CN109042289B - Breeding method of black endosperm high-anthocyanin rice - Google Patents
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- A—HUMAN NECESSITIES
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Abstract
The invention discloses a breeding method of black endosperm high anthocyanin rice, which comprises the following steps: taking rice rich in starch-fat complex as a parent A, taking rice with black seed coats as a parent B, taking the parent B as a female parent, and hybridizing with the parent A to obtain F1Seed generation; backcrossing with parent A to obtain BC1F1Generation; planting BC in sequence1F1～BC1F6Performing generation selfing, and harvesting black endosperm high anthocyanin rice by taking light yellow seed coats, black endosperm, crude fat content of more than or equal to 2% and more than 70%, existence of starch-fat complex form and anthocyanin content as screening indexes. The rice bred and obtained by the invention not only has the advantages of light yellow skin, black endosperm, high content of starch-fat complex and high content of anthocyanin, but also the anthocyanin is effectively expressed in the edible endosperm part, and natural high anthocyanin products such as black rice noodles and the like can be developed without using food coloring agents, thereby particularly meeting the total requirements of people on the nature, safety and health of foods.
Description
Technical Field
The invention relates to the technical field of rice breeding, in particular to a breeding method of black endosperm high anthocyanin rice.
Background
Anthocyanins, also known as anthocyanidins, are natural pigments widely present in plants in nature and widely used in food coloring, dyes, pharmaceuticals, cosmetics, and the like. The anthocyanin belongs to plant nutrients of bioflavonoids, not only has strong free radical scavenging capacity and oxidation resistance, but also has active molecules such as enzyme activity reduction and mutation resistance, and has important health care function for human health.
The special color rice such as black rice, red rice, purple rice and the like is rich in anthocyanin, is a main material of the traditional food supplement in China, and is researched and dedicated to the breeding of special color rice.
The invention patent application with application publication number CN102524052A discloses a breeding method of rice with the function of producing colored rice, which comprises the following steps: (1) selecting a three-line maintainer line B as a female parent to be hybridized with a black rice male parent, backcrossing the first hybrid generation with the maintainer line B for multiple generations, and breeding a maintainer line black B which contains black seed coats and has other characters consistent with the maintainer line B; (2) hybridizing the sterile line A corresponding to the three-line maintainer line B with the maintainer line black B to produce a sterile line black A containing heterozygous black seed coat genes; (3) hybridizing sterile black A and restorer line giant embryo glutinous R to produce hybrid F1 generation seed 'hybrid F1', which contains hybrid black seed coat, giant embryo and glutinous gene; (4) hybrid F1 seed "hybrid F1" is used as production seed.
The invention patent application with application publication number CN102630557A discloses a breeding method of a functional formula rice variety; the invention selects rice varieties with larger production popularization area and good quality as recurrent parents, respectively hybridizes with black rice, purple rice, red rice, glutinous rice and giant embryo rice, progeny and recurrent parents are backcrossed for many generations, near isogenic lines with agronomic characters similar to those of the recurrent parents and the characters of the black rice, the purple rice, the red rice, the glutinous rice and the giant embryo rice are respectively bred, and the production seeds of the formula rice are mixed according to a certain proportion.
Unfortunately, natural anthocyanin and the like of so-called colored rice are generally only present in seed coats, that is, the anthocyanin-rich rice which is eaten daily is brown rice, and therefore, the taste and the cooking characteristics are poor.
In daily life, people eat polished rice which mainly contains endosperm and does not contain anthocyanin; namely, after the rice is polished to remove seed coats (bran) and become polished rice, the nutritional ingredients such as anthocyanins contained in the seed coats are basically completely lost. Therefore, if anthocyanin can be produced by functional nutrition enrichment in polished rice, the above problems can be solved, and the nutritional quality and economic value of polished rice can be increased.
In order to achieve the aim, a Liu dazzling subject group of academies of China's academy of sciences firstly realizes the specific synthesis of anthocyanin in endosperm in japonica rice (ZH11) and indica rice (HG1) through agrobacterium-mediated rice genetic transformation, and creates transgenic black endosperm rice ' purple rice ' with high antioxidant activity for the first time. Unfortunately, in the present context, the commercialization of transgenic black endosperm rice "purple rice" is difficult.
Disclosure of Invention
The invention aims to provide a breeding method of black endosperm high anthocyanin rice, the rice obtained by breeding by the method not only has the advantages of light yellow epidermis, black endosperm, high starch-fat complex content and high anthocyanin content, but also the anthocyanin is effectively expressed in edible endosperm, natural high anthocyanin products such as black rice noodles and the like can be developed without using food coloring agents, and the method particularly meets the general requirements of people on the nature, safety and health of foods.
