capacity and thermal conductivity of an intact plant leaf in about one minute. The method involves the application of a short laser pulse of defined length and intensity to a small area of the leaf sample, causing a temperature increase that is measured using a near infrared sensor. The temperature increase is combined with known leaf properties (thickness and density) to determine the specific heat capacity. The thermal conductivity is then calculated based on the profile of the subsequent temperature decline, taking thermal radiation and convective heat transfer into account. The associated calculations and critical aspects of sample handling are discussed.

A method was developed to determine the specific heat capacity and thermal conductivity of leaf tissue by non-invasive, contact-free near infrared laser probing, which requires less than 1 min per sample.

迅速なレーザー法は葉熱特性の非侵襲的および非接触決意を容易にプロービング

Plants can produce valuable substances such as secondary metabolites and recombinant proteins. The purification of the latter from plant biomass can be ...

生化学

Identification of Plant Ice-binding Proteins Through Assessment of Ice-recrystallization Inhibition and Isolation Using Ice-affinity Purification

Ice-binding proteins (IBPs) belong to a family of stress-induced proteins that are synthesized by certain organisms exposed to subzero temperatures. In plants, freeze damage occurs when extracellular ice crystals grow, resulting in the rupture of plasma membranes and possible cell death. Adsorption of IBPs to ice crystals restricts further growth by a process known as ice-recrystallization inhibition (IRI), thereby reducing cellular damage. IBPs also demonstrate the ability to depress the freezing point of a solution below the equilibrium melting point, a property known as thermal hysteresis (TH) activity. These protective properties have raised interest in the identification of novel IBPs due to their potential use in industrial, medical and agricultural applications. This paper describes the identification of plant IBPs through 1) the induction and extraction of IBPs in plant tissue, 2) the screening of extracts for IRI activity, and 3) the isolation and purification of IBPs. Following the induction of IBPs by low temperature exposure, extracts are tested for IRI activity using a &#39;splat assay&#39;, which allows the observation of ice crystal growth using a standard light microscope. This assay requires a low protein concentration and generates results that are quickly obtained and easily interpreted, providing an initial screen for ice binding activity. IBPs can then be isolated from contaminating proteins by utilizing the property of IBPs to adsorb to ice, through a technique called &#39;ice-affinity purification&#39;. Using cell lysates collected from plant extracts, an ice hemisphere can be slowly grown on a brass probe. This incorporates IBPs into the crystalline structure of the polycrystalline ice. Requiring no a priori biochemical or structural knowledge of the IBP, this method allows for recovery of active protein. Ice-purified protein fractions can be used for downstream applications including the identification of peptide sequences by mass spectrometry and the biochemical analysis of native proteins.

This paper outlines the identification of ice-binding proteins from freeze-tolerant plants through the assessment of ice-recrystallization inhibition activity and subsequent isolation of native IBPs using ice-affinity purification.

アイスアフィニティー精製を用いた氷再結晶阻害とアイソレーションの評価による植物の氷結合タンパク質の同定

Ice-binding proteins (IBPs) belong to a family of stress-induced proteins that are synthesized by certain organisms exposed to subzero temperatures. In ...

RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop cultivars against environmental stresses. An experimental protocol is presented for RGB and hyperspectral imaging of root systems. The approach uses rhizoboxes where plants grow in natural soil over a longer time to observe fully developed root systems. Experimental settings are exemplified for assessing rhizobox plants under water stress and studying the role of roots. An RGB imaging setup is described for cheap and quick quantification of root development over time. Hyperspectral imaging improves root segmentation from the soil background compared to RGB color based thresholding. The particular strength of hyperspectral imaging is the acquisition of chemometric information on the root-soil system for functional understanding. This is demonstrated with high resolution water content mapping. Spectral imaging however is more complex in image acquisition, processing and analysis compared to the RGB approach. A combination of both methods can optimize a comprehensive assessment of the root system. Application examples integrating root and aboveground traits are given for the context of plant phenotyping and plant physiological research. Further improvement of root imaging can be obtained by optimizing RGB image quality with better illumination using different light sources and by extension of image analysis methods to infer on root zone properties from spectral data.

