Name: high-throughput, microscale protocol for the analysis of processing parameters and nutritional qualities in maize (zea mays l.)
Uploaded: 2018-06-16T15:00:00.000+00:00
Duration: 5 min 55 s
Description: Watch this Scientific Journal Video about High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize Zea mays L. at JoVE.com

作者要感谢汤姆帕特森和陶氏 AgroSciences 的分析技术小组使用他们的实验室设施和他们的导师。这项工作的资金部分是通过凯洛格公司和陶氏 AgroSciences 的礼物, 并通过美国农业部舱口赠款, ILLU-802-354 奖。学生支持 CJBW 是由伊利诺伊州杰出的奖学金和威廉 B 和南茜·刘汉铨的作物科学奖学金。

1. 生产煮熟的面粉
<ol><li>在电加热板上放置一个15升罐装压力锅。</li>
	<li>将自来水1升放入罐装压力锅, 加热至100摄氏度。</li>
	<li>当水加热时, 在1夸脱罐装罐中放置100克工业剥落粒或剥落砂砾材料 (12% 含水量, 湿基)7的样品。 注: 本研究的代表性结果是基于威严 macke等的剥落砂砾材料。9采用 Rausch等的实验室尺度干铣工艺规程。7
</li>
	<li>添加糖盐溶液, 由200毫升蒸馏水、2克盐、6克粒状白糖和2克液态麦芽萃取物组成。 注: 可以一次分析多个样品, 虽然样品的确切数量将取决于罐头压力锅的大小。</li>
	<li>用玻璃搅拌棒将溶液与剥落砂砾材料混合。</li>
	<li>在罐装压力锅中的水开始沸腾后, 添加1升的自来水以冷却罐装压力锅中的水。</li>
	<li>把罐头罐放在罐装压力锅里, 这样它们就能彼此等距, 从罐装压力锅的墙上。</li>
</ol><img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57809/57809fig1.jpg"> 图 1: 在罐装压力锅中放置罐装罐.罐头罐应该被放置相等从彼此和从罐头压力锅的边, 保证甚而烹调和避免损坏罐头罐。<a href="https://www.jove.com/files/ftp_upload/57809/57809fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="8"><li>让水沸腾。把盖子放在罐装压力锅上。</li>
	<li>在 15 psi 上烹调大的剥落的面粉或剥落的砂砾材料为一 h. 允许罐装压力锅在打开前完全冷却和减压。</li>
	<li>使用耐热手套从罐装压力锅上取下盖子。</li>
	<li>使用钳子从罐装压力锅中取出罐装罐。把罐子放在耐热的表面上。 注: 所产生的中间产品在这一点上是熟的面粉。</li>
	<li>如果使用 Rausch 所概述的协议所产生的剥落砂砾材料 。7,烹调后用刮刀除去非胚乳材料。如果使用工业剥落磨料, 跳过这一步。</li>
</ol><img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57809/57809fig2.jpg"> 图 2: 去除非胚乳材料.(a) 在烹调过程中, 在去除非胚乳材料之前, 煮熟的砂砾样品。(b) 去除非胚乳材料后的熟砂样品。<a href="https://www.jove.com/files/ftp_upload/57809/57809fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="13"><li>将30克熟的面粉 (每加工样品) 放在一条重量船上, 在烤箱中干燥65摄氏度, 12 小时。烘干后, 用咖啡机研磨熟砂样品, 并将其贮存在阴凉干燥处进行酚类分析。</li>
</ol>2. 生产烘烤的面粉
<ol><li>
把剩下的熟的面粉放在铝箔衬里的烘烤板上。
	<ol>
<li>要提高吞吐量, 请同时烘烤两个样本。要做到这一点, 在烘烤板上创建两个箔船。这消除了样品间交叉污染的可能性。</li>
	</ol>
</li>
</ol><img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57809/57809fig3.jpg"> 图 3: 在烘烤板上放置熟的面粉.在烘烤前, 两个不同的熟砂样品放在单独的箔船上烘烤托盘上。照片上的小船上贴着绿胶带。这增加了议定书的吞吐量, 同时确保没有发生交叉污染。<a href="https://www.jove.com/files/ftp_upload/57809/57809fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="2"><li>
将两个样品放在预热的对流烤箱中, 放置在107.2 摄氏度 (225°F) 50 分钟的烘烤盘中。
	<ol>
<li>搅拌样品25分钟后烘烤, 以确保甚至烘烤。</li>
	</ol>
</li>
	<li>在50分钟的结束时间内, 取出含有前两个样品的烘烤盘, 并允许在室温下冷却30分钟。</li>
	<li>在冷却期结束时, 从烤粒中间产品中取30克样品。把这个样品放在一个重量船在一个烤箱在65°c 12 小时。烘干后, 用咖啡机和商店进行植物分析, 将烘烤后的砂砾试样研磨成细粉。</li>
</ol>3. 生产最终烤玉米片产品
<ol><li>
用玉米饼印刷机把烤过的面粉卷起来。
	<ol>
<li>从铝箔衬里的烤盘中取出剩余的烤粒, 将这些面粉放在大约1米长的羊皮纸纸上。 注: 为了增加吞吐量, 将羊皮纸纸长度折叠成袋是有帮助的。这将减少在随后滚动阶段丢失的样本量。</li>
		<li>通过玉米饼印刷机慢慢地在袋中喂出被烘烤的砂砾样。注意不要在印刷机上捏手指。</li>
	</ol>
</li>
</ol><img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57809/57809fig4.jpg"> 图 4: 羊皮纸纸袋.(a) 羊皮纸纵向折叠。(b) 邮袋的长而开的一侧折叠起来。(c) 长边以10°的角度再次折叠。(d) 邮袋的短、开边被折叠。这将是通过玉米饼印刷机喂养的邮袋的一侧。<a href="https://www.jove.com/files/ftp_upload/57809/57809fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="2"><li>将轧制的烤粒切成2.5 厘米 x 2.5 厘米 (1 英寸2) 方块。使用一个工具, 如比萨饼切割机, 以削减/评分的滚动面团通过羊皮纸纸。</li>
</ol><img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/57809/57809fig5.jpg"> 图5。把卷成的面团切成薄片.轧面团是通过羊皮纸的纸。<a href="https://www.jove.com/files/ftp_upload/57809/57809fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="3"><li>
打开羊皮纸, 并允许轧制, 切割, 烘烤的面粉在室温下干燥12小时。
	<ol>
<li>为了提高吞吐量, 在室温下将干燥样品储存在箔覆盖的重量船上, 直到多个样品 (通常为24或更多) 准备好进行烘烤。</li>
	</ol>
</li>
	<li>预加热对流烤箱到204.4 °c (400 °F)。将干燥的 untoasted 鳞片样品放在平烤板上。传播样本, 使样本的最小重叠发生。这甚至可以保证烘烤。</li>
	<li>把样品放在烤箱里 60–90, 直到它达到合适的颜色 (见图 6)。</li>
</ol><img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/57809/57809fig6.jpg"> 图 6: 最终烤玉米片的正确颜色.图片左侧的玉米片被烘烤了适当的时间。图片右侧的玉米片烤得太久了。<a href="https://www.jove.com/files/ftp_upload/57809/57809fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="6"><li>允许样品在室温下冷却约5分钟。这就产生了最后的烤玉米片。</li>
	<li>用咖啡机将烤好的玉米片样品研磨成细粉。</li>
</ol>4. 植物和统计分析
注意: 根据植物的确切兴趣和实验室设备, 这些分析协议可能会发生变化。
<ol><li>使用植物中概述的协议 (如 Wilmsmeyer等) 来确定内容。3遵守议定书规定的所有安全程序。</li>
	<li>使用适当的统计模型分析数据。 注: 这些示例数据是用一个 RCBD 的分割图来分析的, 其中整个地块单元是收获谷物的场图, 次要单元是加工阶段。对 SAS (9.3 版) 混合过程进行了分析, 并在 R 中生成了数据。</li>
</ol>

