Name: analysis of effect of compound salt stress on seed germination and salt tolerance analysis of pepper (capsicum annuum l.)
Uploaded: 2022-11-30T15:00:00
Description: Watch this Scientific Journal Video about Analysis of Effect of Compound Salt Stress on Seed Germination and Salt Tolerance Analysis of Pepper Capsicum annuum L. at JoVE.com

这项工作得到了江西省科技厅（20203BBFL63065）和江西省教育厅科技研究项目（GJJ211430）的支持。我们要感谢意得辑（www.editage.cn）的英文编辑。

注意：本文提出了一种评估不同混合盐胁迫下辣椒种子萌发和幼苗生长的响应特征和内部机制的协议，可作为种子耐盐性评估的参考方法。
1. 实验准备
<ol><li>为栽培品种准备作物种子——耐盐性强的红天湖101和低耐盐的新乡8号。</li>
	<li>制备0.2%KMnO4 溶液作为种子消毒试剂。首先，称取 4.0 g KMnO4，然后加入 2，000 mL 蒸馏水。 注意：高锰酸钾由?...

<ol>
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S., Rossaneis, A. C., Pinho-Ribeiro, F. A., Verri, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capsaicin:+Current+understanding+of+its+mechanisms+and+therapy+of+pain+and+other+pre-clinical+and+clinical+uses.">Capsaicin: Current understanding of its mechanisms and therapy of pain and other pre-clinical and clinical uses.</a> Molecules. 21 (7), 844-878 (2016).</li><li>Zhao, Z., 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=Investigation,+collection+and+identification+of+pepper+germplasm+resources+in+Guangxi.">Investigation, collection and identification of pepper germplasm resources in Guangxi.</a> Journal of Plant Genetic .Resources. 21 (4), 908-913 (2020).</li><li>Zhang, J., 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=Biochar+alleviated+the+salt+stress+of+induced+saline+paddy+soil+and+improved+the+biochemical+characteristics+of+rice+seedlings+differing+in+salt+tolerance.">Biochar alleviated the salt stress of induced saline paddy soil and improved the biochemical characteristics of rice seedlings differing in salt tolerance.</a> Soil and Tillage Research. 195, 104372-104381 (2019).</li><li>Ashraf, M., Foolad, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-sowing+seed+treatment-A+shotgun+approach+to+improve+germination,+plant+growth,+and+crop+yield+under+saline+and+non-saline+conditions.">Pre-sowing seed treatment-A shotgun approach to improve germination, plant growth, and crop yield under saline and non-saline conditions.</a> Advances in Agronomy. 88, 223-271 (2005).</li><li>Esra, K. O. &. #. 1. 9. 9. ;., &#220;st&#252;n, A. S., &#304;&#351;lek, C., Arici, Y. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+exogenously+applied+spermine+and+putrescine+on+germination+and+in+vitro+growth+of+pepper+(Capsicum+annuum+l.)+seeds+under+salt+stress.">Effect of exogenously applied spermine and putrescine on germination and in vitro growth of pepper (Capsicum annuum l.) seeds under salt stress.</a> Anadolu University Journal of Science and Technology C-Life Sciences and Biotechnology. 3 (2), 63-71 (2014).</li><li>Demir, I., Mavi, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+salt+and+osmotic+stresses+on+the+germination+of+pepper+seeds+of+different+maturation+stages.">Effect of salt and osmotic stresses on the germination of pepper seeds of different maturation stages.</a> Brazilian Archives of Biology and Technology. 51 (5), 897-902 (2008).</li><li>Khan, H. A., 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=Effect+of+seed+priming+with+NaCl+on+salinity+tolerance+of+hot+pepper+(Capsicum+annuum+L.)+at+seedling+stage.">Effect of seed priming with NaCl on salinity tolerance of hot pepper (Capsicum annuum L.) at seedling stage.</a> Soil and Environment. 28 (1), 81-87 (2009).</li><li>Zhou, L. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+salinity+stress+on+cotton+(Gossypium+hirsutum+L.)+root+growth+and+cotton+field+soil+micro-ecology.">Effects of salinity stress on cotton (Gossypium hirsutum L.) root growth and cotton field soil micro-ecology.</a> Nanjing Agricultural University. , (2010).</li><li>Ding, D. X., 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=Exogenous+zeaxanthin+alleviates+low+temperature+combined+with+low+light+induced+photosynthesis+inhibition+and+oxidative+stress+in+pepper+(Capsicum+annuum+L.)+plants.">Exogenous zeaxanthin alleviates low temperature combined with low light induced photosynthesis inhibition and oxidative stress in pepper (Capsicum annuum L.) plants.</a> Current Issues in Molecular Biology. 44 (6), 2453-2471 (2022).</li><li>Liu, Z. B., Yang, B. Z., Ou, L. J., Zou, X. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+different+Ca2++spraying+period+on+alleviating+pepper+injury+under+the+waterlogging+stress.">The impact of different Ca2+ spraying period on alleviating pepper injury under the waterlogging stress.