Molecular markers closely linked with dwarf gene and application of molecular markers

Abstract

The invention belongs to the field of crop genetic breeding, and in particular to molecular markers closely linked with a dwarf gene and an application of the molecular markers. The molecular markersare umc1383 and umc2241, wherein the sequence of the umc1383 is shown in SEQ ID No.1, and the sequence of the umc2241 is shown in SEQ ID No.2. According to the invention, genetic analysis is carried out on an EMS-induced maize dwarf mutant (Zheng 58d), and it finds that the phenotype of the dwarf mutant is controlled by a recessive single gene. Polymorphism screening is performed on Zheng 58, Zheng 58d, a hybrid pool and a homozygous pool by using SSR primers covering a whole genome, and a target gene is positioned on a first chromosome, so that the two SSR molecular markers umc1383 and umc2241 which are tightly linked with the target gene are screened out. The Zheng 58d contains the dwarf gene and thus has excellent rust resistance and comprehensive characteristics.

Description

technical field

[0001] The invention belongs to the field of crop genetics and breeding, in particular to molecular markers closely linked with dwarf genes and applications thereof. Background technique

[0002] Breeding practice has proved that every breakthrough in breeding depends on the discovery and utilization of favorable genes. The first revolution in rice breeding occurred in the 1960s. Based on the discovery of the dwarf gene sd and its application in dwarf breeding, the yield, resistance and variety of rice have been greatly improved. Corn is an important food crop and an important source of animal feed and industrial raw materials. Zhengdan 958 is the variety with the largest planting area in my country. It has ranked first in the planting area for 15 consecutive years, and has created great economic, social and ecological benefits. In view of its high yield, high density, stable yield, wide adaptability, multi-resistance and suitable for full-process mechanizat...

Examples

Embodiment 1

[0040] Acquisition of SSR molecular markers:

[0041] 1. Analysis of the genetic effect of the dwarf gene

[0042] Take dwarf Zheng 58d ( figure 1) were reciprocally crossed with Zheng 58 and B73, and it was found that the F1 phenotypes were all normal plants, indicating that the mutation belonged to a gene mutation in the nucleus and was a recessive gene; the corresponding BC1 and F2 segregation populations were constructed, and 258 BC1 strains The plant heights of the segregation population and 326 F2 segregation populations were investigated, and it was found that the proportions of normal plants and dwarf plants in the BC1 and F2 segregation populations were 136:122 and 252:74, respectively, and the chi-square test was completely consistent with 3 The segregation ratio of :1 or 1:1 indicated that Zheng 58d was controlled by the recessive monogenic gene (Table 1).

[0043] Table 1 Genetic analysis of dwarf mutant genes

[0044]

[0045] 2. Preliminary mapping of Zheng...

Embodiment 2

[0048] The application of molecular marker of the present invention in corn breeding, concrete steps:

[0049] In Hainan in the winter of 2013, the dwarf inbred line "Zheng 58d" was used as the donor, and the backbone self-selected lines Zheng 63, Zheng 36, Zheng 754, and Zheng P6 were used as the recipients, and the dwarf gene was introduced into the recipients through hybridization , get F1;

[0050] F1 plants were planted in Zhengzhou in the summer of 2014, and the pollen of Zheng 63, Zheng 36, Zheng 754, and Zheng P6 was used to backcross pollinate the F1 plants, and the BC1 population corresponding to Zheng 63, Zheng 36, Zheng 754, and Zheng P6 (containing AA and Aa two genotypes);

[0051] Use the "alkali cooking method" to quickly extract DNA: cut a small part of the endosperm and put it into a 96-well PCR plate, add 100ul of 0.1M NaOH, cover the PCR plate and heat bath at 100°C for 10 minutes on the PCR amplification instrument ; Take out and add an equal volume of 1...

technical field

[0001] The invention belongs to the field of crop genetics and breeding, in particular to molecular markers closely linked with dwarf genes and applications thereof. Background technique

[0002] Breeding practice has proved that every breakthrough in breeding depends on the discovery and utilization of favorable genes. The first revolution in rice breeding occurred in the 1960s. Based on the discovery of the dwarf gene sd and its application in dwarf breeding, the yield, resistance and variety of rice have been greatly improved. Corn is an important food crop and an important source of animal feed and industrial raw materials. Zhengdan 958 is the variety with the largest planting area in my country. It has ranked first in the planting area for 15 consecutive years, and has created great economic, social and ecological benefits. In view of its high yield, high density, stable yield, wide adaptability, multi-resistance and suitable for full-process mechanizat...

Examples

Embodiment 1

[0040] Acquisition of SSR molecular markers:

[0041] 1. Analysis of the genetic effect of the dwarf gene

[0042] Take dwarf Zheng 58d ( figure 1) were reciprocally crossed with Zheng 58 and B73, and it was found that the F1 phenotypes were all normal plants, indicating that the mutation belonged to a gene mutation in the nucleus and was a recessive gene; the corresponding BC1 and F2 segregation populations were constructed, and 258 BC1 strains The plant heights of the segregation population and 326 F2 segregation populations were investigated, and it was found that the proportions of normal plants and dwarf plants in the BC1 and F2 segregation populations were 136:122 and 252:74, respectively, and the chi-square test was completely consistent with 3 The segregation ratio of :1 or 1:1 indicated that Zheng 58d was controlled by the recessive monogenic gene (Table 1).

[0043] Table 1 Genetic analysis of dwarf mutant genes

[0044]

[0045] 2. Preliminary mapping of Zheng...

Embodiment 2

[0048] The application of molecular marker of the present invention in corn breeding, concrete steps:

[0049] In Hainan in the winter of 2013, the dwarf inbred line "Zheng 58d" was used as the donor, and the backbone self-selected lines Zheng 63, Zheng 36, Zheng 754, and Zheng P6 were used as the recipients, and the dwarf gene was introduced into the recipients through hybridization , get F1;

[0050] F1 plants were planted in Zhengzhou in the summer of 2014, and the pollen of Zheng 63, Zheng 36, Zheng 754, and Zheng P6 was used to backcross pollinate the F1 plants, and the BC1 population corresponding to Zheng 63, Zheng 36, Zheng 754, and Zheng P6 (containing AA and Aa two genotypes);

[0051] Use the "alkali cooking method" to quickly extract DNA: cut a small part of the endosperm and put it into a 96-well PCR plate, add 100ul of 0.1M NaOH, cover the PCR plate and heat bath at 100°C for 10 minutes on the PCR amplification instrument ; Take out and add an equal volume of 1...

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