P23681-B16 :이 연구는 오스트리아 과학 기금 (FWF)에 의해 투자되었다.

식물 재료의 준비 (1) <ol><li> 대상 식물 재료 중 하나 잎 또는 전체 식물 (HOSTA 종. 또는 위 Phaseolus 심상)를 사용합니다. </li></ol> 얼음 핵 활동 (INA) 박테리아를 함유하는 물 솔루션 2. 준비 <ol><li> 문화는 INA 세균, 슈도모나스 시린 개 (Pseudomonas syringae)는 25 ° C 슈도모나스에 한천 (F)에서 배양 접시에 (주 CIT의-7) 제조업체의 방향을 100 % 글리세롤의 10g / ℓ로 준비했다. </li><li> 배양은 4 ℃에서 충분히 성장한 후 장소 필요하지만 얼음 핵 활성의 수준을 높일 이전 이틀 동안 4 ℃에서 유지 될 때까지. </li><li> 일회용 플라스틱과 한천의 표면으로부터 하나의 플레이트로부터 긁어 박테리아 또는 재사용 가능한 금속 주걱을 사용하여 장소시 탈 이온수 10 내지 15 ㎖에 25 ㎖ 일회용 큐벳에서. · 농도는 1 × 107 1 × 109의 범위 내에 있어야 </sup&gt; ml의 -1. 이 솔루션은 흐린 나타납니다. 농도가 단지 근사 될 필요성 혈구 또는 분광 광도계 농도를 확인할 필요는 없다. </li><li> 박테리아를 분배하는 10 초 이상 동안 큐벳을 소용돌이. 주의 : 수득 INA 혼합물의 특정한 농도는 중요하지 않으며 기술 된 프로토콜은 얼음 핵 활동의보다 적절한 레벨을 제공 할 것이다. INA 박테리아와 물의 혼합물은 핵 실험에서 나중에 사용될 것이다. </li></ol> 3. 냉동 실험 설정하기 <ol><li> 보호 아크릴 상자 안에 고해상도 적외선 카메라 (SC-660)를 올려되도록 상자 전면의 개구부를 통해 렌즈 프로젝트 및 박스의 후방 개구를 통하여 랩탑 또는 기록 장치 출구에 카메라를 연결하는 와이어 . 상자의 뚜껑을 고정하고있는 위치에 환경 챔버 또는 냉장고 내부의 상자를 배치 할 모든 것아야 대상 식물 재료를 볼 수 있습니다. <ol><li> 반사 된 적외선 에너지로부터의 간섭을 방지하기 위해 흑색 도화지와 챔버의 벽을 라이닝 식물 물질 주위의 어두운 배경을 제공한다. </li><li> 가시 파장에서의 화상 기록이 요구 될 때 광원으로부터의 가열을 최소화하기 위해 LED 조명 챔버를 장착한다. 식물이 카메라로 표시하는 등의 배터리로 작동 옷장 빛 또는 다른 작은 LED 장치로 조명 만 최소가 필요합니다. <ol><li> 대상 식물 재료의 볼 이미지가 촬영되면, LED 조명을 끄십시오. 포트 또는 챔버 내의 다른 개구부를 통해 카메라에 모든 외부 유선 연결 (컴퓨터 FireWire 연결, 전원 코드 등)를 분배. </li></ol></li><li> 피하거나 챔버 내에서 온도 기울기를 감소시키기 위해 발포 단열재와 포트 또는 개구에 여분의 공간을 채우기. 1 ° C에서 챔버의 초기 온도를 설정합니다. </li></ol></li><li> 식물 재료는 카메라의 시야에 있고, 식물 재료가 원격보기 화면 또는 선택된 소프트웨어 내에 표시되도록 식물 또는 식물 파트를 정렬. </li><li> 식물 전에 동결 제어 실험을 시작으로, 식물 재료의 크기에 따라 1 시간 30 분 동안 0 ° C에서 평형을 허용. 이 실험은 동결 시작되면 식물의 온도가 몇도만큼 공기 온도 뒤쳐하지 않을 것을 보장한다. 