Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia.
Jožef Stefan International Postgraduate School, Ljubljana, Slovenia.
Plant Biotechnol J. 2024 Sep;22(9):2518-2529. doi: 10.1111/pbi.14365. Epub 2024 May 10.
Protein complexes from edible oyster mushrooms (Pleurotus sp.) composed of pleurotolysin A2 (PlyA2) and pleurotolysin B (PlyB) exert toxicity in feeding tests against Colorado potato beetle (CPB) larvae, acting through the interaction with insect-specific membrane sphingolipid. Here we present a new strategy for crop protection, based on in planta production of PlyA2/PlyB protein complexes, and we exemplify this strategy in construction of transgenic potato plants of cv Désirée. The transgenics in which PlyA2 was directed to the vacuole and PlyB to the endoplasmic reticulum are effectively protected from infestation by CPB larvae without impacting plant performance. These transgenic plants showed a pronounced effect on larval feeding rate, the larvae feeding on transgenic plants being on average five to six folds lighter than larvae feeding on controls. Further, only a fraction (11%-37%) of the larvae that fed on transgenic potato plants completed their life cycle and developed into adult beetles. Moreover, gene expression analysis of CPB larvae exposed to PlyA2/PlyB complexes revealed the response indicative of a general stress status of larvae and no evidence of possibility of developing resistance due to the functional inactivation of PlyA2/PlyB sphingolipid receptors.
来自食用蘑菇（香菇属）的蛋白质复合物，由 pleurotolysin A2（PlyA2）和 pleurotolysin B（PlyB）组成，在针对科罗拉多马铃薯甲虫（CPB）幼虫的饲养试验中表现出毒性，通过与昆虫特异性膜神经酰胺的相互作用发挥作用。在这里，我们提出了一种基于植物体内生产 PlyA2/PlyB 蛋白复合物的新作物保护策略，并以 Désirée 品系转基因马铃薯植物的构建为例说明了这一策略。将 PlyA2 定向到液泡而 PlyB 定向到内质网的转基因植物有效地免受 CPB 幼虫的侵害，而不影响植物的性能。这些转基因植物对幼虫的取食率表现出明显的影响，取食转基因植物的幼虫比取食对照的幼虫平均轻五到六倍。此外，只有一小部分（11%-37%）取食转基因马铃薯植物的幼虫完成了它们的生命周期并发育成成年甲虫。此外，暴露于 PlyA2/PlyB 复合物的 CPB 幼虫的基因表达分析显示出幼虫普遍应激状态的反应迹象，并且由于 PlyA2/PlyB 神经酰胺受体的功能失活，没有出现产生抗性的可能性的证据。
