Application of receptor-like kinase gene MdMRLK2 in improving water utilization efficiency of plants
A kind of receptor kinase, receptor kinase technology, applied in the field of genetic engineering, can solve the problems of poor stability repeatability, large error, slow detection speed, etc., to achieve the effect of improving water use efficiency, high root system vitality, and improving WUE
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Embodiment 1
[0041] Example 1 Analysis of expression patterns of apple receptor-like kinase MdMRLK2 in different tissues and under various stress treatments
[0042] Samples of 'Pinova' apple stems, leaves, flowers, pulp, peel, leaf buds, flower buds and 'Pingyi Sweet Tea' apple roots, stems, and leaves were extracted and reverse transcribed for tissue-specific expression analysis. The annual "Pingyi sweet tea" seedlings with consistent growth were picked. It was sprayed with 80 μM ABA solution (abscisic acid) respectively, and samples were taken at 0h, 1h, 3h, 6h, 12h, and 24h; , 36h sampling; 45°C high temperature stress and sampling at 0h, 4h, 6h, 8h, 12h, 24h; , 24h sampling; RNA was extracted from the above samples and reverse transcribed. Design specific quantitative primers, and detect the expression of MdMRLK2 under different stress conditions by fluorescent quantitative PCR. MdMRLK2 quantitative primer sequence is:
[0043] q MdMRLK2-F: 5'-CCAGATCAATGGGGCCAAATGCACA-3', SEQ ID ...
Embodiment 2
[0046] Example 2 Transformation and Identification of Transgenic Apple Plants Overexpressing Apple Receptor-like Kinase MdMRLK2
[0047] (1) Construction of plant overexpression vector
[0048] The vector used for genetic transformation is pBI121, which exhibits kanamycin resistance in plants. The coding region of the target gene MdMRLK2 was connected to the pBI121 vector through two restriction sites of XbaI and SacI on the vector. The method includes the following steps: clone the apple gene MdMRLK2 by using the 'Golden Delicious' apple cDNA as a template, and identify the PCR recovery product of the apple gene MdMRLK2 that has been digested with XbaI and SacI by agarose gel, and use T4 to linearize the pBI121 vector Ligase performs the ligation. The ligation product was transformed into Escherichia coli DH5α competent, screened with LB petri dishes containing Kan antibiotics, the monoclonal colonies on the petri dish were taken for amplification, and PCR detection was per...
Embodiment 3
[0059] Example 3 Carry out long-term moderate water deficit treatment to wild-type WT (gala variety), MdMRLK2 overexpression apple plants
[0060] Wild-type WT and MdMRLK2 overexpressed apple plants OE-1 and OE-2 were subcultured, multiplied and rooted. Rooted WT and transgenic plants were transplanted into plastic pots (8 cm × 8 cm) filled with nutrient soil, vermiculite and perlite (3: 1: 1; v: v: v) and placed in 14 h light: 10 h dark placed in a light incubator for growth. After one month of growth, the plants with consistent growth were transplanted into flowerpots (30cm×18cm) filled with an equal amount of sand and soil mixture, and placed in the horticultural field of Northwest A&F University in Yangling, Shaanxi, China (34°20′N, 108 °24'E) in the semi-open greenhouse cultivation, when the plants grew to 50cm, the seedlings were divided into the well-watered control group CK (field water holding capacity of 75wt%-85wt%, test sample size n=45) and moderate Water defici...
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