State Key Laboratory of Tree Genetics and Breeding, Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, 210037, China.
Key Laboratory of Forest Genetics and Biotechnology of Ministry of Education, Nanjing Forestry University, Nanjing, 210037, China.
BMC Plant Biol. 2023 Oct 9;23(1):480. doi: 10.1186/s12870-023-04496-1.
Suspension culture is widely used in the establishment of efficient plant regeneration systems, as well as in the mass production of plant secondary metabolites. However, the establishment of a suspension culture system of Cunninghamia lanceolata is genotype-dependent given that proembryogenic masses (PEMs) are prone to browning during this process in recalcitrant genotypes. Previously, we reported that the plant peptide hormone phytosulfokine (PSK) can tremendously decrease the hydrogen peroxide (HO) level and help to initiate somatic embryogenesis (SE) in recalcitrant C. lanceolata genotypes. However, to date, no studies have revealed whether or how PSK may contribute to the establishment of a suspension culture system in these recalcitrant genotypes.
Here, we demonstrated that exogenous application of PSK effectively inhibited PEM browning during suspension culture in a recalcitrant genotype of C. lanceolata. Comparative time-series transcriptome profiling showed that redox homeostasis underwent drastic fluctuations when PEMs were cultured in liquid medium, while additional PSK treatment helped to maintain a relatively stable redox homeostasis. Interestingly, PSK seemed to have a dual effect on peroxidases (PRXs), with PSK simultaneously transcriptionally repressing ROS-producing PRXs and activating ROS-scavenging PRXs. Furthermore, determination of HO and MDA content, as well as cell viability, showed that exogenous PSK treatment inhibited PEM browning and safeguarded PEM suspension culture by decreasing the HO level and increasing PEM activity.
Collectively, these findings provide a valuable tool for the future establishment of large-scale C. lanceolata PEM suspension culture without genotype limitations.
悬浮培养广泛应用于建立高效的植物再生系统,以及大量生产植物次生代谢产物。然而,由于在顽固基因型中,胚性愈伤组织(PEM)在悬浮培养过程中容易褐变,因此建立杉木悬浮培养体系是依赖于基因型的。此前,我们报道植物肽激素植物磺基高丝氨酸(PSK)可以极大地降低过氧化氢(HO)水平,并有助于启动顽固杉木基因型的体细胞胚胎发生(SE)。然而,迄今为止,尚无研究表明 PSK 如何有助于在这些顽固基因型中建立悬浮培养体系。
在这里,我们证明了外源性 PSK 的应用可以有效抑制杉木顽固基因型悬浮培养中 PEM 的褐变。比较时间序列转录组谱分析表明,当 PEM 在液体培养基中培养时,氧化还原稳态发生剧烈波动,而额外的 PSK 处理有助于维持相对稳定的氧化还原稳态。有趣的是,PSK 似乎对过氧化物酶(PRXs)有双重作用,PSK 同时转录抑制 ROS 产生的 PRXs 并激活 ROS 清除的 PRXs。此外,HO 和 MDA 含量以及细胞活力的测定表明,外源性 PSK 处理通过降低 HO 水平和提高 PEM 活性来抑制 PEM 褐变并保护 PEM 悬浮培养。
总的来说,这些发现为未来建立不受基因型限制的大规模杉木 PEM 悬浮培养提供了有价值的工具。