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Sedum lineare salt-tolerant gene SlWRKY and application thereof

A salt-tolerant gene, the technology of S. japonica, applied in the field of molecular biology and biology, can solve the problems of limiting crop types and yields, low utilization efficiency, and differences in salt tolerance.

Inactive Publication Date: 2019-12-13
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

my country is one of the countries with large saline-alkali land in the world, and the current salinized land area is about 30 million hm 2 , accounting for more than 20% of my country's arable land, and widely distributed, low utilization efficiency, seriously hindering the development of agricultural production
Soil salinity also limits the variety and yield of crops, and there are differences in salt tolerance between different crops or different varieties of the same crop

Method used

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  • Sedum lineare salt-tolerant gene SlWRKY and application thereof
  • Sedum lineare salt-tolerant gene SlWRKY and application thereof
  • Sedum lineare salt-tolerant gene SlWRKY and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Cloning of the S1WRKY gene of Sedum linearare (S1 for short)

[0029] Total RNA was extracted from S. japonica (taken from Binhai New District, Tianjin) which was treated with salt tolerance in 150mM NaCl aqueous solution, using the plant RNeasy Plant Mini Kit (Transgene Code#E101-0150rxns), and using EasyScript Frist-Strand cDNA SynSgesis SuperMix (Transgene Code#AE301-03100rxns) was reverse transcribed into cDNA. High-throughput sequencing of cDNA was performed to obtain 78407 transcripts (Novogene performed high-throughput sequencing), and the 3' end sequence of the SlWRKY gene was obtained through comparative analysis with the GO database, using RACE technology (Takara-RACE kit) The full-length cDNA sequence of the SlWRKY gene was obtained, and the amplified SlWRKY gene was sequenced and analyzed to obtain a complete SlWRKY gene with a full-length of 1014bp (SEQ ID No.1). The protein encoded by the S. japonica S1WRKY gene is the amino acid sequence shown in SEQ ...

Embodiment 2

[0080] 1. Transformation of Arabidopsis

[0081] (1) Transform Arabidopsis.

[0082] Specific steps for transforming Arabidopsis:

[0083] Nutrient soil (vermiculite: sterile mixed at a ratio of 1:3), sow Arabidopsis seeds (commercial) vernalized in a refrigerator at 4°C for 72 hours on the soil surface, cover with plastic wrap for the first 7 days, and put them in tissue culture on the shelf. The culture conditions are as follows: the light is 10000 lux; the daytime is 16 hours, 22°C; the night is 8 hours, the temperature is 18°C. Cut off the fruit pods and pollinated flowers of Arabidopsis thaliana that have grown for 1.5 months and bolted about 15 cm. Pick a single colony containing gene-positive clones into 50 mL of YEB liquid medium containing 30 μg / mL gentamicin, 50 μg / mL rifampicin, and 50 μg / mL kanamycin (5 g peptone, 1 g yeast extract, 5 g beef extract , 5 g of sucrose, dilute to 1 L, pH = 7), 28 ° C, 180 rpm, until the OD600 value is 0.6-0.8. Centrifuge the bact...

Embodiment 3

[0089] 1. Transform poplar

[0090] The poplars transformed by the positive clones were Populus alba × P. alba INRA clone N7171-B4 (hereinafter referred to as 717 poplars) tissue culture seedlings. (commodity)

[0091] (1) Put the axillary buds or terminal buds of 717 poplar on the basic medium (MS salt 2.2g, sucrose 30g, constant volume to 1L, pH=5.7, agar 7.2g) for subculture, and culture for 6 weeks to obtain tissue culture seedlings ; Cut off the 1cm stem section without axillary buds of the tissue cultured seedlings, and pre-cultivate them under dark conditions at 24°C for 3 days after scratching the wound;

[0092](2) The positive clone bacterial liquid (OD600=0.8) obtained by the selected embodiment 1 is centrifuged at room temperature and 4000rpm for 10min, the supernatant is discarded, and an equal volume of M solution (M solution: MS salt 4.4g) is used for the precipitation , sucrose 30g, auxin NAA1.86mg, cytokinin 2ip1.02mg, acetosyringone As19.86mg, dilute to 1L,...

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Abstract

The invention discloses a sedum lineare salt-tolerant gene SlWRKY and an application thereof. The nucleotide sequence of the sedum lineare salt-tolerant gene SlWRKY is shown in SEQ ID NO. 1; experiments prove that the tolerance of arabidopsis thaliana and poplar transfected by the SlWRKY gene to salt is enhanced, the sedum lineare provided by the invention is indicated to play an important role inimproving the salt tolerance of crops by enhancing the salt-tolerant gene SlWRKY.

Description

technical field [0001] The invention relates to a Sedum lineare salt-tolerant gene S1WRKY and its application, belonging to the fields of molecular biology and biotechnology. Background technique [0002] Soil salinization has become one of the main factors restricting the sustainable development of agriculture. Plants in a high-salt environment for a long time not only inhibit their growth, but also cause crop yield reduction and even death. About 100 million hm worldwide 2 Land is affected by soil salinity, which accounts for 6% to 7% of the world's total cultivated land, and the area is still expanding. Sea level rise due to global warming may aggravate the problem. my country is one of the countries with large saline-alkali land in the world, and the current salinized land area is about 30 million hm 2 , accounting for more than 20% of my country's cultivated land area, and widely distributed, low utilization efficiency, seriously hindering the development of agricult...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 杨少辉白婧平岳靖杨合宇王洁华
Owner TIANJIN UNIV
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