WO2014026312A1 - Isopentényl-transférase de coton, gène codant pour celle-ci et ses utilisations - Google Patents

Isopentényl-transférase de coton, gène codant pour celle-ci et ses utilisations Download PDF

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WO2014026312A1
WO2014026312A1 PCT/CN2012/080001 CN2012080001W WO2014026312A1 WO 2014026312 A1 WO2014026312 A1 WO 2014026312A1 CN 2012080001 W CN2012080001 W CN 2012080001W WO 2014026312 A1 WO2014026312 A1 WO 2014026312A1
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plant
seq
gene
expression vector
tobacco
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PCT/CN2012/080001
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English (en)
Chinese (zh)
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何云蔚
陈淼
崔洪志
王建胜
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创世纪转基因技术有限公司
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Priority to PCT/CN2012/080001 priority Critical patent/WO2014026312A1/fr
Priority to CN201280004133.5A priority patent/CN103748223B/zh
Publication of WO2014026312A1 publication Critical patent/WO2014026312A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)

Definitions

  • the present invention relates to a prenyltransferase and a gene encoding the same, and particularly to a cotton-derived isopentenyl transferase IPT1-1 and Its coding gene, and its application in the cultivation of transgenic plants with improved drought tolerance.
  • BACKGROUND OF THE INVENTION Abiotic stresses, such as drought, salt, extreme temperature, chemical pollution and oxygen damage, can cause serious damage to plant growth and development, and cause great losses to crop yield.
  • the impact of drought on crop yield is It takes the first place in natural adversity, and its harm is equivalent to the sum of other disasters. It is the bottleneck of agricultural development in many areas.
  • the world's dry and semi-dry areas account for 34% of the land area; China's dry and semi-arid areas account for 52% of the country's land area, and the annual area is 20 to 2.7 million hectares.
  • genes and their expression products can be divided into three categories: (1) genes and products involved in signal cascade amplification and transcriptional control; (2) genes and their expression products that directly contribute to the protection of biofilms and proteins; ) Proteins associated with the uptake and transport of water and ions.
  • genes and products involved in signal cascade amplification and transcriptional control (2) genes and their expression products that directly contribute to the protection of biofilms and proteins; ) Proteins associated with the uptake and transport of water and ions.
  • studies on the ability of plants to improve stress tolerance through transgenic techniques, as well as studies on stress-tolerant crops, xerophytes and halophytes, have yielded significant results for stress-related genes and signal transduction.
  • the system has further advanced the corner army (Liu Q.1998.
  • the present inventors cloned the DNA sequence of a gene encoding a isopentenyltransferase (designated herein as IPT1-1) of cotton using SSH in combination with RACE. It was found that the introduction of the transgenic plants significantly improved the drought tolerance of the transgenic plants, and these traits were stably inherited.
  • a first aspect of the invention provides a gene encoding a prenyltransferase IPT1-1 of cotton, the sequence of which is SEQ ID NO: 2.
  • a second aspect of the invention provides a recombinant expression vector comprising the gene of the first aspect of the invention, and the nucleotide sequence of the gene is operably linked to an expression control sequence of the expression vector; preferably, The vector is the rd29A-GMPT1-1-2300 carrier shown in Fig. 2.
  • a third aspect of the invention provides a recombinant cell comprising the gene of the first aspect of the invention or the recombinant expression vector of the second aspect of the invention; preferably, the recombinant cell is a recombinant Agrobacterium cell.
