CN103262790A - Method for selectively breeding Honglian type rice blast-resistant sterile lines - Google Patents
Method for selectively breeding Honglian type rice blast-resistant sterile lines Download PDFInfo
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Abstract
The invention discloses a method for selectively breeding Honglian type rice blast-resistant sterile lines, belonging to the technical field of crop breeding. The method for selectively breeding Honglian type rice blast-resistant sterile lines disclosed by the invention comprises the following steps of: collecting Honglian type sterile lines, maintainer lines and rice blast-resistant rice materials, and combining the conventional hybridization and backcross breeding technology of Honglian type sterile maintainer lines and rice blast-resistant rice materials with the auxiliary selection technology marked by rice blast molecules, specifically: firstly, breeding the maintainer lines into rice blast-resistant maintainer lines, and then, carrying out multi-generation backcross with the sterile lines to selectively breed Honglian type sterile lines having rice blast-resistant characteristics. The method for selectively breeding Honglian type rice blast-resistant sterile lines disclosed by the invention has the advantages of being clear in target, precise and high-efficiency in technology and free from ecological and food safety risks and public concerns of transgenic rice, and is capable of reducing investment and use of pesticide, realizing agricultural efficiency and farmer income and providing support for restoring farmland ecology.
Description
Technical field
The invention belongs to the plant breeding technique field, relate to a kind of method of seed selection red lotus type blast resisting male sterile line.
Background technology
Rice blast, a kind of fungoid disease is one of important disease of paddy rice, can cause the underproduction significantly, the underproduction 40~50% when serious, even No kernels or seeds are gathered, as in a year of scarcity.All there is generation in each rice district, the world, also is one of important content of rice disease research.Rice blast is sick plants variously, wherein takes place in the majorityly with leaf portion, joint portion, can cause the underproduction in various degree after the generation, and especially panicle blast or joint pest take place early and weigh, can cause dead ears so that has no harvest.In recent years, rice blast year area takes place and rise year by year, and local great outburst is much, the disease area is no less than 4,000,000 hectares every year on average.The biological strain of rice blast is numerous, and variation is fast, therefore cultivates resistant variety at certain regional better resistance, but shows relatively poor, even not anti-in another ecotope.The occurrence cause of rice blast is comparatively complicated, and outside the Pass having with the decline of rice varieties resistance, also relevant with extraneous weather conditions and field management level, rice blast can occur in any days, any season.At present, the peasant often strengthens the rich water consumption, and inclined to one side nitrogen fertilizer application of later stage causes the paddy rice overgrowth, and branches and leaves are luxuriant, and ventilation condition is poor, and these all can encourage the generation of rice blast.
Just because of the seriousness of rice blast, a lot of province ,city and areas have classified rice blast resistance as the important indicator of breed breeding, have right of veto by one vote, and namely high sense rice blast kind will not be passed through without exception.Current breeder's seed selection anti-rice blast rice male sterile line method is various, but great majority are still taked the method for field observation.The modern biology technology, especially the molecular marker assisted selection technology has been brought into play important function in breeding.
Summary of the invention
The object of the present invention is to provide a kind of method of seed selection red lotus type blast resisting male sterile line, be that non-transgenic blast resisting red lotus type three is the method that the male sterile line molecular marker assisted selection combines with traditional breeding method, can realize the sustainable development of green, environmental protection, nuisanceless high-quality hybrid rice.The method of the invention is combined closely molecular mark technology and traditional breeding method exactly, utilizes technological means such as serial of methods such as field resistance evaluation, indoor inoculation evaluation to cultivate the blast resisting kind simultaneously.
Purpose of the present invention is achieved through the following technical solutions:
A kind of method of seed selection red lotus type blast resisting male sterile line comprises the steps:
(1) the anti-rice blast rice material of collection red lotus type male sterile line, maintenance system, known molecular mark;
(2) will keep system and anti-rice blast rice material to plant in the rice field; Treating this two kinds of paddy rice heading, is that female parent carries out artificial emasculation, is that male parent is hybridized with the anti-rice blast rice material to keep, and collects to keep system and F1 seed;
(3) being seed (maintenance of step (2) collection is seed) with F1 and corresponding the maintenance, planting in the rice field, when to be eared, is that female parent carries out artificial emasculation with F1, and maintenance is that recurrent parent is backcrossed, and collects corresponding system and the BC1F1 seed of keeping;
(4) be that seed (maintenance of step (3) collection is seed) is planted in the rice field with BC1F1 and corresponding the maintenance, BC1F1 is carried out molecular markers for identification, and to filter out the plant that contains blast resistant gene be that female parent carries out artificial emasculation, system carries out pairing backcrossing with corresponding maintenance, collects corresponding system and the BC2F1 seed of keeping;
(5) repeating step (4) is up to the economical character stable and consistent, so far, backcrosses and breeds the new maintenance system of containing the blast resisting target gene;
(6) the maintenance system of containing the blast resisting target gene with red lotus type male sterile line and seed selection success carries out test cross, backcrosses, and becomes the stable red lotus type male sterile line with blast resisting up to backcross breeding.
