WO2021190139A1

WO2021190139A1 - Method for breeding colored rice two-line hybrid rice

Info

Publication number: WO2021190139A1
Application number: PCT/CN2021/074605
Authority: WO
Inventors: 吴殿星; 章思妍; 陈霄雨
Original assignee: 浙江大学
Priority date: 2020-03-23
Filing date: 2021-02-01
Publication date: 2021-09-30
Also published as: CN111264380A; CN111264380B

Abstract

The present invention relates to the technical field of rice breeding, and provides a method for breeding colored rice two-line hybrid rice. The method comprises: carrying out hybridization by using colored rice C as a male parent and a two-line sterile line S as a female parent, and carrying out backcrossing with the female parent to obtain BCnF2 generation seeds; selecting male sterile plants, performing a paired test cross on the male sterile plants and a two-line restoring line R, and selecting and maintaining homozygous sterile line plants having colored seed coats, as well as corresponding hybrid seeds; detecting seed heterosis, and selecting and maintaining hybrid plants and sterile line plants having heterosis so as to obtain colored rice sterile line seeds; identifying the fertility, yield, quality and resistance of the sterile line to obtain excellent colored rice sterile line plants; and breeding and producing seeds to obtain a colored rice two-line hybrid system.

Description

Breeding method for two-line hybrid rice of colored rice

This application claims the priority of a Chinese patent application filed with the Chinese Patent Office on March 23, 2020, the application number is 202010206105.8, and the invention title is "a method for breeding two-line hybrid rice with colored rice", the entire content of which is incorporated by reference Incorporated in this application.

Technical field

The invention relates to the technical field of rice breeding, in particular to a method for selecting and breeding colored rice two-line hybrid rice.

Background technique

Colored rice refers to rice with brown rice color, mainly black, purple, red, yellow, green, etc. Colored rice is brown rice and contains seed coat, rice embryo and aleurone layer, so its protein, fat, amino acid, vitamin, mineral and trace elements content is significantly higher than that of white rice (polished rice) eaten daily, and it is rich in natural pigments And a variety of physiologically active substances, very in line with my country's traditional food and medicine culture. For example, in the folk, black rice is called "longevity rice", and it is said that "every blackness must be supplemented"; red rice is often used to "tonify blood", it can nourish yin and kidney, invigorate the spleen and appetite, and is a health food that delays aging.

In the context of the current deterioration of the ecological environment, the markedly different lifestyles of people, and the markedly faster pace, the dietary structure has undergone tremendous changes. Various chronic diseases such as the "three highs", atherosclerosis, coronary heart disease and cancer have occurred frequently year by year. Society and family have brought suffering. Eating well, eating full, and eating healthy has become a fashionable pursuit of health. Colored rice has more complete nutrition and contains physiologically active substances that can prevent various chronic diseases, making it a healthy choice for many consumers.

China has rich germplasm resources of non-ferrous rice, especially in Yunnan, Guizhou, Sichuan, Hubei, Hubei, Guangxi, Jiangsu and other places where there are many local varieties. However, including the germplasm of individual varieties approved by the province, the non-ferrous rice varieties are mainly conventional rice varieties, which have problems such as high plant type, poor tillering ability, low seed setting rate, insufficient grain, and poor resistance. Since hybrid rice has become a widely used variety in rice production in my country, the yield of colored rice is obviously relatively low compared with ordinary conventional rice, especially hybrid rice, which is popularized in production.

