Molecular marker GSSR187 closely linked with pepper anther yellow gene and application thereof

Abstract

The invention discloses a molecular marker GSSR187 closely linked with pepper anther yellow gene and application thereof. The invention provides a primer pair for authenticating or assisting to authenticate whether to-be-tested pepper has a character of anther yellow, so as to perform amplification to obtain the following primer pairs of DNA segments: pepper genomic DNA is utilized as a template, and primers as shown in a sequence 1 and a sequence 2 are utilized to perform PCR amplification, thereby obtaining DNA segments, specifically primer pairs formed by two single-stranded DNAs as shown in the sequence 1 and the sequence 2. According to the molecular marker GSSR187 closely linked with pepper anther yellow gene and the application thereof, the existing excellent varieties of pepper materials on the market are selected to perform hybridization, haploid cultivation and doubling, and then perform anther color screening and authentication, thereby knowing that the anther yellow is controlled by a single recessive gene; based on further research, the single recessive gene is closely linked with SSR marker GSSR187, and the GSSR187 can predict whether the anther is yellow before the pepper flowers, so that the GSSR187 can be applied to pepper seed breeding.

Description

technical field

[0001] The invention belongs to the field of genetic engineering and relates to a molecular marker GSSR187 closely linked with pepper anther yellow gene and its application. Background technique

[0002] Pepper (Capicum annuum L.) is an important vegetable crop widely cultivated worldwide. Pepper heterosis is obvious, and the use of heterosis can effectively solve the pepper market demand. At present, the main application in the pepper market is F 1 generation hybrids. The level of seed purity directly affects the yield and product quality of the first generation of hybrids. During hybrid seed production of pepper, the generation of selfed seeds of the female parent will significantly reduce the purity of hybrids. In the process of producing seeds using male sterile dual-purpose lines of pepper, if 50% of the fertile plants cannot be completely pulled out, the quality of hybrid seeds will also be greatly reduced. Moreover, the fertility of the sterile li...

Examples

Embodiment 1

[0062] Example 1. Breeding and identification of pepper material DH121 with the trait of "anther yellow" controlled by a single recessive gene

[0063] 1. Breeding of pepper material DH121 with the trait of "anther yellow" controlled by a single recessive gene

[0064] A method for breeding pepper material DH121 with the trait of "anther yellow" controlled by a single recessive gene, such as figure 1 shown.

[0065] (1) Selection of hybrid materials: four excellent pepper varieties "Baou", "Marilyn", "Boshou No. 1" and "Boshou No. 2" for hybridization.

[0066] (2) Hybridization: "Baou" was crossed with "Boshou No. 1" to obtain single-cross hybrid 1; "Marilyn" was crossed with "Boshou No. 2" to obtain single-cross hybrid 2. Then cross the single-cross hybrid 1 and the single-cross hybrid 2 to obtain a double-cross hybrid.

[0067] (3) Haploid cultivation and doubling: the double-cross hybrids are haploid-cultivated to obtain several naturally doubled doubled haploids (DH st...

Embodiment 2

[0095] Example 2. Acquisition of the SSR marker GSSR187 closely linked to the ayw gene

[0096] 1. Experimental method

[0097] Adopt Tiangen Plant Genomic DNA Extraction Kit to extract parents, F in Example 1 2 Genomic DNA of each individual plant in the population. First, 1200 pairs of SSR primers (labeled genSSR, GSSR, ZS, PS) and 2400 pairs of InDel primers (labeled GI, CIDH) distributed evenly on 12 chromosomes of pepper were used to screen the polymorphisms of the parents of pepper. Then, in F 2 The DNA of 7 individual plants of yellow anthers and purple anthers were selected in the population, and the gene pools of yellow anthers and non-yellow anthers were constructed, and the polymorphic primers screened in the previous step were used to perform BSA analysis on the DNA of the yellow and non-yellow anther gene pools of peppers. Further screening of polymorphic primers. Finally, using the polymorphic primer pair F screened out in the previous step 2 The genotypes o...

Embodiment 3

[0106] Example 3, the application of the SSR marker GSSR187 closely linked to the ayw gene in the identification of pepper anther color

[0107] 1. Experimental materials

[0108] The BC constructed from the pepper material DH121 and the pepper inbred line 12-Z65 as parents in Example 1 1 96 individual plants (Table 3) were randomly selected from the population as experimental materials, and the SSR marker GSSR187 closely linked to the ayw gene obtained in Example 2 was verified, and it was determined to use this marker to screen pepper varieties with the trait of "yellow anthers" , for the accuracy of marker assisted breeding (Marker assisted selection, MAS).

