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Gene SLF participating in controlling intra-specific and inter-species reproductive disorder of solanaceae plants and application of gene SLF

A reproductive disorder, Solanaceae technology, applied in the field of plant genetic engineering and breeding, which can solve problems such as unclear

Active Publication Date: 2014-01-01
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pollen S factor of tomato is still unclear

Method used

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  • Gene SLF participating in controlling intra-specific and inter-species reproductive disorder of solanaceae plants and application of gene SLF
  • Gene SLF participating in controlling intra-specific and inter-species reproductive disorder of solanaceae plants and application of gene SLF
  • Gene SLF participating in controlling intra-specific and inter-species reproductive disorder of solanaceae plants and application of gene SLF

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Embodiment 1: the cloning of tomato SLF candidate gene

[0118] 1. Tomato material

[0119] In terms of tomato cultivation, the present invention uses the self-compatible tomato sequencing species Heinz (LA4345) (purchased from Tomato Genetic Resource Centre, TGRC, California, USA) as research material and transgenic recipient. In terms of wild tomato, the present invention uses self-incompatible tomato hairy (LA1777) (purchased from Tomato Genetic Resource Centre, TGRC, California, USA) as a research material. Heinz was not compatible with LA1777 pollination; on the contrary, LA1777 was compatible with Heinz pollination and could obtain mature seeds, showing a typical one-way incompatibility.

[0120] 2. Cloning of candidate SLF genes in cultivated tomato

[0121] The present invention uses the expanded petunia pollen S factor PiSLF2 amino acid sequence in the cultivated tomato genome database (SGN Tomato Combined-WGS, BAC and unigene sequences model, version 2.40, ...

Embodiment 2

[0124] Embodiment 2: the functional research of tomato SLF gene:

[0125] 1. Yeast two-hybrid results of the carboxy-terminal of 5 ShSLF gene coding products and PhSv-RNase

[0126] In order to verify the uniqueness of the function of ShSLF genes, that is, whether the carboxy-terminal of their encoded products can interact with S-RNase, we constructed the sequences of five ShSLF genes without the F-box region (the initial 150 bases) into The yeast two-hybrid vector pGBK-T7 (purchased from Clonetech Company), and the PhSv-Rnase sequence was constructed into the yeast two-hybrid vector pGAD-T7 (purchased from Clonetech Company). The above-mentioned vectors were combined in pairs to co-transform yeast AH109 (purchased from Clonetech Company), and the co-transformants were streaked on yeast four-deficient medium to detect whether there was interaction between AD and BD fusion proteins. The results of growing at 30°C for 72 hours were as follows: Figure 14 shown.

[0127] 2. Sh...

Embodiment 3

[0147] Example 3: Verification of whether the tomato SLF gene functions as ui1.1

[0148] 1. Build of Cultivated Tomato with functional SLF and ui6.1

[0149] (1) Cultivation of Agrobacterium

[0150] The previously mentioned pBI101-PhSLF-S3A promoter-ShSLF recombinant plasmid was transformed into LBA4404 Agrobacterium competent cells (prepared by conventional methods in our laboratory, referring to "Plant Genetic Engineering", Wang Guanlin, Fang Hongyun, Science Publishing House, 2nd edition in 2004), cultured on YEB plate medium containing 50mg / L kanamycin and 25mg / L rifampicin at 28°C for 48 hours. Pick a single clone in 2ml YEB (50mg / 1 Kan, 25mg / 1 Rif) liquid medium containing kanamycin and rifampicin, culture overnight (28°C, 220rpm) to saturation, and inoculate 100ml YEB at a ratio of 1:100 Mass culture in medium (28°C, 220rpm) to OD600=0.6-0.8, 4000rpm, centrifuge at 4°C for 20min to collect the bacteria, discard the supernatant, resuspend in MS liquid medium and dilu...

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Abstract

The invention provides a gene SLF participating in controlling intra-specific and inter-species reproductive disorder of solanaceae plants and application of the gene SLF, and discloses the gene SLF participating in controlling intra-specific and inter-species reproductive disorder of the solanaceae plants and egg white of codes of the gene SLF. A nucleotide sequence of the gene SLF is shown in the SEQ ID No:1-5, and an amino acid sequence of the egg white of the codes is shown in the SEQ ID No:6-10. The invention discloses a method that the gene SLF is implanted in petunia, tomato plants and other solanaceae plants through the genetic engineering technique, and therefore the intra-specific and inter-species reproductive disorder is eliminated. The invention further discloses the egg white utilizing polynucleotide and the codes thereof, and genetic breeding is conducted between planted tomato plants and wild tomato plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering and breeding. Specifically, the present invention relates to a gene family SLF ( S - l ocus f -box), this family contains 5 SLF genes. The present invention also relates to the protein encoded by the SLF gene and its functional analogs, as well as the vector containing the SLF gene nucleotide sequence and the host cell containing the gene nucleotide sequence or the vector; in addition, the present invention also relates to A method for using the vector to transform plant cells to break the reproductive barriers within and between species of Solanaceae plants and related applications in Solanaceae genetics and breeding. Background technique [0002] Reproductive barriers restrict the improvement of crop genetic diversity and the utilization of hybrid vigor. For example, during the process of artificial domestication and breeding, the genetic diversity of cultivated tomato is ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/63C12N5/10C07K14/415A01H5/00
Inventor 薛勇彪郭晗李群张玉娥孔昭胜
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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