Breeding Methods for Three-Line Male Sterile Lines of Rice with Low Gluten

Abstract

The present invention discloses a breeding method for a three-line sterile line of rice with low gluten protein, and belongs to the technical field of special rice breeding. It comprises the following steps: using a japonica rice conventional rice variety 04DP containing the low gluten protein gene lgc-1 as the male parent, hybridizing with a japonica rice variety Xiushui 128 with excellent agronomic traits, breeding offspring and a maintenance line Chunjiang 23B with excellent economic traits as the female parent for hybridization, pedigree breeding, molecular markers and trait identification and other technologies, breeding a maintenance line B11 with low gluten protein gene polymerization, and then breeding a cytoplasmic male sterile line L909A with excellent economic traits and low gluten protein characteristics by hybridization and backcrossing with the sterile line variety 81A. The sterile line is used to prepare hybrid rice with low gluten protein properties, which can be suitable for patients with kidney disease and diabetes who increase the burden on the kidneys due to excessive intake of gluten protein, and meet the hybrid rice with low gluten protein content suitable for this type of people on the market.

Description

Technical Field

[0001] The invention belongs to the field of rice breeding, and in particular relates to a method for breeding a low-gluten three-line sterile rice line. Background Art

[0002] Gluten is a key storage protein in rice grains, accounting for approximately 60%-80% of the grain's protein content. It is easily absorbed by the human body. However, for patients with kidney disease and diabetes, excessive gluten intake can burden the kidneys, disrupt protein metabolism, and worsen their condition. Therefore, breeding functional rice with low gluten content is particularly important for these patients.

[0003] Research institutions such as Nanjing Agricultural University, Jiangsu Academy of Agricultural Sciences, and Jiaxing Academy of Agricultural Sciences have conducted research on low-gluten rice, resulting in the development of conventional low-gluten rice lines such as W3660, 2054, 2059, and 2084. However, these lines suffer from low yields and poor resistance. Combining low-gluten rice with indica-japonica hybrid rice could further enhance its nutritional value and achieve high yields and efficiency.

[0004] Therefore, it is of great significance to breed a three-line hybrid rice sterile line with low gluten characteristics, which maintains its low gluten function while having high yield, high resistance and high efficiency. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a method for breeding rice sterile lines with low gluten characteristics.

[0006] The technical solution of the present invention is: a method for breeding a three-line maintainer line of low-gluten rice, comprising the following steps:

[0007] (1) The rice 04DP, which carries the lgc-1 gene, was crossed with Xiushui 128. A progeny containing the lgc-1 gene and exhibiting excellent economic traits was selected based on pedigree analysis and molecular testing. The progeny was named L9037.

[0008] (2) L9037 was hybridized with the maintainer line Chunjiang 23B to obtain F1. Based on the pedigree method and molecular detection, a stable plant containing the low-gluten gene lgc-1 and excellent economic traits was selected and named maintainer line B11;

[0009] (3) hybridizing the maintainer B11 with the sterile line 81A to produce the hybrid MF1;

[0010] (4) The hybrid MF1 was self-pollinated and tested for pollen fertility to select sterile plants with excellent economic traits;

[0011] (5) backcrossing the sterile plants selected in step (4) with the maintainer line B11 to select sterile plants with excellent economic traits;

[0012] (6) Repeat steps (4) and (5) until the sterile line is stable, and combine molecular marker-assisted technology to screen out sterile lines with excellent economic traits and containing the low-gluten gene lgc-1.

[0013] Furthermore, in step (1), the method of breeding based on pedigree method combined with molecular detection is:

[0014] (a) Hybrid seeds F1 were obtained by crossing rice 04DP carrying the gluten gene lgc-1 with Xiushui 128;

[0015] (b) Planting hybrid seeds F1 and harvesting self-pollinated seeds F2;

[0016] (c) Planting self-pollinated F2 seeds and selecting F3 strains with excellent economic traits from the population;

[0017] (d) Plant the F3 line and self-pollinate for 3-5 generations. Each generation is screened for economic traits and tested for the gluten protein lgc-1 molecule to obtain the japonica rice material L9037 containing the gluten protein gene lgc-1 and excellent economic traits.