[0109] 2. Experimental method

[0110] For the BC constructed from the pepper material DH121 and the pepper inbred line 12-Z65 as parents in Example 1 1 Randomly selected 96 individual plants (Table 3) in the population, after the seedling stage single plant listing mark sampling, extract its genomic DNA and use it ...

technical field

[0001] The invention belongs to the field of genetic engineering and relates to a molecular marker GSSR187 closely linked with pepper anther yellow gene and its application. Background technique

[0002] Pepper (Capicum annuum L.) is an important vegetable crop widely cultivated worldwide. Pepper heterosis is obvious, and the use of heterosis can effectively solve the pepper market demand. At present, the main application in the pepper market is F 1 generation hybrids. The level of seed purity directly affects the yield and product quality of the first generation of hybrids. During hybrid seed production of pepper, the generation of selfed seeds of the female parent will significantly reduce the purity of hybrids. In the process of producing seeds using male sterile dual-purpose lines of pepper, if 50% of the fertile plants cannot be completely pulled out, the quality of hybrid seeds will also be greatly reduced. Moreover, the fertility of the sterile li...

Examples

Embodiment 1

[0062] Example 1. Breeding and identification of pepper material DH121 with the trait of "anther yellow" controlled by a single recessive gene

[0063] 1. Breeding of pepper material DH121 with the trait of "anther yellow" controlled by a single recessive gene

[0064] A method for breeding pepper material DH121 with the trait of "anther yellow" controlled by a single recessive gene, such as figure 1 shown.

[0065] (1) Selection of hybrid materials: four excellent pepper varieties "Baou", "Marilyn", "Boshou No. 1" and "Boshou No. 2" for hybridization.

[0066] (2) Hybridization: "Baou" was crossed with "Boshou No. 1" to obtain single-cross hybrid 1; "Marilyn" was crossed with "Boshou No. 2" to obtain single-cross hybrid 2. Then cross the single-cross hybrid 1 and the single-cross hybrid 2 to obtain a double-cross hybrid.

[0067] (3) Haploid cultivation and doubling: the double-cross hybrids are haploid-cultivated to obtain several naturally doubled doubled haploids (DH st...

Embodiment 2

[0095] Example 2. Acquisition of the SSR marker GSSR187 closely linked to the ayw gene

[0096] 1. Experimental method

[0097] Adopt Tiangen Plant Genomic DNA Extraction Kit to extract parents, F in Example 1 2 Genomic DNA of each individual plant in the population. First, 1200 pairs of SSR primers (labeled genSSR, GSSR, ZS, PS) and 2400 pairs of InDel primers (labeled GI, CIDH) distributed evenly on 12 chromosomes of pepper were used to screen the polymorphisms of the parents of pepper. Then, in F 2 The DNA of 7 individual plants of yellow anthers and purple anthers were selected in the population, and the gene pools of yellow anthers and non-yellow anthers were constructed, and the polymorphic primers screened in the previous step were used to perform BSA analysis on the DNA of the yellow and non-yellow anther gene pools of peppers. Further screening of polymorphic primers. Finally, using the polymorphic primer pair F screened out in the previous step 2 The genotypes o...

Embodiment 3

[0106] Example 3, the application of the SSR marker GSSR187 closely linked to the ayw gene in the identification of pepper anther color

[0107] 1. Experimental materials

[0108] The BC constructed from the pepper material DH121 and the pepper inbred line 12-Z65 as parents in Example 1 1 96 individual plants (Table 3) were randomly selected from the population as experimental materials, and the SSR marker GSSR187 closely linked to the ayw gene obtained in Example 2 was verified, and it was determined to use this marker to screen pepper varieties with the trait of "yellow anthers" , for the accuracy of marker assisted breeding (Marker assisted selection, MAS).

[0109] 2. Experimental method

[0110] For the BC constructed from the pepper material DH121 and the pepper inbred line 12-Z65 as parents in Example 1 1 Randomly selected 96 individual plants (Table 3) in the population, after the seedling stage single plant listing mark sampling, extract its genomic DNA and use it ...

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