Institute of Biochemistry and Physiology of Plants and Microorganisms, Saratov Scientific Centre of the Russian Academy of Sciences (IBPPM RAS), 13 Prospekt Entuziastov, 410049, Saratov, Russia.
Vavilov Saratov State Agrarian University, 1 Teatralnaya Ploshchad, 410012, Saratov, Russia.
World J Microbiol Biotechnol. 2022 Feb 24;38(4):62. doi: 10.1007/s11274-022-03247-y.
In vitro somatic callus culturing is used widely in plant biotechnology, but its effectiveness depends largely on the donor plant genotype. Bacteria or components of their cells are rarely used to activate morphogenesis. In this work, inoculation of explants from immature wheat (Triticum aestivum L.) embryos with a suspension of living cells of the bacterium Azospirillum brasilense Sp7 resulted in callus death after 7 days of growth, in contrast to explant treatment with a suspension of heat-killed whole cells of Sp7. The experiments used two wheat lines, LRht-B1a and LRht-B1c, which differ in morphogenic activity. Growing calluses with the lipopolysaccharide of A. brasilense Sp7 increased the yield of regenerated plants 2- to 3.5-fold in both lines. This increase was through the activation of regenerant formation from morphogenic calluses. We have demonstrated for the first time the effects of bacterial flagellin on plant tissue culture. The polar-flagellum flagellin of A. brasilense Sp7 leveled the genotypic differences in the morphogenic ability of callus tissue. Specifically, it increased the yield of morphogenic calluses in the weakly morphogenic line LRht-B1a to the yield value in the highly morphogenic line LRht-B1c but lowered the yield of regenerants in the highly morphogenic line LRht-B1c to the yield value in the weakly morphogenic line LRht-B1a. Thus, bacterial lipopolysaccharides and flagellins can be used to regulate the formation of morphogenic calluses and regenerants in plant tissue culturing in vitro.
植物生物技术中广泛应用体外体细胞愈伤组织培养，但它的效果在很大程度上取决于供体植物基因型。细菌或其细胞成分很少被用于激活形态发生。在这项工作中，用活细胞悬浮液接种未成熟小麦（Triticum aestivum L.）胚外植体的悬浮液导致愈伤组织在生长 7 天后死亡，而用热杀死的 Sp7 全细胞悬浮液处理外植体则相反。实验使用了两个小麦品系 LRht-B1a 和 LRht-B1c，它们在形态发生活性上有所不同。用 Azospirillum brasilense Sp7 的脂多糖培养愈伤组织，在两条品系中均将再生植物的产量提高了 2 到 3.5 倍。这种增加是通过从形态发生愈伤组织中激活再生体形成实现的。我们首次证明了细菌鞭毛蛋白对植物组织培养的影响。A. brasilense Sp7 的极性鞭毛鞭毛蛋白抹平了愈伤组织形态发生能力的基因型差异。具体来说，它增加了弱形态发生品系 LRht-B1a 中形态发生愈伤组织的产量，达到了高度形态发生品系 LRht-B1c 的产量值，但降低了高度形态发生品系 LRht-B1c 中再生体的产量，达到了弱形态发生品系 LRht-B1a 的产量值。因此，细菌脂多糖和鞭毛蛋白可用于调节植物组织培养中形态发生愈伤组织和再生体的形成。