Material described in the step (1) can be China's various places paddy rice red lotus type male sterile line, maintenance system and other germ plasm resource; Preferably, described red lotus type is sterile to be the red 3A of red lotus male sterile line Lopa Nationality, maintenance is the red 3B of Lopa Nationality, and the anti-rice blast rice material of known molecular mark is that RGD-7S(contains two rice blast resistance genes of Pi1, Pi2), described molecular labeling is RM224 and AP5930.
Filtering out the plant that contains blast resistant gene in the step (4) also screens by identifying further combined with the evaluation of rice blast indoor inoculation and field resistance; The rice blast indoor inoculation is identified and is specially: the bacterial strain of selecting the rice blast advantage pathological race ZA group of Hubei Province for use, bacterial strain is cultivated 7 days (26 ℃ of constant temperature) with Yeast Cultivation base (PDA medium) purifying, cut the mycelia piece that diameter is 2mm with card punch, some mycelia pieces are moved to product spore medium (culture medium prescription: rice bran 20g, yeast 2.5g, agar 20g, distilled water 1000mL) on, cultivated 8~10 days under 26 ℃ of constant temperature, early 7:00 is to late 19:00 for light application time 12h(, and fluorescent lamp is auxiliary to be shone); Cultivate conidial suspension, its concentration is about 2 * 10
5Individual/mL, in inoculum, add 0.5~0.8%(V/V) Tween20 and carry out spray inoculation, with atomizing (volume median diameter 50~100 μ m of droplet) sprayer inoculum is sprayed on the rice seedling, per 100 young plants spray 30mL approximately; Preserve moisture (RH=95%) 24 hours in 26~28 ℃ of following dark, then seedling-cultivating tray is moved to 25~30 ℃, RH cultivate under 95% the environment, the inoculation back was investigated in 7~10 days, and the occurring degree of susceptible variety is not less than sense (S2) level can think that test is effective; Field resistance is identified to be preferably in evaluation base, the sick garden of national level rice blast secondary, enshi and is carried out.
The rice blast resistance of the plant that among the present invention screening is obtained identifies that carry out in evaluation base, the sick garden of national rice blast secondary in the enshi, and this area is national experiment base, and the gained test data is reliable.
People's such as the molecular mark Technical Reference Liu Wu leather of using among the present invention, Wang Feng method (Acta Agronomica Sinica, 2008,34 (07): 1128-1136).
Beneficial effect of the present invention is:
By with clearly defined objective, directional transformation, technology path accurately modern molecular mark technology combine the red lotus type blast resisting male sterile line of seed selection environment-friendly high-efficiency with conventional breeding.The present invention takes the non-transgenic technology, does not exist the ecology of transgenic paddy rice and food safety risk and the public to worry problem, to realize the sustainable development of green, environmental protection, nuisanceless high-quality hybrid rice.Meanwhile, the male sterile line of cultivating blast resisting is conducive to select the hybrid rice of blast resisting, reduces input and the use of agricultural chemicals, can realize growth of agricultural efficiency, increasing peasant income, provides support for recovering agricultural ecological.
Embodiment
Be described in further detail below in conjunction with the present invention of embodiment, but embodiments of the present invention are not limited thereto.If do not specialize the conventional means that used technological means is well known to those skilled in the art among the embodiment.
Embodiment 1
(1) collect the red 3A of red lotus male sterile line Lopa Nationality, to keep be that the anti-rice blast rice material RGD-7S(of the red 3B of Lopa Nationality, known molecular mark (molecular labeling RM224 and AP5930) contains two rice blast resistance genes of Pi1, Pi2).