To this end, breeding workers have carried out research on the selection and breeding of colored rice hybrid varieties, such as Shi Bangzhi and others (A preliminary study on the restoration of 23 red rice varieties in different ecological rice farming areas in Guizhou, Southwest Agricultural Journal, 2013 04). To find out the restoring ability of red rice in different cytoplasmic three-line sterile lines in different ecological rice farming areas in Guizhou, and to screen the red rice resources with restorer genes for heterosis utilization, using 23 red rice as materials and 3 different cytoplasms The three-line sterile line has been cross-tested and matched to screen the red rice restorer line. Qiao Zhongying et al. (Research and Utilization of Black Hybrid Japonica Rice, Jiangsu Agricultural Sciences, 2009 04) used backcrossing, crosses, etc. to determine the restoration relationship of some colored rice resources with better traits and at the same time transform them, and carried out colored rice In the breeding of new sterile lines, the experiment found that the black rice three-line hybrid rice formulated with black rice A has good plant shape, good resistance and high yield, which can improve the related undesirable traits of black rice. Zhang Juncai and Xiao Yeqing (Preliminary Report on Three-line Matching Breeding of Black Rice, Jiangxi Journal of Agriculture, 2000 01) adopted the methods of artificial protection and restoration, and bred 2 black rice sterile lines (Dahei A and Dahei A and Xuehei A) and 3 restoration lines (Heihui No. 1, Hehui No. 2 and Hehui No. 3), and the group test was carried out. Huang Ping (Japonica-type purple rice nucleocytoplasmic sterile line Jin 315A, seed, first issue of 2017) using self-selected sterile line (Chuanxiang 29A/JingRuan//IRRI2B/qu3) and independent selection The bred new line (72B-2/Zimi //XR 11) was further crossed, and different excellent genes were aggregated to enrich the genetic background, and a new japonica-type purple rice nucleocytoplasmic interaction new sterile line Jin 315A was obtained. At present, only a small number of colored rice hybrid rice has passed provincial certification or experimental reports. The main red rices are Dianza 701 (sterile line D5A, red rice restorer line Diankunxiang 4), Jinnong 3 You 3 (Sterile line Jinnong 3A, red rice restorer line Jinhui 3), Jinyouhong (sterile line Jin23A, cold-resistant red rice restorer line Hongzero-4), Gaoyouhong 88 (sterile line Gaofeng A , Red rice restorer line R Hong 88), Yu You Hong 9 (sterile line Yu 802A, red rice restore line Yuhui 9341), Ⅱ You 132 (sterile line Ⅱ-32A is the female parent, red rice restore line), Teyouhong 1256 (sterile line Longtepu A, red rice restorer line R1256), there is only one black rice variety Heiyou 1 (sterile line Zhenshan 97A, black rice restorer line 3350) and the reported test combination Heiyou 906 (sterile line 4D983, black rice restorer line 4D906) and Heiyou 703 (sterile line 4D983, black rice restorer line 4D703). For the above-mentioned varieties, whether they are red rice or black rice hybrids, the sterile lines are ordinary non-colored rice, while the restoring lines are red rice or black rice. Due to genetic characteristics, the seed coats of these varieties are not obvious enough. For example, the brown rice of red rice hybrid rice is reddish-brown, atypically red; the brown rice of black rice hybrid rice is black or dark brown. Although the yield and resistance are higher than that of conventional colored rice varieties, the functional nutrition has decreased. Compared with hybrid rice super varieties, there is no advantage in yield and quality, which restricts production and consumer applications.

Summary of the invention

Based on the above reasons, the present invention provides a method for breeding two-line hybrid rice with colored rice. The colored hybrid rice obtained by the method not only has colored seed coats, but also has the advantages of high yield, high quality and disease resistance. The present invention aims to provide a colored hybrid rice with higher yield, better quality and dual use.

The specific technical solutions are as follows:

A method for breeding colored two-line hybrid rice includes the following steps:

(1) Crossing the colored rice C as the male parent and the two-line sterile line S as the female parent. The obtained offspring are backcrossed with the female parent for n times to obtain BC _n F ₁ generation and BC _n F ₁ selfing. Afterwards, harvest the BC _n F _second generation seeds;

(2) Observation BC _n F ₂ generations grain, selected from the left and homozygous seed coat colored seeds, plant cultivation by selecting male sterile plants flowering, the male sterile plants obtained S _'1; the male sterile plants S _'1 and R restorer line two pairs of test cross, the F ₁ hybrid is obtained;

S again male sterile plant _{'. 1} for cut pile reproduction, fertility Switch to choose to obtain sterile line fertile plant S' _2, S observe sterile line plants' seed ₂ seed coat color, selected from the left to give pure together and seed coat colored seeds S'3 of CMS plants, and selecting plants sterile line S _'3 and R hybrid F restorer lines of the pair of _1a obtained from the measured cross the generation F ₁ hybrids;

