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Method for guiding exogenous gene into cleistogamous indica rice by using PMI (phosphomannose isomerase) selection marker

A technology of exogenous gene and closed-growth pollination, applied in the field of genetic transformation, can solve the problem of screening marker genes without the genetic transformation method of closed-growth pollinated indica rice

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

[0005] Therefore, there is currently no screening marker gene and corresponding transformation method suitable for the genetic transformation method of closed-growth pollinated indica rice varieties.

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  • Method for guiding exogenous gene into cleistogamous indica rice by using PMI (phosphomannose isomerase) selection marker
  • Method for guiding exogenous gene into cleistogamous indica rice by using PMI (phosphomannose isomerase) selection marker
  • Method for guiding exogenous gene into cleistogamous indica rice by using PMI (phosphomannose isomerase) selection marker

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Embodiment Construction

[0054] Unless otherwise specified, the operations in the following specific embodiments are performed 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.

[0055] Example——Genetic Transformation Method of Clover-pollinated Indica Rice 9m90 Using Mannose as Screening Agent

[0056] 1. Induction and pre-culture of mature embryo callus

[0057] The mature seeds of the closed glume pollinated indica rice 9m90 (preserved by the Rice Research Institute of the Anhui Academy of Agricultural Science...

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Abstract

The invention provides a method for guiding an exogenous gene into cleistogamous indica rice by using a PMI (phosphomannose isomerase) selection marker. Specifically, the exogenous gene is guided into a cleistogamous indica rice variety 9m90 by using mannose as a selecting agent through mediation of a pCAMBIA1381-PMI carrier-containing agrobacterium tumefaciens strain EHA105. After guiding is ended, PCR (polymerase chain reaction) detection and identification prove that the exogenous gene is guided into the cleistogamous indica rice variety 9m90. The method successfully realizes the genetic transformation of the cleistogamous indica rice variety and does not need to use antibiotics or weed killers for screening; the gene drift caused by external spreading of pollen is avoided while the character improvement is accelerated; and the environmental protection degree is increased.

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 clover-pollinated indica rice variety 9m90 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 the rapid ...

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Application Information

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Patent Type & Authority Applications(China)
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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