When ear in Lingshui, Hainan in March, (2) 2009 with the red 3B of Lopa Nationality serve as maternal be that male parent is carried out artificial hybridization through artificial emasculation, with RGD-7, collect the red 3B of Lopa Nationality and F1 seed.
The red 3B of Lopa Nationality and the F1 that collect in Wuhan, Hubei plantation step (2) in May, (3) 2009 when to be eared, are that maternal the red 3B of Lopa Nationality is that male parent is backcrossed through artificial emasculation with F1, collect the red 3B of Lopa Nationality and BC1F1 seed.
BC1F1 and the red 3B of Lopa Nationality that collect in Lingshui, Hainan plantation step (3) in November, (4) 2009, transplant behind the land for growing field crops and BC1F1 to be decided strain carry out the plant that molecular labeling and rice blast indoor inoculation evaluation and screening have Pi1, Pi2 gene, castrate with recurrent parent (the red 3B of Lopa Nationality that step (3) is collected) selected BC1F1 individual plant and backcross in March, 2010, collects BC2F1 and the red 3B seed of Lopa Nationality;
The rice blast indoor inoculation is identified and is specially: the bacterial strain of selecting the rice blast advantage pathological race ZA group of Hubei Province for use, bacterial strain is cultivated 7 days (26 ℃ of constant temperature) with PDA medium purifying, cut the mycelia piece that diameter is 2mm with card punch, some mycelia pieces are moved to product spore medium (culture medium prescription: rice bran 20g, yeast 2.5g, agar 20g, distilled water 1000mL) on, cultivated 8~10 days under 26 ℃ of constant temperature, early 7:00 is to late 19:00 for light application time 12h(, and fluorescent lamp is auxiliary to be shone); Cultivate conidial suspension, its concentration is about 2 * 10
5Individual/mL, in inoculum, add 0.5~0.8%(V/V) Tween20 and carry out spray inoculation, with atomizing (volume median diameter 50~100 μ m of droplet) sprayer inoculum is sprayed on the rice seedling, per 100 young plants spray 30mL approximately; Preserve moisture (RH=95%) 24 hours in 26~28 ℃ of following dark, then seedling-cultivating tray is moved to 25~30 ℃, RH cultivate under 95% the environment, the inoculation back was investigated in 7~10 days, and the occurring degree of susceptible variety is not less than sense (S2) level can think that test is effective.
BC2F1 and the recurrent parent (the red 3B of Lopa Nationality) that collect in bestow favour (base is identified in the sick garden of national rice blast secondary, enshi) and Wuhan plantation step (4) in May, (5) 2010 respectively, decide strain behind the transplanting land for growing field crops and carry out molecular labeling and rice blast indoor inoculation evaluation and screening, simultaneously according to the resistance performance of bestowing favour, BC2F1 carries out artificial emasculation and recurrent parent is backcrossed to being selected in, and collects BC3F1 and the red 3B seed of Lopa Nationality; Winter in 2010 is at Lingshui, Hainan plantation BC3F1, transplants BC3F1 to be decided strain behind the land for growing field crops and carry out molecular labeling and rice blast indoor inoculation evaluation and screening, and the maintenance that obtain the economical character stable and consistent, contains the blast resisting target gene is BC3F1.
Be that male parent and Lopa Nationality red 3A carry out test cross with selected BC3F1 in March, (6) 2011, collect test cross kind F1 with the selfed seed BC3F2 of BC3F1; In late May, 2011 is bestowing favour and Wuhan plantation test cross F1 and BC3F2 respectively, after transplanting the land for growing field crops, BC3F2 is carried out molecular labeling and rice blast indoor inoculation evaluation and screening, simultaneously according to the resistance performance of bestowing favour, pollen microscopic examination is carried out to test cross F1 in the heading back, select sterile material completely to carry out pairing backcrossing for maternal with selected BC3F2, collect the seed BC1F1 that backcrosses, and collect the selfed seed BC3F3 of BC3F2; In early December, 2011 is at Lingshui, Hainan plantation first backcross generation BC1F1 and BC3F3, after transplanting the land for growing field crops BC3F3 is carried out molecular marker screening, in March, 2012, pollen microscopic examination was carried out to first backcross generation in the heading back, select sterile individual plant completely to obtain BC2F1 for maternal with selected BC3F3 pairing backcrossing, and collect the selfed seed BC3F4 of BC3F3; In early April, 2012 carries out resistance and identifies after receiving and planting in rice blast epidemic-stricken area, enshi; Continue to repeat to screen the work of backcrossing in Wuhan, breeding the new red lotus type of blast resisting three up to 2013 is male sterile line, the red 5A of the red lotus male sterile line of called after Lopa Nationality.The rice blast resistance weighted mean of the red 5A of red lotus male sterile line Lopa Nationality is that 1 grade of resistance rice blast percent of loss is 1.5 grades.