(3) The step (2) to obtain hybrid plants and Sterile F _1a S _'3 pairs seed planting, plants were observed and F _1a hybrid plants sterile line S' ₃ heterosis selecting hybrids with heterosis CMS plants and the corresponding plants, plants and F _1b obtain hybrid plants sterile line S _'4; then observe plants sterile line S' fertility _4, selected from the left to give complete sterility and stable homozygous sterile plant-based S _'5;

Then, the plants were sterile line S _'5 with R retest restorer lines, when the plants mature, _1c obtained hybrid F, F hybrid seed coat color was observed _1c and detects the quality of grain, selected from the left to give homozygous quality and hybrid seed coat colored F _1d, and selecting hybrid plants sterile line F _1d corresponding to obtain plants sterile line S _'6, the plants were sterile line S' ₆ for cut pile regeneration, selected from the left to give Fertility fertile plants into sterile line S _'7, the plants were harvested sterile line S' seed _7;

(4) planting the plants sterile line S _'7 seed, the plants were obtained to identify indoor and outdoor fertility, sterility choose to stay in line with the characteristics and seed homozygous sterile line, high-quality, colored seed coat CMS plants obtain plants sterile line S _'8; redetection plants sterile line S' ₈ seed coat color and hybrid seed after recovery system R retest obtained, yield, quality and resistance to rice, selected to give a colored seed coat, yield high, good quality rice strong resistance and excellent colored rice plants two Sterile S _'9;

(5) the use of colored rice plants Two Sterile S _'9 and R restorer Two Sterile be colored rice breeding and seed, rice obtained sterile line of a colored two line S ^{+ C;} establishing two colored rice The two-line hybrid system of colored rice is composed of the sterile line S ^+C , the restorer line R, and the hybrid rice variety S ^+C ^{×R, which is a hybrid between the sterile line S +C and the restorer line R.}

_{_{"S '1 ~ S' 9}} " are obtained during the breeding population of plants sterile line, number of plants> 1 the above methods involve breeding; and the "S ^{+ C"} is obtained from a breeding is not complete Breeding plants.

Further, the colored rice C is any one of black rice, red rice and purple rice.

Further, the colored rice C is Zhongze Heizhen.

Zhongze Heizhen is a black rice japonica rice variety with huge embryos jointly bred by Zhejiang University and Zhongze Biological Technology Co., Ltd.; this variety has been applied to the Plant New Variety Protection Office of the Ministry of Agriculture for new variety protection, application number: 20150962.5, available for purchase in China Ze Biological Technology Co., Ltd.

Further, the two-line sterile line S and the restorer line R are conventional high-quality, high-yield two-line hybrid rice combinations.

Further, the two-line sterile line S is Guangzhan 63-4S; the restorer line R is Yangdao 6.

Guangzhan 63-4S, which is selected by the Northern Hybrid Japonica Rice Research Center and Hefei Fengle Seed Industry Co., Ltd., can be obtained as a gift or purchased, and can also be requested from the National Germplasm Bank.

The restoration line R Yangdao No. 6 is a public material, which was selected and bred by Jiangsu Lixiahe Academy of Agricultural Sciences. It can be obtained or purchased as a gift, and can also be requested from the National Germplasm Bank.

According to the rice breeding technology and process, the present invention selects and ^{breeds the two-line sterile line S +C of} colored rice, the restorer line R and the new hybrid rice variety S ^{+ C} × R, the specific requirements include: 1) S ^{+ C} line and S have similar sterile characteristics, with high outcrossing seed setting rate and high breeding yield, and meet the identification standards of two lines of sterile lines; 2) S ^{+ C} The yield, quality and resistance levels of ×R are similar to the hybrid rice varieties of sterile line S and restorer line R.

Further, in step (1), the n is 3 to 5 times.

Further, in step (3), the evaluation indicators for heterosis include: yield, rice quality, and pest resistance.

Furthermore, the yield is ≥500kg/mu, the rice quality reaches the national third-level and above standards, and the medium resistance to bacterial blight is used as the criterion for heterosis.

Further, in step (4), the characteristics of the sterile line are that the outcrossing seed setting rate is ≥40%; the seed production yield is ≥200kg.

