CN115104528B - Method for improving rice blast resistance - Google Patents
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- A01H1/12—Processes for modifying agronomic input traits, e.g. crop yield
- A01H1/122—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- A01H1/1245—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, e.g. pathogen, pest or disease resistance
- A01H1/1255—Processes for modifying agronomic input traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, e.g. pathogen, pest or disease resistance for fungal resistance
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Abstract
The invention discloses a method for improving rice blast resistance, which belongs to the technical field of agricultural cultivation, and specifically comprises the following steps: step one: screening parents: respectively selecting three rice breeding materials carrying Pia, pikm and (Pizt, pi9 and Pigm) genes as donors, and selecting rice breeding materials needing improvement as recurrent parents; step two: hybridization: hybridizing the three donors in the first step with recurrent parent respectively to obtain first filial generation (F) 1 ‑a、F 1 -km and F 1 The invention can greatly reduce the harm of rice blast, has wider gene range for resisting rice blast, can stabilize the rice blast resistance, has wider resistance range, can ensure that the growth of rice is not limited by seasons and geographic positions, shortens the improvement period of the rice, and can avoid the north-south running of improvement personnel.
Description
Technical Field
The invention relates to the technical field of agricultural cultivation, in particular to a method for improving rice blast resistance.
Background
The rice blast is also called rice blast, fire blast and click blast, is a disease caused by rice blast pathogenic bacteria and occurs on rice, the rice blast can occur in the whole growth period of the rice, is respectively called seedling, leaf, ear and node blast, is distributed in a world rice area, is a main disease in rice crop production, wherein the disease in Asia and African rice areas is heavy, the general mountain area is heavier than plain, the polished round-grained nonglutinous rice is heavier than indica rice in China, other rice areas are heavier than early rice except early rice in south China, the epidemic year is generally reduced by 10-20%, the weight is 40-50%, and local field blocks and even grains are not harvested, so that the improvement of the rice blast resistance is particularly necessary.
Through searching, chinese patent No. CN201510175308.4 discloses a method for improving rice blast resistance by utilizing middle and early 39, and the aim of improving the rice blast resistance is fulfilled by polymerizing 5 rice blast resistance genes of the middle and early 39, but the improvement period is long, a large amount of manpower and physics are needed to be input, the cost consumption is increased, and the problem of method defects is brought.
Disclosure of Invention
The invention aims to solve the defects in the prior art, and provides a method for improving rice blast resistance.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
a method for improving rice blast resistance, which comprises the following specific steps:
step one: screening parents: respectively selecting three rice breeding materials carrying Pia, pikm and (Pizt, pi9 and Pigm) genes as donors, and selecting rice breeding materials needing improvement as recurrent parents;
step two: hybridization: hybridizing the three donors in the first step with recurrent parent respectively to obtain first filial generation (F) 1 -a、F 1 -km and F 1 -(9、zt、gm)；
Step three: first backcross: planting the first filial generation obtained in the second step, backcrossing the flowering period of the first filial generation with the recurrent parent in the first step to obtain a first-generation backcross hybrid, planting the first-generation backcross hybrid, selecting the first-generation backcross hybrid carrying Pia, pikm and (Pizt, pi9 and Pigm) genes by utilizing a pyrosequencing quantitative genetic analysis system, selecting plants with the same agronomic characters as the recurrent parent in the selected first-generation backcross hybrid, and marking the plants as BC respectively 1 F 1 -a、BC 1 F 1 -km and BC 1 F 1 -(9、zt、gm)；
Step four: multiple backcross screening: at BC 1 F 1 -a、BC 1 F 1 -km and BC 1 F 1 The flowering period of the- (9, zt, gm) plant is back crossed with the recurrent parent again, the third back crossed m (m is more than 2) times is repeated to obtain an mth generation back crossed hybrid, the mth generation back crossed hybrid is planted, the pyrosequencing quantitative genetic analysis system is used again to select the homozygous plant carrying Pia, pikm and (Pizt, pi9, pigm) genes, and the homozygous plant with the same agronomic characters as the recurrent parent is selected from the homozygous plant and marked as BC m F 1 -a、BC m F 1 -km and BC m F 1 -(9、zt、gm)；
Step five: crossbreeding: in BC m F 1 -a and BC m F 1 The Km is taken as parent, hybridization is carried out to obtain a hybrid generation, the hybrid generation is planted, a pyrosequencing quantitative genetic analysis system is utilized to select the hybrid generation carrying Pia and Pikm genes simultaneously as a second generation parent, and the second generation parent is combined with BC m F 1 Crossing the homozygous plants of- (9, zt and gm) to obtain a hybrid second generation, planting the hybrid second generation, using the hybrid second generation as an inbred parent, carrying out pedigree breeding to a seventh generation, and selecting the homozygous plants with the same agronomic characters as the recurrent parent from the stable strain plants to obtain the pre-improved rice;
step six: infection test: planting the pre-improved rice, and then carrying out resistance identification on each stage of the pre-improved rice by using rice blast pathogenic bacteria with strong pathogenicity;
step seven: post-treatment: harvesting the qualified mature rice in the step six, and airing and storing the rice to obtain the improved rice.
