MiRNA, namely ghr-miR01, related to plant's response to drought stress and application of miRNA
A drought stress, plant technology, applied in the field of new miRNA, can solve the problems of increased shedding of buds, reduced yield, and the impact of photosynthesis
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Embodiment 1
[0027] Example 1: Solexa sequencing analysis verifies the role of ghr-miR01 in the drought resistance mechanism of cotton
[0028] 1. Material cultivation and drought treatment: Shizao No. 1 cotton seeds were sown in culture pots with a 3:1 mixture of vermiculite and nutrient soil at a temperature of 28 °C and a relative humidity of 70%. The control materials were watered on the 7th, 14th, 21st, and 28th days after sowing, respectively, and watered 150 mL each time. On the 35th day, young leaves were taken, frozen in liquid nitrogen, and stored at -80 ℃; Water on 7, 14, and 21, 150 mL each time, take the young leaves on the 35th day, freeze in liquid nitrogen, and store at -80 ℃.
[0029] 2. Solexa sequencing analysis and identification of new miRNA: The samples obtained in the previous step were sent to Shanghai Meiji Biological Company for solexa sequencing and analysis. see figure 1 , the results showed that the expression number of ghr-miR01 was 12 under normal growth co...
Embodiment 2
[0030] Example 2: Real-time PCR analysis verifies the role of ghr-miR01 in the drought resistance mechanism of cotton
[0031] 1. Material cultivation and NaCl treatment
[0032]Shizao No. 1 cotton seeds were sown in culture pots mixed with vermiculite and nutrient soil at a ratio of 3:1, the temperature was 28 °C, and the relative humidity was 70%. The control materials were watered on the 7th, 14th, 21st, and 28th days after sowing, respectively, and watered 150 mL each time. On the 35th day, young leaves were taken, frozen in liquid nitrogen, and stored at -80 ℃; 7, 14, 21 watering, watering 150mL each time, on the 35th day, young leaves were taken, frozen in liquid nitrogen, and stored at -80°C.
[0033] 2. Isolation of RNA
[0034] Total RNA was extracted with the polysaccharide polyphenol plant total RNA extraction kit (catalogue number: DP441) from GIANGEN, and the extraction method was as follows:
[0035] 1. Homogenization treatment. Grind 50-100 mg leaf samples q...
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