Cloning and application of tobacco alkaloid synthesis negative regulation gene NtARF6

• Main Authors: ZHANG HONGBO, ZOU CONGMING, YUAN CHENG, HU MENGYANG, SUI XUEYI, ZENG JIANMIN, WU XINGFU, LI YONGPING, WANG BINGWU, GAO YULONG, SHI JUNLI, KONG GUANGHUI, LI WENZHENG, SONG ZHONGBANG, HUANG CHANGJUN, ZHAO LU, LIU YONG, LI MEIYUN
• Format: Patent
• Language: Chinese; English
• Published: 08.05.2020
• Subjects: AGRICULTURE; ANIMAL HUSBANDRY; BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FISHING; FORESTRY; HUMAN NECESSITIES; HUNTING; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; NEW PLANTS OR PROCESSES FOR OBTAINING THEM; ORGANIC CHEMISTRY; PEPTIDES; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TRAPPING; VINEGAR; WINE

The invention discloses a tobacco alkaloid synthesis negative regulation gene NtARF6 and a cloning method and application thereof. The nucleotide sequence of the tobacco alkaloid synthesis negative regulation gene NtARF6 is shown as SEQ ID: NO: 1, and the coded amino acid sequence is shown as SEQ ID: NO: 2. The invention further discloses the cloning method of the tobacco alkaloid synthesis negative regulation gene NtARF6. The method comprises the following specific steps: A, determining an NtARF6 gene sequence; B, extracting tobacco RNA, and performing reverse transcription to obtain a firstchain cDNA; C, designing and synthesizing a specific primer according to the NtARF6 gene sequence, and performing PCR amplification by taking the cDNA as a template; D, recovering and purifying a PCRproduct; and E, constructing an overexpression vector containing the NtARF6 gene. The overexpression vector is overexpressed in nicotiana tabacum through agrobacterium-mediated transformation, and a transgenic plant is prepare