CN109618917B - Breeding method of long grain odor type disease-resistant high-quality black rice three-line sterile line - Google Patents
Breeding method of long grain odor type disease-resistant high-quality black rice three-line sterile line Download PDFInfo
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Abstract
The invention discloses a breeding method of a long grain fragrant disease-resistant high-quality black rice three-line sterile line, which belongs to the technical field of special rice breeding. The sterile line is utilized to prepare the high-quality three-line black rice hybrid rice with strong dominance specificity, the appearance quality of the black rice can be improved, and the requirements of the market on the long, thin, fragrant and high-quality black rice products are met.
Description
Technical Field
The invention belongs to the technical field of breeding of special rice, and particularly relates to a breeding method of a three-line sterile line of long grain odor type disease-resistant high-quality black rice.
Background
Along with the improvement of the living standard of people and the adjustment of grain structure, the precious special rice resources are fully excavated and utilized, the innovation of high-quality rice resources is strengthened, the planting area of famous, special and high-quality rice is enlarged, the increasing needs of domestic and foreign markets are met, and the method is a great trend of the current rice production. Black rice is a very characteristic type in rice seed resources in China, and integrates natural color, fragrance, taste, nutrition and food therapy. The black rice is rich in nutrition, and contains protein, carbohydrate, B vitamins, vitamin E, calcium, phosphorus, potassium, magnesium, iron, zinc and other nutrient elements. The composition proportion is superior to that of common rice, and the amino acid pattern is closer to that of a human body, so that the black rice is more nutritious than common rice. However, the yield of the black rice is low, the leaf morphology and the resistance of the black rice are poor, the further development and the utilization of the black rice are limited, and the breeding of the high-quality black rice sterile line is an effective way for improving the quality of the black rice hybrid rice by using the successful practice of the heterosis utilization of the rice for the purpose of improving the yield, the quality and the resistance of the black rice variety. The southern high-quality long-grain rice is favored by consumers, and the market lacks the black rice products with long, thin, fragrant and high-quality rice grains, so that the cytoplasmic male sterile line of the high-quality, long-grain, fragrant and disease-resistant black rice is bred, and the high-quality trilinear black rice hybrid rice with strong specificity of superiority is prepared, and has important significance for the production and application of special rice.
Disclosure of Invention
The invention aims to provide a breeding method of a three-line sterile line of long grain odor type disease-resistant high-quality black rice, which is capable of breeding the three-line sterile line variety of the long grain odor type disease-resistant high-quality black rice with good leaf morphology and moderate growth period.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for selectively culturing the three-line sterile line of long-grain fragrant disease-resistant high-quality black rice features that the black rice variety A with high anthocyanin content, the rice variety B containing Pigm rice blast-resistant gene, the rice variety C with long grains, big grains and low content of fragrant amylose, the cytoplasmic male sterile maintainer line variety D with high association power as parent, selecting a black rice line of a cytoplasmic male sterile maintainer line with good leaf morphology, moderate growth period, long and thin rice grains, odor type, disease resistance and high quality through hybridization and double crossing, adopting the technologies of pedigree method breeding, character identification and the like to obtain a long-grain odor type disease-resistant high-quality black rice maintainer line variety with stable inheritance, then the corresponding cytoplasmic male sterile line variety is used as female parent to make saturation backcross, finally the cytoplasmic male black rice sterile line whose rice grain is long and thin, fragrant, disease-resistant and high-quality and whose character is identical to that of male parent is bred.
The black rice variety A with high anthocyanin content is Nantan84, and the color value of the brown rice anthocyanin is more than 20; the rice variety B containing the Pigm rice blast resistant gene is 'Gumei No. 4'; the rice variety C with long grains, large grains and low content of the fragrant amylose is 'corn needle fragrance', the grain length of the rice variety C is 9.0 mm, and the length-width ratio of the rice variety C is 4.8; the general cytoplasmic male sterile maintainer line variety D with good combining ability is 'Guangdong 8B' or 'Hengfeng B'.