식물 재료의 온도는 대기 온도의 0.5 ° C 내에있을 때 평형이 달성된다. <ol><li> 화분에 심은 식물을 사용하는 경우 화분의 흙 위에 스티로폼 단열재의 층을 놓습니다. 식물이 평형 한 후, 챔버의 냉각 개시. 주 : 냄비의 토양 표면에 절연 층이 식물 주변 공기 포트로부터 계속 열 손실의 양을 감소시키고, FRE으로부터 뿌리를 방지ezing이 전형적으로 인해 토양에 잔류 열 존재의 대규모 저수지에 자연 서리 이벤트 기간 동안 발생하지 않을 것입니다. </li></ol></li><li> 3.4.1-3.4.4에서 논의 된 바와 같이, 원하는 카메라 매개 변수 (색상 팔레트, 온도 범위, 관심의 특정 영역 등)을 설정합니다. <ol><li> 라이브 영상을 보는 동안 온도 변화를 표시하는 무지개 색상 표를 선택합니다. </li><li> 단지 소프트웨어의 이미지 아래에있는 온도 막대를 조정하여 5 ° C의 온도 범위를 설정합니다. </li><li> 선택된 팔레트 (무지개)에 의해 정의 된 거짓 컬러 화상에 적외선 데이터를 변환하기위한 리니어 스케일 (알고리즘)를 선택하고, 5 ℃까지 온도 범위를 설정하고, 화상에 기초하여 자동으로 추적. 실험을 수행하는 동안 또는 수동으로 설정 범위를 조정합니다. <ol><li> 소프트웨어가 제공하는 관심의 정의 된 영역 내에서 특정 지점의 온도 또​​는 평균 온도를 사용하여이다. . 기록 된 비디오 시퀀스로부터 혹은 이미지 파일에 포함 된 정보로부터 모든 화소의 온도 데이터를 검색 ResearchIR 3 소프트웨어 내에서 일반적인 화면을 보여준다. </li></ol></li><li> 관심의 특정 지점을 나타내는 식물 조직의 위치에 커서를 놓습니다. 포인트 (1-3 사이즈 화소), 박스, 선, 타원, 또는 원으로 관심 영역을 정의. 점 또는 도형의 여러 조합이 이미지 위에 위치 될 수있다. </li></ol></li><li> 비디오 시퀀스 녹화 <ol><li> 60 Hz에서와 촬영 수동으로 중지 할 기록하기 위해 카메라를 설정합니다. </li><li> 녹화 된 비디오 파일이 배치 될 컴퓨터 또는 외부 드라이브의 위치를​​ 나타냅니다. </li><li> 녹화 시작. 참고 : 큰 비디오 파일이 생성되기 때문에 외장 하드 드라이브에 기록하는 것이 좋습니다. 녹화 된 비디오 파일은 나중에 NE 함유 부분만을 포함하도록 편집 될 수cessary 정보. 이것은 크게 파일 크기를 감소시킬 것이다. </li><li> 0.5 -1.0 ° C에 의해 점진적으로 챔버의 온도를 낮 춥니 다. 식물 온도가 대기 온도와 평형을 때까지 기다린 후 0.5 ~ 1.0 ° C로 다시 온도를 낮 춥니 다. 식물 관찰되는 조직과 형태의 질량에 따라, 평형은 10 분 (15)에 걸릴 수 있습니다. 따라서, 약 4 ° C / 시간의 냉각 속도를 제공. </li><li> 식물 얼과 관측이 완료 될 때까지 이러한 방식으로 계속. 냉동 처리가 완료되면, 기록 종료. 참고 : 그들은 동일한 온도에서 때문에 식물 재료 및 배경이 동색 인 경우에 식물 조직은 공기 온도로 평형화 하였다. 배경 온도 및 식물 조직의 온도가 동일하게되어 있기 때문에, 다시 온도를 낮추고 식물 조직과 간의 온도 차이가있을 때까지 식물 재료를 시각화하기 어려울 수있다IR 온도. </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>