[0018] Furthermore, in step (2), the method of breeding based on pedigree method combined with molecular detection is:

[0019] (a) hybridizing L9037 with the maintainer line Chunjiang 23B to obtain hybrid seeds F1;

[0020] (b) Planting F1 seeds and harvesting self-pollinated F2 seeds with excellent economic traits;

[0021] (c) Planting self-pollinated F2 seeds and selecting F3 strains with excellent economic traits from the population;

[0022] (d) Plant the F3 line, self-pollinate for 3-5 generations, screen for economic traits and detect the gluten protein lgc-1 molecule in each generation, and obtain the maintainer line B11 containing the gluten protein gene lgc-1 and excellent economic traits.

[0023] Furthermore, in step (6), the stability of the sterile line means that the pollen abortion rate is ≥99.99% and the bagging fruit setting rate is ≤0.01%.

[0024] Furthermore, the excellent economic traits refer to a moderate growth period, strong tillering, many effective ears, a large number of grains per ear, a high fruit set rate and a thousand-grain weight.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The breeding method of the present invention, through a pedigree method combined with molecular marker detection, can rapidly produce a three-line sterile rice line with low gluten content. This sterile line can be used to formulate low-gluten hybrid rice suitable for patients with kidney disease and diabetes, who experience increased kidney burden from excessive gluten intake, thus meeting the market demand for low-gluten hybrid rice suitable for these patients. DETAILED DESCRIPTION

[0027] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are purchased from commercial channels unless otherwise specified.

[0028] Description of the biological materials used in the present invention:

[0029] Rice 04DP: Publicly used in the non-patent document "Breeding of Low-Gluten Rice Lines Using Molecular Markers," Journal of Zhejiang Agricultural Sciences, 2015, 27(9): 1505-1509. The public may obtain this biological material from the applicant, who guarantees that it will be released to the public within 20 years from the date of application.

[0030] Xiushui 128: An approved variety with the approval number Zhejiang Rice 2007003, selected and bred by the Jiaxing Academy of Agricultural Sciences in Zhejiang Province.

[0031] Chunjiang 23B: This material is publicly used in the non-patent document "Indica-Japonica Super Hybrid Rice Breeding Technology Innovation and Variety Development," Chinese Agricultural Science, 2016, 49(2): 207-217. The public may obtain this biological material from the applicant, who guarantees that it will be released to the public within 20 years from the date of application.

[0032] Sterile line 81A: an approved variety, approved number Zhejiang Rice (sterile line) 2021019, a sterile line bred by Jinhua Academy of Agricultural Sciences and Jinhua Sancai Seed Company, with parents from Jiahe 212A and 81B.

[0033] The materials used in the present invention are all from Jiaxing Academy of Agricultural Sciences and are available to the public.

[0034] Example 1

[0035] (1) The rice 04DP carrying the gluten gene lgc-1 was hybridized with Xiushui 128 to obtain hybrid seeds F1;

[0036] (2) Planting hybrid seeds F1 and harvesting self-pollinated seeds F2;

[0037] (3) Planting self-pollinated F2 seeds and selecting F3 strains with excellent economic traits from the population;

[0038] (4) The F3 plant line was planted and self-pollinated for four generations. In each generation, its economic traits were screened and the low-gluten protein lgc-1 marker was molecularly detected to obtain the japonica rice material L9037 with excellent economic traits and low-gluten protein marker.

[0039] (5) hybridization between L9037 and the rice maintainer line Chunjiang 23B to obtain hybrid seeds F1;

[0040] (6) Planting F1 seeds and harvesting self-pollinated F2 seeds with excellent economic traits;

[0041] (7) Planting self-pollinated F2 seeds and selecting F3 strains with excellent economic traits from the population;

[0042] (8) Plant line F3 was self-pollinated for five generations. Starting from F3, molecular testing for economic traits and the low-gluten lgc-1 marker was performed in each generation. A maintainer line B11 with excellent economic traits and the low-gluten marker was obtained.

[0043] Excellent economic traits refer to moderate growth period, strong tillering, many effective ears, many grains per ear, high fruit set rate and thousand-grain weight.