Further, the planting in the first step to the sixth step is processed by adopting a seedling artificial vernalization technology, and the specific steps are as follows:
s1, enabling the abdominal furrows of rice seeds to face downwards and inoculating the rice seeds into a culture dish, enabling the rice seeds to be induced artificially to obtain young rice plants, and transplanting the young rice plants into an incubator when the plant height of the young rice plants is 4-6 cm;
s2, placing the incubator in a manual climate chamber, simulating the vernalization process of the rice under the natural condition of the field, and performing manual vernalization.
Further, the transplanting depth of the young rice plants in the step S1 is 3-4 cm.
Further, in the step S2, the parameters of the artificial climate chamber are that the temperature is 16-26 ℃ when the artificial climate chamber is illuminated, the temperature is 8-12 ℃ when the artificial climate chamber is not illuminated, the humidity is 50-70%, and the illumination time is 8-10 hours when the artificial climate chamber is illuminated every 24 hours.
Further, in the first step, the rice breeding material carrying the Pia gene is Aichi Asahi variety.
Further, in the first step, the rice breeding material carrying the Pikm gene is Tsuyuake variety.
Further, in the first step, the rice breeding materials carrying (Pizt, pi9 and Pigm) genes are obtained by cross breeding of Toride1, 75-1-127 and variety Chimonanthus praecox 4.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention ensures that the improved rice has Pia, pikm, pizt, pi and Pigm genes simultaneously, ensures that the improved rice has obvious rice blast resistance, can greatly reduce the damage of rice blast, has wider gene range for resisting rice blast, can ensure that the rice blast resistance is stable, has wider resistance range, and can ensure that the rice is planted by adopting a method of artificial vernalization treatment, so that the growth of the rice is not limited by seasons and geographical positions, the improvement period of the rice is shortened, and the north-south running of improvement personnel can be avoided.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate the invention and together with the embodiments of the invention, serve to explain the invention.
FIG. 1 is a schematic flow chart of a method for improving rice blast resistance according to the present invention.
Detailed Description
Example 1:
referring to fig. 1, the present invention provides a technical solution: a method for improving rice blast resistance, which comprises the following specific steps:
step one: screening parents: respectively selecting two rice breeding materials carrying Pia and (Pizt, pi9 and Pigm) genes as donors, selecting a rice breeding material to be improved as recurrent parent, wherein the rice breeding material carrying Pia genes is Aichi Asahi variety, and the rice breeding material carrying (Pizt, pi9 and Pigm) genes is Toride1, 75-1-127 and Chimonanthus praecox No. 4 variety;
step two: hybridization: hybridizing the two donors in the first step with recurrent parent respectively to obtain first filial generation (F) 1 -a and F 1 -(9、zt、gm)；
Step three: first backcross: planting the first filial generation obtained in the second step, carrying out backcross on the flowering period of the first filial generation and the recurrent parent in the first step to obtain a first-generation backcross hybrid, planting the first-generation backcross hybrid, selecting the first-generation backcross hybrid carrying Pia and (Pizt, pi9 and Pigm) genes by utilizing a pyrosequencing quantitative genetic analysis system, selecting plants with the same agronomic characters as the recurrent parent in the selected first-generation backcross hybrid, and marking the plants as BC respectively 1 F 1 -a and BC 1 F 1 -(9、zt、gm)；
Step four: multiple backcross screening: at BC 1 F 1 -a and BC 1 F 1 The flowering period of the- (9, zt, gm) plant is backcrossed with the recurrent parent again, the third backcross m (m is more than 2) times is repeated to obtain an mth generation backcross hybrid, the mth generation backcross hybrid is planted, the pyrosequencing quantitative genetic analysis system is used for selecting a homozygous plant carrying Pia and (Pizt, pi9, pigm) genes again, and the homozygous plant with the same agronomic characters as the recurrent parent is selected from the homozygous plants and marked as BC m F 1 -a and BC m F 1 -(9、zt、gm)；
Step five: crossbreeding: in BC m F 1 -a and BC m F 1 - (9, zt, gm) as parent, hybridizing to obtain hybrid generation, planting the hybrid generation, selecting the hybrid generation carrying Pia and (Pizt, pi9, pigm) genes simultaneously as second generation parent by utilizing a pyrosequencing quantitative genetic analysis system, carrying out selfing, pedigree breeding to sixth generation, and selecting homozygous plants with the same agronomic characters as recurrent parent from stable strain plants to obtain the pre-improved rice;
step six: infection test: planting the pre-improved rice, and then carrying out resistance identification on each stage of the pre-improved rice by using rice blast pathogenic bacteria with strong pathogenicity;
step seven: post-treatment: harvesting qualified mature rice in the step six, airing and storing the rice to obtain improved rice, and planting in the step one to the step six by adopting a seedling artificial vernalization treatment technology, wherein the specific steps are as follows:
s1, enabling the abdominal furrows of rice seeds to face downwards and inoculating the rice seeds into a culture dish, enabling the rice seeds to be induced artificially to obtain young rice plants, and transplanting the young rice plants into an incubator when the plant height of the young rice plants is 4cm, wherein the transplanting depth of the young rice plants is 3-4 cm;
s2, placing the incubator in a manual climate chamber, wherein the parameters of the manual climate chamber are that the temperature is 16 ℃ when illumination is carried out, the temperature is 8 ℃ when no illumination is carried out, the humidity is 50%, the illumination time is 8 hours when illumination is carried out every 24 hours, and the artificial vernalization is carried out by simulating the vernalization process of rice under the natural condition of the field.