</a> Acta Horticulturae Sinica. 42 (8), 1487-1494 (2015).</li><li>Aloui, H., Souguir, M., Latique, S., Hannachi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Germination+and+growth+in+control+and+primed+seeds+of+pepper+as+affected+by+salt+stress.">Germination and growth in control and primed seeds of pepper as affected by salt stress.</a> Cercet&#259;ri agronomice &#238;n Moldova. 47 (3), 83-95 (2014).</li><li>Zhani, K., Elouer, M. A., Aloui, H., Hannachi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selection+of+a+salt+tolerant+Tunisian+cultivar+of+chili+pepper+(Capsicum+frutescens).">Selection of a salt tolerant Tunisian cultivar of chili pepper (Capsicum frutescens).</a> EurAsian Journal of Biosciences. 6, 47-59 (2012).</li><li>Patan&#232;, C., Saita, A., Sortino, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+effects+of+salt+and+water+stress+on+seed+germination+and+early+embryo+growth+in+two+cultivars+of+sweet+sorghum.">Comparative effects of salt and water stress on seed germination and early embryo growth in two cultivars of sweet sorghum.</a> Journal of Agronomy and Crop Science. 199 (1), 30-37 (2013).</li><li>Smith, P. T., Cobb, 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=Accelerated+germination+of+pepper+seed+by+priming+with+salt+solutions+and+water.">Accelerated germination of pepper seed by priming with salt solutions and water.</a> Hortscience. 26 (4), 417-419 (2019).</li><li>Mirosavljevi&#263;, M., 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=Maize+germination+parameters+and+early+seedlings+growth+under+different+levels+of+salt+stress.">Maize germination parameters and early seedlings growth under different levels of salt stress.</a> Ratarstvo i Povrtarstvo. 50 (1), 49-53 (2013).</li><li>Khan, H. A., 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=Hormonal+priming+alleviates+salt+stress+in+hot+pepper+(Capsicum+annuum+L.).">Hormonal priming alleviates salt stress in hot pepper (Capsicum annuum L.).</a> Soil and Environment. 28 (2), 130-135 (2009).</li><li>Zhang, B. B., 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=Effects+of+simulated+salinization+on+seed+germination+and+physiological+characteristics+of+muskmelon+seedlings.">Effects of simulated salinization on seed germination and physiological characteristics of muskmelon seedlings.</a> Chinese Journal of Tropical Crops. 41 (5), 912-920 (2020).</li><li>Guzm&#225;n-Murillo, M. A., Ascencio, F., Larrinaga-Mayoral, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Germination+and+ROS+detoxification+in+bell+pepper+(Capsicum+annuum+L.)+under+NaCl+stress+and+treatment+with+microalgae+extracts.">Germination and ROS detoxification in bell pepper (Capsicum annuum L.) under NaCl stress and treatment with microalgae extracts.</a> Protoplasma. 250 (1), 33-42 (2013).</li><li>Slama, I., Abdelly, C., Bouchereau, A., Flowers, T., Savoure, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diversity,+distribution+and+roles+of+osmoprotective+compounds+accumulated+in+halophytes+under+abiotic+stress.">Diversity, distribution and roles of osmoprotective compounds accumulated in halophytes under abiotic stress.</a> Annals of Botany. 115 (3), 433-447 (2015).</li><li>Muchate, N. S., Nikalje, G. C., Rajurkar, N. S., Suprasanna, P., Nikamd, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiological+responses+of+the+halophyte+Sesuvium+portulacastrum+to+salt+stress+and+their+relevance+for+saline+soil+bio-reclamation.">Physiological responses of the halophyte Sesuvium portulacastrum to salt stress and their relevance for saline soil bio-reclamation.</a> Flora. 224, 96-105 (2016).</li><li>Javed, S. A., 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=Can+different+salt+formulations+revert+the+depressing+effect+of+salinity+on+maize+by+modulating+plant+biochemical+attributes+and+activating+stress+regulators+through+improved+N+supply.">Can different salt formulations revert the depressing effect of salinity on maize by modulating plant biochemical attributes and activating stress regulators through improved N supply.</a> Sustainability. 13 (14), 8022-8037 (2021).</li><li>Chen, J., 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=Effects+of+salt+stress+on+form+of+polyamine+and+antioxidation+in+germinating+tomato+seed.">Effects of salt stress on form of polyamine and antioxidation in germinating tomato seed.</a> Acta Pedologica Sinica. 58 (6), 1598-1609 (2021).</li></ol>