  • a fourth aspect of the present invention provides a method for improving drought tolerance of a plant, comprising: introducing the gene of the first aspect of the invention or the recombinant expression vector of the second aspect of the invention into a plant or plant tissue and causing the gene Expression;
  • the plant is tobacco.
  • a fifth aspect of the invention provides a method for producing a transgenic plant, comprising: cultivating a plant or plant comprising the gene of the first aspect of the invention, the recombinant expression vector of the second aspect of the invention under conditions effective to produce a plant Tissue;
  • the plant is tobacco.
  • a sixth aspect of the present invention provides the gene according to the first aspect of the present invention, the recombinant expression vector of the second aspect of the present invention or the recombinant cell of the third aspect of the present invention for improving drought tolerance of a plant and for use in plant breeding Use;
  • the plant is tobacco.
  • the seventh aspect of the present invention provides the amino acid sequence encoded by the gene of the first aspect of the present invention, as shown in SEQ ID NO: 1.
  • Fig. 1 is a construction flow of a plant expression vector (rd29A-GMPT1-1-2300) of GhIPT1-1.
  • Figure 2 is a plasmid map of the plant expression vector (rd29A-GMPT1-1-2300) of GhIPTl-1.
  • Figure 3 shows the drought tolerance simulation results of GhIPTl-1 T1 transgenic tobacco plants (in the figure, T1A1; right, T1A5) and non-transgenic tobacco plants (left, CK).
  • Figure 4 shows the results of protein expression verification at the transcriptional level of transgenic T1 tobacco plants and non-transgenic control plants.
  • M is Marker
  • 1_5 is the control tobacco
  • 6_18 is the drought-tolerant transgenic tobacco T1 plant
  • 19-24 is the drought-tolerant transgenic tobacco T1 plant.
  • BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be further described below in conjunction with non-limiting examples.
  • Example 1. Cotton SSH library construction under drought stress:
  • a subtractive library was constructed by inhibition subtractive hybridization using the method shown by Clontech's PCR-selectTM cDNA Subtraction Kit.
  • the mRNA of the leaves of the drought-treated cotton seedlings was used as a tester during the experiment, and the mRNA of the leaves of the untreated cotton seedlings was used as a control.
  • the specific steps are as follows:
  • test seedlings were divided into two groups, each with 4 pots and 1 pot per pot.
  • the first group was the control group, cultured at 25 ° C, lighted, and normally watered.
  • the second group was the drought treatment group, 25 °C, light culture, stop watering, treatment for 10 days. After the treatment, the leaves of the top 1/3 of the two seedlings were cut out in time, and then rapidly frozen with liquid nitrogen at -70 °C. Store in the refrigerator.
  • the cotton leaves of the control group and the drought treatment group were respectively taken 0.5 g, and the plant ⁇ extraction kit was used.
  • RNA (invitrogen) Extracts total RNA from cotton. 0% ⁇ Using HITACHI's UV spectrophotometer U-2001 to determine the total RNA absorbance at 260 nm and 280 nm, 0D260/0D280 ratio of 1. 8-2 ⁇ 0, indicating that the total RNA purity is higher, with 1.0% The integrity of total RNA was detected by agarose gel electrophoresis, and the brightness of the 28S band was approximately twice that of the 18S band, indicating good RNA integrity. The mRNA was isolated using Qiagen's Oligotex mRNA purification assay [JP (purification of polyA+ RNA from total RNA).
  • the gene In order to gain the effectiveness of the Expressed Sequence Tag (EST) (unigene), the gene is not cleavable and the obtained sequence is in the untranslated region.
