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Tobacco strigolactones transport protein NtPDR6 and interference expression vector and application thereof

A technology of strigolactone and transporter protein is applied in the field of tobacco strigolactone transporter NtPDR6, which can solve the problem that tobacco strigolactone transport-related genes have not been reported, and achieve the proliferation of stem axillary buds and lateral buds. Obviously, the effect of promoting tobacco branching and reducing gene expression

Active Publication Date: 2014-10-08
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the existing tobacco research reports, there is no report on the transport-related genes of tobacco strigolactone

Method used

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  • Tobacco strigolactones transport protein NtPDR6 and interference expression vector and application thereof
  • Tobacco strigolactones transport protein NtPDR6 and interference expression vector and application thereof
  • Tobacco strigolactones transport protein NtPDR6 and interference expression vector and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, clone tobacco NtPDR6 gene

[0026] According to the sequence of the petunia strigolactone transporter PhPDR1, the primers for amplifying the coding region of the tobacco strigolactone transporter gene (NtPDR6) were designed, and the specific primers were as follows:

[0027] NtPDR6-fw: 5'-atggagggtggtgaagacag-3' (SEQ ID NO.1);

[0028]NtPDR6-rev: 5'-atggagggtggtgaagacag-3' (SEQ ID NO. 2);

[0029] Then, using the root tissue cDNA of tobacco safflower dajinyuan as a template, PCR amplification was carried out with the primer pairs shown in SEQ ID NO.1 and SEQ ID NO.2 respectively. The PCR amplification conditions were: 94°C pre-denaturation for 4 minutes . Denaturation at 94°C for 30s, annealing at 56°C for 30s, extension at 72°C for 240s, a total of 30 cycles. Finally, extend at 72°C for 10 min and store at 4°C. After sequencing the amplified sequence, the sequence shown in SEQ ID NO.3 was obtained, which contained 4482 bp nucleotides, which was a comp...

Embodiment 2

[0030] Example 2. Induced expression analysis of NtPDR6 under hormone and low phosphorus treatment

[0031] Tobacco seedlings grown to the "big cross" stage were transferred from the nutrient soil to deionized water, and after 3 days of recovery, phosphorus deficiency, hormone and high salt induction treatments were performed.

[0032] Phosphorus deficiency treatment: the restored tobacco seedlings were transferred to MS medium without phosphorus for cultivation, and the control group was cultured in normal MS medium. Quick-frozen and stored at -80°C for later use.

[0033] Hormone treatment: Put the restored tobacco seedlings into the nutrient solution supplemented with 50 μM ABA, 100 μM MeJA, and 30 μM NAA, respectively. At the same time, the nutrient solution added with the same amount of water was used as the control. After 24 hours of treatment, the seedlings were taken into a 1.5 mL centrifuge tube medium, quick-frozen in liquid nitrogen, and stored at -80°C for later u...

Embodiment 3

[0036] Embodiment 3, NtPDR6 gene interference vector construction

[0037] According to the obtained NtPDR6 gene sequence, a pair of primers (including forward fragment primer and reverse fragment primer) were designed at the N-terminus to amplify the truncated fragment (SEQ ID NO.9) with a length of 315bp, and according to the enzyme cleavage position Restriction sites were designed at the upstream and downstream of the primers, and the insertion direction of the sequence was controlled by the restriction sites, so as to obtain the interference sequence of the RNAi interference vector.

[0038] The specific primer sequences are as follows:

[0039] Forward Fragment Primer:

[0040] NtPDR6-if-fw:5'-ccg ctcgag taactaaacttgatttggtggaaag-3' (SEQ ID NO.10), the underline indicates the XhoI restriction site;

[0041] NtPDR6-if-rew:5'-cgg ggtacc ctggtttgatgattccactgactt-3' (SEQ ID NO.11), the underline indicates the KpnI restriction site;

[0042] Reverse Fragment Primer:

...

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Abstract

The invention discloses tobacco strigolactones transport protein NtPDR6 and an interference expression vector and application thereof. The tobacco strigolactones transport protein NtPDR6 has an amino acid sequence shown in SEQ ID NO.4 or obtained through substituting, deleting or adding at least one amino acid on SEQ ID NO.4; and a nucleotide sequence for coding the protein is shown in SEQ ID NO.3 or is obtained through substituting, deleting or adding at least one nucleotide on SEQ ID NO.3. The protein has the effect of inhibiting lateral branch, axillary bud or lateral bud proliferation, the lateral branch growth of tobacco subjected to interference expression is obviously active, axillary bud or lateral bud proliferation is obvious, and the protein has significance for controlling the plant type of tobacco and improving the yield of tobacco.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and specifically relates to tobacco strigolactone transporter NtPDR6, and also relates to a gene encoding tobacco strigolactone transporter NtPDR6, a carrier for interfering with the expression of the gene, and an application thereof. Background technique [0002] Phytohormones are secondary metabolites that are induced by specific environmental signals and can regulate plant physiological responses. play an important regulatory role in the process. Common plant hormones include auxin, gibberellin (GA), cytokinin (CTK), abscisic acid (ABA), ethylene (ETH), salicylic acid ( salicylic acid (SA), jasmonic acid (JA) and brassinosteroid (brassinosteroid, BR). Among them, auxin and cytokinin are two hormones traditionally considered to control the development of plant branches. However, recent studies have found that , there is another hormone that can regulate the development of plant branches, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84C12N15/66A01H5/00
CPCC07K14/415C12N15/8261
Inventor 王根洪夏庆友谢小东高军平熊海军
Owner SOUTHWEST UNIV
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