该研究方法包括影响实验结果准确性的四个关键步骤。首先，由于高盐浓度溶液中溶质含量增加导致混合盐溶解不良，以及氯化钙等试剂在水中更难溶解的溶解度低，因此称量的试剂必须在研钵中充分研磨。此外，在确定容量之前，必须 通过超声波溶解 试剂。其次，每次必须完全摇动配置的盐溶液，并将其添加到培养皿中以供使用。第三，培养皿加入盐溶液后必须保留合适的水层，每个?...

盐度是全球作物生产力的主要限制因素1.目前，世界上近19.5%的灌溉土地和2.1%的旱地受到盐碱化的影响，每年约有1%的农田退化为盐碱地。到2050年，预计50%的耕地将受到盐碱化的影响2，3。除了自然因素，如天然岩石风化和沿海附近或周围的咸雨水，地表蒸发快，降雨量少，农业管理方法不合理，加剧了土壤盐碱化的过程。土壤盐碱化抑制了植物根系的生长，并减少了水分和养分从植物根部到叶子的吸收和运输。这种抑制导致生理性缺水、营养失衡和离子毒性，导致作物产量下降和作物产量完全丧失。耕地盐碱化正逐渐成为影响全球农业粮食生产的最关键的非生物胁迫因素之一4.盐胁迫减少了可用于农业的耕地，这可能导致未来农产品供需之间的严重不平衡。因此，探讨土壤盐渍化对作物生长的影响及生理生化机制，有利于培育耐盐品种、盐渍土可持续利用和农产品安全。
辣椒（辣椒 L.）因其高营养和药用价值而在世界各地种植。例如，辣椒素是一种生物碱，负责胡椒的辛辣味。辣椒素可用于缓解疼痛， 减肥， 改善心血管， 胃肠道， 和呼吸系统， 以及其他几种应用.胡椒还富含生物活性物质，尤其是不同的抗氧化化合物（类胡萝卜素、酚类和类黄酮）和维生素C 6。目前，据报道，辣椒是我国种植面积最大的蔬菜作物，年种植面积超过1.5×106 公顷，占我国蔬菜种植总面积的8%-10%。辣椒产业已成为我国最大的蔬菜产业之一，产值最高7。然而，辣椒栽培经常受到多种生物（害虫和真菌）和非生物胁迫，特别是盐胁迫，对种子萌发、生长和发育有直接的负面影响，导致辣椒果实产量和品质下降8.
种子萌发是植物与环境相互作用的第一阶段。种子萌发对周围介质的波动高度敏感，特别是土壤盐胁迫可能对生理和代谢产生逆转作用，最终扰乱作物的正常生长、发育和形态发生9。在以往的研究中，广泛研究了盐胁迫下辣椒种子的萌发和幼苗生长;然而，大多数研究使用NaCl作为诱导应力的唯一盐10，11，12。然而，土壤盐害主要是由于钠盐、钙盐和镁盐解离产生的Na+、Ca2+、Mg2+、Cl-、CO3 2-和SO4 2-离子毒性。由于离子之间的协同作用和拮抗作用，混合盐和单一盐对作物生长发育的影响可能大不相同。然而，辣椒种子在混合盐中萌发和生长的相应特征尚不清楚。因此，本研究以耐盐性差异显著的2个辣椒品种为材料。分析7种盐等摩尔混合后不同盐浓度对辣椒种子萌发、生长及生理生化指标的影响，可以揭示辣椒种子萌发对盐胁迫的响应机制。还可为强壮辣椒幼苗的栽培以及盐碱耕地的高产优质栽培提供理论依据。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Calcium chloride</td>
			<td>Shanghai Experiment Reagent Co., Ltd.,China</td>
			<td>Analytical reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifugal machine</td>
			<td>Shanghai Luxianyi Centrifuge Instrument Co., Ltd., China</td>
			<td>TGL-16M</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge tube</td>
			<td>None</td>
			<td>None</td>
			<td></td>
		</tr>
		<tr>
			<td>Conductivity meter</td>
			<td>Shanghai Instrument&#38;Electronics Science Instrument Co., Ltd., China</td>
			<td>DDSJ-308F</td>
			<td></td>
		</tr>
		<tr>
			<td>Constant temperature and humidity box</td>
			<td>Ningbo Laifu Technology Co., Ltd.,China</td>
			<td>PSX-280H</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital display vernier caliper</td>
			<td>Deli Group Co., Ltd.,China</td>
			<td>DL90150</td>
			<td></td>
		</tr>
		<tr>
			<td>Electronic balance</td>
			<td>Mettler Toledo Instruments (Shanghai) Co., Ltd.,China</td>
			<td>ME802E/02</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>Hangzhou Fuyang North Wood Pulp and Paper Co., Ltd.,China</td>
			<td>GB/T1914-2017</td>
			<td></td>
		</tr>
		<tr>
			<td>Grinding rod</td>
			<td>None</td>
