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Genetic transformation method for transferring exogenous gene into closed-husk pollination nonglutinous rice species

A genetic transformation method and exogenous gene technology, applied in biochemical equipment and methods, horticultural methods, genetic engineering, etc., can solve the problem of no genetic transformation method for closed-growth pollinated rice varieties, reduce ecological risks, and be suitable for Qualcomm The effect of large-scale operation and convenient large-scale application

Inactive Publication Date: 2014-09-10
RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no report on the genetic transformation method of closed glume pollinated rice varieties

Method used

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  • Genetic transformation method for transferring exogenous gene into closed-husk pollination nonglutinous rice species
  • Genetic transformation method for transferring exogenous gene into closed-husk pollination nonglutinous rice species
  • Genetic transformation method for transferring exogenous gene into closed-husk pollination nonglutinous rice species

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1——Genetic Transformation Method of Close-grained Indica Rice 9m90

[0063] 1. Induction and pre-culture of mature embryo callus

[0064] The mature seeds of the closed glume pollinated indica rice 9m90 (preserved by the Rice Research Institute of the Anhui Academy of Agricultural Sciences, whose genetic background is the indica variety Zaoxian 788) were dehulled, and the seeds with normal appearance, clean and mildew-free were selected, shaken for 90 sec with 70% alcohol, Pour off the alcohol; then wash the seeds with a solution of 50% sodium hypochlorite containing Tween20 (the effective chlorine concentration of the stock solution is greater than 4%, add 1 drop of Tween20 per 100 ml) solution, and shake on the shaker for 45min (180r / min). Pour off the sodium hypochlorite, wash with sterile water 5-10 times until there is no smell of sodium hypochlorite, finally add sterile water, soak overnight at 30°C. Use a scalpel to separate the embryos along the aleuron...

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Abstract

The invention provides a genetic transformation method for transferring an exogenous gene into a closed-husk pollination nonglutinous rice species. Specifically, the exogenous gene is transferred into the closed-husk pollination nonglutinous rice species by agrobacterium mediation. The inventor finds out that the conventional mediation method is not applicable to the closed-husk pollination rice (especially rice species 9m90), or the conversion efficiency is very low. Therefore, the invention provides a novel conversion method which greatly enhances the conversion efficiency. The PCR (polymerase chain reaction) detection on the plant proves that the exogenous gene has been transferred into the 9m90. The method successfully implements the genetic transformation of the closed-husk pollination nonglutinous rice species, and can avoid the gene drift caused by pollen outflow on the premise of accelerating the character improvement process.

Description

technical field [0001] The present invention relates to the technical fields of biotechnology and plant genetic engineering, in particular, the present invention relates to a genetic transformation method for introducing exogenous genes into indica rice varieties pollinated by clams, especially 9m90 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] However, with the rapid exp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H4/00A01H5/00
Inventor 倪大虎杨剑波魏鹏程李浩杨亚春李莉倪金龙陆徐忠宋丰顺马卉秦瑞英
Owner RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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