Molecular marker linked with drought-resistant rice gene qLRI9, and application thereof

Abstract

The invention discloses a molecular marker linked with a drought-resistant rice gene qLRI9, and application thereof in rice drought resistance identification. Two sets of rice-dry rice recombinant inbred lines (RIL) with different genetic backgrounds are utilized for successfully positioning a genetic locus qLRI9 relevant to rice drought resistance onto a rice No.9 chromosome, drought-resistant beneficial alleles come from a drought-resistant parent IAPAR-9 and are linked with the molecular marker RM24598, and the method for identifying or assistant identifying rice drought resistance is provided based on the molecular marker RM24598 and can be used for identifying a rice drought resistance variety and molecular breeding.

Description

technical field

[0001] The invention belongs to agricultural biotechnology engineering, and specifically relates to a molecular marker linked with rice drought tolerance gene qLRI9 and its application. Background technique

[0002] The problem of land drought is widespread in the world, and the arable land area in arid and semi-arid regions has accounted for 42.9% of the global arable land area. According to statistics, more than 60 million hectares of rice planting areas in the world are under drought stress every year, and the resulting yield loss is staggering. In recent years, droughts that have spread widely and lasted for a long time often occur in my country, and the number of years affected by drought is also increasing year by year. Drought has gradually become a prominent problem affecting agricultural production, and it has a tendency to worsen. The resulting reduction in production is far greater than the sum of the reductions caused by other environmental factor...

Examples

Embodiment 1

[0048] Example 1. Acquisition of Rice Drought Tolerance Gene qLRI9 and Its Closely Linked Molecular Markers

[0049] 1. Mapping of Drought Tolerance QTL and Acquisition of Rice Drought Tolerance Gene qLRI9

[0050] 1. Test materials

[0051]The F 10 The recombinant inbred line population (RIL) was used as the targeting population.

[0052] 2. Genotype identification

[0053] The test materials were planted in the Beijing Changping Experimental Base, and the young rice leaves were clipped, and the genome-wide DNA extraction was carried out according to Doyle and Dickson (1987) with a slightly improved method of cetyltriethylammonium bromide (CTAB). 178 pairs of SSR primers with good amplification effect, polymorphism between parents and uniform distribution in 12 linkage groups were screened out for genotype identification of the mapping population.

[0054] The PCR reaction system is as follows: 10×PCR buffer (containing Mg 2+ ) 1.0μl, 10mM dNTP Mixture 0.25μl, 10pM / μl pr...

Embodiment 2

[0085] Embodiment 2, rice drought tolerance identification method

[0086] The method for identifying the drought tolerance of rice to be tested provided by the invention is as follows:

[0087] 1. Extract the genomic DNA of the rice to be tested, use the genomic DNA as a template, and perform PCR amplification with the RM24598 molecular marker to obtain a PCR product;

[0088] The PCR reaction system is as follows: 10×PCR buffer (containing Mg 2+ ) 1.0μl, 10mM dNTP Mixture 0.25μl, 10pM / μl primer 0.25μl, 0.5U / μl Taq polymerase 0.25μl, 40ng / μl genomic DNA 1.0μl, ddH 2 O to make up to 10 μl.

[0089] The PCR reaction conditions were as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 40 s, 36 cycles; extension at 72°C for 10 min.

[0090] 2. Identify the drought tolerance of the rice to be tested according to the size of the PCR product:

[0091] If the size of the PCR product is 110bp, the rice to ...

Embodiment 3

[0096] Example 3, Application of RM24598 Molecular Marker in the Identification of Rice Variety Drought Tolerance

[0097] 1. Test materials

[0098] The tested materials were Huakewagu, Dalisha, Yunchuanbai, Xiaomizao, Taipei 8, Huangbansuo, Heidagu, Jinxing 1, Guichao 2 and Manpixianghong glutinous rice.

[0099] 2. Identification of Drought Tolerance

[0100] According to the method in Step 3 of Example 1, the drought tolerance of the test materials in Step 1 was identified.