<ol>
	<li>United States Department of Agriculture. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> National Agricultural Statistics Service. , (2017).</li><li>USDA. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> ERS. , (2017).</li><li>Butts-Wilmsmeyer, C. J., Mumm, R. H., Bohn, M. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Concentration+of+Beneficial+Phytochemicals+in+Harvested+Grain+of+U.S.+Yellow+Dent+Maize+(Zea+mays+L.)+Germplasm.">Concentration of Beneficial Phytochemicals in Harvested Grain of U.S. Yellow Dent Maize (Zea mays L.) Germplasm.</a> Journal of Agricultural and Food Chemistry. 65 (38), 8311-8318 (2017).</li><li>Butts-Wilmsmeyer, C. 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=Changes+in+phenolic+acid+content+in+maize+during+food+product+processing.">Changes in phenolic acid content in maize during food product processing.</a> Journal of Agricultural and Food. , (2018).</li><li>Dewanto, V., Wu, X. Z., Liu, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Processed+sweet+corn+has+higher+antioxidant+activity.">Processed sweet corn has higher antioxidant activity.</a> Journal of Agricultural and Food Chemistry. 50 (17), 4959-4964 (2002).</li><li>Fast, R. B., Caldwell, E. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breakfast+cereals+and+how+they+are+made.">Breakfast cereals and how they are made.</a> Breakfast cereals and how they are made. , (2000).</li><li>Rausch, K. D., 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=Laboratory+measurement+of+yield+and+composition+of+dry-milled+corn+fractions+using+a+shortened,+single-stage+tempering+procedure.">Laboratory measurement of yield and composition of dry-milled corn fractions using a shortened, single-stage tempering procedure.</a> Cereal Chemistry. 86 (4), 434-438 (2009).</li><li>Kandhola, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Processing and Genetic Effects on Resistant Starch in Corn Flakes. , (2015).</li><li>Macke, J. A., Bohn, M. O., Rausch, K. D., Mumm, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+factors+underlying+dry-milling+efficiency+and+flaking-grit+yield+examined+in+us+maize+germplasm.">Genetic factors underlying dry-milling efficiency and flaking-grit yield examined in us maize germplasm.</a> Crop Science. 56 (5), 2516-2526 (2016).</li><li>Somavat, P., 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=A+new+lab+scale+corn+dry+milling+protocol+generating+commercial+sized+flaking+grits+for+quick+estimation+of+coproduct+yield+and+composition.">A new lab scale corn dry milling protocol generating commercial sized flaking grits for quick estimation of coproduct yield and composition.</a> Industrial Crops and Products. 109, 92-100 (2017).</li><li>Adam, 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=The+bioavailability+of+ferulic+acid+is+governed+primarily+by+the+food+matrix+rather+than+its+metabolism+in+intestine+and+liver+in+rats.">The bioavailability of ferulic acid is governed primarily by the food matrix rather than its metabolism in intestine and liver in rats.</a> Journal of Nutrition. 132 (7), 1962-1968 (2002).</li><li>Rumpagapom, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Structural Features of Cereal Bran Arabinoxylans Related to Colon Fermentation Rate. , (2011).</li><li>Wong, J. M. W., de Souza, R., Kendall, C. W. C., Emam, A., Jenkins, D. 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=Colonic+health:+Fermentation+and+short+chain+fatty+acids.">Colonic health: Fermentation and short chain fatty acids.</a> Journal of Clinical Gastroenterology. 40 (3), 235-243 (2006).</li></ol>

作者没有什么可透露的。

在整个加工过程中, 玉米基食品的营养含量的变化很可能是由于去除了增益成分和热应力5,10。然而, 在本议定书4、8的发展之前, 对处理如何影响各种营养素的确切方法进行了比较详细的研究。此外, 由于大多数实验室处理方案的规模较大, 对玉米籽粒感官和营养特性的遗传基础的研究往往是不可能的.8。在这里, 我们提出了一个微型实验室方法来研究玉米的营养和感官性状在整个食品加工。
该协议允许取样发生在剥落的砂砾阶段, 烹调后, 烘烤后, 并在轧制过程中遇到的剪切力。因此, 通过对收获玉米籽粒的补充分析, 该议定书有利于分析初始阶段基质, 以及最终的食品和加工中间阶段, 以阐明与营养成分的变化。该协议的这一关键功能使营养和感官特征在整个处理过程中得到分析, 同时也使研究人员能够选择用于这些特定分析的分析化学协议。该协议的另一个主要特点是这种微型协议的效率。首先, 这个协议使用一个小样本, 这是适当的植物育种设置 (表 3)。一公斤谷物往往产生约0.3 公斤大的剥落砂砾成分, 大约有三个大的剥落砂成分生产需要处理。其次, 该协议允许实验室处理每天约16个样本, 这比以前的要求大样本量6的协议效率高得多。
该协议可以很容易地修改, 以模仿其他加工的玉米食品产品的生产。例如, 除了即食早餐谷物9, 大型剥落的面粉还用于生产各种快餐食品。生产这些快餐食品的实验室协议可以可以预见包括调整烹调时间和烹调方法或调整烘烤时间。还有可能将本议定书的修改版本用于研究其他谷物及其各自加工的产品。加工过的谷物产品通常包括烹调、烘烤或烘烤加工阶段, 可以使用此处介绍的协议修改版本进行模仿。
此协议的一个重要限制是它的停止点很少,即一旦处理步骤开始, 它和后续步骤必须完成 (表 3)。有一个单一的停止点后, 生产的熟粒从剥落的面粉。只有在必要的时候, 熟的面粉可以放在一个密封的容器 (如密封罐) 和冷藏至多两天。然而, 储存熟的面粉较长的时间内似乎改变样品。此外, 一旦烘焙开始, 就没有停止点, 直到烤砂面团已轧制, 切割和干燥。
结论
通过这些例子结果 (见 Wilmsmeyer等。4有关更多信息), 我们证明在整个加工过程中可以监测营养成分。此外, 确定了营养变化发生的关键处理阶段。此外, 该处理协议所需的小样本大小使得在植物育种计划的范围内对多种混合动力进行了研究。利用这些杂交种, 我们确定了在整个加工过程中, 哪组混合体保持了最高浓度的不溶性的阿魏酸和 p-香豆酸。这些特征是最终烤玉米片生命起源前潜力的重要标志。11,12,13这些结果可直接用于帮助植物育种者建立繁殖种群, 以改善加工玉米产品的生命起源前潜力。
这种处理协议的主要优点之一是它不限制可以进行的营养分析。如果植物协议存在于谷物分析中, 则可用于对加工产品进行研究。此外, 由于这种处理协议能够独立进行实验室规模的食品加工和营养分析, 因此可以研究多种植物化学物质。然而, 由于使用实验室尺度处理协议产生的中间和最终加工产品很少, 因此研究植物含量的分析协议应该使用小样本量。