			<td>None</td>
			<td></td>
		</tr>
		<tr>
			<td>Hongtianhu&#160; 101</td>
			<td>Seminis Seed (Beijing) Co., Ltd.,China</td>
			<td>11933955/100147K1-137</td>
			<td></td>
		</tr>
		<tr>
			<td>Ice machine</td>
			<td>Shanghai Kehuai Instrument Co., Ltd., China</td>
			<td>IM150G</td>
			<td></td>
		</tr>
		<tr>
			<td>Liquid nitrogen</td>
			<td>None</td>
			<td>None</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium chloride</td>
			<td>Tianjin Kermel Chemical Reagent Co., Ltd.,China</td>
			<td>Analytical reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium sulfate</td>
			<td>Tianjin Kermel Chemical Reagent Co., Ltd.,China</td>
			<td>Analytical reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish</td>
			<td>Jiangsu Yizhe Teaching Instrument Co., Ltd.,China</td>
			<td>I-000163</td>
			<td></td>
		</tr>
		<tr>
			<td>Pocket knife</td>
			<td>None</td>
			<td>None</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium permanganate (KMnO4</td>
			<td>Xilong Scientific Co.,Ltd.,China</td>
			<td>Analytical reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Pure water equipment</td>
			<td>Sichuan Youpu Ultrapure Technology Co., Ltd.,China</td>
			<td>UPT-I-20T</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium bicarbonate</td>
			<td>Xilong Scientific Co.,Ltd.,China</td>
			<td>Analytical reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium carbonate</td>
			<td>Xilong Scientific Co.,Ltd.,China</td>
			<td>Analytical reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium chloride</td>
			<td>Xilong Scientific Co.,Ltd.,China</td>
			<td>Analytical reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium sulfate&#160;</td>
			<td>Xilong Scientific Co.,Ltd.,China</td>
			<td>Analytical reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Test kit</td>
			<td>Suzhou Keming, Biotechnology Co., Ltd, Suzhou.,China</td>
			<td>Spectrophotometer method</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultra-low temperature freezer</td>
			<td>SANYO Techno Solution TottoriCo.,Ltd.</td>
			<td>MDF-382</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultraviolet visible spectrophotometer</td>
			<td>Shanghai Precision Scientific Instrument Co., Ltd., China</td>
			<td>&#160;760CRT</td>
			<td></td>
		</tr>
		<tr>
			<td>Xinxiang 8</td>
			<td>Jiangxi Nongwang High Tech Co., Ltd.,China</td>
			<td>GPD Pepper 2017(360013)</td>
			<td></td>
		</tr>
	</tbody>
</table>