  • the laboratory uses Rsal, Haelll to digest the double-stranded cDNA (according to the protocol described in the subtractive kit, obtained by reverse transcription of mRNA), and performs two sets of suppression subtraction. Other steps and methods are based on Clontech's PCR-selectTM. The method shown in the cDNA Subtraction Kit was used for suppression subtractive hybridization, and finally the second PCR product of the two groups of forward subtracted hybrid cDNA fragments was combined.
  • Kit purified, purchased from Qiagen and pGEM-T Easy (purchased from Promega kit) vector, according to the procedure of pGEM-T Easy kit, the specific steps are as follows: Add the following components in sequence with 200 ⁇ 1 PCR tube: Purified positive The second PCR product of the subtractive hybridization cDNA fragment was 3 ⁇ l, ⁇ 4 ligase buffer 5 ⁇ l , pGEM-T Easy vector 1 ⁇ l, ⁇ 4 DNA ligase 1 ⁇ , and ligated overnight at 4 °C. Take 10 linked reaction products and add to 100 competent E. coli JMI09 (purchased from In TAKARA), ice bath for 30 min, heat shock for 60 s, ice bath for 2 min, plus 250 L LB medium
  • LB (ibid.) /X-gal/IPTG (X-gal/IPTG purchased from TAKARA) of 50 ⁇ g/mL ampicillin was cultured on a culture plate at 37 °C. 18 counts of clear white and blue with a diameter > 1 mm in the culture plate. The number of colonies was collected, and 360 white colonies were randomly picked (number: Gh-D001 to Gh-D360). All white clones were picked in 96-well cell culture plates (CORNING) containing LB liquid medium containing 50 ⁇ g/mL ampicillin, and cultured overnight at 37 °C, glycerol was added to a final concentration of 20%, and stored at -80 °C. spare.
  • CORNING 96-well cell culture plates
  • the nested PCR primers Primer 1 and Primer 2R (Clontech's PCR-se ectTM cDNA Subtraction Ki t kit) were used for PCR amplification, and 292 positive clones were obtained, and all positive clones were sent to the UK. ⁇ Jieji (Shanghai) Trading Co., Ltd. sequencing
  • the GhIPTl-1 gene fragment that has been obtained already has a stop codon TAA, and only 5 ' RACE is required.
  • three specific primers were designed as reverse transcription primers and 3'-end specific primers for 5' RACE.
  • GhIPTl-1 GSP1 SEQ ID NO : 4:
  • GhIPTl-1 GSP2 SEQ ID NO: 5:
  • GhIPTl-1 GSP3 SEQ ID NO: 6:
  • the experimental procedure was performed according to the kit instructions (5, RACE System for Rapid Amplification of cDNA Ends kit was purchased from Invitrogen).
  • the first round of PCR amplification was carried out using SEQ ID NO: 5 and 5 ' universal primer AAP (provided with the kit), and mRNA reverse transcription cDNA (reverse transcription primer SEQ ID NO: 4) as a template, the specific steps are as follows: Ex Taq was purchased from TAKARA, 50 ⁇ 1 PCR reaction system: 5 ⁇ 1 ⁇ ⁇ ⁇ Buffer, 3 ⁇ 1 2. 5 mM dNTP, 2. 0 ⁇ 1 mRNA reverse transcribed cDNA, 1. 0 ⁇ 1 Ex Taq 10 ⁇ M of primers SEQ ID NO: 5 and AAP each of 2.0 ⁇ l, and 35 ⁇ l of double distilled water.
  • PCR conditions 94 ° C denaturation for 5 min, 94 ° C denaturation 30 s, 55 ° C annealing 30 s, 72 ° C extend 2min, after 33 cycles, 72 ° C extend 10 min 0
  • the resulting PCR product was double After diluting 50 times of distilled water, take 2.0 ⁇ l as template and carry out the second round of PCR amplification with SEQ ID NO: 6 and 3' primer AUAP.
  • the specific steps are as follows: 50 ⁇ 1 PCR reaction system: 5 ⁇ 1 ⁇ ⁇ ⁇ Buffer, 3 ⁇ 1 2. 5 mM dNTP, 2. 0 ⁇ 1 diluted first round PCR product, 1.
  • PCR reaction conditions predenaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, Anneal at 58 °C for 30 s, 72 °C for 2 min, after 33 cycles, and at 72 °C for 10 min.
  • the second PCR product was recovered from a fragment of about 700 bp (Gel Extraction Kit was purchased from OMEGA). It was ligated to pGEM-T Easy Vector, transformed into JM109 (specifically the same as above), and 10 white colonies were randomly picked up to contain 50 g/mL.
  • Ampicillin was cultured in LB liquid medium, cultured at 37 ° C overnight, and glycerin was added to a final concentration of 20%, and stored at -80 ° C until use.
