Gene and protein encoded by rice root growth and development control gene OsSPR1

A protein and rice root technology, applied in genetic engineering, plant genetic improvement, fermentation, etc., can solve problems such as root length shortening

Inactive Publication Date: 2010-11-24
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ge et al. (2004) reported OsRAA1, enhanced expression of this gene resulted in shorter root length

Method used

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  • Gene and protein encoded by rice root growth and development control gene OsSPR1
  • Gene and protein encoded by rice root growth and development control gene OsSPR1
  • Gene and protein encoded by rice root growth and development control gene OsSPR1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, screening and phenotype of mutants

[0026] Using the Kasalath mutant library induced by EMS as the mutant screening object, M 2 Seeds were rinsed with distilled water, 0.6% dilute HNO 3 Dormancy-breaking treatment for 16 hours, germination in the dark at 37°C until white. Sow the lubai seeds on the nylon mesh floating on the rice culture solution (the formula of the culture solution is the standard formula of the International Rice Institute), and cultivate it for 7 hours at a temperature of about 30 / 22°C (day / night) and 12 hours of light. The next day, the mutants were screened based on the change of root configuration (adventitious root length, lateral root length, adventitious root number, lateral root number, etc.), and a mutant with short adventitious roots and lateral roots was screened. There was no significant difference between the shoot length of the mutant and the wild type, and the main root length was slightly shorter, but the adventitious ...

Embodiment 2

[0030] Embodiment 2, gene localization

[0031] f 2 The mapping population was obtained by crossing the homozygous (OsSPR1) with the japonica rice variety Nipponbare, and a total of 1511 F 2 Individuals, using the rapid extraction method of rice trace DNA to extract genomic DNA for gene mapping from rice leaves. Take about 0.1 g of young rice leaves, freeze them with liquid nitrogen, grind the leaves into powder in a 1.5 ml centrifuge tube, extract the total DNA, and dissolve the obtained DNA in 200 μl sterile water. 2 μl of DNA sample was used for each SSR and STS reaction.

[0032] In the preliminary mapping experiment of OsSPR1 gene, 100 F 2 Individuals were subjected to SSR analysis. According to the published molecular genetic maps created by japonica and indica rice, select SSR primers that are approximately evenly distributed on each chromosome, perform PCR amplification according to known reaction conditions, and then separate by 7% acrylamide gel electrophoresis t...

Embodiment 3

[0034] Example 3, gene prediction and sequence analysis

[0035] According to the results of fine mapping, the Osspr1 gene is located within the 18.1 kb range between the STS1 and STS2 markers on the BAC clone P0506A10 ( figure 1 Middle A). According to the rice gene annotation information of TIGR (http: / / www.tigr.org / tdb / e2k1 / osa1 / ) and the EST database (http: / / www.ncbi.nlm.nih.gov / ), the mapped chromosomal regions In-segment gene prediction analysis, there are 2 unknown genes in this interval. The two genes were amplified using the cDNA of Kasalath wild-type and OsSPR1 mutant roots as templates, and the amplified products were sequenced separately, and the sequencing results were compared and analyzed. In the mutant, a single base mutation occurred in the ORF (LOC Os01g67290) of one of the genes. C was changed to T at 2533bp of the gene, and a premature stop codon appeared, resulting in the deletion of 136 amino acids at the amino acid terminal. Then we sequenced the gene...

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Abstract

The invention relates to the plant gene engineering field, aiming at providing protein encoded by a rice root growth and development control gene OsSPR1 and a gene for encoding the protein. The protein has an amino acid sequence shown by SEQ ID NO:2 and a nucleotide sequence shown by SEQ ID NO:1. In the invention, a rice mutant with a short adventitious root and a short branch root is taken as a research material to determine that the gene is involved in regulating and controlling growth and development of the rice root; the functional deletion mutant of the gene shows reduced ferric content of blades, and the ferric content in the root is in accordance with that of wild rice; and recovery of expression of the gene can recover the ferric content in the blades, thus showing that the gene can regulate and control the ferric content in the blades. The invention provides a molecular regulation and control mechanism for rice root growth and development and ferric ion in-vivo transport, and provides a foundation for regulating and controlling a rice root structure and the ferric content by a genetic engineering method.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to the rice root length regulation gene OsSPR1 and its application. Background technique [0002] The root system of plants plays a very important role in the growth of higher plants. It not only provides physical support for the aboveground parts, but also absorbs nutrients and water from the soil to maintain the growth and development of plants. The root system of plants can be divided into two categories, the tap root system of most dicotyledonous plants and the fibrous root system of most monocotyledonous plants. The root system of dicots is mainly composed of tap roots and lateral roots; while the root system of monocots is mainly composed of tap roots, adventitious roots and lateral roots. As one of the most important food crops in the world, rice has a root system consisting of seed roots, a large number of adventitious roots and latera...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
Inventor 吴平毛传澡贾立强吴忠长陈婕妤
Owner ZHEJIANG UNIV
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