An experimental protocol is presented for assessment of soil grown plant root systems with RGB and hyperspectral imaging. Combination of RGB image time series with chemometric information from hyperspectral scans optimizes insights into plant root dynamics.

RGB とスペクトルのルート画像植物表現型解析および生理学的研究: 実験概要およびイメージ投射プロトコル

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop ...

環境学

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

Freezing in plants can be monitored using infrared (IR) thermography, because when water freezes, it gives off heat. However, problems with color contrast make 2-dimensions (2D) infrared images somewhat difficult to interpret. Viewing an IR image or the video of plants freezing in 3 dimensions (3D) would allow a more accurate identification of sites for ice nucleation as well as the progression of freezing. In this paper, we demonstrate a relatively simple means to produce a 3D infrared video of a strawberry plant freezing. Strawberry is an economically important crop that is subjected to unexpected spring freeze events in many areas of the world. An accurate understanding of the freezing in strawberry will provide both breeders and growers with more economical ways to prevent any damage to plants during freezing conditions.
The technique involves a positioning of two IR cameras at slightly different angles to film the strawberry freezing. The two video streams will be precisely synchronized using a screen capture software that records both cameras simultaneously. The recordings will then be imported into the imaging software and processed using an anaglyph technique. Using red-blue glasses, the 3D video will make it easier to determine the precise site of ice nucleation on leaf surfaces.

Here, we present a protocol to image a strawberry plant freezing in 3 dimensions. Two infrared cameras positioned at slightly different angles are used to produce a red-blue anaglyph video to observe the freezing of the plant in 3 dimensions.

植物の凍結の三次元赤外線ビデオを制作するためのプロトコル

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

Simulating Impacts of Ice Storms on Forest Ecosystems

Ice storms can have profound and lasting effects on the structure and function of forest ecosystems in regions that experience freezing conditions. Current models suggest that the frequency and intensity of ice storms could increase over the coming decades in response to changes in climate, heightening interest in understanding their impacts. Because of the stochastic nature of ice storms and difficulties in predicting when and where they will occur, most past investigations of the ecological effects of ice storms have been based on case studies following major storms. Since intense ice storms are exceedingly rare events it is impractical to study them by waiting for their natural occurrence. Here we present a novel alternative experimental approach, involving the simulation of glaze ice events on forest plots under field conditions. With this method, water is pumped from a stream or lake and sprayed above the forest canopy when air temperatures are below freezing. The water rains down and freezes upon contact with cold surfaces. As the ice accumulates on trees, the boles and branches bend and break; damage that can be quantified through comparisons with untreated reference stands. The experimental approach described is advantageous because it enables control over the timing and amount of ice applied. Creating ice storms of different frequency and intensity makes it possible to identify critical ecological thresholds necessary for predicting and preparing for ice storm impacts.

Ice storms are important weather events that are challenging to study because of difficulties in predicting their occurrence. Here, we describe a novel method for simulating ice storms that involves spraying water over a forest canopy during sub-freezing conditions.

森林生態系に対する氷嵐の影響のシミュレーション

Ice storms can have profound and lasting effects on the structure and function of forest ecosystems in regions that experience freezing conditions. ...

Near-Infrared Temperature Measurement Technique for Water Surrounding an Induction-heated Small Magnetic Sphere

A technique to measure the temperature of water and non-turbid aqueous media surrounding an induction-heated small magnetic sphere is presented. This technique utilizes wavelengths of 1150 and 1412 nm, at which the absorption coefficient of water is dependent on temperature. Water or a non-turbid aqueous gel containing a 2.0-mm- or 0.5-mm-diameter magnetic sphere is irradiated with 1150 nm or 1412 nm incident light, as selected using a narrow bandpass filter; additionally, two-dimensional absorbance images, which are the transverse projections of the absorption coefficient, are acquired via a near-infrared camera. When the three-dimensional distributions of temperature can be assumed to be spherically symmetric, they are estimated by applying inverse Abel transforms to the absorbance profiles. The temperatures were observed to consistently change according to time and the induction heating power.

