BHLH transcription factor for promoting anthocyanin synthesis in lotus and application thereof

A technology of transcription factors and anthocyanins, applied in the field of molecular biology, can solve the problems of insufficient improvement of lotus flower color molecular breeding, achieve the effect of enriching theoretical understanding and promoting molecular breeding

Active Publication Date: 2020-12-01
金天国际医疗科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the bHLH transcription factors that regulate anthocyanin biosynthesis have not been reported in lotus, and the study of MYB transcription factors alone is far from enough to improve the molecular breeding of lotus flower color

Method used

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  • BHLH transcription factor for promoting anthocyanin synthesis in lotus and application thereof
  • BHLH transcription factor for promoting anthocyanin synthesis in lotus and application thereof
  • BHLH transcription factor for promoting anthocyanin synthesis in lotus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Cloning of NnTT8 Gene from Lotus Flower

[0023] Through comparative transcriptomics methods, a bHLH transcription factor gene whose expression level was up-regulated during the flower bud coloring process of the safflower cultivar 'Yehonglian' at different developmental stages was named NnTT8. Total RNA (ribonucleic acid) was extracted from the fresh petals of 'Yehonglian' and reverse-transcribed into cDNA. The CDS sequence of NnTT8 was obtained by PCR amplification with specific primer pair NnTT8-F: ATGGCTGCACCAGTGAGTAATCGCC and NnTT8-R: CATGAATTCATTTATAATTTGGTGT. The PCR reaction system is 50 μL, and the components include: 2×PrimeSTAR Max DNA Polymerase 25 μL, upstream and downstream primers 2 μL each, cDNA 1 μL, ddH 2 O 20 μL. The PCR program was: 35 cycles of 98°C for 10s, 60°C for 15s, and 72°C for 15s. The cloned NnTT8 sequence was sequenced to determine its complete CDS sequence.

Embodiment 2

[0025] Preparation of recombinant plasmid containing NnTT8 gene and recombinant genetically engineered bacteria.

[0026] Using the product obtained in Example 1 as a template, using a pair of specific primers: (The underlined part is the homologous sequence at the end of the upstream vector, and the italic part is the AscI restriction site) and (The underlined part is the homologous sequence at the end of the downstream vector, and the italicized text part is the SpeI restriction site). Perform PCR amplification. The PCR reaction system and procedure are consistent with those in Example 1. After recovering the PCR product, the II One Step Cloning Kit, using homologous recombination to connect to the PMDC83 vector that has been digested with AscI and SpeI, and sequenced to verify that the recombinant plasmid containing the NnTT8 gene was obtained. Then, the plasmid was introduced into the GV3101 Agrobacterium strain, thereby obtaining an engineering bacterium containing a...

Embodiment 3

[0028] Verification of NnTT8 gene function

[0029] The Arabidopsis tt8 mutant was selected to verify the function of NnTT8 gene.

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Abstract

The invention relates to the field of molecular biology, and discloses a bHLH transcription factor for promoting anthocyanin synthesis in lotus and application thereof in lotus flower color breeding.The CDS sequence of the bHLH transcription factor NnTT8 for promoting anthocyanin biosynthesis in the lotus is as shown in SEQ ID No. 1. The bHLH transcription factor NnTT8 for regulating anthocyaninbiosynthesis in lotus is researched for the first time, functional verification is performed on the bHLH transcription factor NnTT8 through an arabidopsis thaliana tt8 mutant recovery experiment, andmolecular breeding of lotus flowers is promoted.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a bHLH type transcription factor promoting anthocyanin synthesis in lotus and application thereof. Background technique [0002] Lotus (Nelumbo nucifera Gaertn.) is one of the ten famous flowers in my country, which has important ornamental and cultural value. Deeply loved by the masses, the lotus industry and lotus exhibitions in various places are in the ascendant. Among them, the color of the lotus plays an important role in determining its ornamental value. Therefore, in lotus breeding, flower color is one of the important target traits. [0003] The breeding work of lotus is mainly through the traditional cross breeding method. After crossing the parents with different phenotypes of the target traits, the phenotypes of the target traits are screened in the offspring to obtain ideal performance of the target traits and stable reproductive phenotypes for many years. plants....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/825
Inventor 祖明艳杨平仿吴倩邓娇韩杰
Owner 金天国际医疗科技有限公司
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