玉米 (玉蜀黍) 是美国最广泛栽培的粮食作物1。在 2016年, 711.2亿公斤 (28亿蒲式耳) 玉米用于人类消费2, 表明玉米在美国饮食中的重要性。玉米谷物的最大好处之一是它是一种相对廉价的商品, 但它也含有有益的植物化学物质, 如酚类物质、不饱和脂肪酸和蛋白质3。因此, 以玉米为基础的食品可能是人类有益植物化学物质的相对廉价来源。
然而, 玉米必须在人类消费之前进行加工。因此, 加工活动往往影响最终加工食品4的营养价值。例如, 在生产快餐食品和即食早餐谷物 (即冷谷物) 的过程中, 玉米籽粒是干磨的, 以生产大型剥落的面粉。在干磨过程中, 麸皮和胚芽被物理去除, 只留下胚乳物质。由于许多植物化学物质主要位于麸皮或胚芽 (例如酚类和不饱和脂肪酸) 中, 这可能导致加工食品4的营养价值显著下降。反之, 下游加工步骤可提高营养价值。例如, 许多食品加工技术包括烹调、烘烤或烘烤。在这些阶段所遇到的热应力可以提高有益植物化学物质的生物利用度5。
从食品科学和人类营养的角度来看, 知道加工如何影响到加工食品的营养价值, 可以预见, 以及如何调整加工参数可能会影响其他感官, 这将是有趣的。品质, 包括颜色, 质地和味道。一项允许在整个加工过程中监测这种品质的协议, 可用于选择玉米品种, 以改善最终加工的玉米食品产品。过去分析这些特征的两个主要障碍是可用协议的规模和吞吐量。例如, 在生产用于实验室分析的早餐谷物时, 快速和考德威尔6建议使用45.4 公斤大的剥落的面粉。这种大量的大剥落的粒度远远超过了大量的大型剥落的面粉或大型剥落砂砾材料7 , 可以生产的小地块现场试验, 是典型的植物育种项目。因此, 制定加工食品生产的微型实验室协议, 可使 (1) 植物育种者改善对食品加工者具有重要意义的营养和感官特性的玉米品种, (2) 加工者有效地设计和测试替代处理策略。
在这篇手稿中, 我们描述了一个高通量修改 Kandohla8所描述的微型处理协议, 用于生产烤玉米片从大的剥落砂砾材料。本文给出了用该处理协议研究玉米中不溶性阿魏酸和对香豆酸变化的实例实验结果。我们在这项具体研究中的目标是确定 (1) 玉米的酚酸含量如何在生产即食早餐谷物时发生变化, (2) 在哪些加工阶段发生这些变化, (3) 是否我们的实验杂交种对加工应力反应不同, 该协议可与高通量分析化学协议结合, 有效地分析营养性状。该议定书还可进行调整, 以模仿其他谷物生产的其他加工玉米食品或加工食品产品。

<table><tbody><tr><td>Canning pressure cooker</td><td>Wisconsin Aluminum Foundry Co.</td><td>Model 921</td><td>Any can be used, but it should be large enough to accommodate multiple canning jars</td></tr><tr><td>Single burner or large hot plate</td><td>Waring Professional</td><td>Model SB30</td><td>Any can be used, but it should be large enough so that canning pressure cooker can securely be placed on burner or hot plate</td></tr><tr><td>1 quart wide mouth canning jars</td><td>Ball</td><td>1440096258</td><td>Any can be used, but they should be wide mouthed quart jars</td></tr><tr><td>1 L Beaker</td><td>Fisher Scientific</td><td>09-841-104</td><td></td></tr><tr><td>Stir plate</td><td>Corning</td><td>6796420D</td><td></td></tr><tr><td>Magnetic stir bar</td><td>Fisher Scientific</td><td>14-513-67</td><td></td></tr><tr><td>1 L Graduated cylinder</td><td>Kimble</td><td>20027500</td><td></td></tr><tr><td>Spatula</td><td>Wal-Mart</td><td>552145280</td><td></td></tr><tr><td>Hot pads</td><td>Wal-Mart</td><td>556501140</td><td></td></tr><tr><td>Scale</td><td>Any</td><td>NA</td><td>Mettler Toledo Model MS105DU or Similar</td></tr><tr><td>Weigh boats</td><td>Fisher Scientific</td><td>08-732-113</td><td></td></tr><tr><td>Sugar</td><td>Wal-Mart</td><td>9259244</td><td></td></tr><tr><td>Salt</td><td>Morton (Purchased at Wal-Mart)</td><td>9244849</td><td></td></tr><tr><td>Liquid malt extract</td><td>By the Cup (Purchased on Amazon)</td><td>NA</td><td>https://www.amazon.com/Barley-Malt-Extract-Syrup-Bottle/dp/B01N4SK72C</td></tr><tr><td>Labeling tape&amp;nbsp;</td><td>Fisher Scientific</td><td>15966</td><td></td></tr><tr><td>Permanent marker</td><td>Wal-Mart</td><td>55529894</td><td></td></tr><tr><td>Convection oven</td><td>Wal-Mart</td><td>1598495</td><td></td></tr><tr><td>Baking pan (usually included with oven)</td><td>Wal-Mart</td><td>1598495</td><td></td></tr><tr><td>Cooking foil</td><td>Wal-Mart</td><td>564264789</td><td></td></tr><tr><td>Tortilla press</td><td>E&amp;amp;A Hotel &amp;amp; Restaurant Equipment and Supplies</td><td>CTM-2000</td><td></td></tr><tr><td>Parchment paper</td><td>Reynolds (Purchased at Wal-Mart)</td><td>551219672</td><td></td></tr><tr><td>Pizza cutter</td><td>Farberware (Purchased at Wal-Mart)</td><td>&amp;nbsp;553012200</td><td></td></tr><tr><td>Cooling racks</td><td>Flytt (Purchased on Amazon)</td><td>NA</td><td>https://www.amazon.com/dp/B075HQY627/ref=sspa_dk_detail_7?psc=1&amp;amp;pd_rd_i=B075HQY627&amp;amp;pd_rd_wg=WaJol&amp;amp;pd_rd_r=SF07KCHMP753WAPG6ED4&amp;amp;pd_rd_w=2BOwf</td></tr><tr><td>SAS Version 9.4</td><td>SAS Institute</td><td>Version 9.4</td><td></td></tr><tr><td>R</td><td>R Foundation for Statistical Computing</td><td>Version 3.4.0</td><td></td></tr></tbody></table>

