Annulohypoxylon stygium is an ascomycete that helps Tremella fuciformis produce the fruiting body in wild state or artificial cultivation. Although the interaction between these two fungi is well known, the underlying molecular mechanism is limited. In this study, the 981 bp and 886 bp promoter sequences of glyceraldehyde-3-phosphate dehydrogenase (gpd) gene and α-tubulin gene have been cloned, respectively. Sequence analysis showed that gpd promoter contained nine CAAT boxes, and single TGACG-motif, TATA box, ABRE motif, STRE motif, MYB motif, and W box. The α-tubulin promoter included eight CAAT boxes, three STRE, two TATA boxes and MYB, single Box 4, CAT-box, CCAAT-box, TGA-element, and ABRE. Subsequently, we evaluated the promoters' function by constructing four vectors pGEH, pGRH, pTEH, and pTRH to drive fused enhanced green fluorescent protein and hygromycin B phosphotransferase (egfp-hph) or red fluorescent protein and hygromycin B phosphotransferase (rfp-hph) expression under the control of gpd or α-tubulin promoters in A. stygium. The integration of the transformed DNA into A. stygium genome was verified by PCR, Southern blot, fluorescence microscopy, and quantitative real-time PCR (qRT-PCR). All the results indicated that the two promoters could drive egfp-hph and rfp-hph expression. This result could provide help in gene functional studies by using gpd and α-tubulin promoters to direct gene over-expression or build dual promoter silencing systems.

环丙氧基苯乙烯是一种伴生菌，有助于银耳在野外或人工栽培下产生子实体。尽管这两种真菌之间的相互作用是众所周知的，但其潜在的分子机制是有限的。在这项研究中，分别克隆了3-磷酸​​甘油醛脱氢酶（gpd）基因和α-微管蛋白基因的981 bp和886 bp启动子序列。序列分析表明，gpd启动子包含9个CAAT框，以及单个TGACG基序，TATA框，ABRE基序，STRE基序，MYB基序和W框。该α微管蛋白启动子包括八个CAAT框，三个STRE，两个TATA框和MYB，单个Box 4，CAT框，CCAAT框，TGA元素和ABRE。随后，我们通过构建四个载体pGEH，pGRH，pTEH和pTRH来驱动融合增强的绿色荧光蛋白和潮霉素B磷酸转移酶（egfp-hph）或红色荧光蛋白和潮霉素B磷酸转移酶（rfp-hph）表达来评估启动子的功能。在触角拟南芥中gpd或α-微管蛋白启动子的控制下。将转化的DNA整合到触角A.通过PCR，Southern印迹，荧光显微镜和定量实时PCR（qRT-PCR）验证基因组。所有结果表明，这两个启动子可以驱动egfp-hph和rfp-hph表达。该结果可通过使用gpd和α-微管蛋白启动子指导基因过表达或构建双启动子沉默系统，为基因功能研究提供帮助。