analysis of effect of compound salt stress on seed germination and salt tolerance analysis of pepper (capsicum annuum l.)

为确定辣椒在萌发期的耐盐性和生理机制，以耐盐性差异较大的红天湖101和新乡8品种为研究材料。使用六种 0、3、5、10、15 和 20 g/L 的混合盐浓度，使用等摩尔比得出的 Na 2 CO 3、NaHCO3、NaCl、CaCl 2、MgCl 2、MgSO4 和 Na 2SO4。为确定其效果，测定种子萌发、幼苗生长和生理等相关指标，并利用隶属函数分析对耐盐性进行综合评价。结果表明：随着混合盐浓度的增加，两个品种的萌发势、发芽指数、发芽率、种子萌发活力指数、根长和根鲜重显著降低，而相对盐率逐渐增加;地上下胚轴长度和鲜重先增加后降低，丙二醛（MDA）、脯氨酸（Pro）含量、过氧化氢酶（CAT）、过氧化物酶（POD）和超氧化物歧化酶（SOD）活性先降低后增加。红天湖101种子的发芽势、发芽指数、萌发率、种子萌发活力指数、根长、根鲜重、MDA和Pro含量以及CAT活性均高于新乡8号。但红天湖101的下胚轴长度、地上鲜重和相对盐率均低于新乡8号。耐盐性综合评价表明，随着混合盐浓度的增加，两个隶属函数指标的总加权值先增大后减小。与隶属函数值最高的5 g/L相比，盐浓度为3 g/L、10 g/L和15 g/L的指数分别降低了4.7%—11.1%、25.3%—28.3%和41.4%—45.1%。本研究为耐盐辣椒品种的选育提供了理论指导，分析了耐盐和耐盐栽培的生理机制。

种子萌发特性 随着混合盐浓度的增加，红天湖101和新乡8的发芽势和发芽指数显著降低。两个品种的盐浓度从0-3 g/L急剧下降，盐浓度从3-20 g/L缓慢而稳定地下降（图1A，B）。随着混合盐浓度的增加，两个品种的发芽率逐渐降低，而品种的相对盐率逐渐增加。在3-15 g/L盐浓度下，发芽率和相对盐率无显著差异。然而，在所有其他盐浓度下，差异是显?...

Watch this Scientific Journal Video about Analysis of Effect of Compound Salt Stress on Seed Germination and Salt Tolerance Analysis of Pepper Capsicum annuum L. at JoVE.com

Analysis of Effect of Compound Salt Stress on Seed Germination and Salt Tolerance Analysis of Pepper Capsicum annuum L.

