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Method for introducing foreign gene into cleistogamy japonica rice by utilizing PMI (phosphomannose isomerase) selection marker

A technology of exogenous gene and closed glume pollination, applied in the field of genetic transformation

Inactive Publication Date: 2014-08-20
RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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  • Description
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Problems solved by technology

[0005] However, there is no report on the genetic transformation method of high-efficiency and simple closed-growth pollinated rice varieties with PMI as a screening marker

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  • Method for introducing foreign gene into cleistogamy japonica rice by utilizing PMI (phosphomannose isomerase) selection marker
  • Method for introducing foreign gene into cleistogamy japonica rice by utilizing PMI (phosphomannose isomerase) selection marker
  • Method for introducing foreign gene into cleistogamy japonica rice by utilizing PMI (phosphomannose isomerase) selection marker

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Embodiment Construction

[0059] Unless otherwise specified, the operations involved in the following specific embodiments are all carried out by common conventional operations in the art. Those skilled in the art can easily obtain teachings about such routine operations from the prior art, for example, with reference to the textbooks Sambrook and David Russell, Molecular Cloning: A Laboratory Manual, 3rd ed., Vols 1, 2; Charles Neal Stewart, Alisher Touraev, Vitaly Citovsky and Tzvi Tzfira, Plant Transformation Technologies, etc. The medicinal raw materials, reagent materials, etc. used in the following examples are all commercially available products unless otherwise specified.

[0060] Example 1——Genetic Transformation Method of Clover-pollinated Japonica Rice 8m30 Using Mannose as Screening Agent

[0061] 1. Induction and pre-culture of mature embryo callus

[0062] Shell the mature seeds of japonica rice 8m30 pollinated by clamgrass (preserved by the Rice Research Institute of Anhui Academy of A...

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Abstract

The invention provides a method for introducing a foreign gene into cleistogamy japonica rice by utilizing a PMI (phosphomannose isomerase) selection marker. Specifically, the method is characterized in that the foreign gene is introduced into the cleistogamy japonica rice variety 8m30 through mediation of an agrobacterium bacterial strain EHA 105 containing a pCAMBIA1381-PMI vector in the presence of mannose serving as a selection agent. PCR (polymerase chain reaction) detection and identification show that the foreign gene is introduced into the cleistogamy japonica rice variety 8m30. The method for introducing the foreign gene into the cleistogamy japonica rice by utilizing the PMI selection marker has the advantages that genetic transformation of the cleistogamy japonica rice variety is successfully realized, no antibiotics or herbicide is needed for selection, a character improvement process is sped up, gene drift caused by outward propagation of pollen is avoided, and further environment friendliness is improved.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and plant genetic engineering. Specifically, the present invention relates to a genetic transformation method for introducing exogenous genes into the japonica rice variety 8m30 pollinated by clamshell by using PMI screening markers. Background technique [0002] Rice (Oryza sativa L.) is the most important food crop in my country and even in the world, and more than 60% of the world's population uses rice as a staple food. In recent years, in the face of increasingly frequent drought, extreme temperature and other abiotic stresses, population growth and ecological environment pressure, the importance of modern biotechnology methods, especially transgenic technology, in crop genetic improvement has become increasingly prominent. Transgenic technology has provided a new path and opened up new space for crop yield improvement, quality improvement and resistance enhancement. [0003] With th...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H4/00A01H5/00
Inventor 李浩杨剑波魏鹏程李莉杨亚春倪大虎宋丰顺倪金龙秦瑞英陆徐忠马卉
Owner RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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