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Regulating Plant Nitrogen Nutrition and Alkali Stress Sensing Gene cml9(q6-1) and Its Application

A nitrogen nutrition, gene technology, applied in the field of plant biology

Active Publication Date: 2020-11-13
INNER MONGOLIA M GRASS ECOLOGY & ENVIROMENT GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the development and utilization of plant stress resistance genes, the current research mainly focuses on the model plants such as Arabidopsis or rice, wheat and other crops, and there is a lack of excavation of the abundant stress resistance genes in wild plants.

Method used

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  • Regulating Plant Nitrogen Nutrition and Alkali Stress Sensing Gene cml9(q6-1) and Its Application
  • Regulating Plant Nitrogen Nutrition and Alkali Stress Sensing Gene cml9(q6-1) and Its Application
  • Regulating Plant Nitrogen Nutrition and Alkali Stress Sensing Gene cml9(q6-1) and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1, Cypress sativa sample collection

[0021] In order to maximize the abundance of RNAs related to stress resistance in the body, we chose to sample the cypress roots in relatively cold December (2013), and the samples were quickly stored in liquid nitrogen for future use after collection.

Embodiment 2

[0022] Embodiment 2, the construction of expression vector

[0023] 1. Since the RNA of cypress plants is difficult to extract, our target gene was synthesized by Nanjing GenScript, and the subsequent expression vector was constructed using the gene synthesized by the company. The cloning vector used for the synthetic gene was pUC57 and the resistance was Amp.

[0024] 2. The completion of the connection and construction of the target carrier

[0025] The expression vector used in the present invention is pRI 101AN, and its resistance is Kan.

[0026] The connection of the target vector is to simultaneously digest the synthesized gene fragment and the target vector with 5':Sal I 3':EcoR I, recover from DNA gel, connect with DNA ligase, then transform Escherichia coli, undergo colony PCR and The method of extracting the plasmid, restriction restriction verification, and finally obtaining the plasmid connected to the vector gene.

[0027] Double enzyme digestion reaction syst...

Embodiment 3

[0070] Embodiment 3, the acquisition of transgenic plants

[0071] In this example, the transgenic plant code-named CML9 (Q6-1) was obtained by the floc infection method.

[0072] 1. Pick the Agrobacterium that has been properly sterilized for activation (the purpose of activation is to ensure the vitality of the bacteria), add the activated bacteria to 5ml LB culture solution containing antibiotics (Rif50mg / ml and Kan50mg / ml), shake at 28°C Cultivate the bed overnight, put the shaken bacterial solution into the centrifuge, 5000x g, 10min, discard the supernatant, add 1ml of infection solution (5% sucrose, 0.02% silwet77 suspension dissolved in water), and use Gently blow and aspirate the plastic tip dropper to suspend the bacteria.

[0073] 2. Select three boxes of wild-type Col-0 plants that are growing well and have bloomed, cut off the pods that have formed on the plants and the flower buds that have completed pollination, and use a rubber dropper to suck the bacteria liq...

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Abstract

The invention relates to a sensing gene CML9 (Q6-1) for adjusting plant nitrogen nutrition and alkaline stress from strong xerophyte Sabina vulgaris. in northwest part of Inner Mongolia. The encoding protein of the gene has a calcium ion binding site. A preparation method comprises the following steps of preparing a material, cloning the gene, building a subsequent plant expression carrier, and further guiding the gene into a mode plant arabidopsis thaliana wild model by an agrobacterium-mediated floc infection method; screening a resistance seedling, so as to obtain a T3 transgenic plant with insertion and purification of multiple single genes. Compared with the wild type, the transgenetic plant can well grow under the low nitrogen condition, and poorly grow under the alkaline stress condition, so that the encoding protein of the gene has the function of adjusting the plant nitrogen nutrient and the alkaline stress. The gene has the advantages that a new direction is provided for the culturing of drought-resistant plants by the gene engineering technique, and the important meaning is realized for the molecular seed breeding under the drought stress.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a nitrogen nutrition and alkali stress sensing gene CML9 (Q6-1) related to windbreak and sand-fixing tree species Sabina vulgaris. in the desertification area of ​​northern my country. In the present invention, the tree species Cypress syringae from the nursery stock base of Inner Mongolia Mengcao Drought Resistant Co., Ltd. is selected as the experimental material, and the nitrogen nutrition and alkali stress induction genes of the tree species are obtained by using the method of high-throughput sequencing of the Illumina Solexa transcriptome. The alkali stress-sensitive gene CML9(Q6-1) was analyzed by bioinformatics, and the phenotype analysis of different elements was carried out at the same time, so as to understand the physiological function of the gene more intuitively. Background technique [0002] Sand cypress (Sabina vulgaris.), also known as Chazi juniper ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
CPCC07K14/415C12N15/8273
Inventor 张林祁智杨红艳杨佳刘亚玲王召明苑峰金悦
Owner INNER MONGOLIA M GRASS ECOLOGY & ENVIROMENT GRP CO LTD
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