Zhang He-He, Zhang Qi-Wen, Idrees Atif, Lin Jia, Song Xue-Sen, Ji Qing-E, Du Ying-Gang, Zheng Min-Lin, Chen Jia-Hua
Institute of Beneficial Insects, Plant Protection College, Fujian Agriculture and Forestry University, Fuzhou 350002, PR China; UN (China) Center for Fruit Fly Prevention and Treatment, Fuzhou 350002, PR China; State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Fuzhou 350002, PR China; Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fuzhou 350002, PR China.
Jia Si Xie Agricultural College, Weifang University of Science and Technology, Shouguang 262700, PR China.
Comp Biochem Physiol B Biochem Mol Biol. 2019 May;231:11-19. doi: 10.1016/j.cbpb.2019.01.017. Epub 2019 Feb 5.
Tyrosine hydroxylase (TH) is the initial enzyme responsible for cuticle sclerotization and pigmentation in many insect species, but to date, no direct functional studies have focused on TH in Zeugodacus tau. Here, the 3336-bp full-length cDNA of TH was isolated from Z. tau, a notorious horticultural pest infesting fruits and vegetables. qRT-polymerase chain reaction revealed that ZtTH transcripts were highly abundant at the time of pupal tanning and during adult emergence and were expressed in the midgut, integument and head of molting larvae. The pupation and eclosion rates gradually decreased when the 1st-instar larvae were fed diets containing higher concentrations of the TH inhibitor 3-iodo-tyrosine (3-IT). Moreover, pupal weights were significantly decreased, and abnormal uncolored phenotypes were observed after 20 mg/g 3-IT was incorporated into the diet. In addition, the suppression of TH function (mediated by RNA interference) led to a decrease in TH mRNAs and eclosion rates, accompanied by less-pigmented phenotypes. There was a severe impairment of larval-pupal cuticle tanning, leading to pupae with less yellowish pigment or that were completely white and transparent, when we injected 2 μL of 24.4 mM or 73.27 mM 3-IT into 3rd-instar larvae or prepupae. These results suggest that TH is an important enzyme for the normal growth and pupal pigmentation of Z. tau and that TH is a potential gene target for use in the control of Z. tau.
酪氨酸羟化酶(TH)是许多昆虫物种中负责表皮硬化和色素沉着的初始酶,但迄今为止,尚未有直接的功能研究聚焦于南瓜实蝇中的TH。在此,从南瓜实蝇(一种臭名昭著的侵害水果和蔬菜的园艺害虫)中分离出了3336bp的TH全长cDNA。实时定量聚合酶链反应显示,ZtTH转录本在化蛹鞣化时以及成虫羽化期间高度丰富,并且在蜕皮幼虫的中肠、体壁和头部表达。当一龄幼虫食用含有较高浓度TH抑制剂3-碘酪氨酸(3-IT)的饲料时,化蛹率和羽化率逐渐降低。此外,当饲料中添加20mg/g的3-IT后,蛹重显著降低,并观察到异常的无色表型。此外,TH功能的抑制(通过RNA干扰介导)导致TH mRNA和羽化率降低,同时伴有色素沉着较少的表型。当我们向三龄幼虫或预蛹注射2μL 24.4mM或73.27mM的3-IT时,幼虫-蛹的表皮鞣化严重受损,导致蛹的黄色色素减少或完全呈白色且透明。这些结果表明,TH是南瓜实蝇正常生长和蛹色素沉着的重要酶,并且TH是用于控制南瓜实蝇的潜在基因靶点。