high-throughput, microscale protocol for the analysis of processing parameters and nutritional qualities in maize (zea mays l.)

玉米是美国和世界上重要的粮食作物。然而, 在人类消费之前必须处理玉米谷物。此外, 玉米杂交种的整体籽粒组成和加工特性各不相同, 对最终加工产品的质量有影响。因此, 要想从玉米生产出更健康的加工食品, 就必须知道如何优化特定种质资源的加工参数, 以此来解释籽粒组成和加工特性的差异。这包括更好地了解当前的加工技术如何影响最终加工食品的营养质量。在这里, 我们描述了一个微型协议, 既模拟处理管道生产玉米片从大的剥落颗粒, 并允许处理多个谷物样品同时。作为高通量分析管道的一部分, 可对薄片颗粒、中间加工产品或最终加工产品以及玉米籽粒本身进行营养成分分析。本程序是专门为纳入玉米育种研究项目而开发的, 可用于其他粮食作物的改造。为分析玉米中不溶性阿魏酸和香豆酸含量提供了一个实例。样品在五个不同的加工阶段被采取了。我们证明, 在微型加工过程中, 取样可以在多个阶段进行, 这种加工技术可以在专门的玉米育种项目的背景下使用, 在我们的例子中, 大部分的营养成分都丢失了。在食品加工过程中。

