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Application of corn gene ZmABCG20 in regulating male fertility of crops

A technology of male fertility and male sterility, which is applied in the fields of application, genetic engineering, and plant genetic improvement, and can solve problems such as unstable fertility of sterile lines, affecting the purity of hybrids, and the withdrawal of cytoplasmic male sterility technology from the market.

Pending Publication Date: 2019-03-08
HAINAN BOLIAN RICE GENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cytoplasmic male sterile lines that have been used in maize have some defects: First, because the cytoplasmic male sterile lines need specific restorer genes to restore fertility, the utilization rate of germplasm resources is very low, which limits the efficiency of breeding of excellent varieties Secondly, the fertility of some sterile lines is unstable, and the fertility can be restored under certain conditions, which affects the purity of the hybrid; finally, due to the single cytoplasmic genotype, it causes a large outbreak of corn spot disease, which directly leads to cytoplasmic male sterility technology. qiut the market
Through genome bioinformatics analysis, Pang et al. (Pang et al., 2013, Inventory and general analysis of the ATP-binding cassette (ABC) gene superfamily inmaize (Zea May L.). Gene, 2013, 526(2):411-428) Fifty-four ABCG genes have been identified from maize species, but no genes related to male fertility have been found so far

Method used

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  • Application of corn gene ZmABCG20 in regulating male fertility of crops
  • Application of corn gene ZmABCG20 in regulating male fertility of crops
  • Application of corn gene ZmABCG20 in regulating male fertility of crops

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0087] Example 2 M 2 Substitute Planting and Character Observation

[0088] in M 2 During the heading and flowering period of the first generation, 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. In the family numbered 3326, 9 plants with abnormal fertility were found, which could not loose powder normally, but had normal fruiting ( figure 1 ). The anthers of the mutant are smaller than the wild type, light yellow in color, and have no visible pollen, but there is no obvious difference from the wild type in vegetative growth, heading stage, and panicle type, so it is selected as a candidate mutant material for further research. According to the finally identified mutated gene (see Examples 6, 7, 8), the mutant was named zmabcg20-1.

Embodiment 3

[0089] Example 3 Sterile Mutant Pollen Microscopic Examination and Genetic Analysis

[0090] The pollen fertility was counted by the ratio of iodine-stained pollen to non-colored pollen. Observing the morphology of zmabcg20-1 male flowers under a stereomicroscope, the anthers were smaller and lighter in color than the wild type ( figure 2 ). 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 them on a glass slide, cover with a cover glass, observe the pollen iodine staining under a microscope and take pictures. The wild type has more pollen and is stained blue-black, while the mutant cannot see the pollen grains ( figure 2 ).

[0091] Mutants can bear fruit normally under open pollination ( figure 1 ), indicating that the mutant is a male sterile mutant, and ear fertility is not affected. Harvest zmabcg20-1 open-pollinated s...

Embodiment 4

[0092] Example 4 Leaf Sampling and DNA Extraction

[0093] In this study, the CTAB method was used to extract DNA from maize leaves. The specific method is as follows: Weigh about 0.1g leaves, put them into a centrifuge tube, add 600 μL CTAB extraction buffer, 5 μL RNase A, shake to disperse, put in a water bath at 65°C for 0.5hr, and shake gently in between 2-3 times; add an equal volume of chloroform / Tris-saturated phenol (1:1, v / v), mix well, and shake gently for 10 min; centrifuge at 10000 rpm at 4°C for 20 min; transfer the supernatant to a new tube, and add 1 / 10 volume of 3M sodium acetate (pH 5.2), 0.6-1 times the volume of cold isopropanol; gently shake and mix until flocculent precipitates appear; centrifuge at 10000rpm at 4°C for 10min; discard the supernatant, and use 70% ethanol by volume Wash the precipitate twice; air dry, add 50 μL 1×TE to dissolve the precipitate, and store at -20°C. The DNA concentration was detected by Nanodrop2000 and diluted to 10ng / L for ...

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Abstract

The invention provides application of a corn gene ZmABCG20 in regulating male fertility of crops. The genomic DNA sequence of the ZmABCG20 in the maize variety B73 is shown as SEQ ID NO: 1, and the encoded protein sequence is shown as SEQ ID NO: 3. The invention also provides a mutant zmabcg20-1 of the gene ZmABCG20, and application thereof, the mutant gene sequence is shown as SEQ ID NO: 7; the invention further provides a molecular marker identification method for the mutant gene. The pollen development control gene, the mutant and a molecular marker of the mutant can be applied to cross breeding and hybrid seed production of the crops.

Description

technical field [0001] The invention belongs to the field of genetic engineering and molecular breeding, and in particular relates to the application of the corn gene ZmABCG20 in regulating the male fertility of crops. 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 spo...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/113C12N15/82A01H5/00A01H6/46C07K14/415
CPCC07K14/415C12N15/8289
Inventor 黄培劲李新鹏李京琳陈磊李燕群陈江淑陈思兰安保光龙湍吴永忠
Owner HAINAN BOLIAN RICE GENE TECH CO LTD
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