下面的论文介绍了一种方案，用于测量两种辣椒品种的种子萌发、幼苗生长和生理指标，这些品种在响应六种混合盐浓度时具有耐盐性差异。该协议可用于评估辣椒品种的耐盐性。

复合盐胁迫对辣椒种子萌发的影响及耐盐性分析

To determine the salt tolerance and physiological mechanism of pepper (Capsicum annuum L.) at the germination stage, the Hongtianhu 101 and Xinxiang 8 ...

本研究为盐碱土耐盐辣椒品种选育、高产品质标定提供理论指导。实验周期为15天，效率高，速度快，重复性好，结果准确。该技术可为其他作物耐盐性评价提供技术参考和理论指导。希望初步专家宁愿评估种子的耐盐性。首先准备耐盐性强的红天湖101品种和低耐盐性的新乡8品种的作物种子。使用前准备2000毫升0.2%高锰酸钾溶液作为种子消毒试剂。使用七种盐制备混合盐溶液，包括碳酸钠，碳酸氢钠，氯化钠，氯化钙，氯化镁，硫酸镁和硫酸钠。然后准备一次性使用，九厘米培养皿和九厘米中速定性滤纸。从每个品种中选择大小一致、颗粒饱满的辣椒种子，红天湖101种子平均直径为4.2毫米，新乡8种子平均直径为3.7毫米。根据测试工作负载计算选择的种子总数。对于种子消毒，将选定的辣椒种子浸泡在0.2%高锰酸钾溶液中。15分钟后，用蒸馏水冲洗种子五次。将灭菌的种子转移到蒸馏水中浸泡24小时。完成后，用蒸馏水冲洗种子几次，然后干燥以备进一步使用。制备六种浓度的混合盐，并使用电导率仪测量盐溶液的电导率。在带有两层滤纸的培养皿中，均匀放置40颗辣椒种子。准备种子进行六次实验处理和五次重复。向培养皿中加入适量的六种混合盐浓度进行发芽，以确保滤纸湿润。将种子放入28摄氏度和80%空气湿度的空气培养箱中，以便在黑暗中发芽。种子发芽后，让幼苗在大约 450 勒克斯的光照强度下生长 14 天。光照周期为12小时黑暗，12小时光照期在培养箱中。每12小时补充培养皿中的溶液以保留湿滤纸，并每24小时用相应浓度的混合盐溶液彻底洗涤滤纸，以保持培养皿中恒定的混合盐浓度。对于种子萌发指标，确定发芽率，每天播种后用自由基破种皮，达到种子直径长度的一半作为发芽标记。为了确定幼苗生长指数，在播种后第14天从每个培养皿中随机选择10个代表性幼苗，并测量根长和下胚轴长度。用刀将辣椒幼苗分成自由基和地面部分。擦拭幼苗上的水分，并分别称量幼苗以确定新鲜重量。为了保存辣椒幼苗，请在播种后第 14 天从每次处理中选择大约 24 克具有代表性的全辣椒幼苗。去除地表水后，立即将幼苗在液氮中冷冻一分钟。并将它们存放在零下 80 摄氏度的冰箱中。从收集的每个处理中取出大约一克幼苗样品，一式三份。立即将幼苗样品放入离心管中。加入液氮，用研磨棒研磨样品，测定幼苗的生理指标，包括幼苗保护酶活性、丙二醛、脯氨酸含量等。随着混合盐浓度的增加，红天湖101和新乡8的发芽势和发芽指数显著降低。随着混合盐浓度的增加，两个品种的发芽率逐渐降低，而品种的相对盐害率增加。红天湖101的种子萌发活力指数在各混合盐浓度水平上均高于新乡8号。混合浓度15克/L时，红天湖101和新乡8根长分别比对照减少89.4%和91.1%，根鲜重分别减少81.7%和71.2%。基于品种差异，新乡8号下胚轴长度和地上鲜重各盐浓度均高于红天湖101号。丙二醛和脯氨酸含量在每升5克和3克浓度时分别达到最低值。在盐浓度为每升3至15克时，新乡8号脯氨酸含量缓慢增加并保持相对稳定，而红天湖101脯氨酸含量迅速增加。随着混合盐浓度的增加，红天湖101和新乡8幼苗的过氧化氢酶、过氧化物酶和超氧化物歧化酶活性先降低后升高，最低值为3克/升。红天湖101的过氧化氢酶和过氧化物酶活性均高于新乡8号，且差异逐渐增大。通过保留湿滤纸并完全清洗滤纸，保持培养皿中混合盐浓度的恒定。研究人员可以在此实验的基础上进行创新，例如改变不同混合盐的比例，以获得更广泛和深入的结果。

