Voorend Wannes, Lootens Peter, Nelissen Hilde, Roldán-Ruiz Isabel, Inzé Dirk, Muylle Hilde
Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Gent, Belgium ; Department of Plant Biotechnology and Bioinformatics, Ghent University, Technologiepark 927, 9052 Gent, Belgium ; Plant Sciences Unit - Growth and Development, Institute for Agricultural and Fisheries Research (ILVO), Caritasstraat 21, 9090 Melle, Belgium.
Plant Sciences Unit - Growth and Development, Institute for Agricultural and Fisheries Research (ILVO), Caritasstraat 21, 9090 Melle, Belgium.
Plant Methods. 2014 Nov 6;10(1):37. doi: 10.1186/1746-4811-10-37. eCollection 2014.
In grasses, leaf growth is often monitored to gain insights in growth processes, biomass accumulation, regrowth after cutting, etc. To study the growth dynamics of the grass leaf, its length is measured at regular time intervals to derive the leaf elongation rate (LER) profile over time. From the LER profile, parameters such as maximal LER and leaf elongation duration (LED), which are essential for detecting inter-genotype growth differences and/or quantifying plant growth responses to changing environmental conditions, can be determined. As growth is influenced by the circadian clock and, especially in grasses, changes in environmental conditions such as temperature and evaporative demand, the LER profiles show considerable experimental variation and thus often do not follow a smooth curve. Hence it is difficult to quantify the duration and timing of growth. For these reasons, the measured data points should be fitted using a suitable mathematical function, such as the beta sigmoid function for leaf elongation. In the context of high-throughput phenotyping, we implemented the fitting of leaf growth measurements into a user-friendly Microsoft Excel-based macro, a tool called LEAF-E. LEAF-E allows to perform non-linear regression modeling of leaf length measurements suitable for robust and automated extraction of leaf growth parameters such as LER and LED from large datasets. LEAF-E is particularly useful to quantify the timing of leaf growth, which forms an important added value for detecting differences in leaf growth development. We illustrate the broad application range of LEAF-E using published and unpublished data sets of maize, Miscanthus spp. and Brachypodium distachyon, generated in independent experiments and for different purposes. In addition, we show that LEAF-E could also be used to fit datasets of other growth-related processes that follow the sigmoidal profile, such as cell length measurements along the leaf axis. Given its user-friendliness, ability to quantify duration and timing of leaf growth and broad application range, LEAF-E is a tool that could be routinely used to study growth processes following the sigmoidal profile.
在禾本科植物中,通常通过监测叶片生长来深入了解生长过程、生物量积累、刈割后的再生等情况。为了研究禾本科植物叶片的生长动态,需要定期测量其长度,以得出随时间变化的叶片伸长率(LER)曲线。从LER曲线中,可以确定一些参数,如最大LER和叶片伸长持续时间(LED),这些参数对于检测基因型间的生长差异和/或量化植物对不断变化的环境条件的生长反应至关重要。由于生长受生物钟影响,特别是在禾本科植物中,还受温度和蒸发需求等环境条件变化的影响,LER曲线显示出相当大的实验变异性,因此通常不遵循平滑曲线。因此,很难量化生长的持续时间和时间点。出于这些原因,应使用合适的数学函数对测量数据点进行拟合,例如用于叶片伸长的β型Sigmoid函数。在高通量表型分析的背景下,我们将叶片生长测量的拟合过程集成到一个基于Microsoft Excel的用户友好型宏中,这个工具称为LEAF-E。LEAF-E允许对叶片长度测量进行非线性回归建模,适用于从大型数据集中稳健且自动地提取叶片生长参数,如LER和LED。LEAF-E对于量化叶片生长的时间点特别有用,这对于检测叶片生长发育差异具有重要的附加价值。我们使用在独立实验中出于不同目的生成的已发表和未发表的玉米、芒草属植物和二穗短柄草的数据集,说明了LEAF-E的广泛应用范围。此外,我们表明LEAF-E还可用于拟合遵循Sigmoid曲线的其他与生长相关过程的数据集,例如沿叶轴的细胞长度测量。鉴于其用户友好性、量化叶片生长持续时间和时间点的能力以及广泛的应用范围,LEAF-E是一种可常规用于研究遵循Sigmoid曲线的生长过程的工具。