The breeding method of the long grain odor type disease-resistant high-quality black rice three-line sterile line specifically comprises the following steps:
(1) respectively hybridizing a black rice variety A with high anthocyanin content, a rice variety B containing Pigm rice blast resistance genes, a rice variety C with long grains, large grains and low fragrance amylose content and a cytoplasmic male sterility maintainer line variety D with good combining ability to obtain F 1（B/A） 、F 1（D/C） Hybrid combination, planting hybrid combination F 1（B/A） 、F 1（D/C） And carrying out compound crossing to obtain a compound crossing combination F 1(D/C//B/A) Planting and backcrossing combination F 1(D/C//B/A) Obtaining F 2 Seed generation;
(2) f is to be 2 Sowing seeds, selecting black rice plant line with good leaf morphology, moderate growth period, long grain, high stigma exsertion rate and brown rice anthocyanin color value greater than 20, and obtaining F 3 Seed generation;
(3) f is to be 3 Seeding for generation seed, extracting leaf DNA, taking rice variety B containing Pigm rice blast resistance gene as contrast, adopting molecular marker InDel587 closely linked with resistance gene to F 3 Each single plant of the strain is marked for auxiliary selection, and then the plant containing resistance gene Pigm is selected to be planted in the field of rice blast disease area for disease-resistant screeningSelecting black rice plant lines with good plant leaf morphology, moderate growth period, long grains, high stigma exposure rate and brown rice anthocyanin color value more than 20;
(4) f selected in step (3) 3 Harvesting seeds F according to line of black rice plant 4 Rough rice is obtained by a small-sized rice huller to obtain F 4 Brown rice is compared with rice variety C, and brown rice with grain length of not less than 8.5 mm, length-width ratio of not less than 4.8, amylose content of less than 16%, anthocyanin color value of more than 20, and fragrance F after cooking is selected from the brown rice 4 Black rice seeds;
(5) f selected in the step (4) 4 Sowing the black rice seeds, and performing single-plant test cross by using the corresponding cytoplasmic male sterile line as female parent in the flowering period of rice to obtain the corresponding test cross first generation and test cross male parent F 5 ；
(6) The first generation of test cross and the male parent F of test cross are obtained 5 Seeding in pairs, test cross male parent F 5 Screening according to the steps (3) and (4), and reserving F meeting the breeding target 5 Instead, 1wt% of I is used 2 The KI performs dyeing microscopic examination on the first test cross generation, selects a strain with the pollen type of complete typical abortion, and performs test cross on the strain and a corresponding male parent F 5 Backcrossing the single plant to obtain corresponding male parent F 6 And backcross the first generation;
(7) subjecting the obtained male parent F 6 Screening according to the steps (3) and (4) and passing through step I 2 And (3) selecting a plant line with the pollen type of complete typical abortion by KI staining microscopic examination, obtaining a male parent which meets the breeding target, carrying out saturation backcross on the male parent and the corresponding plant line with complete typical abortion, carrying out backcross for 5-6 generations, and finally breeding the high-quality black rice cytoplasmic male sterile line with stable sterility, long grain, fragrance and disease resistance.
Compared with the prior art, the invention has the following advantages:
the invention relates to the breeding of the black rice three-line sterile line variety with long and thin rice grain, fragrance, disease resistance and rich anthocyanin, the sterile line is utilized to prepare the high-quality black rice hybrid rice with strong dominant specificity and three lines, which has important significance for the production and application of special rice and has the following advantages: (1) creating a high-quality specific black rice three-line sterile line, and enriching the existing black rice three-line sterile line; (2) solves the problem that most of the existing black rice three-line sterile lines have no disease resistance; (3) the bred black rice three-line sterile line variety has the characteristics of fragrance, excellent rice quality, rich anthocyanin and the like; (4) the bred black rice three-line sterile line is adopted for preparation, and the long grain fragrant disease-resistant high-quality black rice hybrid combination can be obtained, so that the market demand on long and thin grain fragrant black rice products is met, and the production economic benefit can be improved.