[0044] The method of breeding a sterile line with low gluten characteristics comprises the following steps:

[0045] (1) Using the sterile line 81A as the female parent and the above-mentioned low-gluten protein maintenance line B11 as the male parent, hybridization and backcrossing were performed m times to obtain BC m F1 generation seeds;

[0046] (2) Planting BC m From the F1 generation seeds, 21 sterile plants were selected and bagged to determine the sterility. Then, 17 sterile lines A'1 with the low-gluten marker lgc-1 were selected based on molecular marker detection. These lines were then backcrossed with the maintainer line B11 and 14 sterile lines A'2 with excellent agronomic traits were selected.

[0047] (3) The sterile line A'2 was planted and the sterile line 81A was used as a control to identify the sterility of the sterile line A'2. Finally, 10 sterile lines A'3 with a pollen abortion rate ≥99.99%, a bagging fruit set rate ≤0.01% and excellent agronomic traits were selected. They were then backcrossed with the low-gluten protein maintainer line B11 to obtain 46 sterile lines A'.

[0048] (4) CMS line A'4 was planted, and CMS line A'5, whose sterility was consistent with the characteristics of CMS line, was selected and backcrossed with low gluten maintainer line B11. 1 CMS line A'61 with low gluten marker lgc-1, low gluten content and good agronomic traits was selected and named L909A.

[0049] Table 1 Analysis of various material properties:

[0050]

[0051] Therefore, the low-gluten sterile line bred by the present invention has the characteristics of low gluten, which is of great significance for promoting the breeding of low-gluten three-line hybrid rice.

Claims

1. A method for breeding a low-gluten three-line sterile rice line, characterized in that: The following steps are involved: (1) Containing gluten gene lgc-1 The rice 04DP was crossed with Xiushui 128, and a rice containing the low-gluten gene was selected based on the pedigree method and molecular detection. lgc-1 The offspring with excellent economic traits was named L9037; (2) L9037 was hybridized with the maintainer line Chunjiang 23B to obtain F1, and the line containing the low-gluten gene was selected based on the pedigree method and molecular detection. lgc-1 The stable individual plants with excellent economic traits were named maintainer line B11; (3) hybridizing the maintainer B11 with the sterile line 81A to produce the hybrid MF1; (4) The hybrid MF1 was self-pollinated and its pollen fertility was identified to screen out sterile plants with excellent economic traits; (5) backcrossing the sterile plants selected in step (4) with the maintainer line B11 to select sterile plants with excellent economic traits; (6) Repeat steps (4) and (5) until the sterile line is stable, and combine molecular marker-assisted technology to screen out the lines with good economic traits and containing the low-gluten gene. lgc-1 of the sterile system.

2. The breeding method according to claim 1, characterized in that: In step (1), the method of breeding based on pedigree method combined with molecular detection is: (a) Hybrid seeds F1 were obtained by crossing rice 04DP carrying the lgc-1 gene with Xiushui 128. (b) Planting hybrid seeds F1 and harvesting self-pollinated seeds F2; (c) Planting self-pollinated F2 seeds and selecting F3 lines with excellent economic traits from the population; (d) Plant F3 lines, self-pollinate for 3-5 generations, and screen for economic traits and low-gluten eggs in each generation. White lgc-1 Molecular testing to obtain the gluten gene lgc-1 The japonica rice material L9037 has excellent economic traits.

3. The breeding method according to claim 1, characterized in that: In step (2), the breeding method based on pedigree method combined with molecular detection is: (a) L9037 was hybridized with the maintainer line Chunjiang 23B to obtain hybrid seeds F1; (b) Planting F1 seeds and harvesting self-pollinated F2 seeds with excellent economic traits; (c) Planting self-pollinated F2 seeds and selecting F3 lines with excellent economic traits from the population; (d) Plant F3 lines, self-pollinate for 3-5 generations, and screen for economic traits and low-gluten eggs in each generation. White lgc-1 Molecular testing to obtain the gluten gene lgc-1 And the maintenance line B11 with excellent economic traits.

4. The breeding method according to claim 1, characterized in that: In step (6), the stability of the sterile line means that the pollen abortion rate is ≥99.99% and the bagging fruit setting rate is ≤0.01%.

5. The breeding method according to any one of claims 1 to 3, characterized in that: The excellent economic traits refer to moderate growth period, strong tillering, many effective ears, many grains per ear, high fruit setting rate and thousand-grain weight.

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