Example 2:
referring to fig. 1, the present invention provides a technical solution: a method for improving rice blast resistance, which comprises the following specific steps:
step one: screening parents: respectively selecting three rice breeding materials carrying Pia, pikm and (Pizt, pi9 and Pigm) genes as donors, selecting a rice breeding material to be improved as recurrent parent, wherein the rice breeding material carrying Pia genes is Aichi Asahi variety, the rice breeding material carrying Pikm genes is Tsuyuake variety, and the rice breeding materials carrying (Pizt, pi9 and Pigm) genes are Tolide 1, 75-1-127 and Chimonanthus praecox No. 4 variety for cross breeding;
step two: hybridization: hybridizing the three donors in the first step with recurrent parent respectively to obtain first filial generation (F) 1 -a、F 1 -km and F 1 -(9、zt、gm)；
Step three: first backcross: planting the first filial generation obtained in the second step, carrying out backcross on the flowering period of the first filial generation and the recurrent parent in the first step to obtain a first-generation backcross hybrid, planting the first-generation backcross hybrid, selecting the first-generation backcross hybrid carrying Pia, pikm and (Pizt, pi9 and Pigm) genes by utilizing a pyrosequencing quantitative genetic analysis system, selecting plants with the same agronomic characters as the recurrent parent in the selected first-generation backcross hybrid, and marking the plants as BC respectively 1 F 1 -a、BC 1 F 1 -km and BC 1 F 1 -(9、zt、gm)；
Step four: multiple backcross screening: at BC 1 F 1 -a、BC 1 F 1 -km and BC 1 F 1 The flowering period of the- (9, zt, gm) plant is back crossed with the recurrent parent again, the third back crossed m (m is more than 2) times is repeated to obtain an mth generation back crossed hybrid, the mth generation back crossed hybrid is planted, the pyrosequencing quantitative genetic analysis system is used again to select the homozygous plant carrying Pia, pikm and (Pizt, pi9, pigm) genes, and the homozygous plant with the same agronomic characters as the recurrent parent is selected from the homozygous plant and marked as BC m F 1 -a、BC m F 1 -km and BC m F 1 -(9、zt、gm)；
Step five: crossbreeding: in BC m F 1 -a and BC m F 1 The Km is taken as parent, hybridization is carried out to obtain a hybrid generation, the hybrid generation is planted, a pyrosequencing quantitative genetic analysis system is utilized to select the hybrid generation carrying Pia and Pikm genes simultaneously as a second generation parent, and the second generation parent is combined with BC m F 1 Crossing the homozygous plants of- (9, zt and gm) to obtain a hybrid second generation, planting the hybrid second generation, using the hybrid second generation as an inbred parent, carrying out pedigree breeding to a seventh generation, and selecting the homozygous plants with the same agronomic characters as the recurrent parent from the stable strain plants to obtain the pre-improved rice;
step six: infection test: planting the pre-improved rice, and then carrying out resistance identification on each stage of the pre-improved rice by using rice blast pathogenic bacteria with strong pathogenicity;
step seven: post-treatment: harvesting qualified mature rice in the step six, airing and storing the rice to obtain improved rice, and planting in the step one to the step six by adopting a seedling artificial vernalization treatment technology, wherein the specific steps are as follows:
s1, enabling the abdominal furrows of rice seeds to face downwards and inoculating the rice seeds into a culture dish, enabling the rice seeds to be induced artificially to obtain young rice plants, and transplanting the young rice plants into an incubator when the plant height of the young rice plants is 5cm, wherein the transplanting depth of the young rice plants is 3-4 cm;
s2, placing the incubator in a manual climate chamber, wherein the parameters of the manual climate chamber are that the temperature is 21 ℃ when the illumination is carried out, the temperature is 10 ℃ when the illumination is not carried out, the humidity is 60%, the illumination time is 9 hours when the illumination is carried out every 24 hours, and the manual vernalization is carried out by simulating the vernalization process of the rice under the natural condition of the field.