Drawings
FIG. 1 is a comparison graph of kernel lengths of the black rice variety "Nantan 84" and the rice variety "Yuzhenxiang" used as parents in example 1 and the selected bred black rice three-line sterile line "Heiguanxiang B".
FIG. 2 is a comparison graph of kernel lengths of the black rice variety "Nantan 84" and the rice variety "Yuzhenxiang" used as parents in example 2 and the selected bred black rice three-line sterile line "Hehengxiang B".
Detailed Description
In order to make the present invention more comprehensible, the technical solutions of the present invention are further described below with reference to specific embodiments, but the present invention is not limited thereto.
Example 1:
(1) hybridizing a black rice variety A 'Nantan 84' (the color value of brown rice anthocyanin is more than 20) with high anthocyanin content serving as a male parent and a rice variety B 'flos Pruni mume No. 4' containing Pigm rice blast resistant gene serving as a female parent to obtain F 1（B/A） Generation; hybridizing a long-grain and large-grain rice variety C with fragrance and low amylose content, namely 'Yuzhenxiang' (the grain length of the rice is 9.0 mm, the length-width ratio is 4.8), serving as a male parent and a cytoplasmic male sterility maintainer line variety D 'Guangdong 8B', serving as a female parent, so as to obtain F 1（D/C） Generation; planting F 1（B/A） 、F 1（D/C） Then through the pair F 1（B/A） 、F 1（D/C） Backcrossing to obtain F 1(D/C//B/A) Planting and backcrossing combination F 1(D/C//B/A) Obtaining F 2 Seed generation;
(2) f is to be 2 Seeding the seeds and selecting the shape of the leavesGood, moderate growth period, long grain, high stigma exsertion rate and brown rice anthocyanin color value > 20 to obtain F 3 Seed generation;
(3) f is to be 3 Sowing seeds, extracting leaf DNA, using rice variety B 'Gumei No. 4' containing Pigm rice blast resistant gene as contrast, adopting molecular marker InDel587 closely linked with resistant gene to F 3 Each individual plant of the strain is marked for auxiliary selection, then a plant containing a resistance gene Pigm is selected to be planted in a field of a rice blast disease area for disease-resistant screening, and a black rice strain with good leaf morphology, moderate growth period, long grains, high stigma exposure rate and brown rice anthocyanin color value larger than 20 is selected;
(4) f selected in step (3) 3 Harvesting seeds F according to line of black rice plant 4 Husking with a small hulling machine to obtain F 4 The brown rice is compared with rice variety C, and brown rice with grain length of more than 8.5 mm, length-width ratio of more than 4.8, amylose content of less than 16%, anthocyanin color value of more than 20, and fragrance after cooking F is selected from the brown rice 4 Black rice seeds;
(5) f selected in the step (4) 4 Sowing the seeds of black rice, and performing single-plant test cross by using cytoplasmic male sterile line 'Guang 8A' as female parent in the flowering period of rice to obtain corresponding test cross first generation and test cross male parent F 5 ；
(6) Subjecting the obtained first test cross generation and test cross male parent F 5 Seeding in pairs, test cross male parent F 5 Screening according to the steps (3) and (4), and reserving F meeting the breeding target 5 Instead, 1wt% of I is used 2 Performing staining microscopic examination on the first test cross generation by KI, selecting a strain with the pollen type of complete typical abortion, and performing test cross on the strain and a corresponding test cross male parent F 5 Backcrossing the single plant to obtain corresponding male parent F 6 And backcross the first generation;
(7) subjecting the obtained male parent F 6 Screening according to the steps (3) and (4) and passing through step I 2 -KI staining microscopy selects a plant line with the pollen type of complete typical abortion to obtain a male parent meeting the breeding target, and then the male parent and the corresponding plant line with complete typical abortion are subjected to saturation backcross and backcrossAnd (5) crossing for 5 generations, and finally breeding a cytoplasmic male sterile line 'black Guangxiang A' of high-quality black rice with stable sterility, slender grains, fragrance and disease resistance, wherein the cytoplasmic male sterile line is a 'black Guangxiang B' corresponding to the recurrent male parent.
