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Rive CYP704B2 gene mutant, and molecular identification method and applications thereof

A technology of CYP704B2 and rice, applied in application, genetic engineering, plant gene improvement, etc., can solve problems such as low breeding efficiency, susceptibility to disease in breeding method, poor quality, etc.

Active Publication Date: 2015-10-28
HAINAN BOLIAN RICE GENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The three-line breeding method has problems such as disease susceptibility, poor quality, and low breeding efficiency
Since this mutation deletes part of the sequence of the upstream gene, there may be potential non-target trait variation in this mutant, which may adversely affect its utilization

Method used

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  • Rive CYP704B2 gene mutant, and molecular identification method and applications thereof
  • Rive CYP704B2 gene mutant, and molecular identification method and applications thereof
  • Rive CYP704B2 gene mutant, and molecular identification method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: Cobalt 60 radiation mutagenesis mutant library

[0039] Indica rice backbone parent variety 93-11 was selected as the material for radiation treatment. The whole genome sequence of this variety has been completed, which is very beneficial for molecular breeding of rice.

[0040] In the summer of 2013, in Changsha Cobalt 60 ( 60 Co) Radiation 93-11 Seed (M 0 generation) 10 kg, planted in Hainan Lingao field in July, and harvested by individual plants. 1 A total of about 6,500 seeds were harvested.

[0041] choose M 1 There are 3200 strains of generation seeds, and 50 individual plants are planted for each strain. In the spring of 2014, it was planted in Lingao Field, Hainan. After transplanting, carefully observe the field traits at the tillering stage, booting stage, heading stage, flowering stage, and filling stage, and screen various types of mutants such as plant type, panicle type, fertility, and yield. A single plant is harvested and stored as a...

Embodiment 2

[0042] Example 2: M 2 Substitute Planting and Character Observation

[0043] in M 2 During the period of heading and flowering, the morphology of anthers was observed in the field, and anthers with abnormalities such as light white color, small shape, and small pollen amount were selected for further microscopic examination under a microscope. A plant with abnormal fertility was found in the family numbered 1907, and the mutant had no obvious difference from the wild type in terms of vegetative growth, heading stage, and panicle type. figure 1 is the plant type of the mutant plant and the wild-type plant ( figure 1 left) and panicle ( figure 1 Right) Comparison photos, but the anthers are smaller than the wild type, the color is light yellow, and the seed setting rate is very low, so it is selected as a candidate mutant material for further research.

Embodiment 3

[0044] Embodiment 3: Microscopic examination of pollen, selfing, outcrossing

[0045] The pollen fertility was counted by counting the ratio of iodine-stained pollen to non-colored pollen. Observing the floret morphology of the 1907 mutant under a stereomicroscope, it was found that the pistil was not significantly different from the wild type, and the anthers were smaller and lighter in color than the wild type ( figure 2 left). Collect florets at the flowering stage in the field, take out the anthers with tweezers, and put them in iodine-potassium iodide solution (0.6% KI, 0.3% I 2 , w / w), gently squeeze the anthers, drop on the glass slide, cover with a cover glass, observe the pollen iodine staining under the microscope and take pictures ( figure 2 Right). The wild type has more pollen and is stained blue-black, while the mutant has no pollen grains.

[0046] The wild-type plants of the same family set fruit normally after bagging selfing, but the 1907 mutant did not...

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Abstract

The invention provides a rive CYP704B2 gene mutant, and applications thereof, and belongs to the technical field of gene engineering. According to the rive CYP704B2 gene mutant, indica rice 93-11 is subjected to cobalt 60 radiation-induced mutation so as to replace three G bases after the 794th base of rice CYP704B2 gene with one T base; the CYP704B2 gene mutant is named as cyp704B2-3; the nucleotide sequence of the CYP704B2 gene mutant is represented by SEQ ID No.1; it is further confirmed that rice recessive genic male sterility is induced by the CYP704B2 gene mutant; the CYP704B2 gene mutant can be used for culturing transgenic rice with recessive genic male sterility, and possesses great importance in rice genetic resource genetic improvement-breeding. The invention also provides a molecular marked identification method and applications of the rive CYP704B2 gene mutant in breeding.

Description

technical field [0001] The invention belongs to the field of plant molecular biology, and in particular relates to a rice CYP704B2 gene mutant cyp704B2-3 and its molecular identification method and application. Background technique [0002] Plant male sterile mutation is a very common phenomenon in nature, at least male sterile mutants have been found in 617 species of 43 families and 162 genera. Genetically, plant male sterility is divided into three categories: nuclear male sterility, cytoplasmic male sterility and nuclear-cytoplasmic interaction male sterility: 1) Nuclear male sterility is produced by nuclear gene mutations, and there are dominant mutations and recessive mutations , There are sporophytic gene mutations and gametophytic gene mutations. Dominant mutations and gametophytic gene mutations can only be inherited through female gametes, recessive mutations can be inherited through both female gametes and male gametes, and follow Mendel's laws. Some sporophytic...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N15/82C12N5/10A01H5/00
Inventor 黄培劲李新鹏李京琳安保光张维龙湍
Owner HAINAN BOLIAN RICE GENE TECH CO LTD
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