A technique utilizing wavelengths of 1150 and 1412 nm to measure the temperature of water surrounding an induction-heated small magnetic sphere is presented.

誘導加熱の小さな磁気圏を取り巻く水の近赤外温度計測

A technique to measure the temperature of water and non-turbid aqueous media surrounding an induction-heated small magnetic sphere is presented. This ...

工学

Ice-Cap: A Method for Growing Arabidopsis and Tomato Plants in 96-well Plates for High-Throughput Genotyping

It is becoming common for plant scientists to develop projects that require the genotyping of large numbers of plants. The first step in any genotyping project is to collect a tissue sample from each individual plant. The traditional approach to this task is to sample plants one-at-a-time. If one wishes to genotype hundreds or thousands of individuals, however, using this strategy results in a significant bottleneck in the genotyping pipeline. The Ice-Cap method that we describe here provides a high-throughput solution to this challenge by allowing one scientist to collect tissue from several thousand seedlings in a single day 1,2. This level of throughput is made possible by the fact that tissue is harvested from plants 96-at-a-time, rather than one-at-a-time.
The Ice-Cap method provides an integrated platform for performing seedling growth, tissue harvest, and DNA extraction. The basis for Ice-Cap is the growth of seedlings in a stacked pair of 96-well plates. The wells of the upper plate contain plugs of agar growth media on which individual seedlings germinate. The roots grow down through the agar media, exit the upper plate through a hole, and pass into a lower plate containing water. To harvest tissue for DNA extraction, the water in the lower plate containing root tissue is rapidly frozen while the seedlings in the upper plate remain at room temperature. The upper plate is then peeled away from the lower plate, yielding one plate with 96 root tissue samples frozen in ice and one plate with 96 viable seedlings. The technique is named "Ice-Cap" because it uses ice to capture the root tissue. The 96-well plate containing the seedlings can then wrapped in foil and transferred to low temperature. This process suspends further growth of the seedlings, but does not affect their viability. Once genotype analysis has been completed, seedlings with the desired genotype can be transferred from the 96-well plate to soil for further propagation. We have demonstrated the utility of the Ice-Cap method using Arabidopsis thaliana, tomato, and rice seedlings. We expect that the method should also be applicable to other species of plants with seeds small enough to fit into the wells of 96-well plates.

Ice-Cap: A Method for Growing Arabidopsis and Tomato Plants in 96-well Plates for High-Throughput Genotyping

The Ice-Cap method allows one to grow plants in 96-well plates and non-destructively harvest root tissue from each seedling. DNA extracted from this root tissue can be used for genotyping reactions. We have found that Ice-Cap works well for Arabidopsis thaliana, tomato, and rice seedlings.

アイスキャップ：成長させる方法シロイヌナズナ</emハイスループットジェノタイピング用96ウェルプレートで>とトマト

It is becoming common for plant scientists to develop projects that require the genotyping of large numbers of plants. The first step in any genotyping ...

生物学

Using Flatbed Scanners to Collect High-resolution Time-lapsed Images of the Arabidopsis Root Gravitropic Response

Research efforts in biology increasingly require use of methodologies that enable high-volume collection of high-resolution data. A challenge laboratories can face is the development and attainment of these methods. Observation of phenotypes in a process of interest is a typical objective of research labs studying gene function and this is often achieved through image capture. A particular process that is amenable to observation using imaging approaches is the corrective growth of a seedling root that has been displaced from alignment with the gravity vector. Imaging platforms used to measure the root gravitropic response can be expensive, relatively low in throughput, and/or labor intensive. These issues have been addressed by developing a high-throughput image capture method using inexpensive, yet high-resolution, flatbed scanners. Using this method, images can be captured every few minutes at 4,800 dpi. The current setup enables collection of 216 individual responses per day. The image data collected is of ample quality for image analysis applications.

This protocol describes a process for rapid collection of images of Arabidopsis seedlings responding to a gravity stimulus using commercially-available flatbed scanners. The method allows for inexpensive, high-volume capture of high-resolution images amenable for downstream analysis algorithms.

