Development of a novel and rapid phenotype-based screening method to assess rice seedling growth
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Plant Methods Open Access
METHODOLOGY
Development of a novel and rapid phenotype‑based screening method to assess rice seedling growth Lena Vlaminck1,2,3, Chananchida Sang‑Aram1,3, Deborah Botterman3, Christine Jewel C. Uy3, Mary Kay Harper4, Dirk Inzé1,2, Godelieve Gheysen5 and Stephen Depuydt1,2,3*
Abstract Background: Rice (Oryza sativa) is one of the most important model crops in plant research. Despite its considerable advantages, (phenotypic) bioassays for rice are not as well developed as for Arabidopsis thaliana. Here, we present a phenotype-based screening method to study shoot-related parameters of rice seedlings via an automated computer analysis. Results: The phenotype-based screening method was validated by testing several compounds in pharmacological experiments that interfered with hormone homeostasis, confirming that the assay was consistent with regard to the anticipated plant growth regulation and revealing the robustness of the set-up in terms of reproducibility. Moreover, abiotic stress tests using NaCl and DCMU, an electron transport blocker during the light dependent reactions of pho‑ tosynthesis, confirmed the validity of the new method for a wide range of applications. Next, this method was used to screen the impact of semi-purified fractions of marine invertebrates on the initial stages of rice seedling growth. Certain fractions clearly stimulated growth, whereas others inhibited it, especially in the root, illustrating the possible applications of this novel, robust, and fast phenotype-based screening method for rice. Conclusions: The validated phenotype-based and cost-efficient screening method allows a quick and proper analysis of shoot growth and requires only small volumes of compounds and media. As a result, this method could potentially be used for a whole range of applications, ranging from discovery of novel biostimulants, plant growth regulators, and plant growth-promoting bacteria to analysis of CRISPR knockouts, molecular plant breeding, genomewide association, and phytotoxicity studies. The assay system described here can contribute to a better understanding of plant development in general. Keywords: Oryza sativa, Phenotype-based screening, Seedling growth, Automated computer analysis, Marine natural products, Plength, RIVA Background Rice (Oryza sativa) is one of the most important crops worldwide. It is also a main model species for plant research. It is an excellent model system for *Correspondence: [email protected] 3 Laboratory of Plant Growth Analysis, Ghent University Global Campus, Incheon 21985, South Korea Full list of author information is available at the end of the article
monocotyledonous crops, because it has many functionally conserved genes and shows synteny with other Poaceae, such as wheat (Triticum sp.), maize (Zea mays), barley (Hordeum vulgare), and sorghum (Sorghum bicolor) [1, 2]. Rice has a relatively short life cycle compared to other cereals and can be maintained via selffertilization with hardly any cross-contamination risk [3]. It
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