Nakashima Jin, Pattathil Sivakumar, Avci Utku, Chin Sabrina, Alan Sparks J, Hahn Michael G, Gilroy Simon, Blancaflor Elison B
Analytical Instrumentation Facility, North Carolina State University, 2410 Campus Shore Drive, Raleigh, NC, 27606, USA.
Mascoma LLC (Lallemand Inc.), 67 Etna Road, Lebanon, NH, 03766, USA.
NPJ Microgravity. 2023 Aug 22;9(1):68. doi: 10.1038/s41526-023-00312-0.
A large and diverse library of glycan-directed monoclonal antibodies (mAbs) was used to determine if plant cell walls are modified by low-gravity conditions encountered during spaceflight. This method called glycome profiling (glycomics) revealed global differences in non-cellulosic cell wall epitopes in Arabidopsis thaliana root extracts recovered from RNA purification columns between seedlings grown on the International Space Station-based Vegetable Production System and paired ground (1-g) controls. Immunohistochemistry on 11-day-old seedling primary root sections showed that ten of twenty-two mAbs that exhibited spaceflight-induced increases in binding through glycomics, labeled space-grown roots more intensely than those from the ground. The ten mAbs recognized xyloglucan, xylan, and arabinogalactan epitopes. Notably, three xylem-enriched unsubstituted xylan backbone epitopes were more intensely labeled in space-grown roots than in ground-grown roots, suggesting that the spaceflight environment accelerated root secondary cell wall formation. This study highlights the feasibility of glycomics for high-throughput evaluation of cell wall glycans using only root high alkaline extracts from RNA purification columns, and subsequent validation of these results by immunohistochemistry. This approach will benefit plant space biological studies because it extends the analyses possible from the limited amounts of samples returned from spaceflight and help uncover microgravity-induced tissue-specific changes in plant cell walls.
利用一个庞大且多样的聚糖导向单克隆抗体（mAb）文库来确定植物细胞壁是否会因太空飞行中遇到的低重力条件而发生改变。这种称为聚糖谱分析（糖组学）的方法揭示了从国际空间站上的蔬菜生产系统中生长的拟南芥幼苗以及配对的地面（1g）对照中从RNA纯化柱回收的根提取物中非纤维素细胞壁表位的整体差异。对11日龄幼苗初生根切片的免疫组织化学分析表明，在通过糖组学显示出太空飞行诱导结合增加的22种单克隆抗体中，有10种对太空生长的根的标记比地面生长的根更强烈。这10种单克隆抗体识别木葡聚糖、木聚糖和阿拉伯半乳聚糖表位。值得注意的是，三种富含木质部的未取代木聚糖主链表位在太空生长的根中的标记比地面生长的根更强烈，这表明太空飞行环境加速了根次生细胞壁的形成。这项研究突出了糖组学仅使用RNA纯化柱中的根高碱性提取物对细胞壁聚糖进行高通量评估的可行性，以及随后通过免疫组织化学对这些结果进行验证的可行性。这种方法将有利于植物空间生物学研究，因为它扩展了从太空飞行返回的有限量样本中可能进行的分析，并有助于揭示微重力诱导的植物细胞壁组织特异性变化。