シロイヌナズナ根重力屈性反応の高分解能時間経過画像を収集するためにフラットベッドスキャナを使用して

Research efforts in biology increasingly require use of methodologies that enable high-volume collection of high-resolution data. A challenge laboratories ...

Identification of Novel Regulators of Plant Transpiration by Large-Scale Thermal Imaging Screening in Helianthus Annuus

Plant adaptation to biotic and abiotic stresses is governed by a variety of factors, among which the regulation of stomatal aperture in response to water deficit or pathogens plays a crucial role. Identifying small molecules that regulate stomatal movement can therefore contribute to understanding the physiological basis by which plants adapt to their environment. Large-scale screening approaches that have been used to identify regulators of stomatal movement have potential limitations: some rely heavily on the abscisic acid (ABA) hormone signaling pathway, therefore excluding ABA-independent mechanisms, while others rely on the observation of indirect, long-term physiological effects such as plant growth and development. The screening method presented here allows the large-scale treatment of plants with a library of chemicals coupled with a direct quantification of their transpiration by thermal imaging. Since evaporation of water through transpiration results in leaf surface cooling, thermal imaging provides a non-invasive approach to investigate changes in stomatal conductance over time. In this protocol, Helianthus annuus seedlings are grown hydroponically and then treated by root feeding, in which the primary root is cut and dipped into the chemical being tested. Thermal imaging followed by statistical analysis of cotyledonary temperature changes over time allows for the identification of bioactive molecules modulating stomatal aperture. Our proof-of-concept experiments demonstrate that a chemical can be carried from the cut root to the cotyledon of the sunflower seedling within 10 minutes. In addition, when plants are treated with ABA as a positive control, an increase in leaf surface temperature can be detected within minutes. Our method thus allows the efficient and rapid identification of novel molecules regulating stomatal aperture.

Identification of Novel Regulators of Plant Transpiration by Large-Scale Thermal Imaging Screening in Helianthus Annuus

We provide a method for identifying modulators of foliar transpiration by large-scale screening of a compound library.

ヘリアンサス・アンヌウスにおける大規模な熱画像スクリーニングによる植物蒸散の新規レギュレーターの同定

Plant adaptation to biotic and abiotic stresses is governed by a variety of factors, among which the regulation of stomatal aperture in response to water ...

シロイヌナズナ実生のクロロフィル生合成の高分解能イメージング

Non-invasive Assay for Chlorophyll Biosynthesis Kinetics Determination during Early Stages of Arabidopsis De-etiolation

Chlorophyll biosynthesis is a hallmark of de-etiolation, one of the most dramatic stages in the plant life cycle. The tightly controlled and highly dynamic process of chlorophyll biosynthesis is triggered during the shift from the dark to the light in flowering plants. At the moment when etiolated seedlings are exposed to the first traces of sunlight, rapid (in order of seconds) conversion of protochlorophyllide into chlorophyllide is mediated by unique light-accepting protein complexes, leading via subsequent metabolic steps to the production of fully functional chlorophyll. Standard techniques for chlorophyll content analysis include pigment extraction from detached plant tissues, which does not apply to studying such fast processes. To investigate chlorophyll kinetics in vivo with high accuracy and spatiotemporal resolution in the first hours after light-induced de-etiolation, an instrument and protocol were developed. Here, we present a detailed procedure designed for statistically robust quantification of chlorophyll in the early stages of Arabidopsis de-etiolation.

著者スポットライト:脱エチオレーション中のクロロフィル合成の非侵襲的高分解能測定

Here, we describe an advanced tool designed for chlorophyll biosynthesis monitoring during the early stages of Arabidopsis seedling de-etiolation. The novel methodology provides non-invasive real-time chlorophyll fluorescence imaging at high spatial and temporal resolution.

シロイヌナズナ脱etiolationの初期段階におけるクロロフィル生合成速度測定のための非侵襲的アッセイ

Chlorophyll biosynthesis is a hallmark of de-etiolation, one of the most dramatic stages in the plant life cycle. The tightly controlled and highly ...