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Rice osms4 mutant-based seed production, seed reproduction and two-line hybrid breeding methods

A mutant and rice technology, applied in the fields of seed production, seed propagation and two-line hybrid breeding based on rice-based osms4 mutants, can solve problems such as application limitations, low reproductive yield, and impact on the application prospects of two-line hybrid rice. Achieve the effect of simplifying the production procedure and solving the drift phenomenon

Inactive Publication Date: 2010-12-08
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The emergence of this problem has seriously affected the application prospects of two-line hybrid rice, and is a major obstacle to the further promotion and application of two-line hybrid rice.
[0005] On the other hand, the temperature-sensitive CMS lines with low sterility threshold temperature are sterile in summer seed production, but the yield of their own reproduction is low and unstable, and sometimes even fails because they return to the fertile temperature. The range is very narrow, generally only about 3°C, so the application of this type of material is also greatly restricted

Method used

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  • Rice osms4 mutant-based seed production, seed reproduction and two-line hybrid breeding methods
  • Rice osms4 mutant-based seed production, seed reproduction and two-line hybrid breeding methods
  • Rice osms4 mutant-based seed production, seed reproduction and two-line hybrid breeding methods

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Experimental program
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Effect test

Embodiment 1

[0023] Obtaining of improved mutant (osms4) plants

[0024] This mutant was used in our laboratory 60 Japonica rice 9522 seeds were induced by Coγ-rays at a dose of 280Gy. A male sterile mutant in the mutagenized F2 generation was backcrossed for three generations, and a stable genetic mutant osms4 regulated by a recessive single gene was obtained. All plant materials were planted at the experimental base of Shanghai Academy of Agricultural Sciences. The osms4 mutant was backcrossed with japonica rice 9522, all the F1 generations were fertile, and segregation occurred in the self-crossed F2 generation, of which 419 were normal plants and 126 were mutant plants, and the ratio of normal plants to mutant plants was close to 3:1 (χ 2 =1.028, P>0.05), indicating that the phenotype of the male sterile mutant was caused by a single nuclear gene mutation.

Embodiment 2

[0026] Study on the relationship between the fertility of the improved mutant (osms4) and the temperature and light environment

[0027] Through years of experiments and research, it is found that under natural conditions, the young panicle differentiation stage is the pollen mother cell formation stage. When the daily light is below 13 hours, the fertility of the sterile line is stable, the anthers are white, and the pollen staining is typical. When the daily light time is more than 14 hours, the minimum temperature is 25°C, and the maximum temperature is above 35°C, it will be transformed from sterile to fertile. The normal pollen grains of pollen staining are 45-80%, and the seed setting rate is 60-80%. Between 80%. Fertility conversion is sensitive to light duration, light intensity and spectral range. When the light time did not meet the fertility conversion conditions, it showed insensitivity to temperature fluctuations.

[0028] The light-temperature type genic male s...

Embodiment 3

[0030] Identify mutation sites and obtain improved genes

[0031] (1) Positioning groups. The osms4 was crossed with the indica rice line Longtefu B, the F2 generation was obtained by selfing, and the male sterile plants were selected as the positioning population.

[0032] (2) Rice DNA extraction. Using the improved CTAB method. The simple steps are as follows: Take 0.1-0.2 grams (about half a piece) of leaves and put them in a small mortar, add an appropriate amount of liquid nitrogen, grind them to powder immediately, put them into a 2ml centrifuge tube, add 700ul of 1.5xCTAB solution preheated at 100℃ in In a centrifuge tube, mix carefully and place in a 56°C water bath. Take out the centrifuge tube after 20 minutes, add an equal volume of chloroform / isoamyl alcohol, mix vigorously, centrifuge (13000rpm) for 10 minutes, take the supernatant into a new tube, add Mix 900ul of absolute ethanol and place at -20°C for more than half an hour. The precipitated DNA was centrif...

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Abstract

The invention discloses rice osms4 mutant-based seed production, seed reproduction and two-line hybrid breeding methods, which belongs to the field of biotechnology. In the invention, the mutation of seeds of japonica rice 9522 are induced by 60Co gamma rays, and a third-generation male sterile mutant in the generation F2 obtained by induction is backcrossed to obtain an OsMS4 mutant which is controlled by a recessive single gene and can be inherited stably. In the invention, a plant which has OsMS4 gene mutation demonstrated by abnormal development of anther, failure of microspores in the normal secondary mitosis, complete pollen abortion and completely male sterility under a short lighting period condition can be used as a male sterile line in crossbreeding, and thus the male sterile plant OsMS4 can be used for two-line hybrid rice breeding; and selfed seeds can be obtained by changing a lighting condition, and thus the sterile line can be retained.

Description

technical field [0001] The invention relates to an application in the field of biotechnology, in particular to a method for seed production, seed propagation and two-line crossbreeding based on the rice osms4 mutant. Background technique [0002] Male sterile line: Refers to a female parent rice material with abnormal development or degeneration of male reproductive organs (mainly degeneration of anthers or pollen) but normal development of pistil reproductive organs. Due to abnormal early development of anthers or pollen, there is no pollen type or no life. Pollen cannot be self-pollinated and fruited, and can only be fertilized and fruited by relying on foreign pollen. Therefore, with the help of this mother rice as a genetic tool, a large number of hybrid seeds can be produced through artificial assisted pollination. From the perspective of breeding strategy, the development of hybrid rice can be divided into three development stages: three-line method, two-line method an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/02A01H1/06
CPCC12N15/8218C07K14/415A01H1/02C12Q1/6895A01H1/00C12Q2600/156A01H1/023A01H1/045C12N15/01C12N15/8225C12N15/8227C12N15/8231C12N15/8233C12N15/8234
Inventor 张大兵张辉梁婉琪袁政
Owner SHANGHAI JIAOTONG UNIV
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