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

(1) The present invention crosses and backcrosses the two-line sterile line of colored rice and the certified hybrid rice, and selects the two-line sterile line with colored seed coat and heterosis through the restorer line, and the obtained colored rice hybrid Rice and its sterile lines not only have colored seed coats, but also have the advantages of high yield, high quality and disease resistance.

(2) The colored rice hybrid rice variety prepared by the two-line sterile line of colored rice obtained in the present invention has a more beautiful and more colorful seed coat compared with the colored rice hybrid rice variety prepared by the colored rice restorer line that has been selected and approved in the past. Obviously, it is not reddish brown like the brown rice of the red rice hybrid rice, which is atypical red; unlike the brown rice of the black rice hybrid rice, which is black or dark brown, although the yield and resistance have been improved, there is functional nutrition instead The problem has declined.

(3) The present invention specifically selects a new target two-line hybrid rice combination with high yield, high quality and disease resistance (hybrid rice is Yangliangyou 6, the sterile line is Guangzhan 63-4S, and the restorer line is Yangdao 6) , Breeding colored hybrid rice similar in yield, quality and resistance to the target variety Yangliangyou 6, which is of high quality and has both uses, which significantly reduces the risk of production extension.

(4) The method of the present invention adopts the color rice breeding method with two-line sterile lines as the core. It is necessary to select maintainer lines and then transfer to sterile lines. Compared with the breeding of three-line sterile lines, the breeding time is significantly shortened. , Can reduce the transfer time of the three-line sterile line for 7-8 consecutive generations.

Detailed ways

The present invention will be further described below in conjunction with specific embodiments. The following are only specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto.

The breeding method provided in the following examples is based on high-yield, high-quality, disease-resistant two-line hybrid rice Yangliangyou 6 (the parents of Yangliangyou 6 are: female parent Guangzhan 63-4S, male parent Yangdao 6 )-Based hybrid rice breeding method for colored rice.

Among them, Yangliangyou 6 is an excellent two-line hybrid rice selected by agricultural scientific research in the Lixiahe area of Jiangsu. Approved by the Crop Variety Approval Committee of Jiangsu Province and Guizhou Province in 2003, approved by the Crop Variety Approval Committee of Henan Province in 2004, approved by the Crop Variety Approval Committee of Hubei Province in 2005, and approved by the National Variety Approval Committee in 2005, the approval number is National Approved Rice 2005024. The rice quality is determined by the Food Quality Supervision, Inspection and Testing Center of the Ministry of Agriculture. The roughness rate is 80.4%, the whole rice rate is 58.8%, the chalky grain rate is 14%, the chalkiness is 2.8%, the amylose content is 15.4%, and the gel consistency is 83 mm. The aspect ratio is 3.1, reaching the national standard three-level high-quality rice quality standard. The average yield per mu in the two-year regional test was 555.80 kg, which was an increase of 4.87% compared to the control Ⅱ You 725. Medium resistant to bacterial blight. Can be purchased from Beijing Golden Nonghua Seed Industry Co., Ltd.

Example 1

A method for breeding two-line hybrid rice with colored rice, the specific steps are as follows:

(1) Population construction: Take the indica black rice line Zhongze Heizhen (i.e. colored rice + C) as the male parent and Guangzhan 63-4S (i.e., the two-line sterile line S) as the female parent. The progeny of and Guangzhan 63-4S were backcrossed 4 times to obtain 2 BC ₄ F ₁ generation plants, BC ₄ F ₁ generation selfed 1 time, and BC ₄ F ₂ generation seeds were harvested;

(2) Selection of testcross: Observe _{1202 seeds of BC 4} F ₂ generation, select the seeds that are homozygous and have colored seed coats, and plant them as a single plant. A total of 322 plants will be planted; 53 fertile plants to give male-sterile plants S _'1; the male-sterile plants S' ₁ and 6 Yangdao (i.e.: restorer line R) paired test cross, the F ₁ hybrid is obtained;

53 and then the male-sterile plants S 'cut _{pile. 1} for reproduction, fertility choose to give CMS into fertile plant S' _2, CMS line plant was observed S _'2 seeds of seed coat color, XuanLiu give homozygous and the seed coat colored plant S sterile line 26 _'3 seeds and selecting the plant S sterile line 26 from the F ₁ hybrids are' paired with _the 3rd Yangdao 6 _{Hybrid F 1a} obtained after testcross;