该协议允许对加工后的玉米食品、玉米片进行抽样和营养分析, 从大剥落的面粉开始, 并通过加工的中间阶段持续到最终产品。该议定书与 Rausch等所概述的议定书相结合。7生产混合颗粒样品中的剥落砂砾成分。介绍了全谷物、大粒砂、熟砂、烤砂、烤玉米片加工阶段对混合样品的营养含量的信息。无论在评价的杂交品种, 大部分不溶性的阿魏酸和 p-香豆酸被删除在干铣削 (图 7)。在烹调过程中, 不溶性的阿魏酸和 p-香豆酸的另一减少。在烹调过程中观察到的不溶性阿魏酸和 p-香豆酸含量的减少, 可能是由于去除了大量的非胚乳材料, 而这些物质仍然存在于大的剥落砂砾材料中。多自由度对比表明, 阿魏酸和 p-香豆酸含量保持稳定, 在其余的处理, 无论混合 (表 1)。
此外, 杂交品种在其不溶性的阿魏酸含量和 p-香豆酸含量方面的初步排名在最后加工阶段没有表明混合动力的排名 (表 2和图 8)。换言之, 在整个内核中的初始内容并不表明在加工结束时, 混合体中哪一种杂交将拥有最不溶性的阿魏酸或 p-香豆酸。因此, 为了研究加工食品营养特性的遗传特征, 必须利用微尺度过程来研究玉米籽粒。
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/57809/57809fig7.jpg"> 图 7: 在整个加工过程中改变不溶性的酚酸含量.(a) 在整个加工过程中改变不溶性的阿魏酸含量。(b) 在整个加工过程中改变不溶性的香豆酸含量。周: 全仁, 成品: 剥落砂砾, CG: 熟的砂砾, BG: 烤的砂砾, 给: 烤玉米片。不同的颜色点代表不同的杂交种。图最初发表在补充信息的烟头-Wilmsmeyer等。4<a href="https://www.jove.com/files/ftp_upload/57809/57809fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/57809/57809fig8.jpg"> 图 8: 交互图的混合处理阶段相互作用.(a) 不溶性的阿魏酸含量的相互作用图。(b) 不溶性香豆酸含量的相互作用图。相交的线表示一个变化的秩相互作用, 这意味着无论是不溶性的阿魏酸和不溶性的香豆酸含量的最终烤玉米片可以预测的基础上, 其中任一的初始内容整个内核中的植物化学物质。周: 全仁, 成品: 剥落砂砾, 对: 烤玉米片。图最初发表在补充信息的烟头-Wilmsmeyer等。4<a href="https://www.jove.com/files/ftp_upload/57809/57809fig8large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td></td>
			<td colspan="2" style="text-align: center;">阿魏酸</td>
			<td colspan="2" style="text-align: center;">对羟基香豆酸</td>
		</tr>
<tr>
<td>混合</td>
			<td style="text-align: center;">F值</td>
			<td style="text-align: center;">p值</td>
			<td style="text-align: center;">F值</td>
			<td style="text-align: center;">p值</td>
		</tr>
<tr>
<td>B73xMO17</td>
			<td style="text-align: center;">0.07</td>
			<td style="text-align: center;">0.93</td>
			<td style="text-align: center;">0.34</td>
			<td style="text-align: center;">0.72</td>
		</tr>
<tr>
<td>B73xPHG47</td>
			<td style="text-align: center;">0.02</td>
			<td style="text-align: center;">0.98</td>
			<td style="text-align: center;">0.61</td>
			<td style="text-align: center;">0.55</td>
		</tr>
<tr>
<td>LH1xMO17</td>
			<td style="text-align: center;">0.08</td>
			<td style="text-align: center;">0.93</td>
			<td style="text-align: center;">0.14</td>
			<td style="text-align: center;">0.87</td>
		</tr>
<tr>
<td>PHJ40xLH123HT</td>
			<td style="text-align: center;">0.32</td>
			<td style="text-align: center;">0.73</td>
			<td style="text-align: center;">0.74</td>
			<td style="text-align: center;">0.48</td>
		</tr>
<tr>
<td>PH207xPHG47</td>
			<td style="text-align: center;">0.15</td>
			<td style="text-align: center;">0.86</td>
			<td style="text-align: center;">0.24</td>
			<td style="text-align: center;">0.79</td>
		</tr>
<tr>
<td>PHJ40xMO17</td>
			<td style="text-align: center;">0.01</td>
			<td style="text-align: center;">0.99</td>
			<td style="text-align: center;">0.31</td>
			<td style="text-align: center;">0.74</td>
		</tr>
<tr>
<td>PHG39xPHZ51</td>
			<td style="text-align: center;">0.06</td>
			<td style="text-align: center;">0.94</td>
			<td style="text-align: center;">0.07</td>
			<td style="text-align: center;">0.93</td>
		</tr>
</tbody></table>表 1: 多自由度对比试验了熟粒、烤粒和烤玉米片中不溶性的酚酸含量的差异。
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td></td>
			<td colspan="2" style="text-align: center;">阿魏酸</td>
			<td colspan="2" style="text-align: center;">对羟基香豆酸</td>
		</tr>
<tr>
<td></td>
			<td style="text-align: center;">F值</td>
			<td style="text-align: center;">p 值</td>
			<td style="text-align: center;">F值</td>
			<td style="text-align: center;">p 值</td>
		</tr>
<tr>
<td>杂交种</td>
			<td style="text-align: center;">7.15</td>
			<td style="text-align: center;">0.001</td>
			<td style="text-align: center;">8。7</td>
			<td style="text-align: center;">< 0.001</td>
		</tr>
<tr>
<td>自交系</td>
			<td style="text-align: center;">4.07</td>
			<td style="text-align: center;">0.007</td>
			<td style="text-align: center;">6.57</td>
			<td style="text-align: center;">< 0.001</td>
		</tr>
<tr>
<td colspan="5">注: 所有按基因分型的按加工阶段 intereactions 在α = 0.05 不显著。</td>
		</tr>
</tbody></table>表 2: 按加工阶段相互作用的基因型的意义。
<table align="center" border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td style="text-align: center;">协议步骤</td>
			<td style="text-align: center;">关键信息</td>
			<td style="text-align: center;">故障 排除</td>
			<td style="text-align: center;">高通量建议</td>
		</tr>
<tr>
<td style="text-align: center;">1.2 和1。6</td>
			<td style="text-align: center;">这两个步骤的耦合, 使水加热, 而不打破罐装罐。</td>
			<td style="text-align: center;">那</td>
			<td style="text-align: center;">在添加罐装罐之前, 加热一半的水会增加吞吐量。</td>
		</tr>
<tr>
<td style="text-align: center;">1。4</td>
			<td style="text-align: center;">那</td>
			<td style="text-align: center;">那</td>
			<td style="text-align: center;">预测量成分。 给定的量是一个罐子, 所以乘以数量或大量的配料所需的罐子的数目, 使用的步骤1.4.2。将产生的混合物均匀地分在罐罐中。</td>
		</tr>
<tr>
<td style="text-align: center;">1.4 注</td>
			<td style="text-align: center;">不要让罐子接触到罐装压力锅的边缘。他们将打破, 样品将丢失。</td>
			<td style="text-align: center;">那</td>
			<td style="text-align: center;">那</td>
		</tr>
<tr>
<td style="text-align: center;">1.9 和1.10</td>
			<td style="text-align: center;">在烹调之前, 水应该达到沸腾的沸点。</td>
			<td style="text-align: center;">如果在一个 h 后未彻底煮熟, 则检查以确保压力重量设置为 15 psi, 水是达到充分煮沸之前, 盖子放在罐头压力锅和定时器设置。</td>
			<td style="text-align: center;">那</td>
		</tr>
<tr>
<td style="text-align: center;">1.13</td>
			<td style="text-align: center;">除去在烹调过程中漂浮到顶端的非胚乳材料。 这将扭曲任何植物结果, 如果在样本中留下。</td>
			<td style="text-align: center;">如果非胚乳物质在烹调过程中不上升到顶端, 那么面粉就不会煮彻底。 请参见有关步骤1.9 和1.10 的信息。</td>
			<td style="text-align: center;">那</td>
		</tr>
<tr>
<td style="text-align: center;">1.15、2.4 和3。7</td>
			<td style="text-align: center;">研磨样品到细粉。</td>
			<td style="text-align: center;">如果植物分析似乎没有工作, 确保样品已研磨到细粉, 这样就有一个更大的表面积暴露在溶剂。</td>
			<td style="text-align: center;">那</td>
		</tr>
<tr>
<td style="text-align: center;">2。1</td>
			<td style="text-align: center;">不要让样品互相接触。 它们将成为交叉污染。</td>
			<td style="text-align: center;">那</td>
			<td style="text-align: center;">同时烘烤两个样品, 在一张烹饪纸上制作各自的箔船。</td>
		</tr>
<tr>
<td style="text-align: center;">2.2。1</td>
			<td style="text-align: center;">搅拌样品后25分钟, 以确保甚至烘烤。</td>
			<td style="text-align: center;">如果样品似乎没有均匀烘烤, 在更频繁的间隔 (例如每15分钟) 搅拌。</td>
			<td style="text-align: center;">那</td>
		</tr>
<tr>
<td style="text-align: center;">3。1</td>
			<td style="text-align: center;">将烘烤的砂砾面团放在羊皮纸纸袋中。 这样可以确保样品在冲压过程中不会丢失。</td>
			<td style="text-align: center;">如果样品开始出来的羊皮纸纸袋的末尾, 使邮袋更长。我们发现1米似乎是足够的。</td>
			<td style="text-align: center;">那</td>
		</tr>
<tr>
<td style="text-align: center;">3。2</td>
			<td style="text-align: center;">把羊皮纸纸袋收起来。</td>
			<td style="text-align: center;">如果切割工具通过羊皮纸, 使用一个更枯燥的工具。</td>
			<td style="text-align: center;">我们发现, 一个比萨切割机是最好的工具, 以削减烘烤的面粉成方形。 我们没有用这个工具剪掉羊皮纸, 但是烤的面粉仍然能够很快地被切成正方形。</td>
		</tr>
<tr>
<td style="text-align: center;">3。5</td>
			<td style="text-align: center;">变得非常舒适的颜色和不烤太久。</td>
			<td style="text-align: center;">如果样品变得太黑, 减少用于敬酒的时间。</td>
			<td style="text-align: center;">在个别箔覆盖的重量船上储存多个干燥的烘烤砂砾样品, 直到多个样品准备好进行烘烤。</td>
		</tr>
</tbody></table>表 3:关键步骤表、疑难解答步骤和建议.