As shown in fig. 1, "Nantan 84" brown rice grain length =6.3 mm, brown rice grain width =2.3 mm, aspect ratio = 2.7; the length of the 'corn fragrant' brown rice grain is =9.1 mm, the width of the brown rice grain is =1.9 mm, and the length-width ratio is = 4.8; the seed-selected 'black Guangxiang B' brown rice has the grain length =9.1 mm, the grain width =1.9 mm and the length-width ratio = 4.8. Compared with the conventional variety of 'Nantan 84' and 'Heiziang B', the brown rice has longer grain length and larger length-width ratio; compared with the contrast white rice high-quality rice 'Yuzhenxiang' and 'Heiguanxiang B', the brown rice has the same length and aspect ratio. (according to the classification standard of the brown rice length of GT/T19557.7-2004 Rice guide for testing specificity, consistency and stability of New plant variety, the brown rice length is more than 7mm, and the brown rice is long grain.)
The anthocyanin color value of the bred 'black Guangxiang B' brown rice reaches 23.5, is 14.3 percent higher than that of a parent Nantan84, has a sharp tip spindle shape, has brown rice grains with the length of 9.1mm and the width of 1.9 mm, has the length-width ratio of 4.8, has moderate variety fragrance, is rice blast resistant grade 2, has the early season full-growth period of about 100 days, the plant height of 96.6cm, 19.3 ten thousand ears per mu of effective ears, the total grain number of each ear of 142, the setting rate of 87.55 percent, the thousand grain weight of 19.48g, the amylose content of 13.80 percent and the glue consistency of 90.3 mm.
The sterile line is used for preparing the three-line high-quality black rice hybrid rice with strong dominance specificity, the quality of the high-quality rice of the sterile line 'Guang 8A' can be maintained, the combining ability is strong, the disease resistance is good, the hybrid combination rice grains are slender, fragrant, and contain nutritional ingredients such as anthocyanin and the like.
Example 2:
(1) hybridizing a black rice variety A 'Nantan 84' (the brown rice anthocyanin color value of which is more than 20) with high anthocyanin content serving as a male parent and a rice variety B 'flos Pruni mume No. 4' containing Pigm rice blast resistant gene serving as a female parent to obtain F 1（B/A） Generation; the rice variety C 'Yuzhenxiang' (grain length 9.0 mm, length-width ratio 4.8) with long grain, large grain and low amylose content is used as male parent, and the cytoplasmic male sterility maintainer line with good combining abilityThe variety D 'Hengfeng B' is used as a female parent and hybridized to obtain F 1（D/C） Generation; planting F 1（B/A） 、F 1（D/C） Then through the pair F 1（B/A） 、F 1（D/C） Double crossing to obtain F 1(D/C//B/A) Planting and backcrossing combination F 1(D/C//B/A) Obtaining F 2 Seed generation;
(2) f is to be 2 Sowing seeds for generation, selecting black rice plant line with good plant leaf morphology, moderate growth period, long grain, high stigma exsertion rate and brown rice anthocyanin color value greater than 20 to obtain F 3 Seed generation;
(3) f is to be 3 Sowing seeds, extracting leaf DNA, using rice variety B 'Gumei No. 4' containing Pigm rice blast resistant gene as contrast, adopting molecular marker InDel587 closely linked with resistant gene to F 3 Each individual plant of the strain is subjected to marker-assisted selection, then a plant containing a resistance gene Pigm is selected to be planted in a field of a rice blast disease area for disease-resistant screening, and a black rice strain with good leaf morphology, moderate growth period, long grains and high stigma exposure rate and brown rice anthocyanin color value larger than 20 is selected;