[0101]The results are shown in Table 4. It can be seen from the table that among the 10 rice varieties, Taipei No. 8 and Huangbansuo have a leaf curl degree of 7, both of which are rice varieties with weak drought tolerance, and Jinxing No. 1 has a leaf curl degree of 5, which indicates drought tolerance. Among the rice varieties, the remaining 7 rice varieties Huakewagu, Dalisha, Yunchuanbai, Xiaomizao, Heidagu, Guichao No. 2 and Manpixianghong glutinous rice are all drought-tolerant. Strong rice...

technical field

[0001] The invention belongs to agricultural biotechnology engineering, and specifically relates to a molecular marker linked with rice drought tolerance gene qLRI9 and its application. Background technique

[0002] The problem of land drought is widespread in the world, and the arable land area in arid and semi-arid regions has accounted for 42.9% of the global arable land area. According to statistics, more than 60 million hectares of rice planting areas in the world are under drought stress every year, and the resulting yield loss is staggering. In recent years, droughts that have spread widely and lasted for a long time often occur in my country, and the number of years affected by drought is also increasing year by year. Drought has gradually become a prominent problem affecting agricultural production, and it has a tendency to worsen. The resulting reduction in production is far greater than the sum of the reductions caused by other environmental factor...

Examples

Embodiment 1

[0048] Example 1. Acquisition of Rice Drought Tolerance Gene qLRI9 and Its Closely Linked Molecular Markers

[0049] 1. Mapping of Drought Tolerance QTL and Acquisition of Rice Drought Tolerance Gene qLRI9

[0050] 1. Test materials

[0051]The F 10 The recombinant inbred line population (RIL) was used as the targeting population.

[0052] 2. Genotype identification

[0053] The test materials were planted in the Beijing Changping Experimental Base, and the young rice leaves were clipped, and the genome-wide DNA extraction was carried out according to Doyle and Dickson (1987) with a slightly improved method of cetyltriethylammonium bromide (CTAB). 178 pairs of SSR primers with good amplification effect, polymorphism between parents and uniform distribution in 12 linkage groups were screened out for genotype identification of the mapping population.

[0054] The PCR reaction system is as follows: 10×PCR buffer (containing Mg 2+ ) 1.0μl, 10mM dNTP Mixture 0.25μl, 10pM / μl pr...

Embodiment 2

[0085] Embodiment 2, rice drought tolerance identification method

[0086] The method for identifying the drought tolerance of rice to be tested provided by the invention is as follows:

[0087] 1. Extract the genomic DNA of the rice to be tested, use the genomic DNA as a template, and perform PCR amplification with the RM24598 molecular marker to obtain a PCR product;

[0088] The PCR reaction system is as follows: 10×PCR buffer (containing Mg 2+ ) 1.0μl, 10mM dNTP Mixture 0.25μl, 10pM / μl primer 0.25μl, 0.5U / μl Taq polymerase 0.25μl, 40ng / μl genomic DNA 1.0μl, ddH 2 O to make up to 10 μl.

[0089] The PCR reaction conditions were as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 40 s, 36 cycles; extension at 72°C for 10 min.

[0090] 2. Identify the drought tolerance of the rice to be tested according to the size of the PCR product:

[0091] If the size of the PCR product is 110bp, the rice to ...

Embodiment 3

[0096] Example 3, Application of RM24598 Molecular Marker in the Identification of Rice Variety Drought Tolerance

[0097] 1. Test materials

[0098] The tested materials were Huakewagu, Dalisha, Yunchuanbai, Xiaomizao, Taipei 8, Huangbansuo, Heidagu, Jinxing 1, Guichao 2 and Manpixianghong glutinous rice.

[0099] 2. Identification of Drought Tolerance

[0100] According to the method in Step 3 of Example 1, the drought tolerance of the test materials in Step 1 was identified.

[0101]The results are shown in Table 4. It can be seen from the table that among the 10 rice varieties, Taipei No. 8 and Huangbansuo have a leaf curl degree of 7, both of which are rice varieties with weak drought tolerance, and Jinxing No. 1 has a leaf curl degree of 5, which indicates drought tolerance. Among the rice varieties, the remaining 7 rice varieties Huakewagu, Dalisha, Yunchuanbai, Xiaomizao, Heidagu, Guichao No. 2 and Manpixianghong glutinous rice are all drought-tolerant. Strong rice...

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