Research

JoVE Journal

Biology

Analysis of Effect of Compound Salt Stress on Seed Germination and Salt Tolerance Analysis of Pepper (Capsicum annuum L.)

Analysis of Physiologic E-Selectin-Mediated Leukocyte Rolling on Microvascular Endothelium

微血管内皮生理E -选择素介导白细胞滚动分析

E-selectin is a type-1 membrane protein on microvascular endothelial cells that helps initiate recruitment of circulating leukocytes to cutaneous, bone ...

科研

生物学

A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity

一个神经元和星形胶质细胞共培养检测神经毒性的高含量分析

High Content Analysis (HCA) assays combine cells and detection reagents with automated imaging and powerful image analysis algorithms, allowing ...

Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets

采用芯片识别亚型特定的靶基因的全基因组分析

Recruitment of transcriptional and epigenetic factors to their targets is a key step in their regulation. Prominently featured in recruitment are the ...

Profiling of Methyltransferases and Other S-adenosyl-L-homocysteine-binding Proteins by Capture Compound Mass Spectrometry (CCMS)

Profiling of Methyltransferases and Other S-adenosyl-L-homocysteine-binding Proteins by Capture Compound Mass Spectrometry CCMS

分析甲基化和其他 S腺苷 L</sub捕捉复合质谱（CCMS）>同型半胱氨酸结合蛋白

There is a variety of approaches to reduce the complexity of the proteome on the basis of functional small molecule-protein interactions such as affinity ...

Metabolic Profile Analysis of Zebrafish Embryos

斑马鱼胚胎的代谢谱分析

A growing goal in the field of metabolism is to determine the impact of genetics on different aspects of mitochondrial function. Understanding these ...

A Seed Coat Bedding Assay to Genetically Explore In Vitro How the Endosperm Controls Seed Germination in Arabidopsis thaliana

A Seed Coat Bedding Assay to Genetically Explore In Vitro How the Endosperm Controls Seed Germination in Arabidopsis thaliana

种皮床品检测到基因探索<em&gt;体外</em&gt;如何控制胚乳种子萌发的<em&gt;拟南芥</em

The Arabidopsis endosperm consists of a single cell layer surrounding the mature embryo and playing an essential role to prevent the germination of ...

Analysis of Translation Initiation During Stress Conditions by Polysome Profiling

翻译起始的分析在应力条件由多聚核糖体剖析

Precise control of mRNA translation is fundamental for eukaryotic cell homeostasis, particularly in response to physiological and pathological stress. ...

Analysis of Oxidative Stress in Zebrafish Embryos

在斑马鱼胚胎氧化应激分析

High levels of reactive oxygen species (ROS) may cause a change of cellular redox state towards oxidative stress condition. This situation causes ...

Measurement and Analysis of Extracellular Acid Production to Determine Glycolytic Rate

测量外产酸和分析，以确定糖酵解率

Extracellular measurement of oxygen consumption and acid production is a simple and powerful way to monitor rates of respiration and glycolysis1. Both ...

Deferred Growth Inhibition Assay to Quantify the Effect of Bacteria-derived Antimicrobials on Competition

递延生长抑制试验来量化竞争性细菌来源的抗菌剂的影响

Competitive exclusion can occur in microbial communities when, for example, an inhibitor-producing strain outcompetes its competitor for an essential ...