Molecular marker related to Oryza sativa L. fertility and floral organ number, and application of molecular marker

A molecular marker, rice technology, applied in the field of plant molecular biology, to achieve the effect of stable fertility

Active Publication Date: 2021-03-19
HAINAN BOLIAN RICE GENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the genetic regulation mechanism of rice flower development cannot yet form a general outline like that of Ar

Method used

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  • Molecular marker related to Oryza sativa L. fertility and floral organ number, and application of molecular marker
  • Molecular marker related to Oryza sativa L. fertility and floral organ number, and application of molecular marker
  • Molecular marker related to Oryza sativa L. fertility and floral organ number, and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Phenotype and genetic analysis of rice nuclear male sterile female hypofertility mutant mil2-4390

[0055] (1) Screening of mil2-4390 mutants

[0056] In June 2013, 10 kg of 93-11 seeds were irradiated with cobalt 60 to obtain M 0 generation. The irradiated seeds were planted in the experimental field of Lingao County, Hainan Province, and harvested by individual plants after maturity. A total of M 1 There are about 6,500 substitute materials. In the spring of 2014, 3617 M with more seeds were selected 1 The generation materials were planted into strains, and 50 individual plants were planted in each strain. Various types of mutants such as plant type, panicle type, fertility, yield, etc. were screened at the tillering stage, booting stage, heading stage, flowering stage, and grain filling stage, and harvested and stored. In the No. 4390 family, a mutant was found to be male sterile, which was named mil2-4390. Wild type and mutant mil2-4390 Whole plant a...

Embodiment 2

[0065] Example 2 Leaf Sampling and DNA Extraction

[0066] Use the CTAB method to extract DNA from rice leaves. The specific method is as follows: Weigh about 0.1g leaves, put them into a 2.0mL centrifuge tube, add 800μL 1.5×CTAB extraction buffer, 5μL RNase A, shake to disperse, and put them in a water bath at 65°C for 0.5h. Shake 2-3 times; add an equal volume of chloroform / Tris-saturated phenol (1:1, v / v), mix well, shake gently for 10 minutes; centrifuge at 10000rpm for 20 minutes; transfer the supernatant to a new tube, add 1 / 10 volume of 3M Sodium acetate (pH 5.2), 0.6 times the volume of isopropanol; gently shake and mix until flocculent precipitate appears; centrifuge at 10000rpm for 10min; discard the supernatant, wash the precipitate twice with 70% ethanol by volume; air dry , add 200 μL 1×TE to dissolve the precipitate, and store at -20°C. The DNA concentration was detected by Nanodrop 2000 and diluted to 10ng / L for use as a PCR template.

Embodiment 3

[0067] Example 3 Preliminary Mapping and Detection of the Male Sterility Gene MIL2-4390

[0068] The MIL2-4390 gene was mapped using bulked segregant analysis and recessive-class analysis for extreme traits. A F 2 population, 305 sterile strains were isolated. Using 301 pairs of SSR markers evenly distributed on 12 chromosomes to screen polymorphic markers between 93-11 and Minghui 63, 64 pairs of polymorphic SSR markers were screened, and the polymorphism was low, indicating that The genome sequences of Hui 63 and 93-11 were slightly different. Build the F respectively 2 The mixed gene pool of fertile strains and sterile strains of the population, using the 64 pairs of polymorphic markers to analyze the fertile gene pool and the sterile gene pool. The results showed that the SSR markers 12.109 and 12.184 on chromosome 12 were linked to the mutant phenotype of mil2-4390. Further use the above linkage markers to analyze the F of Minghui 63 and mil2-4390 one by one 2 The g...

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Abstract

The invention provides a molecular marker related to Oryza sativa L. fertility and a floral organ number, and application of the molecular marker. After an indica type Oryza sativa L. variety 93-11 issubjected to cobalt 60 irradiation mutagenesis, screening is carried out to obtain a male nuclear sterile mutant; through a map-based cloning method, the mutant is positioned on the No.12 chromosomelong arm of Oryza sativa L., 314.070Kb bases, including an MIL2 gene, from a base site 16702239 to the base site 17016311 are subjected to deletion; and the deleted mutant is named mil2-4390, the nucleotide sequences of two ends of a deletion fragment are disclosed by SEQ ID No.1, and the mutant can be authenticated through the molecular marker M1. The mil2-4390 mutant causes Oryza sativa L. recessive male nuclear sterility and female low fertility and also causes variation of the Oryza sativa L. floral organ number, and therefore, the mil2-4390 mutant can be used for preparing transgenosis Oryza sativa L. with recessive male nuclear sterility or floral organ variation, and performs an important function in genetic improving and breeding of Oryza sativa L. germplasm resources.

Description

technical field [0001] The invention relates to the field of plant molecular biology, in particular to a gene mutant related to rice nuclear male sterility, low female fertility, and development of flower organ numbers, and its molecular marker and application. Background technique [0002] Rice is not only a model plant for monocot genome research, but also an important food crop. Rice is the staple food of one-third of the world's humans, and its flowers are not only reproductive organs, but also the basis for grain formation. The normal development of rice flowers directly affects rice yield and rice quality, so the research on rice flower development has always been the focus of rice genetics and breeding workers. Studying rice flower development not only has important theoretical significance, but also has important guiding significance for rice yield and quality genetics and breeding. [0003] Through the study of rice molecular genetics, it was found that the "ABCDE...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46C12Q1/6895C12N15/11
CPCC07K14/415C12N15/8289C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 唐杰吴春瑜李佳林曾翔吴永忠黄培劲
Owner HAINAN BOLIAN RICE GENE TECH CO LTD
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