  • SEQ ID NO: 6 and 3' primer AUAP were used for PCR amplification (reaction system and reaction conditions are the same as above), and three positive clones were obtained and sent to Yingji Jieji (Shanghai) Trading Co., Ltd. for sequencing and sequencing. The 5' end of the cDNA.
  • the obtained 5' RACE product was cloned and sequenced, and spliced with the cloned Gh-D211 sequencing result.
  • CTATGTCTGC CAGCAAACTA GTACAACCCC GAGTGAATTT GGGGCTCAAC CTGGAGACTA
  • GhIPTl-lF SEQ ID NO: 7:
  • GhIPTl-lR SEQ ID NO: 8:
  • PCR reaction was carried out using TaKaRa's PrimeSTAR HS DNA polymerase and cotton cDNA as a template.
  • 50 ⁇ 1 PCR reaction system 10 ⁇ 5XPS Buffer, 3 ⁇ 1 2.5 mM dNTP, 2.0 ⁇ 1 cDNA, 1.0 ⁇ 1 PrimeSTAR, 10 ⁇ M primer SEQ ID NO: 7 and SEQ ID NO: 8 each 2.0 ⁇ 1, and 30 ⁇ l of double distilled water.
  • PCR reaction conditions pre-denaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 58 °C for 30 s, extension at 72 °C for 2 min, after 33 cycles, extension at 72 °C for 10 min.
  • the PCR amplification product was added with A tail: The PCR product was added 2.5 times of absolute ethanol, placed at -20 ° C for 10 minutes, centrifuged, and the supernatant was removed, dried, and dissolved in 21 ⁇ l of double distilled water. Add 2.5 ⁇ l 10 X Ex Buffer, 0.5 ⁇ l 5 mM dATP, 2.5 ⁇ 1 ⁇ Taq. Reaction conditions: The reaction was carried out at 70 ° C for 30 minutes. A DNA fragment of about 900 bp was recovered (Omega Recovery Kit), ligated into the pGEM T-easy vector, transformed into JM109 (method as above), and 10 white colonies were randomly picked from LB containing 50 ⁇ g/mL ampicillin.
  • SEQ ID NO: 7 and SEQ ID NO: 8 were used for bacterial liquid PCR amplification (reaction system and reaction conditions are the same as above), and 4 positive clones were obtained and sent to Yingjie Jieji (Shanghai) Trading Co., Ltd. for sequencing.
  • the sequence is SEQ ID. NO: 2.
  • CTGGAGACTA TCTTCCGCAG AAAGGATAAG GTGGTGTTTG TAATGGGAGC
  • the plant binary expression vector PCAMBIA2300 (purchased from Beijing Dingguo Changsheng Biotechnology Co., Ltd.) was selected as the plant expression vector, and the 35S promoter of the ⁇ gene containing the double enhancer was replaced with the Pnos promoter to reduce the expression of prion protein in plants. .
  • the inducible promoters rd29A and Tnos were selected as promoters and terminators of the GhIPTl-1 gene.
  • Pnos were amplified using the plant expression vector PBI 121 (purchased from Beijing Huaxia Ocean Technology Co., Ltd.) using primers SEQ ID NO: 9 and SEQ ID NO: 10, using TaKaRa's PrimeSTAR HS DNA polymerase.
  • SEQ ID NO: 10 each of 2.0 ⁇ l, and 31 ⁇ of double distilled water.
  • PCR reaction conditions pre-denaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 56 °C for 30 s, extension at 72 °C for 30 s, after 33 cycles, extension at 72 °C for 10 min.
  • PCAMBIA2300-1 was obtained by EcoRI, Bglll digestion with pCAMBIA2300 (promega, T4 ligase cassette).
  • SEQ ID NO: 11 and SEQ ID NO: 12 amplify Tnos using PBI 121 as a template, using PrimeSTAR HS DNA polymerase of TaKaRa.
  • 50 ⁇ 1 PCR reaction system 10 ⁇ 1 5 X PS Buffer, 3 ⁇ l 2.5 mM dNTP, 1.0 ⁇ l PBI121, 1.0 ⁇ l PrimeSTAR, 10 ⁇ primers SEQ ID NO: 11 and SEQ ID NO: 12 each 2.0 ⁇ l, and 31 ⁇ l of double distilled water.
  • PCR reaction conditions predenaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 58 °C for 30 s, extension at 72 °C for 30 s, after 33 cycles, extension at 72 °C for 10 min.
  • pCAMBIA2300-2 was obtained by restriction enzyme digestion with pCAMBIA2300-l (promega T4 ligase cassette) by Sacl and EcoRI
  • SEQ ID NO: 13 and SEQ ID NO: 14 The Arabidopsis rd29A promoter was amplified using Arabidopsis thaliana (Columbia type, available from www. arabidopsis.org) DNA as a template (see Zeng J., et L. 2002, Preparation). Of total DNA from "recalcit rant plant taxa", Acta Bot. Sin., 44(6): Method 694-697 for extracting Arabidopsis DNA). PrimeSTAR HS DNA polymerase from TaKaRa was used.
