Application of paddy rice genes OsMADS25 to promoting tap root system plant lateral root growth
A gene and rice technology, applied in the application field of rice OsMADS25 gene in promoting the growth of lateral roots of taproot plants, can solve problems such as internal factors that have no profound explanation for changes, and achieve improved nitrogen use efficiency, important theoretical significance and potential applications Value, the effect of promoting early growth and development
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Embodiment 1
[0027] Example 1 Preparation of transgenic Arabidopsis
[0028] (1) Take the roots of rice seedlings, grind them with liquid nitrogen, add TRIzol reagent (Invitrogen Company), and extract RNA. 0.5 μg RNA was taken for reverse transcription reaction to obtain cDNA.
[0029] (2) Design specific primers according to the OsMADS25 gene sequence on Genbank, add SalI and NotI restriction enzyme sites to the 5' ends of the primers respectively, and the sequences are as follows:
[0030] Upstream primer: 5'-GCGTCGACATGGGGAGAGGGAAGATTG-3' (SEQ ID No.2);
[0031] Downstream primer: 5'-TTGCGGCCGCTGTAGTAGGCGTGATGCTG-3' (SEQ ID No. 3).
[0032] (3) Perform PCR amplification using the full-length cDNA as a template.
[0033] high fidelity enzyme HS DNA Polymerase (TaKaRa Code: DR010A) system:
[0034] 5×PrimeSTAR Buffer (Mg 2+ plus) 10 μl,
[0035] dNTP Mixture (2.5mM each) 4μl,
[0036] Upstream primer (10 μM) 1 μl,
[0037] Downstream primer (10 μM) 1 μl,
[0038] Template cDNA ...
Embodiment 2
[0059] The lateral root growth observation statistics of embodiment 2 transgenic Arabidopsis
[0060] Arabidopsis is a taproot plant. The main root grows first, and the lateral roots grow after the main root grows for a period of time. To the obtained transgenic Arabidopsis (T of embodiment 1 0 T obtained after generation seed planting 3 Homozygous) in different concentrations of KNO 3 The growth of the root system on the B5 modified medium was counted.
[0061] Put the plate with transgenic Arabidopsis in the refrigerator at 4°C for two days, then put it vertically into the growth room, check the root system every day, mark the time when the first lateral root occurs and the time when the main root reaches 2cm, and calculate the transgenic Arabidopsis Growth promotion rate of first lateral root and main root relative to wild-type Arabidopsis.
[0062] The calculation formula of Advancement (growth promotion rate, %) is:
[0063] Advancement (growth promotion rate, %) = (...
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