Molecular characterization and functional analysis of akt-1 in pinewood nematode, Bursaphelenchus xylophilus
Molecular characterization and functional analysis of akt-1 in pinewood nematode, Bursaphelenchus xylophilus
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松材线虫 松材线虫 akt-1 的分子表征和功能分析
DOI：
10.1111/efp.12647
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发表时间：
2021-01-17
期刊：
影响因子：
1.4
通讯作者：
Hu，Jiafu
中科院分区：
文献类型：
--
作者：
Zhou，Lifeng;Ji，Jingjing;Hu，Jiafu
松材线虫是松树萎蔫病的病原体，导致全球数百万公顷松树的毁灭。蛋白激酶B（AKT）是一种结构保守的丝氨酸/苏氨酸激酶，具有普列克底物蛋白同源结构域和激酶结构域，在不同的动物界中发挥着多种生物学作用。因此，我们对嗜木芽孢杆菌（Bxy-akt-1）中fakt-1的分子特性和生物学功能进行了研究。在繁殖阶段，反转录实时定量PCR显示Bxy ‐akt‐ 1的表达在胚胎发生过程中逐渐增加，在第二阶段的幼鱼（J2）达到最高水平，然后从J3到成虫逐渐下降。有趣的是，Bxy ‐akt‐ 1的表达在男性中显著高于女性。此外，Bxy ‐akt‐ 1在分散的第三期幼虫（DL 3）中的表达水平高于J2，而在DL 4中则降至正常水平。利用构建于秀丽隐杆线虫中的Bxy ‐akt‐ 1启动子：：gfp报告基因模拟Bxy ‐akt‐1的时空表达模式。结果表明，Bxy ‐akt‐ 1在胚胎发生期广泛表达于头部周围的皮下组织，在胚后主要表达于咽部、外阴部、尾部的肌细胞和神经系统。敲除Bxy ‐akt‐ 1基因后，小菜蛾出现异常胚胎，孵化率显著降低，但未观察到幼虫的结构性发育停滞。结果表明，Bxy ‐akt‐ 1是胚胎发生所必需的，但可能不像秀丽隐杆线虫那样在控制dauer进入方面发挥重要作用。
Bursaphelenchus xylophilusis the causal agent of pine wilt disease, which leads to the devastation of millions of hectares of pine trees worldwide. Protein kinase B (AKT) is a structurally conserved serine/threonine kinase with a pleckstrin homology domain and a kinase domain, and it plays versatile biological roles across different animal kingdoms. We, therefore, investigated the molecular characteristics and biological function ofakt‐1inB. xylophilus(Bxy‐akt‐1). In propagative stages, reverse transcription quantitative real‐time PCR revealed thatBxy‐akt‐1expression gradually increased during embryogenesis, reaching the highest level at the second‐stage juvenile (J2), and then steadily decreased from J3 to adults. Intriguingly, the expression ofBxy‐akt‐1was significantly higher in males than in females. Moreover, the expression level ofBxy‐akt‐1in dispersal third‐stage dauer larvae (DL3) was higher than that in J2, but it decreased to normal level in DL4.Bxy‐akt‐1promoter::gfp reporter constructed inCaenorhabditis eleganswas utilized to mimic the spatio‐temporal expression pattern ofBxy‐akt‐1. The results showed thatBxy‐akt‐1was broadly expressed in the hypodermis around the head during embryogenesis and was mainly expressed in the muscle cells of pharynx, vulva, tail and nerve systems in post‐embryonic stages. Abnormal embryos were observed, and the hatching rate was significantly decreased afterBxy‐akt‐1was knocked down inB. xylophilus, but no constitutive developmental arrest of dauer larvae was observed. The results indicated thatBxy‐akt‐1is indispensable for embryogenesis, but might not play a significant role in controlling dauer entry, as inC. elegans.