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A method for reproducing sporophyte recessive male sterility

A technology for reproducing spores and male nuclei, applied in the field of genetic engineering, can solve problems such as the production of difficult hybrids, and achieve the effect of reducing concerns about transgenics and reducing losses

Active Publication Date: 2022-04-29
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the recessive sterility insensitive to temperature and light in the natural state cannot produce a sterile population with a sterile plant rate of 100%, it is difficult to apply it to the production of hybrids

Method used

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  • A method for reproducing sporophyte recessive male sterility
  • A method for reproducing sporophyte recessive male sterility

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Experimental program
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Effect test

Embodiment 1

[0023] Embodiment 1, vector construction process

[0024] 1) OsMYB76R gene loaded into pCAMBIA1301

[0025] According to previous studies, the OsMYB76 gene is an essential gene for anthocyanin synthesis in rice. In rice ZhongjiuB and other materials, due to the loss-of-function mutation of the OsMYB76 gene, all organs do not show anthocyanin color (Zhang Yi, 2009). After optimizing the OsMYB76 gene and removing the commonly used enzyme cutting sites, the OsMYB76R gene (SEQ ID NO.11) was artificially synthesized. Using OsMYB76R as a template, the primers JCF1 (SEQ ID NO.12) / JCR2 (SEQ ID NO.13) were used The target fragment was amplified and recovered for use. The GUS gene in pCAMBIA1301 was completely excised with NcoI and BstEII, and the backbone was recovered for use; OsMYB76R was loaded into pCAMBIA1301 by homologous recombination to obtain vector 1301-JC.

[0026] 2) Modification of 1301-JC multiple cloning sites

[0027] In order to facilitate the loading of various regu...

Embodiment 2

[0036] Embodiment 2, the method for propagating sporophyte recessive male sterility

[0037] 1. Breeding of nine B-osabcg15 in sporophyte sterile material

[0038] A male sterile mutant (Wu et al., 2014) naturally produced by the OsABCG15 gene was used as a sterile donor, Zhongjiu B was used as a recurrent parent, and Zhongjiu B-osabcg15 was produced through multiple backcrosses.

[0039] 2. pA R -Xi-Y-Bi-C transferred to S9 B-osabcg15

[0040] In the fertility-separated population, select sterile Zhongjiu B-osabcg15 plants, take their young ears to induce callus, and use the method of Agrobacterium tumefaciens to introduce the final vector pA R -Xi-Y-Bi-C transfects the aforementioned callus, differentiates into seedlings, and after rooting and transplanting, 8 strains showing anthocyanin color are selected as positive strains, all of which show male fertility ( figure 2 ).

[0041] 3. Propagation and purity control of B-osabcg15 in sporophytic male sterility

[0042] 1...

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Abstract

The invention discloses a method for propagating sporophyte recessive male sterility, the steps are as follows: sporophyte male fertility gene Y, endogenous gametophyte male fertility gene down-regulation expression element Xi, herbicide A resistance gene A R , the herbicide B resistance gene down-regulation element Bi and the anthocyanin gene C or its down-regulation element Ci are linked into the sterile mutant of the sporophyte fertility gene Y, and the positive strain is used to pollinate the sterile strain to reproduce the sporophyte nuclear sterile line. The method is universal and applicable to all plants, and can obtain spore-forming male sterile line with 100% sterile plant rate; and can reduce the cost of separating sterile line and maintainer line; can be early, intuitive, fast, accurate and Economically carry out purity identification; CMS lines with unqualified purity can be improved early, quickly, economically and thoroughly, and continue to be used for seed production; all relevant regulatory elements can be selected from endogenous nucleic acid fragments.

Description

technical field [0001] The invention belongs to the field of genetic engineering and relates to a method for propagating genital male sterility. The method can reproduce a genital male sterile line with a sterile plant rate of 100%, which is beneficial to the application of genital sterility in Hybrid seed production takes full advantage of heterosis. Background technique [0002] Heterosis is a phenomenon in which the first generation of hybrids is superior to their parents in terms of yield, quality and / or resistance, and exists widely in the biological world. Hybrids are the first generation of hybrids produced by crossing two different parents, and are breeds selected by using heterosis. Seed production in hybrid species requires outcross-pollination of the two parents, whereas conventional species does not require outcross-pollination for seed production. In crops such as rice, corn and rapeseed, the use of heterosis is an important strategy to achieve high yield, hig...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29A01H1/02A01G7/06
CPCA01H1/02A01G7/06C07K14/415
Inventor 张毅陈云杜双林
Owner YUNNAN UNIV
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