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Production method of light-sensitive male sterility rice

A production method and male sterility technology, applied in the field of genetic engineering and rice cultivation, can solve the problems of inability to be randomly matched, hybrid sterility between rice subspecies, etc., and achieve the effect of expanding the scope

Inactive Publication Date: 2009-05-20
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are two bottlenecks in these two development directions: because the selection of three-line hybrid rice sterile lines and restorer lines is restricted by genotype, they cannot be matched randomly; Difficult to use in practice

Method used

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  • Production method of light-sensitive male sterility rice
  • Production method of light-sensitive male sterility rice
  • Production method of light-sensitive male sterility rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Construction of expression vector pCAMBIA1304 / anti-osPDCD

[0054] The full-length cDNA of the rice programmed death gene OsPDC was cloned with primers with restriction sites, and the cDNA and the plant expression vector pCAMBIA1304 were digested with BsmFI and BstEII and ligated, so that OsPDCD was reversely inserted downstream of the 35s promoter. The antisense sequence expression vector pCAMBIA1304 / anti-osPDCD5 of OsPDCD was constructed. refer to figure 1 .

Embodiment 2

[0055] Example 2 Callus Culture and Transformation

[0056] 2.1 Culture medium for inducing rice callus

[0057] 2.1.1 Induction and subculture medium: MS+2mg / L 2,4-D.

[0058] 2.1.2 Hyperosmotic medium: MS+2mg / L 2,4-D+46.67g / L sorbitol+46.67g / L mannitol.

[0059] 2.1.3 The first round of screening medium: MS+2mg / L 2,4-D+30mg / L hygromycin.

[0060] 2.1.4 The second round of screening medium: MS+2mg / L 2,4-D+50mg / L hygromycin.

[0061] 2.1.5 Differentiation medium: MS+3mg / L 6-BA+0.5mg / L NAA+50mg / L hygromycin.

[0062] 2.1.6 Rooting and strong seedling medium: 1 / 2 MS+0.1mg / L NAA

[0063] Note: 1. The above media all contain 30g / L sucrose + 2.5g / L agar, pH5.8

[0064] 2. The culture conditions for callus induction, subculture, and screening are dark culture at 26-28°C, and 26-28°C and 16-hour photoperiod for differentiation and rooting of seedlings.

[0065] 2.2 Callus induction and treatment

[0066] 2.2.1 Take immature seeds 12-15 days after pollination, soak them with 70...

Embodiment 3

[0088] Example 3 Screening and detection of rice anti-osPDCD gene transformed plants

[0089] The surviving calli were cultured on the differentiation medium for 30 days under light. After the small plants are differentiated, the small plants are transferred to the medium for rooting and strong seedlings, and then moved to the greenhouse after growing up. The candidate resistant plants were detected by PCR amplification and genomic Southern hybridization respectively, and a total of 7 positive plants containing anti-osPDCD were obtained.

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Abstract

The invention belongs to the fields of genetic engineering and rice cultivation, particularly relates to a production method of photosensitive rice with male sterility. The production method of the photosensitive rice with male sterility creatively and inversely inserts gene into a rice expression carrier, imports the recombinant expression carrier into rice callus cells, and cultivates the photosensitive rice with male sterility through inductive differentiation. The production method of the photosensitive rice with male sterility builds a system that can produce hybrid rice by random grouping and substitutes the dual-functional photosensitive and temperature-sensitive natural mutant systems that are applied in existing production but have lots of problems. The production method creates a hybrid seed production technique in the aspect of hybrid rice cultivation and enlarges the range of excellent hybrid rice combinations.

Description

technical field [0001] The invention belongs to the fields of genetic engineering and rice cultivation, and in particular relates to a production method of photosensitive male sterile rice. Background technique [0002] The popularized area of ​​hybrid rice in my country has reached 220 million mu, accounting for about 50% of the total rice area, and the output accounts for 60% of the total rice output. The annual increased rice production of hybrid rice can feed about 75 million people, making a huge contribution to the development of the national economy. [0003] Hybrid rice has been popularized for 35 years and is currently facing difficulties in further development. The further development of hybrid rice has two directions: 1) develop a seed production technology that can be randomly matched to expand the range of excellent hybrid rice combinations; 2) exploit the potential of hybrid rice to increase yield by utilizing the strong heterosis among rice subspecies. At pr...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H1/00A01H4/00A01H5/00
Inventor 杨金水王玉锋
Owner FUDAN UNIV