Regulation of Chitinase in Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) During Infection by Heliothis virescens ascovirus 3h (HvAV-3h)
Regulation of Chitinase in Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) During Infection by Heliothis virescens ascovirus 3h (HvAV-3h)
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甜菜夜蛾 (Hübner)（鳞翅目：夜蛾科）在烟夜蛾子囊病毒感染 3 小时 (HvAV-3h) 期间几丁质酶的调节
DOI：
10.3389/fphys.2020.00166
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发表时间：
2020-03
影响因子：
4
通讯作者：
Guo-Hua Huang
中科院分区：
文献类型：
--
作者：
Lei He;Yi-Yi Ou-Yang;Ni Li;Ying Chen;Shuang-Qing Liu;Guo-Hua Huang
昆虫几丁质酶在昆虫的蜕皮和变态过程中起着重要的作用。烟芽夜蛾（Heliothis virescens）病毒3 h（HvAV-3 h）能延长寄主幼虫的幼虫期。本研究对甜菜夜蛾Spodoptera exigua（Hubner）幼虫几丁质酶和几丁质结合结构域（CBD）的分子特征和功能进行了分析。对模拟感染和HvAV-3 h感染的幼虫全身和4种不同组织的几丁质酶活性进行了测定，结果表明：感染后48 h，幼虫的几丁质酶活性显著下降;感染后144和168 h，幼虫脂肪体和角质层的几丁质酶活性显著下降。S.甜菜几丁质酶7（SeCHIT 7）在6、9、12、48、72和96 hpi取样时间下调，S.甜菜几丁质酶11（SeCHIT 11）在3-96 hpi时表达下调，而S.甜菜几丁质酶（SeCHIT）在120-168 hpi上调。对SeCHIT 7和SeCHIT 11转录的进一步组织特异性检测表明，SeCHIT 7在脂肪体和角质层中在144和168 hpi下调。SeCHIT 11在168 hpi时在脂肪体、中肠和角质层中下调。此外，S.在HvAV-3 h感染的幼虫中未检测到甜菜几丁质结合域（SeCBD）。对SeCHIT 7 N、SeCHIT 11和SeCBD的体外分析表明，SeCHIT 7 N和SeCHIT 11是典型的几丁质酶。相反，没有检测到几丁质酶活性与SeCBD。而SeCBD则能显著提高SeCHIT 7 N和SeCHIT 11的活性。HvAV-3 h不仅干扰SeCHITs的转录和表达，而且影响SeCBD的正常转录和表达，从而影响宿主幼虫几丁质酶的活性。这些结果将有助于为进一步研究HvAV-3 h的发病机制提供基础。
Insect chitinases play essential roles in the molting and metamorphosis of insects. The virus Heliothis virescens ascovirus 3h (HvAV-3h) can prolong the total duration of the larval stage in its host larvae. In this study, the molecular character and function of chitinase and chitin-binding domain (CBD) were analyzed in larvae of Spodoptera exigua (Hubner) (Lepidoptera: Noctuidae). In detecting the chitinase activity of mock-infected and HvAV-3h-infected larval whole bodies and four different larval tissues, the results showed that larval chitinase activity was significantly decreased at 48 h post infection (hpi) and that the chitinase activity of HvAV-3h-infected larval fat body and cuticle was notably decreased at 144 and 168 hpi. The transcription level of S. exigua chitinase 7 (SeCHIT7) was down-regulated at the 6, 9, 12, 48, 72, and 96 hpi sample times, the S. exigua chitinase 11 (SeCHIT11) was down-regulated at 3-96 hpi, while both S. exigua chitinases (SeCHITs) were up-regulated at 120-168 hpi. Further tissue-specific detection of SeCHIT7 and SeCHIT11 transcription showed that SeCHIT7 was down-regulated at 144 and 168 hpi in the fat body and cuticle. SeCHIT11 was down-regulated at 168 hpi in the fat body, midgut, and cuticle. Additionally, the transcription and expression of S. exigua chitin-binding domain (SeCBD) could not be detected in HvAV-3h-infected larvae. The in vitro analyses of SeCHIT7N, SeCHIT11, and SeCBD showed that SeCHIT7N and SeCHIT11 were typical chitinases. Conversely, no chitinase activity was detected with SeCBD. SeCBD, however, could significantly increase the activity of SeCHIT7N and SeCHIT11. In conclusion, HvAV-3h not only interfered with the transcription and expression of SeCHITs but also affected the normal transcription and expression of SeCBD and, in doing so, influenced the host larval chitinase activity. These results will aid in providing a foundation for further studies on the pathogenesis of HvAV-3h.
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