(4) f selected in the step (3) 3 Harvesting seeds F according to line of black rice plant 4 Rough rice is obtained by a small-sized rice huller to obtain F 4 The brown rice is compared with rice variety C, and brown rice with grain length of more than 8.5 mm, length-width ratio of more than 4.8, amylose content of less than 16%, anthocyanin color value of more than 20, and fragrance after cooking F is selected from the brown rice 4 Black rice seeds;
(5) f selected in the step (4) 4 Sowing the seeds of black rice, and performing single-plant test cross by using cytoplasmic male sterile line 'Hengfeng A' as female parent in the flowering period of rice to obtain corresponding test cross first generation and test cross male parent F 5 ；
(6) Subjecting the obtained first test cross generation and test cross male parent F 5 Seeding in pairs, test cross male parent F 5 Screening according to the steps (3) and (4), and reserving F meeting the breeding target 5 Instead, 1wt% of I is used 2 Performing staining microscopic examination on the first test cross generation by KI, selecting a strain with the pollen type of complete typical abortion, and performing test cross on the strainMale parent F 5 Backcrossing the single plant to obtain corresponding male parent F 6 And backcross first generation;
(7) mixing the obtained male parent F 6 Screening according to the steps (3) and (4) and passing through the step I 2 And (3) selecting a plant line with the pollen type of complete typical abortion by KI staining microscopic examination, obtaining a male parent which meets the breeding target, carrying out saturation backcross on the male parent and the corresponding plant line with complete typical abortion, carrying out backcross for 5 generations, and finally breeding a high-quality black rice cytoplasmic male sterile line 'Hehenxiang A' with stable sterility, slender grains, fragrance and disease resistance, wherein the corresponding recurrent male parent is 'Hehenxiang B'.
As shown in fig. 2, "Nantan 84" brown rice grain length =6.3 mm, brown rice grain width =2.3 mm, aspect ratio = 2.7; the length of the 'corn fragrant' brown rice grain is =9.1 mm, the width of the brown rice grain is =1.9 mm, and the length-width ratio is = 4.8; the bred 'Hehenxiang B' brown rice has the advantages that the length of the brown rice is =8.7 mm, the width of the brown rice is =1.8 mm, and the aspect ratio is = 4.8. Compared with the conventional variety of 'Nantan 84' and 'Hehenxiang B', the brown rice grains are longer and the length-width ratio is larger; compared with the contrast white rice high-quality rice 'Yuzhenxiang' and 'Hehengxiang B', the brown rice has the advantages that the length and the length-width ratio are not greatly different. (according to GT/T19557.7-2004 ' guide rice for testing specificity, consistency and stability of new plant variety ' brown rice ' in the grading standard of brown rice length, the brown rice length is more than 7mm, long grain.)
The bred 'Hehenxiang B' rice grains are slender, fragrant and disease-resistant, the anthocyanin color value of the brown rice reaches 24.16, the rice grain is 17.3 percent higher than that of a parent Nantan84, the 'Hehenxiang B' brown rice is sharp-pointed spindle-shaped, the brown rice grain is 8.7mm in length, the grain width is 1.8 mm, the length-width ratio is 4.8, the variety fragrance is medium, the rice blast resistance is 2 grade, the early season whole growth period is about 94 days, the plant height is 92.1cm, each mu of effective spike is 18.8 ten thousand spikes, the total number of grains per spike is 156, the setting rate is 84.55%, the thousand kernel weight is 18.44g, the amylose content is 14.50%, and the glue consistency is 92.2 mm.