The specific technical scheme is as follows:
a breeding method of black endosperm high anthocyanin rice comprises the following steps:
(1) taking rice rich in starch-fat complex as a parent A, taking rice with black seed coats as a parent B, taking the parent B as a female parent, and hybridizing with the parent A to obtain F1Seed generation;
(2) planting F1Generation, backcrossing with parent A to obtain BC1F1Generation;
(3) planting BC1F1Performing selfing to obtain BC of black seed coat1F2Generation;
(4) planting BC1F2Performing inbreeding, and harvesting BC by taking the content of black seed coat and crude fat as screening indexes1F3Generation;
(5) planting BC1F3Performing selfing, and taking light yellow seed coat, black endosperm, crude fat content more than or equal to 2% and more than 70%, and starch-fat complex form as screening index to obtain BC1F4Generation;
(6) planting BC1F4Generation and BC1F5Performing selfing, namely harvesting BC by taking light yellow seed coats, black endosperm, more than or equal to 2 percent of crude fat content and more than 70 percent of crude fat content and taking the starch-fat complex form and the yield and the agronomic characters as screening indexes1F6Generation;
(7) planting BC1F6And performing selfing, namely obtaining black endosperm high anthocyanin rice by taking light yellow seed coats, black endosperm, more than or equal to 2 percent of crude fat and more than 70 percent of crude fat, existence of starch-fat complex, anthocyanin content, yield and agronomic characters as screening indexes.
Wherein, the crude fat refers to the total content of the fat combined with starch and the non-starch combined fat in the endosperm, and the starch-fat complex refers to a complex formed by combining the fat and the starch, namely the fat combined with the starch; the crude fat content and the starch-fat complex content are based on the mass of polished rice.
The anthocyanin-based bioflavonoid substance in rice has excellent dissolution and accumulation properties in oil (fat). The fat in rice includes two types of fat combined with starch and non-starch-combined fat. Anthocyanin in rice is effectively accumulated in endosperm and still stored in rice products, and the formation of accumulation of anthocyanin in a starch-fat complex pathway is key. The invention fully considers the fat solubility characteristic of the anthocyanin, utilizes the effective accumulation of the anthocyanin in the endosperm under the compound form of high starch and fat, provides a breeding method of black endosperm high anthocyanin rice, and lays a foundation for the breeding and utilization of non-transgenic high anthocyanin health rice.
Preferably, the crude fat content of the parent A is more than or equal to 2% and more than 70% in the form of a starch-fat complex.
Preferably, the parent A is ZDR-hal-01.
ZDR-hal-01 is a special purpose rice bred by the nuclear agricultural science institute of Zhejiang university and Hainingkui Biotechnology GmbH, and can be purchased from the nuclear agricultural science institute of Zhejiang university. The rice has high fat content (2.3%), 80% of fat exists in the form of starch-fat complex, rice seed coat is normally light yellow, endosperm is normally white, and anthocyanin is not contained basically.
Preferably, the parent B is Zheda Heizhen.
Zhejiang black delicacies are special black glutinous rice bred by nuclear agricultural science institute of Zhejiang university and can be purchased from the institute. The rice seed coat is black, the endosperm is normally white, the rice seed coat is basically free of anthocyanin, the fat content is low (0.2%), and the fat mainly exists in a non-starch-fat complex form.
The rice with high anthocyanin in black endosperm bred by selecting ZDR-hal-01 and Zhejiang black pearl hybrid is an effective combination of normal light yellow seed coat, black endosperm, high starch-fat complex and high anthocyanin content, is rich in anthocyanin, and is expressed in the endosperm which is eaten daily.
Preferably, in the step (4), the crude fat content is not less than 2% by mass of polished rice.
Preferably, in the step (8), the anthocyanin content is not less than 0.5mg/gdw based on the dry weight of the polished rice.
Preferably, in the steps (6) to (8), the yield is more than or equal to 400 kg/mu.
Preferably, in steps (6) to (8), the agronomic traits include full-growth period, plant height, tillering, panicle length, grain per panicle, seed set percentage and thousand kernel weight.
More preferably, the whole growth period is 125-135 days, the plant height is 100-110 cm, the maturing rate is not less than 90%, and the thousand seed weight is 24-26 g.
The bred rice variety has very high fat content (2%) which is 4-5 times of that of the common rice variety, the yield is influenced to a certain extent, and the yield is controlled to be more than or equal to 400 kg/mu in the screening process.