(3) Detection verification: the step (2) obtained in hybrid F _1a CMS line plant 26 and S _'3 pairs of seed planting, plants were observed and F _1a hybrid plants sterile line S' ₃ of heterosis, Selection of 16 plants whose heterosis is similar to the heterosis of Yangliangyou 6 (with yield ≥500kg/mu, rice quality up to the national level 3 or above, medium resistance to bacterial blight as the criterion for heterosis) hybrid plants and corresponding CMS plants, plants and F _1b obtain hybrid plants sterile line S _'4; then observe plants sterile line S' fertility _4, selected from the left and complete sterility sterile homozygous stable plant-based S _'5 13 strains;

Then, the 13 plants were sterile line S _'5 and retest Yangdao No. R 6 restorer line, when the plants mature, _1c obtained hybrid F, F hybrid seed coat color was observed _1c and detects the quality of grain, selected from homozygous left, and the quality of seed coat colored six hybrids F _1d, and selecting hybrids F _1d corresponding to CMS plants, plants obtained sterile line S _'6, the plants were sterile line S' ₆ for cut pile reproduction, fertility XuanLiu obtained sterile line into fertile plant S _'7, the plants were harvested sterile line S' seed ₇ 6, i.e., six male sterile lines colored rice seeds;

(4) fertility, yield, quality and identification of resistance: planted six plants sterile line S _'7 seed, the plants obtained are indoor and outdoor sterility identification, elected to stay in line with sterility sterile line characteristics (iso seeds rate ≥40%; yield seed propagation ≥200kg) and homozygous seed, high-quality, colored seed coat CMS plants, plants were obtained four sterile line S _'8; redetection plants sterile line S' ₈ The seed coat color, yield, rice quality and resistance of the hybrid seeds obtained after retesting with the restorer line R. Three excellent colored rice two-line sterile lines with colored seed coat, high yield, good rice quality and strong resistance were selected. plant-based S _'9;

(5) Production Seed propagation: the three colored rice plants Two Sterile S _'9 seed breeding, restorer lines with R 6 Yangdao seed, seed propagation and finally choose a high yield, low cost Colored rice two-line sterile line S ^{+ C} 1 (ZD-br-1S).

(6) Two-line hybrid system of colored rice two-line hybrid rice: establish colored rice two-line sterile line ZD-br-1S, restorer line Yangdao 6, and sterile line ZD-br-1S and restorer line Yangdao 6 The new hybrid rice variety ZD-br-1S×Yangdao 6 constitutes a two-line hybrid system of colored rice.

The combined variety test showed that the selected black rice hybrid rice variety Guangzhan 63-4S ^+C /Yangdao 6 has the same population yield, eating quality and disease resistance as Yangliangyou 6, but its blackness As high as 93%, the black rice pigment E content is 2.62, the nutritional quality is significantly better, and in the end use, it can be used as both special rice and high-quality high-yield rice. At the same time, potential risks can be significantly avoided; the yield is significantly higher than that of Zhongze Heizhen, and the resistance is significantly better, and the nutrition and eating quality are similar; that of hybrid varieties Guangzhan 63-4S/ Zhongze Heizhen, quality and resistance And yield are significantly better, indicating that the effect of using this method is obvious.

Compared with the original parents and hybrid rice varieties prepared, the black rice hybrid rice variety Guangzhan 63-4S ^+C /Yangdao 6 has the heavy metal cadmium content, which highlights the superiority of the super parent and the combination, that is, the cadmium content is all To be low. Cadmium is the most typical harmful heavy metal, and it is also the most worried public nutrition. The low cadmium content indicates that the cultivars selected in the present invention are more edible and safer.

See Table 1 for the specific comparison of seed coat color, blackness, black rice pigment, yield, quality and disease resistance of each variety.

Table 1 Comparison of the index characters of the target improved black hybrid rice varieties of Yangliangyou 6

The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications are also It should be regarded as the protection scope of the present invention.

