AaCaMKs Positively Regulate Development, Infection Structure Differentiation and Pathogenicity in Alternaria alternata, Causal Agent of Pear Black Spot.
AaCaMKs Positively Regulate Development, Infection Structure Differentiation and Pathogenicity in Alternaria alternata, Causal Agent of Pear Black Spot.
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AaCaMKs 正向调控梨黑斑病病原 Alternaria alternata 的发育、感染结构分化和致病性
DOI：
10.3390/ijms24021381
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发表时间：
2023-01-10
影响因子：
5.6
通讯作者：
Prusky, Dov B. B.
中科院分区：
文献类型：
--
作者：
Jiang, Qianqian;Li, Yongcai;Mao, Renyan;Bi, Yang;Liu, Yongxiang;Zhang, Miao;Li, Rong;Yang, Yangyang;Prusky, Dov B. B.
钙/钙调蛋白依赖性蛋白激酶（CaMK）是真核生物Ca2+信号通路的关键下游靶蛋白，在植物真菌的生长发育和致病性中起着重要的调控作用。从互花草中克隆到3个具有保守的PKC_like超家族结构域、ATP结合位点和ACT位点的AaCaMKs (AaCaMK1、AaCaMK2和AaCaMK3)，但其在互花草中的调控机制尚不清楚。在本研究中，通过靶向基因破坏阐明了AaCaMKs在a.a alternata的发育、感染结构分化和致病性中的功能。单次破坏AaCaMKs对营养生长和孢子形态无明显影响，但对菌丝生长、产孢、生物量积累和黑色素合成有显著影响。进一步的表达分析表明，AaCaMKs在疏水和梨蜡基质上的侵染结构分化过程中表达上调。体外和体内分析进一步发现，单个AaCaMKs基因的缺失显著降低了刺花蒿孢子萌发、附着胞形成和侵染菌丝形成。此外，药理学分析证实，CaMK特异性抑制剂KN93抑制了生豆孢子萌发和附着胞的形成。同时，AaCaMKs基因缺失显著降低了互花霉的致病性。这些结果表明，AaCaMKs调控了褐花霉的发育、感染结构分化和致病性，为开发新的有效杀菌剂提供了潜在的靶点。
Calcium/calmodulin-dependent protein kinase (CaMK), a key downstream target protein in the Ca2+ signaling pathway of eukaryotes, plays an important regulatory role in the growth, development and pathogenicity of plant fungi. Three AaCaMKs (AaCaMK1, AaCaMK2 and AaCaMK3) with conserved PKC_like superfamily domains, ATP binding sites and ACT sites have been cloned from Alternaria alternata, However, their regulatory mechanism in A. alternata remains unclear. In this study, the function of the AaCaMKs in the development, infection structure differentiation and pathogenicity of A. alternata was elucidated through targeted gene disruption. The single disruption of AaCaMKs had no impact on the vegetative growth and spore morphology but significantly influenced hyphae growth, sporulation, biomass accumulation and melanin biosynthesis. Further expression analysis revealed that the AaCaMKs were up-regulated during the infection structure differentiation of A. alternata on hydrophobic and pear wax substrates. In vitro and in vivo analysis further revealed that the deletion of a single AaCaMKs gene significantly reduced the A. alternata conidial germination, appressorium formation and infection hyphae formation. In addition, pharmacological analysis confirmed that the CaMK specific inhibitor, KN93, inhibited conidial germination and appressorium formation in A. alternata. Meanwhile, the AaCaMKs genes deficiency significantly reduced the A. alternata pathogenicity. These results demonstrate that AaCaMKs regulate the development, infection structure differentiation and pathogenicity of A. alternata and provide potential targets for new effective fungicides.
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