Horticulture Department, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District, 110866, PR China; National & Local Joint Engineering Research Center of Northern Horticultural Facilities Design & Application Technology (Liaoning), No. 120 Dongling Road, Shenhe District, 110866, PR China; Key Laboratory of Protected Horticulture (Shenyang Agricultural University), Ministry of Education, No. 120 Dongling Road, Shenhe District, 110866, PR China.
Horticulture Department, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District, 110866, PR China; Key Laboratory of Protected Horticulture (Shenyang Agricultural University), Ministry of Education, No. 120 Dongling Road, Shenhe District, 110866, PR China; Key Laboratory of Fruit Postharvest Biology of Liaoning Province, No. 120 Dongling Road, Shenhe District, 110866, PR China.
Plant Sci. 2022 Aug;321:111305. doi: 10.1016/j.plantsci.2022.111305. Epub 2022 May 6.
Tomato is often exposed to high temperature stress during summer cultivation. Stomatal movement plays important roles in photosynthesis and transpiration which restricts the quality and yield of tomato under environmental stress. To elucidate the mechanism of stomatal movement in high temperature tolerance, SlSnRK2s (sucrose non-fermenting 1-related protein kinases) silenced plants were generated in tomato with CRISPR-Cas 9 gene editing techniques. Through the observation of stomatal parameters, SlSnRK2.3 regulated stomatal closure which was responded to ABA (abscisic acid) and activated signaling pathway of ROS (reactive oxygen species) in high temperature stress. Based on the positive functions of SlSnRK2.3, the cDNA library was generated to investigate interaction proteins of SlSnRK2s. The interaction between SlSnRK2.3 and SlSUI1 (protein translation factor SUI1 homolog) was employed by Yeast two hybrid assay (Y2H), Luciferase (LUC), and Bimolecular fluorescence complementation (BiFC). Finally, the specific interactive sites between SlSnRK2.3 and SlSUI1 were verified by site-directed mutagenesis. The consistent mechanism of SlSnRK2.3 and SlSUI1 in stomatal movement, indicating that SlSUI1 interacted with SlSnRK2.3 through ABA-dependent signaling pathway in high temperature stress. Our results provided evidence for improving the photosynthetic capacity of tomato under high temperature stress, and support the breeding and genetic engineering of tomato over summer facility cultivation.
番茄在夏季种植时经常受到高温胁迫。气孔运动在光合作用和蒸腾作用中起着重要作用，这限制了番茄在环境胁迫下的品质和产量。为了阐明气孔运动在高温耐受性中的机制，利用 CRISPR-Cas9 基因编辑技术在番茄中沉默 SlSnRK2s（蔗糖非发酵 1 相关蛋白激酶）。通过观察气孔参数，SlSnRK2.3 调节气孔关闭，这是对 ABA（脱落酸）的反应，并在高温胁迫下激活 ROS（活性氧）信号通路。基于 SlSnRK2.3 的积极功能，生成了 cDNA 文库以研究 SlSnRK2s 的相互作用蛋白。SlSnRK2.3 和 SlSUI1（蛋白质翻译因子 SUI1 同源物）之间的相互作用通过酵母双杂交测定（Y2H）、荧光素酶（LUC）和双分子荧光互补（BiFC）进行。最后，通过定点突变验证了 SlSnRK2.3 和 SlSUI1 之间的特定相互作用位点。SlSnRK2.3 和 SlSUI1 在气孔运动中的一致机制表明，SlSUI1 通过 ABA 依赖的信号通路在高温胁迫下与 SlSnRK2.3 相互作用。我们的研究结果为提高番茄在高温胁迫下的光合能力提供了证据，并支持番茄夏季设施栽培的选育和遗传工程。