Above-described embodiment is preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spiritual essence of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (6)
1. the method for a seed selection red lotus type blast resisting male sterile line is characterized in that comprising the steps:
(1) the anti-rice blast rice material of collection red lotus type male sterile line, maintenance system, known molecular mark;
(2) will keep system and anti-rice blast rice material to plant in the rice field; Treating this two kinds of paddy rice heading, is that female parent carries out artificial emasculation, is that male parent is hybridized with the anti-rice blast rice material to keep, and collects to keep system and F1 seed;
(3) being seed with F1 and corresponding the maintenance, planting in the rice field, when to be eared, is that female parent carries out artificial emasculation with F1, and maintenance is that recurrent parent is backcrossed, and collects corresponding system and the BC1F1 seed of keeping;
(4) be that seed is planted in the rice field with BC1F1 and corresponding the maintenance, BC1F1 is carried out molecular markers for identification, and to filter out the plant that contains blast resistant gene be that female parent carries out artificial emasculation, system carries out pairing backcrossing with corresponding maintenance, collects corresponding system and the BC2F1 seed of keeping;
(5) repeating step (4) is up to the economical character stable and consistent, so far, backcrosses and breeds the new maintenance system of containing the blast resisting target gene;
(6) the maintenance system of containing the blast resisting target gene with red lotus type male sterile line and seed selection success carries out test cross, backcrosses, and becomes the stable red lotus type male sterile line with blast resisting up to backcross breeding.
2. the method for seed selection red lotus type blast resisting male sterile line according to claim 1, it is characterized in that: the described red lotus type of step (1) is sterile to be the red 3A of Lopa Nationality, and described maintenance is the red 3B of Lopa Nationality.
3. the method for seed selection red lotus type blast resisting male sterile line according to claim 1, it is characterized in that: the anti-rice blast rice material of the described known molecular mark of step (1) is RGD-7S, and described molecular labeling is RM224 and AP5930.
4. the method for seed selection red lotus type blast resisting male sterile line according to claim 1 is characterized in that: filter out the plant that contains blast resistant gene in the step (4) and also screen by identifying further combined with the evaluation of rice blast indoor inoculation and field resistance.
5. the method for seed selection red lotus type blast resisting male sterile line according to claim 4, the method that it is characterized in that described rice blast indoor inoculation evaluation is as follows: the bacterial strain of selecting the rice blast advantage pathological race ZA group of Hubei Province for use, bacterial strain was cultivated 7 days with Yeast Cultivation base purifying, cut the mycelia piece that diameter is 2mm with card punch, some mycelia pieces are moved to produce the spore medium; Cultivate conidial suspension, its concentration is 2 * 10
5Individual/mL, in inoculum, add 0.5~0.8% Tween20 and carry out spray inoculation, inoculum is sprayed on the rice seedling per 100 young plants spray 30mL with the atomizing sprayer; Preserved moisture 24 hours in 26~28 ℃ of following dark, then seedling-cultivating tray is moved to 25~30 ℃, RH cultivate under 95% the environment, the inoculation back was investigated in 7~10 days, and the occurring degree of susceptible variety is not less than sense (S2) level can think that test is effective.
6. the method for seed selection red lotus type blast resisting male sterile line according to claim 4 is characterized in that: described field resistance is accredited as in the enshi the sick garden of national rice blast secondary and identifies that the base carries out.
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CN108157165A (en) * | 2017-12-29 | 2018-06-15 | 中国科学院上海生命科学研究院 | A kind of method using Rice Genomics technology fast accurate selection and breeding three lines paddy sterile line |
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CN112362803A (en) * | 2020-09-22 | 2021-02-12 | 武汉大学 | Application of LY9348 in high-throughput screening of high NUE rice varieties |
CN112362803B (en) * | 2020-09-22 | 2022-04-22 | 武汉大学 | Application of LY9348 in high-throughput screening of high NUE rice varieties |
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