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Genetic transformation method of Eichhornia crassipes

A technology of water hyacinth and exogenous genes, which is applied in the field of genetic transformation of water hyacinth, can solve the problems of in-depth development and utilization limitations, difficult transgenic research, etc., and achieve the effects of easy mastery, popularization and application, simple operation and simple process

Inactive Publication Date: 2012-04-04
PEKING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to its special physiological characteristics, it is difficult to conduct transgenic research, and its in-depth development and utilization are limited

Method used

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  • Genetic transformation method of Eichhornia crassipes
  • Genetic transformation method of Eichhornia crassipes
  • Genetic transformation method of Eichhornia crassipes

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0050] Embodiment 1, transfer VP1 gene into water hyacinth

[0051] 1. Construction of plasmid pGII 0229-VP1

[0052] The construction process of plasmid pGII 0229-VP1 includes: artificially synthesizing the VP1 gene shown in Sequence 1, and making its two ends with the restriction site sequences of BamHI and SacI; digesting the synthetic gene and vector pBI121 with BamHI and SacI, and inserting The digested products were ligated to obtain the recombinant vector PBI121-VP1, in which the VP1 gene forms an expression cassette with the promoter 35S and the terminator nos. PBI121-VP1 and pGII 0229 were simultaneously digested with HindIII and EcoRI, ligated to obtain recombinant plasmids, transformed into Escherichia coli, resistance screening, liquid culture, plasmid extraction, enzyme digestion and sequencing identification. The result showed that the enzyme digestion result was correct; the measured VP1 gene sequence was shown in sequence 1, indicating that the constructed rec...

Embodiment 2

[0145] Implementation Example 2. Plant expression plasmid pGII 0229-GFP transforms water hyacinth.

[0146] 1. Construction of plasmid pGII 0229-GFP

[0147]The construction method of plasmid pGII 0229-GFP: the GFP gene shown in Sequence 2 is artificially synthesized, and its two ends are provided with XbaI and SacI restriction site sequences; the synthesized gene and vector pBI121 are digested with XbaI and SacI, and the enzyme The cut products were ligated to obtain the recombinant vector PBI121-GFP, in which the GFP gene forms an expression cassette with the promoter 35S and the terminator nos. Simultaneous digestion of pBI121-GFP and pGII 0229 with HindIII and EcoRI, ligation to obtain recombinant vector, transformation of Escherichia coli, resistance screening, liquid culture, extraction of plasmid, enzyme digestion and sequencing identification, the result of enzyme digestion was correct, and the measured GFP The gene sequence is shown in sequence 2, indicating that the...

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Abstract

The invention discloses a genetic transformation method of Eichhornia crassipes, comprising the following steps: after the germination of a pollen tube in the pistil of a pollinated pontederiaceae plant and before the pollen tube closes, adding a solution of recombinant plasmids containing exogenous genes or a solution of recombinant strains containing exogenous genes dropwisely on a pollinated column cap to introduce the exogenous genes in a zygote cell through the pollen tube and obtain a seed having transferred exogenous genes along with the growth of the zygote cell. Experiments prove that the method has the advantages of high transformation efficiency, the average conversion rate can reach 1.44 %, and the obtained transferred seed has high seedling rate. Thus the method has a wide application prospect in the field of genetic improvement of Eichhornia crassipes.

Description

technical field [0001] The invention relates to a method for genetic transformation of water hyacinth. Background technique [0002] Water hyacinth (Eichhornia crassipes) belongs to the genus Eichhornia crassipes of the genus Hyacinthaceae, commonly known as water water lily and water hyacinth. Water hyacinth was introduced into China as an ornamental plant in 1901, and was promoted as pig feed in the 1950s and 1960s. Since then, water hyacinth has been out of control in China. Because water hyacinth is very fertile, especially nitrogen and phosphorus fertilizers, it is widely present in many eutrophic waters such as Dianchi Lake, Chaohu Lake, and Taihu Lake in my country. Water hyacinth has a strong purification effect of enriching N / P and other nutrient elements, various heavy metals, absorbing and degrading phenols and cyanide, and at the same time, the water hyacinth extract itself has the effect of inhibiting the growth of algae. In addition, water hyacinth also has m...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H1/02A01G1/00A01C1/00C12N15/42
Inventor 安成才黄萍张起涛付力文任娇宋莲芬杨扬孔双蕾高超何善平黄国维李艳东张磊于祥春张旭邬倩
Owner PEKING UNIV