Das Shouvik, Bhati Vikrant, Dewangan Bhagwat Prasad, Gangal Apurva, Mishra Gyan Prakash, Dikshit Harsh Kumar, Pawar Prashant Anupama Mohan
Laboratory of Plant Cell Wall Biology, Regional Centre for Biotechnology, NCR Biotech Science, Cluster 3rd Milestone, Faridabad-Gurgaon Expressway, Faridabad, 121001, Haryana, India.
Division of Genetics, Indian Agricultural Research Institute, New Delhi, 110012, India.
Plant Methods. 2024 Sep 2;20(1):135. doi: 10.1186/s13007-024-01260-w.
Dissection of complex plant cell wall structures demands a sensitive and quantitative method. FTIR is used regularly as a screening method to identify specific linkages in cell walls. However, quantification and assigning spectral bands to particular cell wall components is still a major challenge, specifically in crop species. In this study, we addressed these challenges using ATR-FTIR spectroscopy as it is a high throughput, cost-effective and non-destructive approach to understand the plant cell wall composition. This method was validated by analysing different varieties of mungbean which is one of the most important legume crops grown widely in Asia.
Using standards and extraction of a specific component of cell wall components, we assigned 1050-1060 cm and 1390-1420 cm wavenumbers that can be widely used to quantify cellulose and lignin, respectively, in Arabidopsis, Populus, rice and mungbean. Also, using KBr as a diluent, we established a method that can relatively quantify the cellulose and lignin composition among different tissue types of the above species. We further used this method to quantify cellulose and lignin in field-grown mungbean genotypes. The ATR-FTIR-based study revealed the cellulose content variation ranges from 27.9% to 52.3%, and the lignin content variation ranges from 13.7% to 31.6% in mungbean genotypes.
Multivariate analysis of FT-IR data revealed differences in total cell wall (600-2000 cm), cellulose (1000-1100 cm) and lignin (1390-1420 cm) among leaf and stem of four plant species. Overall, our data suggested that ATR-FTIR can be used for the relative quantification of lignin and cellulose in different plant species. This method was successfully applied for rapid screening of cell wall composition in mungbean stem, and similarly, it can be used for screening other crops or tree species.
剖析复杂的植物细胞壁结构需要一种灵敏且定量的方法。傅里叶变换红外光谱（FTIR）经常被用作一种筛选方法来识别细胞壁中的特定键合。然而，对特定细胞壁成分进行定量以及将光谱带分配给这些成分仍然是一项重大挑战，尤其是在农作物物种中。在本研究中，我们使用衰减全反射傅里叶变换红外光谱（ATR - FTIR）解决了这些挑战，因为它是一种高通量、经济高效且无损的方法，用于了解植物细胞壁组成。通过分析不同品种的绿豆（亚洲广泛种植的最重要的豆类作物之一）对该方法进行了验证。
通过使用标准品以及提取细胞壁成分中的特定成分，我们确定了1050 - 1060厘米和1390 - 1420厘米的波数，它们可分别广泛用于定量拟南芥、杨树、水稻和绿豆中的纤维素和木质素。此外，使用溴化钾（KBr）作为稀释剂，我们建立了一种能够相对定量上述物种不同组织类型中纤维素和木质素组成的方法。我们进一步使用该方法对田间种植的绿豆基因型中的纤维素和木质素进行定量。基于ATR - FTIR的研究表明，绿豆基因型中的纤维素含量变化范围为27.9%至52.3%，木质素含量变化范围为13.7%至31.6%。
对FT - IR数据的多变量分析揭示了四种植物物种的叶片和茎中总细胞壁（600 - 2000厘米）、纤维素（1000 - 1100厘米）和木质素（1390 - 1420厘米）存在差异。总体而言，我们的数据表明ATR - FTIR可用于不同植物物种中木质素和纤维素的相对定量。该方法成功应用于快速筛选绿豆茎中的细胞壁组成，同样，它也可用于筛选其他作物或树种。
