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Isolation and cloning of paddy rice photoperiod sensitive genic male sterility gene pms1 and application thereof

A light-sensitive nuclear sterility, pms1 technology, applied in applications, genetic engineering, plant genetic improvement, etc., can solve the problems of complex breeding procedures and production links, application limitations, and high costs

Active Publication Date: 2017-12-01
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The three-line hybrid rice technology is mature and widely planted in my country, but there are still some application limitations
Due to the single sterile cytoplasm of three-line hybrid rice, limited by the relationship between recovery and conservation, the combination of groups is not free enough, so the cycle of breeding new combinations is relatively long
In production, its breeding procedures and production links are more complicated, resulting in higher cost of seed production

Method used

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  • Isolation and cloning of paddy rice photoperiod sensitive genic male sterility gene pms1 and application thereof
  • Isolation and cloning of paddy rice photoperiod sensitive genic male sterility gene pms1 and application thereof
  • Isolation and cloning of paddy rice photoperiod sensitive genic male sterility gene pms1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Isolation clones comprise the DNA fragment of the pms1 gene segment

[0051] 1. Using map-based cloning technology to determine the control gene pms1 of rice light-sensitive sterility

[0052] In the present invention, two parents were used when preparing the positioning population: the photosensitive GMS line Nongken 58S (Li Zebing et al., "Photosensitive GMS Rice Fertility Transformation Mechanism and Application Research", published by Hubei Science and Technology Press, Wuhan, Hubei , 1995 edition) and Minghui 63 (a rice variety selected by the Sanming Academy of Agricultural Sciences for public use). Minghui 63 is a typical indica variety. Nongken 58S was used as the recurrent female parent to cross with Minghui 63, and Nongken 58S / Minghui 63BC were obtained after multiple backcrosses 5 f 2 A random population of 6848 strains was used for fine mapping of the photosensitive genic male sterile gene pms1. These individual plants were planted in the expe...

Embodiment 2

[0068] Embodiment 2: Obtaining of full-length cDNA of pms1 gene

[0069] Example 1 The rice light-sensitive sterility gene pms1 was located between the molecular markers P4 and P6, the physical distance in the Minghui 63 genome was 4.0kb, and genetic transformation experiments also confirmed that the candidate pms1 gene was indeed contained in this interval . Check the gene annotation results in this interval on the Rice Genome Annotation Project website (http: / / rice.plantbiology.msu.edu / ), and find that this interval does not contain complete predicted genes, and the nearest left and right sides Both genes are retrotransposon proteins ( figure 1 ), indicating that the pms1 gene probably has a completely new transcript with unknown function.

[0070] In order to comprehensively annotate the genes existing in this interval, a series of primers covering this interval were designed, and the reverse transcription products of young panicle RNA of Nongken 58S and NIL (MH) young pa...

Embodiment 3

[0075] Embodiment 3: the design of light sensitive male sterility gene pms1 molecular marker

[0076] Comparative analysis of the sequences between molecular markers P4 and P7 revealed that there were four sequence differences between Nongken 58S and Minghui 63. Except for the co-segregated molecular marker P5 located in the promoter region of PMS1T, the other three were located on the transcript of PMS1T . There is an AT repeat sequence in the promoter region of PMS1T, the number of repeats in Nongken 58S is 14, and the number of repeats in Minghui 63 is 40. Based on this difference, a co-segregated SSR molecular marker P5 can be designed, which has also been used and has been Named Rssr (Liu N et al. Identification of an 85-kb DNA fragment containing pms1, a locus for photoperiod-sensitive genic male sterility in rice. Mol Gen Genomics 2001, 266(2): 271-275). The primers used for this marker detection are P5-F and P5-R.

[0077] Comparative analysis of the PMS1T nucleotide...

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Abstract

The invention relates to isolation and cloning of paddy rice photoperiod sensitive genic male sterility gene pms1 and applications thereof. The pms1 gene has only one transcript PMS1T, which is a non-coding long-chain RNA. There are insertion / deletion variation as long as 65 bp and two mononucleotide mutations between different alleles of the gene, wherein the sequence of a dominant allele of the pms1 is represented as the SEQ ID No.4 and the sequence of a recessive allele is represented as the SEQ ID No.3. By means of the insertion / deletion variation and one mononucleotide mutation, two molecular markers of the paddy rice photoperiod sensitive genic male sterility gene pms1 are obtained. The dominant allele of the pms1 can reduce the setting rate of NIL (MH) under long-day condition, while excessive expression on total-length or shortcut PMS1T also can reduce the setting rate of NIL (MH) under long-day condition too, so that by inhibiting the expression of the PMS1T in a photoperiod sensitive genic male sterile line Nongken 58S, the fertility of the Nongken 58S can be recovered. The isolation and cloning of the gene has important utilization value for selective breeding and improvement of a novel paddy rice photoperiod sensitive genic male sterile line.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering. It specifically relates to the isolation and cloning, functional verification and application of a rice light-sensitive nuclear sterile gene pms1 in rice improvement. [0002] The pms1 gene isolated in the present invention has only one transcript, PMS1T, which is a non-coding long-chain RNA and does not code protein. There is a 65bp insertion / deletion variation and two single nucleotide mutations between different alleles of this gene. The sequence of the dominant allele of pms1 is shown in SEQ ID NO:4, and the sequence of the recessive allele is shown in SEQ ID NO:3. Using the insertion / deletion variation and a single nucleotide mutation, two molecular markers of rice photosensitive genic male sterile gene pms1 were obtained. The dominant allele of pms1 can reduce the seed setting rate of NIL(MH) under long day, but has no effect on the rate of NIL(MH) under short day. Overe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12Q1/68C12N15/11
Inventor 张启发范优荣
Owner HUAZHONG AGRI UNIV
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