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Protein coding sequence for controlling rice tapetum degradation

A technology for degrading proteins and coding sequences, applied in plant peptides, biochemical equipment and methods, applications, etc., can solve the problem of no description and report of OsTDR gene found

Inactive Publication Date: 2006-12-27
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No description and report of the OsTDR gene relevant to the present invention has been found so far

Method used

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  • Protein coding sequence for controlling rice tapetum degradation
  • Protein coding sequence for controlling rice tapetum degradation
  • Protein coding sequence for controlling rice tapetum degradation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: Obtaining and morphological observation of Ostdr mutant plants

[0041] This mutant was used in our laboratory 60 The japonica rice 9522 seeds were mutated with Co ray, and the treatment dose was 280Gy. A male sterile mutant in the F2 generation was backcrossed for three generations, and a stable genetic mutant Ostdr controlled by a recessive single gene was obtained. All plant materials were planted at the experimental base of Shanghai Academy of Agricultural Sciences. The Ostdr mutant was backcrossed with japonica rice 9522, all the F1 generations were fertile, and segregation occurred in the self-crossed F2 generation, of which 189 were normal plants and 55 were mutant plants, and the ratio of normal plants to mutant plants was close to 3:1 (χ 2 =0.79χ 0.05 3.84), indicating that the phenotype of the male sterile mutant was caused by a single nuclear gene mutation.

[0042] Morphological observation of Ostdr mutant plants. Such as figure 1 , The p...

Embodiment 2

[0043] Embodiment 2: Location and cloning of OsTDR gene

[0044] (1) Positioning groups. Ostdr was crossed with the indica rice line Longtefu B, the F2 generation was obtained by selfing, and the male sterile plants were selected as the positioning population.

[0045] (2) Rice DNA extraction. Using the improved CTAB method. The simple steps are as follows: Take 0.1-0.2 grams (about half a piece) of leaves and put them in a small mortar, add an appropriate amount of liquid nitrogen, grind them to powder immediately, put them into a 2ml centrifuge tube, add 700ul of 1.5xCTAB solution preheated at 100°C to centrifuge After 20 minutes, take out the centrifuge tube, add an equal volume of chloroform / isoamyl alcohol, mix vigorously, centrifuge (13000rpm) for 10 minutes, take the supernatant into a new tube, add 900ul After mixing with absolute ethanol, put it at -20°C for more than half an hour. The precipitated DNA was centrifuged at 14000 rpm (10 minutes). The supernatant wa...

Embodiment 3

[0051] Example 3: Functional analysis of OsTDR gene

[0052] The sequence analysis of the cloned OsTDR gene was carried out on the Internet. The function of the AMS gene in Arabidopsis thaliana, which has a high amino acid sequence homology with the OsTDR gene, has just been reported, and it controls the development of the pollen tapetum. This is similar to the phenotype of the Ostdr mutant, implying that the OsTDR gene also has a similar function ( Figure 4 ). In order to further confirm that the cloned OsTDR gene is the gene that causes the phenotype of the Ostdr mutant, the present invention cuts the BAC clone AP005851 (comprising 2391bp of ORF, 2430bp upstream region and 1857bp downstream region) with BamHI and SalI to construct a 6.6kb The pCAMBIA1301 plasmid was transformed into Agrobacterium tumefaciens EHA105, and transformed into the Ostdr mutant using genetic means to observe whether it would cause the plants to restore wild-type traits. Figure 5 Slices of mature...

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Abstract

This is the coding sequence of degradable protein which can control rice tapetum. This invention belongs to the gene engineering field. The extracted DNA molecule includes: nucleotide sequence that it's coding has the peptides with the rice OsTDR protein activity. The above sequence has the peptides which with the amino acid sequence indicated by the SEQ ID NO. 2, as well as the Nucleotide sequence of 1-1659 of nucleotide indicated by the SEQ ID NO.1. By the degradable protein which can control rice tapetum, what can be filtered through the normal filtration is the substance that interactioned with degradable protein which control rice tapetum or receptor, inhibitor or anatognist etc. This invention can obviously control the degradation of rice tapetum, produce new male sterile rice sterile line, generate hiberdization seeds, perform recurrent selection and create gene pool, so it is very important in agriculture.

Description

technical field [0001] The invention relates to a protein coding sequence, in particular to a protein coding sequence for controlling the degradation of rice tapetum, and belongs to the field of genetic engineering. Background technique [0002] Rice (Oryza sativa L.) is an important food crop. At the same time, because it has a small genome (389Mb) and has high collinearity with other grasses, rice has become a kind of plant functional gene research. Important monocot model organisms. Rice male sterility is of great significance in agricultural production and can be used to produce hybrid seeds, carry out recurrent selection and create gene pools. The genetic mechanism of male sterility has always been an important topic in developmental biology, and male sterility mutations are also suitable materials for studying the development of male gametophytes in plants. In recent years, genetics, cytology, anatomy, physiology and biochemistry have been studied on male sterility i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415
Inventor 张大兵李娜刘海生张大生袁政梁婉琪
Owner SHANGHAI JIAO TONG UNIV
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