Fecha de publicación: 10/01/2020 Fuente: WIPO "tobacco agriculture" The invention discloses a tobacco neonicotinoid synthesis regulation gene NtERF91 and a cloning method and application thereof. The nucleotide sequence of the nicotine synthesis regulation gene NtERF91 is shown as SEQ ID: No.1, and the coded amino acid sequence is shown as SEQ ID: No.2. The invention further discloses a cloning method of the neonicotinoid synthesis regulation gene NtERF91. The cloning method comprises the following specific steps: A, determining the sequence of the NtERF91 gene; B, extracting tobacco RNA, and performing reverse transcription to obtain a first chain cDNA; C, designing and synthesizing a specific primer according to the NtERF91 gene sequence, and performing PCR amplification by taking the cDNA as a template; D, recovering and purifying a PCR product; and E,constructing an overexpression vector containing the NtERF91 gene. The overexpression vector is overexpressed in tobacco through agrobacterium-mediated transformation, and a transgenic plant is prepared. The neonicotinoid content of the obtained transgenic plant is more than 4 times that of a control group. The result shows that the tobacco NtERF91 gene has a great application prospect in the aspect of cultivating tobacco with high neonicotinoid content.