  • PCR reaction system 10 ⁇ 15 ⁇ PS Buffer, 3 ⁇ 1 2 ⁇ 5 mM dNTP, 1.0 ⁇ 1 Arabidopsis DNA, 1.0 ⁇ 1 PrimeSTAR, 10 ⁇ ⁇ primers SEQ ID NO: 13 and SEQ ID NO: 14 each of 2.0 ⁇ 1, and 31 ⁇ 1 of double distilled water.
  • PCR reaction conditions predenaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 58 °C for 30 s, extension at 72 °C for 30 s, after 33 cycles, extension at 72 °C for 10 min by HindIII, Pstl digestion Connected to (connection method is the same as above)
  • pCAMBIA2300-2 obtained pCAMBIA2300_3
  • SEQ ID NO: 15 and SEQ ID NO: 16 amplify GhIPTl-1 (template is GhIPTl-1 obtained in Example 2), using PrimeSTAR HS DNA polymerase of TaKaRa. 50 ⁇ 1 PCR reaction system: 10 ⁇ 15 ⁇ PS Buffer, 3 ⁇ 12 ⁇ 5 mM dNTP, 1.0 ⁇ 1 GhlPTl- 1- pGEM, 1.0 ⁇ 1 PrimeSTAR, 10 ⁇ M primers SEQ ID NO: 15 and SEQ ID NO: 16 each of 2 ⁇ 0 ⁇ 1, and 31 ⁇ 1 of double distilled water.
  • Agrobacterium LBA4404 (available from Biovector Science Lab, Inc) Competent preparation: Agrobacterium LBA4404 was plated on LB solid medium containing 50 ⁇ g/ml rifampicin and 50 ⁇ g/ml streptomycin 1-2 days in advance Single spot inoculation, culture at 28 ° C for 1 to 2 days. Single colonies were picked and inoculated into 5 ml of LB liquid medium containing 50 ⁇ g/ml rifampicin and 50 ⁇ g/ml streptomycin. Incubate overnight at 28 °C (about 12-16 h) to 0D600 value. 0. 4, forming a seed bacterial liquid.
  • Transformation of Agrobacterium Thaw competent cells on ice, add 1 ⁇ l of plasmid to 40 ⁇ l of competent cells, mix and mix with ice bath for about 10 mino. Transfer the mixture of competent and DNA to pre-cooled by gun.
  • the electric shock cup tap to bring the suspension to the bottom, taking care not to have air bubbles.
  • the program of MicroPulser purchased from bio_rad
  • the program of MicroPulser purchased from bio_rad
  • the electric shock cup was immediately taken out and the pre-warmed LB medium at 28 ° C was added. Quickly and gently spread the cells with a gun. The suspension was transferred to a 1.5 ml centrifuge tube and incubated at 28 ° C, 225 rpm for 1 h. Take 100 ⁇ 200 ⁇ ⁇ of the bacterial solution and plate on the corresponding resistant screening medium (LB solid medium containing 50 ⁇ g/ml rifampicin, 50 ⁇ g/ml streptomycin, 50 ⁇ g/ Ml Kanamycin), cultured at 28 °C.
  • LB solid medium containing 50 ⁇ g/ml rifampicin, 50 ⁇ g/ml streptomycin, 50 ⁇ g/ Ml Kanamycin
  • the leaves of the sterile seedlings were cut into 5 mm ⁇ 5 hidden leaf discs, and the leaf discs were inoculated with the Agrobacterium containing the expression vector rd29A-GhIPTl-l-2300 in the logarithmic growth phase for 10 min, and the bacterial liquid was sucked in the dark condition.
  • the cells were cultured for 2 days (MS medium).
  • the obtained transgenic tobacco leaves were taken, DNA was extracted (the Arabidopsis thaliana DNA extraction method in Example 3), and SEQ ID NO: 7 and SEQ ID NO: 8 (50 ⁇ l PCR reaction system: 5 ⁇ 1 ⁇ ⁇ ⁇ Buffer) , 3 ⁇ 1 2 ⁇ 5 mM dNTP, 2. 0 ⁇ 1 DNA, 1. 0 ⁇ 1 Ex Taq 10 ⁇ M primers SEQ ID NO: 9 and SEQ ID NO: 10 each 2. 0 ⁇ 1, and 35 Double distilled water of ⁇ 1.
  • A20 and control tobacco seeds were sown on vermiculite, 15 seeds per pot, 25 ° C, 10 hours light culture / 14 hours dark culture cycle, 1/2 MS every 5 days, 25 days after culture, each pot was kept 4_5 seedlings of uniform size, drought test, transgenic tobacco, control tobacco drought for 14 days (no watering), 25 ° C, 10 hours light culture / 14 hours dark culture cycle.
  • PCR was performed using TaKaRa's PrimeSTAR HS DNA polymerase with reverse transcribed cDNA as a template.
  • 50 ⁇ 1 PCR reaction system 10 ⁇ 1 5 X PS Buffer, 3 ⁇ 1 2. 5 mM dNTP, 2. 0 ⁇ 1 cDNA, 1. 0 ⁇ 1 PrimeSTAR, 10 ⁇ ⁇ primer SEQ ID NO: 7 and SEQ ID NO: 8 each of 2.0 ⁇ l, and 30 ⁇ l of double distilled water.
  • M is DNA Ladder Marker (DL2000, purchased from Shenzhen Ruizhen Biotechnology Co., Ltd.), 1-5 is control tobacco, and 6-18 is drought-tolerant transgenic tobacco T1 plant, 19-24 It is a T1 generation plant that is not tolerant to transgenic tobacco.
  • the strip size shown in the figure is the same as the size of GhIPTl-1.