저자는 더 경쟁 재정적 이해 관계 나 이해 관계의 충돌이 없습니다.

물 우물 0 ° C 이하의 온도와 물이 상당히 가변적 일 수 동결되는 온도로 과냉각하는 기능을 갖는다. 순수 과냉각 36 내열 온도는 약 -40 ° C이며 균질 핵 생성 지점으로 정의된다. 따뜻한 온도에서 물이 동결보다 -40 ° C는 다음 얼음 형성과 성장을위한 촉매 역할을하는 형성하는 작은 얼음 배아를 사용하는 것이 핵형 성제 이종의 존재에 의해 초래됩니다. (37) 자연 속에서 분자의 다?...

식물이 활발하게 성장하는 때 발생하는 온도를 동결하는 것은 식물이 거의 또는 전혀 동결 내성이 경우 특히, 치명적이 될 수 있습니다. 이러한 서리 이벤트는 종종 농업 생산에 파괴적인 영향을 가지고 있으며, 특히 고산, 서브 북극과 북극 생태계 1-6, 식물의 자연 집단에서 사회 구조 형성에 중요한 역할을 할 수있다. 심한 봄 서리의 에피소드는 최근 7-9 년 미국과 남미에서 과일 생산에 큰 영향이 있고 전형적인 평균 저온 다음 따뜻한 날씨의 초기 발병에 의해 악화되고있다. 초기의 따뜻한 날씨는 더 서리 허용 1,3,10-12 매우 작은이 모두 새로운 싹, 잎, 꽃의 성장을 활성화, 휴식 싹을 유도한다. 이러한 불규칙적 인 기상 패턴 지속적인 기후 변화의 직접적인 반사 것으로보고되었으며 FORES위한 공통 날씨 패턴이 될 것으로 예상된다eeable 미래 (13). 증가 된 서리 내성을 제공 할 수있다, 경제적 효과 및 환경 친화적 인 경영 기법이나 농약을 제공하기위한 노력이 이유 호스트의 제한적인 성공이 있었다 그러나이 부분적 내성을 동결 식물 회피 메커니즘을 동결 복잡한 성질에 기인 할 수있다. (14) 식물에서 서리 생존과 관련된 적응 메커니즘은 전통적으로 내성을 동결 방지 동결, 두 가지 범주로 구분하고있다. 전 분류는 식물의 존재와 그 조직에서 얼음의 탈수 효과와 연관된 응력을 견딜 수 있도록하는 유전자의 특정 세트에 의해 조절 생화학 적 메커니즘과 연관된다. 후자는 통상적으로, 그러나 오직, 플랜트 (14)의 구조를 결정 식물의 측면 및 얼음 형성과 연관된 상태. 광고로 동결 방지의 보급에도 불구하고aptive 메커니즘 연구는 거의 기본 메커니즘 및 동결 방지의 규정을 이해하는데 최근에 전념하고있다. 독자는이 주제에 대한 더 자세한 사항은 최근의 리뷰 (15)로 지칭된다. 저온에서 얼음의 형성이 간단한 방법처럼 보일 수도 있지만, 많은 요인은 공장 내에서 확산 식물 조직에있는 얼음 핵이 방법에서 온도를 결정하는 것에 기여한다. 같은 핵형 성제 외적인 얼음의 존재하에, 이종 대 균질 핵 생성 이벤트, 열 이력 (부동액) 단백질, 특정 당 및 다른 osmolytes의 존재, 및 식물의 구조적 측면 호스트로 파라미터 수있는 것은 중요한 플레이 식물 동결 과정에서 역할. 종합적으로, 이러한 매개 변수는 얼음이 시작하고 성장하는 방법 어디 공장이 정지되는 온도에 영향을 미친다. 또한 생성 된 얼음 결정의 형태에 영향을 미칠 수있다.다양한 방법 LTSEM (핵 자기 공명 분광법 (NMR) (16), 자기 공명 영상 (MRI) (17), 크라이 현미경 18-19, 저온 주사 전자 현미경을 포함 실험실 조건 하에서 식물 동결 과정을 연구하기 위해 사용되어왔다 ). 실험실 및 필드 설정에서 전체 식물의 20 냉동 그러나, 주로 열전대로 모니터링하고있다. 동결을 연구하는 열전대의 사용은 물이 액체로부터 고체로 상 변화를 겪는다 열 해방 (융해열)에 기초한다. 동결 후, 발열 이벤트로서 기록된다. 21-23 열전대 식물 동결 공부 선택 전형적인 방법이더라도, 이들의 사용은 이벤트 동안 동결 얻어진 정보의 양을 제한하는 많은 한계가있다. 예를 들어, 열전대와 그것이 어떻게 전파 얼음, 식물에서 개시되는 위치를 결정하는 것은 거의 불가능에 어렵고,그것은 더 속도로 전파하는 경우, 일부 조직은 얼음의 무료 유지합니다. 고해상도 적외선 체열 (HRIT) 24-27의 발전은, 그러나, 상당히 차동 촬상 모드에서 사용될 때, 특히 식물 전체에 동결 과정에 대한 정보를 획득 할 수있는 능력을 증가했다. 28-33을 본 보고서에서는 냉동 과정의 다양한 양태 및 어디에 어떤 온도에서 얼음 식물에서 개시되는 다양한 매개 변수에 영향을 연구하기 위해이 기술의 사용을 설명한다. 프로토콜은 높은, 영하의 온도에서 초본 식물에 동결 개시 외부 핵제 역할을 얼음 핵 활동 (INA) 박테리아의 능력, 슈도모나스 시린 개 (Pseudomonas syringae) (CIT의-7)를 보여 것이다 표시됩니다. 고해상도 적외선 카메라 이 보고서에 설명 된 프로토콜과 예는 적외선 높은 해상도를 사용비디오 복사계. 복사계 (도 1)는 적외선과 가시 광선 이미지 및 온도 데이터의 조합을 제공한다. 카메라의 스펙트럼 응답은 7.5-13.5 μm의 범위 인 640 × 480 화소 해상도를 제공한다. 가시 스펙트럼에 의해 생성 된 이미지 카메라 내장 복잡한 열적 이미지의 해석을 용이하게 실시간에서 IR-이미지로 융합 될 수있다. 카메라 렌즈의 범위는 확대 현미경 관찰을 할 수 있습니다. 카메라는 독립 실행 형 모드로 사용, 또는 인터페이스와 propietary 소프트웨어를 사용하여 노트북을 제어 할 수 있습니다. 소프트웨어가 기록 된 비디오 데이터에 포함 된 열 다양한를 얻기 위해 사용될 수있다. 이는 적외선 복사계의 다양한 시판 것을 주목하는 것이 중요하다. 따라서, 연구자 제품 엔지니어링 지식을 그들의 의도 된 적용을 논의하고 있음을보고 특정 연구자가 임의의 능력을 시험하는 것이 중요C 복사계 필요한 정보를 제공한다. 설명 된 프로토콜에 사용 촬상 복사계는 아크릴 상자 나 스티로폼 N 온난화 및 냉각 프로토콜 동안 축합 노출을 방지하기 위해,로 절연 (도 2)에 배치된다. 이 보호는 모든 카메라 또는 응용 프로그램에 필요하지 않습니다.

