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一种适用于番茄早期发育阶段耐盐性比较体外研究的简单有效生物测定方法。

A Simple and Effective Bioassay Method Suitable to Comparative In Vitro Study of Tomato Salt Tolerance at Early Development Stages.

作者信息

Khaliluev Marat R, Bogoutdinova Liliya R, Raldugina Galina N, Baranova Ekaterina N

机构信息

Laboratory of Plant Cell Engineering, All-Russia Research Institute of Agricultural Biotechnology, Timiryazevskaya 42, 127550 Moscow, Russia.

Department of Biotechnology, Institute of Agrobiotechnology, Russian State Agrarian University-Moscow Timiryazev Agricultural Academy, Timiryazevskaya 49, 127550 Moscow, Russia.

出版信息

Methods Protoc. 2022 Jan 19;5(1):11. doi: 10.3390/mps5010011.

Abstract

In vitro evaluation of tomato seeds and seedlings for salt tolerance has undoubted advantages (high productivity, as well as stability and reproducibility of the obtained experimental data due to the maintenance of constant controlled conditions) in comparison with open-field system and pot experiments. However, even high-quality seeds greatly differ in the uniformity of germination capacity and germination energy. Heterogeneous germination in the habit and developmental stage of plant material significantly distorts the obtaining of relevant experimental data suitable for correct interpretation. In our study, we propose a simple and effective bioassay method suitable to comparative in vitro study of tomato salt tolerance using shoot apex of seedlings at the early first-true-leaf stage. Shoot apexes cultured the on the root induction medium (RIM) supplemented with 0.2 mg/L indole-3-butyric acid (IBA) and NaCl at different concentrations (0-250 mM NaCl) revealed significant differences between two tomato genotypes (line YaLF and cv. Rekordsmen) at the organismal (measurements of CO gas exchange), organ (rhizogenesis frequency; number and length of de novo regenerated roots; root fresh (RFW) and dry (RDW) weights; shoot fresh (SFW) and dry (SDW) weights), tissue (the average cross-sectional area of epidermal and mesophylls cotyledonary cells) and cellular (ultrastructure of chloroplast and nuclear compartments) development levels. In addition, a quantitative comparison of proline and photosynthetic pigments contents under 75 and 150 mm NaCl treatments showed a different response between two tomato genotypes. The proposed methodological approach can be used for other plants with a high response to auxin-induced rhizogenesis in vitro, as well as for the comparative in vitro assessment of other abiotic stresses.

摘要

与露天种植系统和盆栽试验相比,对番茄种子和幼苗进行耐盐性的体外评估具有明显优势(由于能维持恒定的受控条件,实验数据具有高生产率、稳定性和可重复性)。然而,即使是高质量的种子,其发芽能力和发芽势的均匀性也存在很大差异。植物材料在习性和发育阶段的异质发芽会显著扭曲适用于正确解释的相关实验数据的获取。在我们的研究中,我们提出了一种简单有效的生物测定方法,适用于使用第一片真叶早期幼苗的茎尖对番茄耐盐性进行比较体外研究。在添加了0.2 mg/L吲哚-3-丁酸(IBA)和不同浓度(0-250 mM NaCl)NaCl的根诱导培养基(RIM)上培养的茎尖,揭示了两种番茄基因型(YaLF品系和Rekordsmen品种)在生物体水平(CO气体交换测量)、器官水平(发根频率;新生根的数量和长度;根鲜重(RFW)和干重(RDW);茎鲜重(SFW)和干重(SDW))、组织水平(表皮和叶肉子叶细胞的平均横截面积)和细胞水平(叶绿体和细胞核区室的超微结构)发育上的显著差异。此外,对75和150 mM NaCl处理下脯氨酸和光合色素含量的定量比较显示,两种番茄基因型有不同的反应。所提出的方法可用于对生长素诱导的体外发根反应较高的其他植物,以及用于其他非生物胁迫的比较体外评估。

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