ERF transcription factor related to plant stress resistance and encoding gene and application thereof
A transcription factor, coding gene technology, applied in the direction of plant genetic improvement, botanical equipment and methods, applications, etc.
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Embodiment 1
[0063] Example 1 Isolation, Identification and Cloning of the Gene Encoding the Transcription Factor of Lotus japonicus LcERF056
[0064] 1 Materials and methods
[0065] 1.1 Materials
[0066] 1.1.1 Plant material cultivation and stress treatment
[0067] Using lotus root "Leo" as the material, select mature and plump seeds that are free from diseases and insect pests, soak them in 2% NaClO for 15-20 minutes, wash them with sterile water for 3 times, dry them, and put the sterilized seeds on the B5 medium , cultured in a light incubator for 30 days, treated with 150mM sodium chloride and 20% PEG for 0h, 3h, 12h, and 24h, collected leaves were immediately frozen in liquid nitrogen, and stored in a -80°C refrigerator for later use.
[0068] Analysis of tissue expression specificity: The roots, stems, leaves, flowers, and seeds of J. japonicus were taken, quick-frozen in liquid nitrogen, and stored at -80°C until use.
Embodiment 2
[0113] Example 2 Construction of plant expression vector containing LcERF056 gene and its salt tolerance function verification in Arabidopsis
[0114] 1 Materials and methods
[0115] 1.1 Materials
[0116] 1.1.1 Plant material
[0117] Arabidopsis thaliana Columbia (Col-0) ecotype: preserved in the inventor's laboratory.
[0118] 1.1.2 Strain and carrier
[0119] Agrobacterium tumefaciens strain GV3101 was preserved in the inventor's laboratory; gateway entry vector pDNOR201 and expression vector pH7WG2D were purchased from Invitrogen.
[0120] 1.1.3 Medium
[0121] 1.1.3.1YEB medium (1L)
[0122] Peptone 5g, yeast extract 1g, sucrose 5g, MgSO 4 ·7H 2 O0.5g, dilute to 1L with deionized water, if it is a solid medium, add 15g agar powder, and sterilize under high temperature and high pressure at 121°C for 15min.
[0123] 1.1.3.2LB medium (1L)
[0124] Tryptone 10g, sodium chloride 10g, yeast extract 5g, deionized water to 1L, if it is a solid medium, add 15g agar powd...
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