Rice male sterility gene OsLDDT1 and molecular marker and application thereof

A male sterility, molecular marker technology, applied in the fields of genetic engineering and plant genetics and breeding

Pending Publication Date: 2022-04-01
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The research on the regulation of rice anther development and male sterility by the α/β hydrolase gene OsLDDT1 in rice has not been reported yet. The rice OsLDDT1 gene was isolated and cloned to explore the cause of rice male sterility caused by the mutation of this gene, and ...

Method used

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  • Rice male sterility gene OsLDDT1 and molecular marker and application thereof
  • Rice male sterility gene OsLDDT1 and molecular marker and application thereof
  • Rice male sterility gene OsLDDT1 and molecular marker and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The lddt1 mutant and the wild-type ZH8015 were photographed with a Nikon D800 digital camera and a Carl Zeiss LumurV12 stereofluorescence stereomicroscope (Marcusari, Germany) to photograph rice plants and flower organ tissues and other materials. in I 2 -KI pollen dyeing process, anthers and pollen grains soaked in a concentration of 1.2% I 2 -KI solution (about 30 seconds), and then photographed and saved under a Leica microscope.

[0060] figure 1 The mutant was derived from the EMS chemical mutagenesis population of indica rice Zhonghui 8015. The mature anthers of the mutant lddt1 were milky white without pollen, and the spikelets were completely aborted.

[0061] Table 1 BC produced by crossing lddt1 mutants with wild type and 02428 respectively 1 f 1 and F 1 、BC 1 f 2 and F 2 The phenotypic statistics of spikelet seed setting rate, inner and outer floral organs and anther development were carried out for each group.

[0062] Table 1 Genetic analysis of l...

Embodiment 2

[0066] Observation of anther development in lddt1 mutant and wild-type ZH8015 by semi-thin section

[0067] (1) Sampling and fixation: Take the florets of wild type and mutant type at different developmental stages on the living body, and immediately place them in freshly prepared FAA fixative solution (V 37％甲醛 :V 冰乙酸 :V 50％乙醇 =5:5:90), use a vacuum pump to slowly pump air, place in a near-vacuum state, slowly deflate until all samples sink to the bottom, replace with new FAA, and fix for 24 hours.

[0068] (2) Gradient alcohol dehydration: put the samples in different vials according to the length of the florets, remove the glumes and carry out alcohol gradient treatment: 50% ethanol, 60% ethanol, 70% ethanol, 85% ethanol, 95% ethanol soaking at various levels 0.5h; then 100% ethanol for 1h, saturated safranin for 2h.

[0069] (3) Pre-permeation: Anhydrous ethanol and Technovit 7100 resin were mixed in equal volumes to prepare a pre-permeation solution, and pre-permeated a...

Embodiment 3

[0075] Scanning electron microscopy observation of lddt1 mutant and wild-type anther development

[0076] 1) Double fixation of samples: fresh florets of lddt1 mutant and wild type were taken and immersed in 2.5% glutaraldehyde solution immediately. The glumes of the samples at the mature stage of the anthers need to be cut to better fix the samples, and the glumes of other samples do not need to be removed, so as not to damage the surface structure of the anthers during sample processing. The subsequent sample washing and acid treatment methods are the same as those for transmission electron microscopy.

[0077] 2) Dehydration: The dehydration treatment is the same as the transmission electron microscope treatment method, the only difference is that the last step of acetone treatment is replaced by absolute ethanol treatment for 20 minutes.

[0078] 3) drying and observation: critical point drying. The auspicious products were dried in a Hiachai HCP-2 critical point dryer. ...

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Abstract

The invention provides a rice male sterility gene OsLDDT1 as well as a molecular marker and application thereof, and belongs to the technical field of genetic engineering and plant genetic breeding. The invention reveals that the rice gene OsLDDT1 has a biological function of regulating and controlling plant male fertility for the first time. The function of the gene is verified in rice based on map-based cloning and functional complementation experiments, and after a mutant lddt1 is transformed by using a pCAMBIA1300-OsLDDT1 functional complementation vector, the fertility of a male gamete of a transgenic plant is recovered, that is, the male is partially aborted due to function loss mutation of the OsLDDT1 gene, and the fertility of the gene is recovered to be normal due to functional complementation. The rice OsLDDT1 gene disclosed by the invention is completely aborted after function loss, part of seeds can be harvested, beneficial supplementation can be formed with other male sterile genes, and a new gene resource is provided for breeding of a rice intelligent restorer line.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and plant genetic breeding, and specifically relates to rice male sterility gene OsLDDT1 and its molecular marker and application. Background technique [0002] Rice is one of the main food crops, and more than two-thirds of my country's population uses rice as a staple food. Since the 1960s and 1970s, rice breeding has experienced two "green revolutions" of dwarf breeding and hybrid breeding, which have greatly increased rice production and made important contributions to solving the problem of food and clothing for people around the world. Among them, the discovery of the male sterile line and the matching of the "three-line method" and "two-line method" hybrid rice seed production technology have made great contributions to the leap in rice yield per unit and the food security of China and the world; among them , the sterile material is the core, and the creation and utilization of...

Claims

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Application Information

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IPC IPC(8): C12N9/14C12N15/55C12N15/82C12Q1/6895A01H5/00A01H5/10A01H5/02A01H6/46
Inventor 孙廉平程式华曹立勇占小登孙志豪陈驰彭泽群张迎信陈代波
Owner CHINA NAT RICE RES INST
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