In the aspect of agronomic characters, because the forming and accumulation time of the starch-fat complex (starch fat) is longer, and the subsequent pigment forming and accumulation are in the later stage of grain filling maturity, the whole growth period of the breeding variety cannot be too long (about 130 days), so as to avoid the influence of low temperature and bad weather on grain filling maturity in the later stage. The plant height should be moderate (about 105 cm) to resist lodging so as not to influence the seed fullness; the tillering is moderate, the ear size is uniform, the ears are not easy to overlong generally, the grain number of each ear is moderate, and the uniform quality and the consistent maturity of the grains are ensured; the setting rate is high and is more than 90 percent; the thousand grains are moderate in weight (about 25 g) so as to avoid the influence of too large and too small grains on accumulation and yield.
Compared with the prior art, the invention has the beneficial effects that:
(1) the black endosperm high anthocyanin rice bred by the invention not only has the advantages of light yellow epidermis, black endosperm, high starch-fat complex content and high anthocyanin content, but also has effective expression of anthocyanin in edible endosperm part, can develop natural high anthocyanin products such as black rice noodles and the like without using food coloring agent, and particularly meets the total requirements of people on the nature, safety and health of food.
(2) The method adopts a strategy of backcrossing with the high starch-fat complex parent, fully considers the fat solubility characteristic of the anthocyanin, and ensures the selection ratio of the high starch-fat complex and the high anthocyanin.
Detailed Description
1. Breeding materials:
(1) the parent A is ZDR-hal-01. ZDR-hal-01 is a rice of special use bred by Zhejiang university and Hainingkui Biotechnology GmbH, and is available from the institute of Nuclear agricultural science of Zhejiang university. The content of fat in the rice is as high as 2.3%, the fat mainly exists in the form of a starch-fat complex, the rice seed coat is normal light yellow, the endosperm is normal white, the anthocyanin is basically not contained, and the content is extremely low.
(2) The parent B is Zheda Heizhen. Zhejiang black delicacies are special black glutinous rice bred by nuclear agricultural science institute of Zhejiang university and can be purchased from the institute. The rice seed coat is black, the endosperm is normally white, the rice seed coat is basically free of anthocyanin, the content of fat is low, and the fat mainly exists in a non-starch-fat complex form.
2. Determination of fat content
Fat content determination reference national standard-determination of fat in food (GB 5009.6-2016); the starch-fat complex is detected by measuring the total fat content of starch by referring to the national standard, namely starch total fat measurement (GB/T22427.3-2008).
3. The breeding process of the black endosperm high anthocyanin rice comprises the following steps:
(1)2013 in spring, Hainan Lingling water, toZhejiang black pearl as parent B, as female parent, and hybridized with parent A (ZDR-hal-01) to obtain F1(Zheda black delicacies/ZDR-hal-01) generation seeds 9;
(2) planting F in summer and autumn, Hangzhou Zhejiang in 20131(Zheda black delicacies/ZDR-hal-01) plant 9 plants, backcrossing with parent A (ZDR-hal-01) as parent to obtain BC1F1(Zheda black delicacies/ZDR-hal-01// ZDR-hal-01) plants 11 plants;
(3) planting backcross BC in the spring of 2014 in Hainan tomb water1F1(Zheda black pearl/ZDR-hal-01// ZDR-hal-01) generation 11 plants, selfing to obtain BC with all black seed coat color1F2Seed generation;
(4) planting BC according to single plant in summer and autumn in 2014, Hangzhou Zhejiang1F2Generation, selfing and harvesting BC1F3For generation, because the color of the seed coat is inherited through a matrix, 1521 black individual plants are firstly shelled and selected to leave the seed coat during harvesting, and then the crude fat content is determined, and 219 individual plants with the fat content more than or equal to 2 percent are selected to leave;
(5) planting BC in 2015 spring in Hainan Ling water1F3Generation, selfing and harvesting BC1F4During harvesting, 46 individual plants with normal light yellow seed coats and black endosperm are firstly shelled and reserved, then the crude fat content and the starch-fat complex content are respectively measured, and 39 individual plants with black endosperm with the fat content of more than or equal to 2 percent and more than 70 percent existing in the starch-fat complex are reserved;
(6) planting BC in summer and autumn in 2015, Hangzhou Zhejiang1F4Generation, selfing and harvesting BC1F5When the normal light yellow seed coat, black endosperm and high starch-fat complex (the fat content is more than or equal to 2 percent and more than 70 percent exists in the form of the starch-fat complex) are continuously selected, 9 excellent strains with good yield, mature period and disease resistance and good agronomic characters are obtained;
(7) planting BC in 2016 summer and autumn in Zhejiang Hangzhou1F5Generation, selfing and harvesting BC1F6Instead, normal light yellow seed coat, black endosperm, and high starch-fat complex (fat content is more than or equal to 2% and more than 70% to starch)Existing in a powder-fat complex form), 5 excellent strains with good yield, mature period and disease resistance and good agronomic characters are obtained at the same time of continuous selection;
(8) in 2017, BC was planted in summer and autumn, Hangzhou Zhejiang1F6And performing selfing, detecting the anthocyanin content, and breeding 1 shaped rice with black endosperm, high starch-fat complex (the fat content is more than or equal to 2 percent and more than 70 percent exists in the form of the starch-fat complex) and high anthocyanin (the content is more than or equal to 0.5mg/g dw) to obtain the black endosperm high anthocyanin rice.