Watch this Scientific Journal Video about High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize Zea mays L. at JoVE.com

High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize Zea mays L.

在这里, 我们提出了处理谷物样品的微型协议, 并将这种微型方法纳入高通量分析管道。这是对当前可用协议的更高吞吐量适应性。

玉米加工参数和营养品质分析的高通量、微尺度协议 (玉蜀黍)

Maize is an important grain crop in the United States and worldwide. However, maize grain must be processed prior to human consumption. Furthermore, whole ...

high-throughput-microscale-protocol-for-analysis-processing

Research

JoVE Journal

Environment

High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize (Zea mays L.)

6.8K 次观看.  University of Illinois at Urbana-Champaign. 在这里, 我们提出了处理谷物样品的微型协议, 并将这种微型方法纳入高通量分析管道。这是对当前可用协议的更高吞吐量适应性。

视频: High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize Zea mays L.

Quantification of Fungal Colonization, Sporogenesis, and Production of Mycotoxins Using Kernel Bioassays

The rotting of grains by seed-infecting fungi poses one of the greatest economic challenges to cereal production worldwide, not to mention serious risks to human and animal health. Among cereal production, maize is arguably the most affected crop, due to pathogen-induced losses in grain integrity and mycotoxin seed contamination. The two most prevalent and problematic mycotoxins for maize growers and food and feed processors are aflatoxin and fumonisin, produced by Aspergillus flavus and Fusarium verticillioides, respectively.
Recent studies in molecular plant-pathogen interactions have demonstrated promise in understanding specific mechanisms associated with plant responses to fungal infection and mycotoxin contamination1,2,3,4,5,6. Because many labs are using kernel assays to study plant-pathogen interactions, there is a need for a standardized method for quantifying different biological parameters, so results from different laboratories can be cross-interpreted. For a robust and reproducible means for quantitative analyses on seeds, we have developed in-lab kernel assays and subsequent methods to quantify fungal growth, biomass, and mycotoxin contamination. Four sterilized maize kernels are inoculated in glass vials with a fungal suspension (106) and incubated for a predetermined period. Sample vials are then selected for enumeration of conidia by hemocytometer, ergosterol-based biomass analysis by high performance liquid chromatography (HPLC), aflatoxin quantification using an AflaTest fluorometer method, and fumonisin quantification by HPLC.

The devastation of cereal crops by seed-infecting fungi has prompted numerous research efforts to better understand plant-pathogen interactions. To study seed-fungal interactions in a laboratory setting, we developed a robust method for the quantification of fungal reproduction, biomass, and mycotoxin contamination using kernel bioassays.

使用内核生物测定真菌孢子发生和霉菌毒素生产的定量

The rotting of grains by seed-infecting fungi poses one of the greatest economic challenges to cereal production worldwide, not to mention serious risks ...

科研

免疫与感染

Quantifying Plant Soluble Protein and Digestible Carbohydrate Content, Using Corn (Zea mays) As an Exemplar

Elemental data are commonly used to infer plant quality as a resource to herbivores. However, the ubiquity of carbon in biomolecules, the presence of nitrogen-containing plant defensive compounds, and variation in species-specific correlations between nitrogen and plant protein content all limit the accuracy of these inferences. Additionally, research focused on plant and/or herbivore physiology require a level of accuracy that is not achieved using generalized correlations. The methods presented here offer researchers a clear and rapid protocol for directly measuring plant soluble proteins and digestible carbohydrates, the two plant macronutrients most closely tied to animal physiological performance. The protocols combine well characterized colorimetric assays with optimized plant-specific digestion steps to provide precise and reproducible results. Our analyses of different sweet corn tissues show that these assays have the sensitivity to detect variation in plant soluble protein and digestible carbohydrate content across multiple spatial scales. These include between-plant differences across growing regions and plant species or varieties, as well as within-plant differences in tissue type and even positional differences within the same tissue. Combining soluble protein and digestible carbohydrate content with elemental data also has the potential to provide new opportunities in plant biology to connect plant mineral nutrition with plant physiological processes. These analyses also help generate the soluble protein and digestible carbohydrate data needed to study nutritional ecology, plant-herbivore interactions and food-web dynamics, which will in turn enhance physiology and ecological research.

Quantifying Plant Soluble Protein and Digestible Carbohydrate Content, Using Corn Zea mays As an Exemplar

The protocols described herein provide a clear and approachable methodology for measuring soluble protein and digestible (non-structural) carbohydrate content in plant tissues. The ability to quantify these two plant macronutrients has significant implications for advancing the fields of plant physiology, nutritional ecology, plant-herbivore interactions and food-web ecology.

以玉米 (玉蜀黍) 为例量化植物可溶性蛋白和可消化碳水化合物含量

Elemental data are commonly used to infer plant quality as a resource to herbivores. However, the ubiquity of carbon in biomolecules, the presence of ...

环境科学

Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy

In plant mitochondria, some steady-state transcripts have 5&#39; triphosphate derived from transcription initiation (primary transcripts), while the others contain 5&#39; monophosphate generated post-transcriptionally (processed transcripts). To discriminate between the two types of transcripts, several strategies have been developed, and most of them depend on presence/absence of 5&#39; triphosphate. However, the triphosphate at primary 5&#39; termini is unstable, and it hinders a clear discrimination of the two types of transcripts. To systematically differentiate and map the primary and processed transcripts stably accumulated in maize mitochondrion, we have developed a circular RT-PCR (cRT-PCR)-based strategy by combining cRT-PCR, RNA 5&#39; polyphoshpatase treatment, quantitative RT-PCR (RT-qPCR), and Northern blot. As an improvement, this strategy includes an RNA normalization step to minimize the influence of unstable 5&#39; triphosphate.
In this protocol, the enriched mitochondrial RNA is pre-treated by RNA 5&#39; polyphosphatase, which converts 5&#39; triphsophate to monophosphate. After circularization and reverse transcription, the two cDNAs derived from 5&#39; polyphosphatase-treated and non-treated RNAs are normalized by maize 26S mature rRNA, which has a processed 5&#39; end and is insensitive to 5&#39; polyphosphatase. After normalization, the primary and processed transcripts are discriminated by comparing cRT-PCR and RT-qPCR products obtained from the treated and non-treated RNAs. The transcript termini are determined by cloning and sequencing of the cRT-PCR products, and then verified by Northern blot.
By using this strategy, most steady-state transcripts in maize mitochondrion have been determined. Due to the complicated transcript pattern of some mitochondrial genes, a few steady-state transcripts were not differentiated and/or mapped, though they were detected in a Northern blot. We are not sure whether this strategy is suitable to discriminate and map the steady-state transcripts in other plant mitochondria or in plastids.