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Abstract

L'invention concerne une isopentényl-transférase IPT1-1 de coton, le gène codant pour celle-ci et ses utilisations pour la culture de plantes transgéniques à tolérance améliorée de la sécheresse.
PCT/CN2012/080001 2012-08-13 2012-08-13 Isopentényl-transférase de coton, gène codant pour celle-ci et ses utilisations WO2014026312A1 (fr)

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PCT/CN2012/080001 WO2014026312A1 (fr) 2012-08-13 2012-08-13 Isopentényl-transférase de coton, gène codant pour celle-ci et ses utilisations
CN201280004133.5A CN103748223B (zh) 2012-08-13 2012-08-13 一个棉花异戊烯基转移酶及其编码基因与应用

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PCT/CN2012/080001 WO2014026312A1 (fr) 2012-08-13 2012-08-13 Isopentényl-transférase de coton, gène codant pour celle-ci et ses utilisations

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CN104894080B (zh) * 2015-05-14 2017-12-19 中国科学院华南植物园 一种类黄酮异戊烯基转移酶AhFDT3及其编码基因和应用
CN108753749B (zh) * 2018-06-20 2021-06-08 西南大学 家蚕tRNA异戊烯基转移酶基因及其重组载体和应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1404723A (zh) * 2001-09-20 2003-03-26 华中农业大学 一种快速棉花转基因的方法
WO2004000015A2 (fr) * 2002-06-19 2003-12-31 Basf Plant Science Gmbh Compositions et procedes pour ameliorer les performances des plantes
US7705201B2 (en) * 2004-06-23 2010-04-27 Monsanto Technology Llc Transgenic plants expressing cytokinin biosynthetic genes and methods of use therefor

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1404723A (zh) * 2001-09-20 2003-03-26 华中农业大学 一种快速棉花转基因的方法
WO2004000015A2 (fr) * 2002-06-19 2003-12-31 Basf Plant Science Gmbh Compositions et procedes pour ameliorer les performances des plantes
US7705201B2 (en) * 2004-06-23 2010-04-27 Monsanto Technology Llc Transgenic plants expressing cytokinin biosynthetic genes and methods of use therefor

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE EMBL 28 March 2011 (2011-03-28), accession no. Q326777 *
DATABASE NCBI 4 December 2009 (2009-12-04), accession no. P 002312745.1 *
HUAZHONG AGRICULTURAL UNIVERSITY: "Genetic Transformation of Cotton with Isopentenyl Transferase Gene and Regeneration of Transgenic Plants", MASTER'S THESES, 2004, pages D047 - 139 *

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