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Method for changing characteristics of plants

A plant trait and plant technology, applied in the fields of biotechnology and botany, can solve the problems of restricting the improvement of leguminous crop varieties, difficulty in obtaining transgenic plants, and health effects

Inactive Publication Date: 2011-10-05
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, it has been found that it is very difficult to obtain transgenic plants using conventional transgenic techniques (such as the Agrobacterium method) on some other plants such as leguminous crops
At present, there is no particularly efficient transgenic technology for legume crops, which greatly limits the variety improvement of legume crops
In addition, some genetically modified plants are not currently acceptable to many people due to possible health effects

Method used

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Experimental program
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Embodiment approach

[0063] As an embodiment of the present invention, the source gene is prenyltransferase gene. Isopentenyltransferase (IPT) is a catalytic enzyme that can guide the synthesis of cytokinins, participates in the regulation of plant development, morphology and physiological processes, and plays an important role in regulating the levels of cytokinins in plant tissues. Its amino acid sequence is shown in GenBank accession number AAK90970.2; its coding gene sequence is shown in GenBank accession number AE007871.2.

[0064] The isopentenyl transferase can synthesize plant cytokinins in rhizobia, especially trans-zeatin, and the trans-zeatin (a small molecule) is a kind of cytokinin, which can regulate cell division And affect a variety of developmental events, such as shoot development, sink strength, root branching, control of shoot apical dominance, leaf development, chloroplast development, and leaf senescence.

[0065] Other exogenous genes that can be used include but are not li...

Embodiment 1

[0116] Embodiment 1, the construction of the rhizobia meliloti engineering bacteria expressing isopentenyl transferase

[0117] Design a pair of oligonucleotide primers targeting the reading frame of the Agrobacterium tumefaciens prenyltransferase gene, the sequence is as follows: forward: 5'-GCTCATATGTTACTCCATCTCATCTACGG-3', and reverse: 5'-CAGTCTAGAGTGCAATACTTGTAACAGGATCCGTAG-3 '.

[0118] Taking Agrobacterium tumefaciens C58 pTiC58 (see Danhorn T, Hentzer M, Givskov M, Parsek MR, Fuqua C. Phosphorus limitation enhances biotilm formation of the plant pathogen Agrobacterium tumefaciens thr

Embodiment 2

[0119] Embodiment 2, the effect of rhizobia engineering bacteria LMG202 on the formation of alfalfa nitrogen-fixing root nodules

[0120] After surface disinfection of alfalfa (Medicago sativa Shangdong, purchased from Shandong Academy of Agricultural Sciences) seeds, rhizobium LMG202 (containing 10 8 Rhizobia / ml) were mixed together and soaked for 0.5 hours, then sowed in sterilized artificial soil of perlite, placed in an artificial climate chamber to germinate and grow until the fourth week, discarded the artificial soil, washed the plants, and observed The number of nodules was counted, and nitrogenase activity was determined. Alfalfa inoculated with an empty vector (Sinorhizobium meliloti Rm 1021 / pvector) was used as a control.

[0121] The method for measuring nitrogenase activity is as follows: collect 10 alfalfa roots (below the cotyledons), put them in a 10-ml glass bottle, cover the bottle tightly, inject 1 ml of acetylene with a syringe, react at 28°C for 30 minute...

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Abstract

The invention relates to a method for changing characteristics of plants. According to the invention, a special property is adopted, that plants which permit root nodules can realize a symbiosis relation with rhizobium; exogenous genes are expressed in the rhizobium colonized in plant root nodule cells; through existing transporting systems in the plants, gene expression products or components are transferred to other tissue organs of the plants. With the method provided by the present invention, characteristics or phenotypes of the plants are changed.

Description

technical field [0001] The invention belongs to the fields of biotechnology and botany; more specifically, the invention relates to a method for changing plant traits. Background technique [0002] In the prior art, transgenic technology is generally used to change the traits or phenotypes of plants and obtain new varieties of plants. Although the plant transgenic technology in the prior art has been widely studied and applied, the classic one is to use Agrobacterium to carry the gene of interest and transform it into plant callus. This technology is useful for many types of plants, such as model Plant Arabidopsis, economic crops rice, wheat, cotton, tobacco, etc. People have found that it is very difficult to obtain transgenic plants by using conventional transgenic techniques (such as the Agrobacterium method) on other plants such as leguminous crops. At present, there is no particularly efficient transgenic technology for legume crops, which greatly limits the variety i...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N1/21A01H5/00A01N63/02A01P21/00C12R1/41
CPCA01H3/00
Inventor 罗利徐霁李晓琳
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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