We present a circular RT-PCR-based strategy by combining circular RT-PCR, quantitative RT-PCR, RNA 5&#39; polyphosphatase-treatment, and Northern blot. This protocol includes a normalization step to minimize the influence of unstable 5&#39; triphosphate, and it is suitable for discriminating and mapping the primary and processed transcripts stably accumulated in maize mitochondrion.

使用基于循环 RT-PCR 的策略对玉米 Mitochondrion 中初级和已处理转录的区分和映射

In plant mitochondria, some steady-state transcripts have 5&#39; triphosphate derived from transcription initiation (primary transcripts), while the ...

遗传学

Inhibition of Aspergillus flavus Growth and Aflatoxin Production in Transgenic Maize Expressing the &#945;-amylase Inhibitor from Lablab purpureus L.

Aflatoxin contamination in food and feed crops is a major challenge worldwide. Aflatoxins, produced by the fungus Aspergillus flavus (A. flavus) are potent carcinogens that substantially reduce crop value in maize and other oil rich crops like peanut besides posing serious threat to human and animal health. Different approaches, including traditional breeding, transgenic expression of resistance associated proteins, and RNA interference (RNAi)-based host-induced gene silencing of critical A. flavus gene targets, are being evaluated to increase aflatoxin resistance in susceptible crops. Past studies have shown an important role of &#945;-amylase in A. flavus pathogenesis and aflatoxin production, suggesting this gene/enzyme is a potential target to reduce both A. flavus growth and aflatoxin production. In this regard, the current study was undertaken to evaluate heterologous expression (under control of the constitutive CaMV 35S promoter) of a Lablab purpureus L. &#945;-amylase inhibitor-like protein (AILP) in maize against A. flavus. AILP is a 36-kDa protein, which is a competitive inhibitor of A. flavus &#945;-amylase enzyme and belongs to the lectin&#8211;arcelin&#8211;&#945;-amylase inhibitor protein family in common bean. In vitro studies prior to the current work had demonstrated the role of AILP in inhibition of A. flavus &#945;-amylase activity and fungal growth. Fungal growth and aflatoxin production in mature kernels were monitored in real time using a GFP-expressing A. flavus strain. This kernel screening assay (KSA) is very simple to set up and provides reliable and reproducible data on infection and the extent of spread that could be quantified for evaluation of germplasm and transgenic lines. The fluorescence from the GFP strain is closely correlated to fungal growth and, by extension, it is well-correlated to aflatoxin values.&#160; The goal of the current work was to implement this previous knowledge in a commercially important crop like maize to increase aflatoxin resistance. Our results show a 35%&#8211;72% reduction in A. flavus growth in AILP-expressing transgenic maize kernels which, in turn, translated into a 62%&#8211;88% reduction in aflatoxin levels.

Inhibition of Aspergillus flavus Growth and Aflatoxin Production in Transgenic Maize Expressing the &#945;-amylase Inhibitor from Lablab purpureus L.

Here we present a protocol to analyze Aspergillus flavus growth and aflatoxin production in maize kernels expressing an antifungal protein.&#160; Using a GFP-expressing A. flavus strain we monitored the infection and spread of the fungus in mature kernels in real time. The assay is rapid, reliable, and reproducible.

转基因玉米中黄曲霉生长和黄曲霉毒素的产生表达表达.

Aflatoxin contamination in food and feed crops is a major challenge worldwide. Aflatoxins, produced by the fungus Aspergillus flavus (A. flavus) are ...

Lignin Down-regulation of Zea mays via dsRNAi and Klason Lignin Analysis

To facilitate the use of lignocellulosic biomass as an alternative bioenergy resource, during biological conversion processes, a pretreatment step is needed to open up the structure of the plant cell wall, increasing the accessibility of the cell wall carbohydrates. Lignin, a polyphenolic material present in many cell wall types, is known to be a significant hindrance to enzyme access. Reduction in lignin content to a level that does not interfere with the structural integrity and defense system of the plant might be a valuable step to reduce the costs of bioethanol production. In this study, we have genetically down-regulated one of the lignin biosynthesis-related genes, cinnamoyl-CoA reductase (ZmCCR1) via a double stranded RNA interference technique. The ZmCCR1_RNAi construct was integrated into the maize genome using the particle bombardment method. Transgenic maize plants grew normally as compared to the wild-type control plants without interfering with biomass growth or defense mechanisms, with the exception of displaying of brown-coloration in transgenic plants leaf mid-ribs, husks, and stems. The microscopic analyses, in conjunction with the histological assay, revealed that the leaf sclerenchyma fibers were thinned but the structure and size of other major vascular system components was not altered. The lignin content in the transgenic maize was reduced by 7-8.7%, the crystalline cellulose content was increased in response to lignin reduction, and hemicelluloses remained unchanged. The analyses may indicate that carbon flow might have been shifted from lignin biosynthesis to cellulose biosynthesis. This article delineates the procedures used to down-regulate the lignin content in maize via RNAi technology, and the cell wall compositional analyses used to verify the effect of the modifications on the cell wall structure.

Lignin Down-regulation of Zea mays via dsRNAi and Klason Lignin Analysis

A double stranded RNA interference (dsRNAi) technique is employed to down-regulate the maize cinnamoyl coenzyme A reductase (ZmCCR1) gene to lower plant lignin content. Lignin down-regulation from the cell wall is visualized by microscopic analyses and quantified by the Klason method. Compositional changes in hemicellulose and crystalline cellulose are analyzed.

木质素下调<em&gt;玉米</em&gt;通过dsRNAi和Klason木质素分析

To facilitate the use of lignocellulosic biomass as an alternative bioenergy resource, during biological conversion processes, a pretreatment step is ...

