Gordon Cameron S, Rajagopalan Nandhakishore, Risseeuw Eddy P, Surpin Marci, Ball Fraser J, Barber Carla J, Buhrow Leann M, Clark Shawn M, Page Jonathan E, Todd Chris D, Abrams Suzanne R, Loewen Michele C
Department of Biochemistry, University of Saskatchewan, 107 Wiggins Rd., Saskatoon, SK, S7N 5E5, Canada.
National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK, S7N 0W9, Canada.
PLoS One. 2016 Oct 18;11(10):e0164996. doi: 10.1371/journal.pone.0164996. eCollection 2016.
Abscisic acid (ABA) is a well-characterized plant hormone, known to mediate developmental aspects as well as both abiotic and biotic stress responses. Notably, the exogenous application of ABA has recently been shown to increase susceptibility to the fungal pathogen Fusarium graminearum, the causative agent of Fusarium head blight (FHB) in wheat and other cereals. However roles and mechanisms associated with ABA's modulation of pathogen responses remain enigmatic. Here the identification of putative ABA receptors from available genomic databases for Triticum aestivum (bread wheat) and Brachypodium distachyon (a model cereal) are reported. A number of these were cloned for recombinant expression and their functionality as ABA receptors confirmed by in vitro assays against protein phosphatases Type 2Cs. Ligand selectivity profiling of one of the wheat receptors (Ta_PYL2DS_FL) highlighted unique activities compared to Arabidopsis AtPYL5. Mutagenic analysis showed Ta_PYL2DS_FL amino acid D180 as being a critical contributor to this selectivity. Subsequently, a virus induced gene silencing (VIGS) approach was used to knockdown wheat Ta_PYL4AS_A (and similar) in planta, yielding plants with increased early stage resistance to FHB progression and decreased mycotoxin accumulation. Together these results confirm the existence of a family of ABA receptors in wheat and Brachypodium and present insight into factors modulating receptor function at the molecular level. That knockdown of Ta_PYL4AS_A (and similar) leads to early stage FHB resistance highlights novel targets for investigation in the future development of disease resistant crops.
脱落酸(ABA)是一种特征明确的植物激素,已知其介导植物发育过程以及非生物和生物胁迫响应。值得注意的是,最近研究表明,外源施加ABA会增加小麦和其他谷物对禾谷镰刀菌的易感性,禾谷镰刀菌是小麦赤霉病(FHB)的病原体。然而,ABA调节病原体反应的作用和机制仍然不明。本文报道了从小麦(普通小麦)和二穗短柄草(一种模式谷物)的现有基因组数据库中鉴定推定的ABA受体。克隆了其中一些受体用于重组表达,并通过针对2C型蛋白磷酸酶的体外试验证实了它们作为ABA受体的功能。对一种小麦受体(Ta_PYL2DS_FL)的配体选择性分析突出了其与拟南芥AtPYL5相比的独特活性。诱变分析表明,Ta_PYL2DS_FL的氨基酸D180是这种选择性的关键因素。随后,采用病毒诱导基因沉默(VIGS)方法在植物体内敲除小麦Ta_PYL4AS_A(及类似物),获得了对FHB进展早期抗性增强且霉菌毒素积累减少的植株。这些结果共同证实了小麦和二穗短柄草中存在一个ABA受体家族,并揭示了在分子水平上调节受体功能的因素。Ta_PYL4AS_A(及类似物)的敲除导致FHB早期抗性,这突出了抗病作物未来开发中值得研究的新靶点。