Some of the trait indicators in each generation are reported in table 1.
TABLE 1 screening description of the selection of traits for each generation
The main indexes of the shaped rice bred in the embodiment are as follows: the normal light yellow seed coat, the black endosperm, the high starch-fat complex and the high anthocyanin content are effectively combined, the excellent properties of the two parents are effectively polymerized, and the anthocyanin content in the endosperm is 0.53 mg/g-dw. While the seed coat of the parent A (ZDR-hal-01) is normal light yellow, the endosperm is normal white, the content of crude fat is 2.3 percent, 70 percent of fat exists in the form of a starch-fat complex, and anthocyanin in the polished rice is extremely low (0.07 mg/g); the seed coat of parent B (Zhejiang black pearl) is black, the endosperm is normally white, the anthocyanin is extremely low (0.03mg/g), and the fat content is 0.2%.
After the shaped product is obtained by screening, various nutritional indexes of the shaped product are measured, and the mapping of the shaped product and the parental indexes thereof is shown in table 2.
The measurement method of each index in table 2 is as follows: lipid: GB/T5009.6-2003; amylose starch: GB/T15683-; anthocyanins: GB/T22244-2008; vitamin A: GB/T5009.82-2003; vitamin B1: GB/T5009.84-2003; vitamin B2: GB/T5009.85-2003; vitamin E: GB/T5009.82-2003; calcium: GB/T5009.92-2003; iron: GB/T5009.90-2003; zinc: GB/T5009.14-2003.
TABLE 2 comparison of the major nutritional ingredients of ZDR-hal-01, Zhejiang black delicacies and the standardized products
As can be seen from the data in table 2, the advantages of 2 parents are gathered in the set product in terms of nutritional ingredients, and the set product is rich in fat, anthocyanin and amylose. The content of VA is higher than that of the parent. The yield of the set rice is 412 kilograms per mu.
Therefore, compared with parent products, the shaped rice has good antioxidant function due to the fact that the shaped rice is rich in anthocyanin, high in amylose and rich in fat, can be used for deeply processing products such as rice noodles and the like, has good flavor, and is beneficial to quality improvement and synergy of the products.
Claims (1)
1. A breeding method of black endosperm high anthocyanin rice is characterized by comprising the following steps:
(1) taking rice rich in starch-fat complex as a parent A, taking rice with black seed coats as a parent B, taking the parent B as a female parent, and hybridizing with the parent A to obtain F1Seed generation; the parent A isZDR-hal-01；The parent B is Zheda black pearl;
(2) planting F1Generation, backcrossing with parent A to obtain BC1F1Generation;
(3) planting BC1F1Performing selfing to obtain BC of black seed coat1F2Generation;
(4) planting BC1F2Performing inbreeding, and harvesting BC by taking the content of black seed coat and crude fat as screening indexes1F3Generation; the content of the crude fat is more than or equal to 2 percent by mass of the polished rice;
(5) planting BC1F3Performing selfing, and taking light yellow seed coat, black endosperm, crude fat content more than or equal to 2% and more than 70%, and starch-fat complex form as screening index to obtain BC1F4Generation;
(6) planting BC1F4Generation and BC1F5Performing selfing, namely harvesting BC by taking light yellow seed coats, black endosperm, more than or equal to 2 percent of crude fat content and more than 70 percent of crude fat content and taking the starch-fat complex form and the yield and the agronomic characters as screening indexes1F6Generation;
(7) planting BC1F6Performing selfing, and taking light yellow seed coats, black endosperm, crude fat content of which is more than or equal to 2 percent and more than 70 percent, starch-fat complex form, anthocyanin content, yield and agronomic characters as screening indexes to obtain black endosperm high anthocyanin rice;
taking the dry weight of the polished rice as a reference, wherein the anthocyanin content is more than or equal to 0.5mg/g dw;
in the steps (6) to (7), the yield is more than or equal to 400 kilograms per mu; the agronomic characters comprise full growth period, plant height, tillering, spike length, grain number per spike, seed setting rate and thousand kernel weight.
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