PHD transcription factor of soybean and its coding gene and usage
A technology for transcription factors and coding genes, which can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve problems such as the lack of research on protein functions.
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Embodiment 1
[0031] Example 1. Screening of soybean GmPDH2 and its coding gene and cloning of cDNA
[0032] Using soybean drought-resistant variety Jindou 23 and drought-sensitive variety Huibuzhi (purchased from Shanxi Academy of Agricultural Sciences) as materials, more than 60 differentially expressed genes were screened by cDNA-AFLP method, including a 296bp fragment, which was subjected to BLAST The soybean EST database indicated that it is a zinc finger protein containing the PHD domain, and the homologous sequence of this gene was found in the drought and salt cDNA libraries of multiple species in the comparison. After comparison in the soybean EST database, 6 members of this family were obtained, and they were named GmPHD1-6. Through EST extension and splicing, the full-length sequences of 4 GmPHD genes were obtained, among which GmPHD2 has the nucleotide sequence of sequence 1 in the sequence list, and encodes the amino acid residue sequence of sequence 2 in the sequence list. Amino a...
Embodiment 2
[0034] Example 2. Expression characteristics of GmPHD2 gene under abiotic stress
[0035] The drought-resistant soybean variety Jindou 23 was treated with drought, NaCl, ABA, and chilling to analyze the expression of soybean GmPHD2 under abiotic stress. The seeds of Jindou 23 were planted in pots, and after two weeks of growth, the seedlings were subjected to the following stress treatments:
[0036] Drought treatment: Take the soybean seedlings out of the soil to absorb the moisture on the roots, place them on dry filter paper, and incubate them under light conditions for 0 hours, 1 hour, 3 hours, 6 hours and 12 hours before sampling.
[0037] Salt treatment: The roots of soybean seedlings were placed in 150mM NaCl solution, and samples were taken after 0 hour, 1 hour, 3 hours, 6 hours and 12 hours of light culture.
[0038]ABA treatment: The roots of soybean seedlings were placed in 20μM ABA, and samples were taken after 0 hours, 1 hour, 3 hours, 6 hours and 12 hours of light cu...
Embodiment 3
[0041] Example 3. Functional identification of GmPHD2 encoded protein
[0042] Insert GmPHD2 into pBin438 (Li Taiyuan, Tian Yingchuan, Qin Xiaofeng, etc., Research on highly effective insect-resistant transgenic tobacco, Science in China (Series B), 1994, 24(3): 276-282) BamHI and KpnI digestion sites of plant expression vector Between points, the constructed plant expression vector containing GmPHD2 is shown in Figure 2 and named pBGmPHD2. pBGmPHD2 was transformed into Arabidopsis thaliana via Agrobacterium tumefaciens GV3101. PCR results showed that 11 transformed plants were obtained. Northern analysis showed that at least three of them had high expression levels of GmPHD2. The three high-expression plants were named G1-3, G1-6, G1-8. The seeds of G1-3, G1-6, G1-8 were planted into MS medium, and the obtained seedlings were named G2-3, G2-6, G2-8, 5 days later and non-transgenic Arabidopsis (WT) The seedlings were transplanted into 1 / 2MS medium containing 50mM, 100mM, 150mM and...
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