The sterile line is utilized to prepare the high-quality three-line black rice hybrid rice with strong dominance specificity, which not only can maintain the quality of high-quality rice of the sterile line Hengfeng A, but also has strong combining ability, good disease resistance, long and thin rice grains, fragrance, nutrient components anthocyanin and the like.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (1)
1. A breeding method of a three-line sterile line of long grain odor type disease-resistant high-quality black rice is characterized in that: the method comprises the steps of taking a black rice variety A with high anthocyanin content, a rice variety B with Pigm rice blast resistance genes, a long-grain and large-grain rice variety C with low amylose content and fragrance, and a cytoplasmic male sterility maintainer line variety D with good combining ability as a parent, selecting a black rice strain with good leaf morphology, moderate growth period and long-grain, fragrant, disease-resistant and high-quality cytoplasmic male sterility maintainer line through hybridization and repeated crossing, adopting a pedigree method breeding and character identification technology to obtain a stably inherited long-grain fragrant and high-quality black rice maintainer line variety, then taking a corresponding cytoplasmic male sterility line variety as a female parent to carry out saturated backcross, and finally breeding a black rice long-grain, fragrant, disease-resistant and high-quality cytoplasmic male sterility line;
which comprises the following steps:
(1) respectively hybridizing a black rice variety A with high anthocyanin content, a rice variety B containing Pigm rice blast resistance genes, a rice variety C with long grains, large grains and low fragrance amylose content and a cytoplasmic male sterility maintainer line variety D with good combining ability to obtain F 1（B/A） 、F 1（D/C） Hybrid combination, planting hybrid combination F 1（B/A） 、F 1（D/C） And carrying out compound cross to obtain a compound cross combination F 1(D/C//B/A) Composite planting and backcrossing F 1(D/C//B/A) Obtaining F 2 Seed generation;
(2) f is to be 2 Sowing seeds, selecting black rice plant line with good leaf morphology, moderate growth period, long grain, high stigma exsertion rate and brown rice anthocyanin color value greater than 20, and obtaining F 3 Seed generation;
(3) f is to be 3 Sowing seeds, extracting leaf DNA, using rice variety B containing Pigm rice blast resistant gene as contrast, adopting molecular marker InDel587 closely linked with resistant gene to F 3 Individual strains of the linePerforming marker-assisted selection, then selecting plants containing resistance genes Pigm, planting the plants in a rice blast disease area field for disease-resistant screening, and selecting black rice plants with good leaf morphology, moderate growth period, long grains, high stigma exposure rate and brown rice anthocyanin color value more than 20;
(4) f selected in the step (3) 3 Harvesting seeds F according to line of black rice plant 4 Rough rice is obtained by a small-sized rice huller to obtain F 4 Brown rice is compared with rice variety C, and brown rice with grain length of not less than 8.5 mm, length-width ratio of not less than 4.8, amylose content of less than 16%, anthocyanin color value of more than 20, and fragrance F after cooking is selected from the brown rice 4 Black rice seeds;
(5) f selected in the step (4) 4 Sowing the seeds of black rice, and performing single-plant test cross by using the corresponding cytoplasmic male sterile line as female parent in the flowering period of rice to obtain the corresponding test cross first generation and test cross male parent F 5 ；
(6) Subjecting the obtained first test cross generation and test cross male parent F 5 Seeding in pairs, test cross male parent F 5 Screening according to the steps (3) and (4), and reserving F meeting the breeding target 5 Instead, 1wt% of I is used 2 Performing staining microscopic examination on the first test cross generation by KI, selecting a strain with the pollen type of complete typical abortion, and performing test cross on the strain and a corresponding test cross male parent F 5 Backcrossing the single plant to obtain corresponding male parent F 6 And backcross the first generation;
(7) subjecting the obtained male parent F 6 Screening according to the steps (3) and (4) to obtain male parents meeting the breeding target, and processing by a first step 2 Performing KI staining microscopic examination to select a plant line with the pollen type of complete typical abortion, performing saturated backcross on a male parent and a corresponding plant line with complete typical abortion, performing backcross for 5-6 generations, and finally breeding a high-quality black rice cytoplasmic male sterile line with stable sterility, long grain size, fragrance and disease resistance;
wherein the black rice variety A with high anthocyanin content is 'Nantan 84', and the color value of the brown rice anthocyanin is more than 20; the rice variety B containing the Pigm rice blast resistant gene is 'Gumei No. 4'; the rice variety C with long grains, large grains and low amylose content and fragrance is 'Yuzhenxiang', the grains are 9.0 mm long, and the length-width ratio is 4.8; the general cytoplasmic male sterile maintainer line variety D with good combining ability is 'Guangdong 8B' or 'Hengfeng B'.
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