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Rice fibril controlling gene OsRHL1 and uses thereof

A rice and genetic technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2009-07-29
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no related reports on the regulation mechanism of rice bHLH transcription factors on root hair development

Method used

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  • Rice fibril controlling gene OsRHL1 and uses thereof
  • Rice fibril controlling gene OsRHL1 and uses thereof
  • Rice fibril controlling gene OsRHL1 and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1, screening and phenotype of mutants

[0034] 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. On the next day, the mutants were screened under the stereoscope with the change of root hair configuration (including length, thickness, etc.) as the screening criteria, and a mutant almost without root hairs was screened ( figure 1 B). There was no significant difference between the mutant's shoot (including plant height, etc.) and root (tap root length, lateral root length and number, adventitious root length and ...

Embodiment 2

[0035] Embodiment 2, gene localization

[0036] f 2 Mapping populations were obtained from crosses between homozygous (Osrhl1) and japonica cultivar Nipponbare, and a total of 700 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.

[0037] In the preliminary mapping experiment of OsRHL1 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 to detec...

Embodiment 3

[0043] Example 3, gene prediction and sequence analysis

[0044] According to the results of fine mapping, the OsRHL1 gene is located within the 13.1 kb range between the STS1 and STS2 markers on the BAC clone P0554A06 ( figure 2 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 Gene prediction analysis within the segment, there are 4 unknown genes and a bHLH transcription factor in this segment. The bHLH gene was amplified using the genomic DNA and cDNA of the Kasalath wild-type and Osrhl1 mutant roots respectively as templates, and the amplified products were sequenced respectively. The sequencing results were compared and analyzed, and it was found that the gene at 593bp after the start codon ATG The base G was mutated to an A, causing the second intron to not be properly spliced. Specific primers (upstream primer sequence: ATGGAGGGCGGCTTC...

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Abstract

The invention discloses a protein of rice root hair growth control gene OsRHL1 coding, which has an amino acid sequence expressed by SEQ ID NO: 2. The invention further simultaneously discloses genes coding the protein, which has a nucleotide sequence expressed by SEQ ID NO: 1. The invention further simultaneously discloses a method for modifying rice root hair, including transforming rice cells by using the gene and cultivating the rice cells after transformation into plants.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a cDNA sequence of the rice OsRHL1 (rice root hairless 1) gene cloned by a map-based cloning technique, the protein encoded by the cDNA belongs to the bHLH family transcription factor, and the gene is identified through a transgene complementation function experiment. Background technique [0002] The root system is an important organ for plants to absorb water and nutrients, and the root hair is an important part of the physiologically active interface of the functional section of the plant root system. The structure of plant root tip includes four parts: root cap, meristematic zone, elongation zone and maturation zone. Morphological differences between root hair cells and non-root hair cells begin to appear in the meristematic zone; root hairs begin to form in the elongation zone; hair cells mature and fully differentiate in the maturation...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N5/10C12N15/82
Inventor 吴平丁沃娜王文峰童艳丽黄炜陈汉民
Owner ZHEJIANG UNIV
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