<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.

얼음 + 박테리아의 얼음 핵 활동, 슈도모나스 시린 개 (Pseudomonas syringae) (주 CIT의-7) 10 μL의 물 방울과 피를 포함하는 물 10 μL syringae (CIT의-7) hosta 리프의 배축 표면에 배치했다 (HOSTA 종.) (그림 4). 도시 된 바와 같이, INA 박테리아 함유 물 방울과 제 동결 잎 표면에 물이 얼지 드롭 유지하면서 동결 잎을 유도 할 책임이었다. <p class="jove_c...

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).

식물의 얼음 핵 얼음 전파의 연구에 대한 고해상도 적외선 체열 (HRIT)의 사용

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 조회수.  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).

비디오: 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.

빠른 레이저 방법은 잎 열적 특성의 비 침습적과 연락이없는 결정을 촉진 프로빙

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

연구

생화학

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.

식물은 아이스 친화 정제를 사용하여 얼음의 재결정 억제 및 절연의 평가를 통해 단백질을 아이스 결합의 식별

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 및 식물 형질 및 생리 적 연구에 대 한 스펙트럼 루트 이미징: 실험적인 체제 및 프로토콜을 이미징

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

환경과학

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.

식물에 동결의 3 차원 적외선 비디오를 제작 하기 위한 프로토콜

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.

산림 생태계에 얼음 폭풍의 영향 시뮬레이션

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.

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

공학

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.

얼음 총액 : 성장을위한 방법 Arabidopsis높은 처리량 Genotyping을위한 96 - 웰 플레이트에와 토마토 식물

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 ...

생물학

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.

애기 장대 루트 Gravitropic 응답의 고해상도 시간 경과 이미지를 수집하는 평판 스캐너를 사용하여

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.

헬리안투스 아누스의 대규모 열화상 스크리닝에 의한 새로운 식물 증발 규제 기관 식별

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 ...

Arabidopsis 묘목에서 엽록소 생합성의 고해상도 이미징

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.

저자 스포트라이트: De-etiolation 중 엽록소 합성의 비침습적 고분해능 측정

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.

애기장대 탈에티올화의 초기 단계 동안 엽록소 생합성 역학 측정을 위한 비침습적 분석

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