Claims

A method for breeding colored two-line hybrid rice, which is characterized in that it comprises the following steps:

(1) Crossing the colored rice C as the male parent and the two-line sterile line S as the female parent. The obtained offspring are backcrossed with the female parent for n times to obtain BC n F 1 generation and BC n F 1 selfing Afterwards, harvest BC n F second generation seeds;

(2) Observation BC n F 2 generations grain, selected from the left and homozygous seed coat colored seeds, plant cultivation by selecting male sterile plants flowering, the male sterile plants obtained S '1; the male sterile plants S '1 and R restorer line two pairs of test cross, the F 1 hybrid is obtained;

S again male sterile plant '. 1 for cut pile reproduction, fertility Switch to choose to obtain sterile line fertile plant S' 2, S observe sterile line plants' seed 2 seed coat color, selected from the left to give pure and co-colored seed coat CMS plants S '3 seeds and plant S sterile line selected from the F 1 hybrids generation' 3 and R hybrid F restorer lines obtained after the test cross pairs. 1A;

(3) The step (2) to obtain hybrid plants and Sterile F 1a S '3 pairs seed planting, plants were observed and F 1a hybrid plants sterile line S' 3 heterosis selecting hybrids with heterosis CMS plants and the corresponding plants, plants and F 1b obtain hybrid plants sterile line S '4; then observe plants sterile line S' fertility 4, selected from the left to give complete sterility and stable homozygous sterile plant-based S '5;

Then, the plants were sterile line S '5 with R retest restorer lines, when the plants mature, 1c obtained hybrid F, F hybrid seed coat color was observed 1c and detects the quality of grain, selected from the left to give homozygous quality and hybrid seed coat colored F 1d, and selecting hybrid plants sterile line F 1d corresponding to obtain plants sterile line S '6, the plants were sterile line S' 6 for cut pile regeneration, selected from the left to give Fertility fertile plants into sterile line S '7, the plants were harvested sterile line S' seed 7;

(4) planting the plants sterile line S '7 seed, the plants were obtained to identify indoor and outdoor fertility, sterility choose to stay in line with the characteristics and seed homozygous sterile line, high-quality, colored seed coat CMS plants obtain plants sterile line S '8; redetection plants sterile line S' 8 seed coat color and hybrid seed after recovery system R retest obtained, yield, quality and resistance to rice, selected to give a colored seed coat, yield high, good quality rice strong resistance and excellent colored rice plants two Sterile S '9;

(5) the use of colored rice plants Two Sterile S '9 and R restorer Two Sterile be colored rice breeding and seed, to give a colored rice Two Sterile S + C; establishing two colored rice line Sterile line S +C , restorer line R, and hybrid rice variety S +C ×R that crosses between sterile line S +C and restorer line R constitute a two-line hybrid system of colored rice.
The method for breeding two-line hybrid rice with colored rice according to claim 1, wherein the colored rice C is any one of black rice, red rice and purple rice.
The method for breeding two-line hybrid rice with colored rice according to claim 2, wherein the colored rice C is Zhongze Heizhen.
The method for breeding two-line hybrid rice with colored rice according to claim 1, wherein the two-line sterile line S and the restorer line R are conventional high-quality, high-yielding two-line hybrid rice combinations.
The method for breeding two-line hybrid rice with colored rice according to claim 4, wherein the two-line sterile line S is Guangzhan 63-4S; and the restorer line R is Yangdao No. 6.
The method for breeding two-line hybrid rice with colored rice according to claim 1, wherein in step (1), the n is 3 to 5 times.
The method for breeding two-line hybrid rice with colored rice according to claim 1, characterized in that, in step (3), the evaluation indicators for having heterosis include: yield, rice quality and disease resistance.
The method for breeding two-line hybrid rice with colored rice according to claim 7, characterized in that the yield is ≥500kg/mu, the rice quality reaches the national level 3 or above standard, and the medium resistance to bacterial blight is regarded as the heterosis. Judging criteria.
The method for breeding two-line hybrid rice with colored rice according to claim 1, characterized in that, in step (4), the characteristic of the sterile line is that the outcrossing seed setting rate is greater than or equal to 40%; the reproduction seed yield is greater than or equal to 200 kg.