Sensing gene CML11 capable of adjusting plant calcium nutrient and drought signal from Sabina vulgaris and application thereof
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A technology for signal sensing and plant calcium, applied in the field of plant biology
Active Publication Date: 2017-05-24
INNER MONGOLIA M GRASS ECOLOGY & ENVIROMENT GRP CO LTD
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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.
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[0025] 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
[0026] Embodiment 2, the construction of expression vector
[0027] 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-Simple, and the resistance was Amp.
[0028] 2. The completion of the connection and construction of the target carrier
[0029] The expression vector used in the present invention is pOREE3, and its resistance is Kan.
[0030] The connection of the target vector is to simultaneously undergo 5': BamH I 3': Kpn I digestion of the synthesized gene fragment and the target vector, recover from DNA gel, connect with DNA ligase, and 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.
[0031] Double enzyme dig...
Embodiment 3
[0073] Embodiment 3, the acquisition of transgenic plants
[0074] In this example, the tidbit infection method is used to obtain the transgenic plant code CML11
[0075] 1. Pick the Agrobacterium that has been properly sterilized for activation (the purpose of activation is to ensure the vitality of the bacteria), and add the activated bacteria to 5ml LB culture solution containing antibiotics (Rif50mg / ml and Kan 50mg / ml), at 28°C Cultivate overnight on a shaking table, put the shaken bacterial solution into a centrifuge, 5000×g, 10min, discard the supernatant, add 1ml of infection solution (5% sucrose, 0.02% silwet77 suspension dissolved in water), Blow gently with a dropper to suspend the bacteria.
[0076] 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 liquid and drop it on the flower bu...
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Abstract
The invention relates to a sensing gene CML11 capable of adjusting plant calcium nutrient and drought signal from Sabina vulgaris of strong xerophytes of northwestern region of Inner Mongolia. The encoded protein of the gene has a calcium ion bonding site. The gene has the advantages that by preparing material, cloning gene and building subsequent plant expression carriers, the gene is delivered into an arabidopsis thaliana wild type of a model plant by an agrobacterium tumefaciens-mediated feature infection method, and a resistant plant is screened, so as to finally obtain multiple single-gene inserted and purified T3 transgenic plants; the transgenic plant and the wild control plant are subjected to different element expression analysis under the sterile culture condition, and compared with the wild control plant, the transgenic plant has the sensitive features of short root, fresh leaf and light weight under the low-calcium condition and drought hormone ABA conditions, so that the encoded protein of the gene has the function of adjusting plant calcium nutrient and drought signal; a new orientation is provided for the culture of anti-drought plants by the gene engineering technique, and the important significance is realized for the molecular seed breeding threatened by the drought.
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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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
CPCC07K14/415C12N15/8273
Inventor 杨红艳祁智张林杨佳王召明刘亚玲苑峰金悦
Owner INNER MONGOLIA M GRASS ECOLOGY & ENVIROMENT GRP CO LTD
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