Soybean pubescence related protein as well as coding gene and application thereof
A gene and soybean technology, which is applied in the fields of soybean fuzz-sparse-related proteins and their coding genes and applications, and can solve the problems of unreported genes related to hair density
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Embodiment 1
[0085] Example 1 Establishment of GmPs Overexpression Transgenic Plants
[0086] On the basis of a large number of sequence analysis and functional verification, a protein was found from soybean variety Williams 82, and it was named GmPs protein (abbreviated as Ps protein). Its amino acid is shown in SEQ ID NO: 1 in the sequence table,
[0087] The gene encoding the GmPs protein is named GmPs gene, its genome sequence is shown in SEQ ID NO: 7 of the sequence listing (including 1 exon), and its cDNA sequence is shown in SEQ ID NO: 8 of the sequence listing.
[0088] 1. Construction of recombinant plasmids
[0089] 1. The apical meristem of soybean variety Williams 82 was separated from the plant, RNA was extracted and then reverse-transcribed to obtain cDNA of the apical meristem of soybean variety Williams 82.
[0090] 2. Using the cDNA synthesized in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product...
Embodiment 2
[0118] Example 2 Changes in hairy traits in GmPs overexpression transgenic plants
[0119] 1. Gene expression identification
[0120] The recipient strain DN50, the GmPs overexpression transgenic strain OE-1 strain and the OE-2 strain were identified as follows:
[0121] (1) Take the apical meristem of the seedling line, extract the total RNA and reverse transcribe it into cDNA.
[0122] (2) Using the cDNA extracted in step (1) as a template, use the primer pair consisting of F4 and R4 to identify the expression level of the GmPs gene, and use F3 and R3 to identify the internal reference gene (the expression level of the Actin gene).
[0123] F3: (SEQ ID NO: 13);
[0124] R3: (SEQ ID NO: 14).
[0125] F4: (SEQ ID NO: 15);
[0126] R4: (SEQ ID NO: 16).
[0127] The conditions for quantitative PCR amplification were: pre-denaturation at 95°C for 5 minutes; amplification (45 cycles) at 95°C for 10s, 59°C for 15s, and 72°C for 20s. The melting curve starts after the end of t...
Embodiment 3
[0132] Embodiment 3, the drought resistance of GmPs overexpression transgenic line
[0133] Drought seriously affects the growth and development of plants, and severe drought can cause serious loss of yield. Therefore, improving the drought resistance of crops plays an important role in the stable yield of crops. We detected the performance of DN50 and GmPs overexpression transgenic plants for drought resistance. The soybean seedlings of the GmPs overexpression transgenic lines prepared above and the DN50 soybean seedlings of the control group were used for drought resistance treatment, and the seedlings used for drought resistance treatment were uniformly controlled. After two weeks of watering, rehydration treatment was carried out, and the survival rate of soybean seedlings after rehydration was counted. During the water control period, no watering was carried out at all, and the rewatering treatment was to restore the normal watering amount. The results showed that the r...
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