生物工程

Kinematic Analysis of Cell Division and Expansion: Quantifying the Cellular Basis of Growth and Sampling Developmental Zones in Zea mays Leaves

Growth analyses are often used in plant science to investigate contrasting genotypes and the effect of environmental conditions. The cellular aspect of these analyses is of crucial importance, because growth is driven by cell division and cell elongation. Kinematic analysis represents a methodology to quantify these two processes. Moreover, this technique is easy to use in non-specialized laboratories. Here, we present a protocol for performing a kinematic analysis in monocotyledonous maize (Zea mays) leaves. Two aspects are presented: (1) the quantification of cell division and expansion parameters, and (2) the determination of the location of the developmental zones. This could serve as a basis for sampling design and/or could be useful for data interpretation of biochemical and molecular measurements with high spatial resolution in the leaf growth zone. The growth zone of maize leaves is harvested during steady-state growth. Individual leaves are used for meristem length determination using a DAPI stain and cell-length profiles using DIC microscopy. The protocol is suited for emerged monocotyledonous leaves harvested during steady-state growth, with growth zones spanning at least several centimeters. To improve the understanding of plant growth regulation, data on growth and molecular studies must be combined. Therefore, an important advantage of kinematic analysis is the possibility to correlate changes at the molecular level to well-defined stages of cellular development. Furthermore, it allows for a more focused sampling of specified developmental stages, which is useful in case of limited budget or time.

Kinematic Analysis of Cell Division and Expansion: Quantifying the Cellular Basis of Growth and Sampling Developmental Zones in Zea mays Leaves

Quantifying cell division and expansion is of crucial importance to the understanding of whole-plant growth. Here, we present a protocol to calculate cellular parameters determining maize leaf growth rates and highlight the use of these data for investigating molecular growth regulatory mechanisms by directing developmental stage-specific sampling strategies.

细胞分裂和扩张的运动学分析:量化增长和采样产业开发区的细胞学基础<em&gt;玉米</em&gt;叶

Growth analyses are often used in plant science to investigate contrasting genotypes and the effect of environmental conditions. The cellular aspect of ...

生物学

Micron-scale Phenotyping Techniques of Maize Vascular Bundles Based on X-ray Microcomputed Tomography

It is necessary to accurately quantify the anatomical structures of maize materials based on high-throughput image analysis techniques. Here, we provide a 'sample preparation protocol' for maize materials (i.e., stem, leaf, and root) suitable for ordinary microcomputed tomography (micro-CT) scanning. Based on high-resolution CT images of maize stem, leaf, and root, we describe two protocols for the phenotypic analysis of vascular bundles: (1) based on the CT image of maize stem and leaf, we developed a specific image analysis pipeline to automatically extract 31 and 33 phenotypic traits of vascular bundles; (2) based on the CT image series of maize root, we set up an image processing scheme for the three-dimensional (3-D) segmentation of metaxylem vessels, and extracted two-dimensional (2-D) and 3-D phenotypic traits, such as volume, surface area of metaxylem vessels, etc. Compared with traditional manual measurement of vascular bundles of maize materials, the proposed protocols significantly improve the efficiency and accuracy of micron-scale phenotypic quantification.

We provide a novel method to improve the X-ray absorption contrast of maize tissue suitable for ordinary microcomputed tomography scanning. Based on CT images, we introduce a set of image-processing workflows for different maize materials to effectively extract microscopic phenotypes of vascular bundles of maize.

基于 X 射线 Microcomputed 层析成像的玉米血管束微米尺度分型技术研究

It is necessary to accurately quantify the anatomical structures of maize materials based on high-throughput image analysis techniques. Here, we provide a ...

Scalable Transfection of Maize Mesophyll Protoplasts

The transfection of maize mesophyll cells often involves digesting the plant cell walls to create protoplasts and then inserting DNA via electroporation or polyethylene glycol (PEG). Previous methods were developed to produce tens of thousands of transfected protoplasts at once. Here,&#160;we describe a straightforward method to isolate and transfect millions of leaf mesophyll protoplasts in maize (Zea mays L.). This streamlined process removes certain common protoplasting steps, such as washing in W5. Additionally, steps such as centrifugation, PEG-mediated transfection, and incubation have been modified to work with a greater number of protoplasts. The ability to express large libraries of plasmid constructs enables genome-scale experiments, such as massively parallel reporter assays in maize.

Here, we present a high-throughput protocol to transiently transfect millions of maize protoplasts for testing large libraries of plasmids in maize mesophyll cells.

玉米叶肉原生质体的可扩展转染

The transfection of maize mesophyll cells often involves digesting the plant cell walls to create protoplasts and then inserting DNA via electroporation ...

改进的玉米叶片原基成像方法

Improved Methods for Preparing Transverse Sections and Unrolled Whole Mounts of Maize Leaf Primordia for Fluorescence and Confocal Imaging

In maize (Zea mays) and other grasses (Poaceae), the leaf primordia are deeply ensheathed and rolled within the leaf whorl, making it difficult to study early leaf development. Here, we describe methods for preparing transverse sections and unrolled whole mounts of maize leaf primordia for fluorescence and confocal imaging. The first method uses a wire stripper to remove the upper portions of older leaves, exposing the tip of the leaf primordium and allowing its measurement for more accurate transverse section sampling. The second method uses clear, double-sided nano tape to unroll and mount whole-leaf primordia for imaging. We show the utility of the two methods in visualizing and analyzing fluorescent protein reporters in maize. These methods provide a solution to the challenges presented by the distinctive morphology of maize leaf primordia and will be useful for visualizing and quantifying leaf anatomical and developmental traits in maize and other grass species.

作者聚焦：用于荧光和共聚焦成像的玉米叶片原基横向切片和展开整片的改进方法  

Maize leaf primordia are deeply ensheathed and rolled, making them difficult to study. Here, we present methods for preparing transverse sections and unrolled whole mounts of maize leaf primordia for fluorescence and confocal imaging.

用于荧光和共聚焦成像的玉米叶原基横向切片和展开整片的制备方法的改进方法

In maize (Zea mays) and other grasses (Poaceae), the leaf primordia are deeply ensheathed and rolled within the leaf whorl, making it difficult to study ...

玉米加工参数和营养品质分析的高通量、微尺度协议 (玉蜀黍)

摘要

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此视频中的章节

使用内核生物测定真菌孢子发生和霉菌毒素生产的定量

以玉米 (玉蜀黍) 为例量化植物可溶性蛋白和可消化碳水化合物含量

使用基于循环 RT-PCR 的策略对玉米 Mitochondrion 中初级和已处理转录的区分和映射

转基因玉米中黄曲霉生长和黄曲霉毒素的产生表达表达.

木质素下调

细胞分裂和扩张的运动学分析:量化增长和采样产业开发区的细胞学基础

基于 X 射线 Microcomputed 层析成像的玉米血管束微米尺度分型技术研究

玉米叶肉原生质体的可扩展转染

用于荧光和共聚焦成像的玉米叶原基横向切片和展开整片的制备方法的改进方法

用于荧光和共聚焦成像的玉米叶原基横向切片和展开整片的制备方法的改进方法