Guillory Ambre, Bonhomme Sandrine
Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin, Versailles, France.
Methods Mol Biol. 2021;2309:143-155. doi: 10.1007/978-1-0716-1429-7_12.
As a bryophyte and model plant, the moss Physcomitrium (Physcomitrella) patens (P. patens) is particularly well adapted to hormone evolution studies. Gene targeting through homologous recombination or CRISPR-Cas9 system, genome sequencing, and numerous transcriptomic datasets has allowed for molecular genetics studies and much progress in Evo-Devo knowledge. As to strigolactones, like for other hormones, both phenotypical and transcriptional responses can be studied, in both WT and mutant plants. However, as in any plant species, medium- to large-scale phenotype characterization is necessary, owing to the general high phenotypic variability. Therefore, many biological replicates are required. This may translate to large amount of the investigated compounds, particularly expensive (or difficult to synthesize) in the case of strigolactones. These issues prompted us to improve existing methods to limit the use of scarce/expensive compounds, as well as to simplify subsequent measures/sampling of P. patens. We hence scaled up well-tried experiments, in order to increment the number of tested genotypes in one given experiment.In this chapter, we will describe three methods we set up to study the response to strigolactones and related compounds in P. patens.
作为一种苔藓植物和模式植物，小立碗藓（Physcomitrium (Physcomitrella) patens，P. patens）特别适合用于激素进化研究。通过同源重组或CRISPR-Cas9系统进行基因靶向、基因组测序以及大量转录组数据集，使得分子遗传学研究以及进化发育生物学知识取得了很大进展。对于独脚金内酯，与其他激素一样，可以在野生型和突变体植物中研究其表型和转录反应。然而，与任何植物物种一样，由于普遍存在较高的表型变异性，有必要进行中到大规模的表型特征分析。因此，需要许多生物学重复样本。这可能意味着需要大量的被研究化合物，尤其是独脚金内酯这种特别昂贵（或难以合成）的化合物。这些问题促使我们改进现有方法，以限制稀缺/昂贵化合物的使用，并简化后续对小立碗藓的测量/取样。因此，我们扩大了经过充分验证的实验规模，以便在一次给定实验中增加测试基因型的数量。在本章中，我们将描述我们建立的三种用于研究小立碗藓对独脚金内酯及相关化合物反应的方法。