Example 3:
referring to fig. 1, the present invention provides a technical solution: a method for improving rice blast resistance, which comprises the following specific steps:
step one: screening parents: respectively selecting two rice breeding materials carrying Pia and Pikm genes as donors, selecting a rice breeding material to be improved as recurrent parent, wherein the rice breeding material carrying Pia genes is Aichi Asahi variety, and the rice breeding material carrying Pikm genes is Tsuyuake variety;
step two: hybridization: hybridizing the two donors in the first step with recurrent parent respectively to obtain first filial generation (F) 1 -a and F 1 -km；
Step three: first backcross: planting the first filial generation obtained in the second step, carrying out backcrossing on the flowering period of the first filial generation and the recurrent parent in the first step to obtain a first-generation backcross hybrid, planting the first-generation backcross hybrid, selecting the first-generation backcross hybrid carrying Pia and Pikm genes by utilizing a pyrosequencing quantitative genetic analysis system, selecting plants with the same agronomic characters as the recurrent parent in the selected first-generation backcross hybrid, and marking the plants as BC respectively 1 F 1 -a and BC 1 F 1 -km；
Step four: multiple backcross screening: at BC 1 F 1 -a and BC 1 F 1 The flowering period of km plants, backcrossing with recurrent parent again, repeating the three backcrossing m (m > 2) times to obtain mth generation backcrossing hybrid, planting mth generation backcrossing hybrid, selecting homozygous plants carrying Pia and Pikm genes again by utilizing a pyrosequencing quantitative genetic analysis system, and selecting homozygous plants with the same agronomic characters as recurrent parent from the homozygous plants, and marking as BC m F 1 -a and BC m F 1 -km；
Step five: crossbreeding: in BC m F 1 -a and BC m F 1 The km is taken as a parent, hybridization is carried out to obtain a hybrid generation, the hybrid generation is planted, a pyrosequencing quantitative genetic analysis system is utilized to select the hybrid generation carrying Pia and Pikm genes simultaneously as a second generation parent, the parent is self-bred, the pedigree is bred to a sixth generation, and homozygous plants with the same agronomic characters as recurrent parent are selected from stable strain plants, so that the pre-improved rice is obtained;
step six: infection test: planting the pre-improved rice, and then carrying out resistance identification on each stage of the pre-improved rice by using rice blast pathogenic bacteria with strong pathogenicity;
step seven: post-treatment: harvesting qualified mature rice in the step six, airing and storing the rice to obtain improved rice, and planting in the step one to the step six by adopting a seedling artificial vernalization treatment technology, wherein the specific steps are as follows:
s1, enabling the abdominal furrows of rice seeds to face downwards and inoculating the rice seeds into a culture dish, enabling the rice seeds to be induced artificially to obtain young rice plants, and transplanting the young rice plants into an incubator when the plant height of the young rice plants is 6cm, wherein the transplanting depth of the young rice plants is 3-4 cm;
s2, placing the incubator in a manual climate chamber, wherein the parameters of the manual climate chamber are that the temperature is 26 ℃ when illumination is carried out, the temperature is 12 ℃ when no illumination is carried out, the humidity is 70%, the illumination time is 10 hours when illumination is carried out every 24 hours, and the artificial vernalization is carried out by simulating the vernalization process of rice under the natural condition of the field.
Comparative example 1:
this comparative example is a rice with resistance to rice blast which is common in the market.
2 parts of each of the modified rice obtained in examples 1 to 3 was sampled and designated A 1 、A 2 、B 1 、B 2 、C 1 And C 2 Then, 2 parts of rice corresponding to comparative example 1 was designated as E 1 And E is 2 The above samples were planted and rice blast infection tests were performed on various parts of the rice at the same time, and the following infection percentage data were measured:
| A 1 | A 2 | B 1 | B 2 | C 1 | C 2 | E 1 | E 2 | |
| seedling of rice | 0.2 | 0.1 | 0 | 0 | 0.3 | 0.5 | 7 | 8 |
| Blade | 0.1 | 0 | 0 | 0 | 0.2 | 0.3 | 5 | 6 |
| Ear of grain | 0.1 | 0.1 | 0 | 0 | 0 | 0.1 | 4 | 3 |
| Node | 0 | 0 | 0 | 0 | 0 | 0.1 | 6 | 5 |
Compared with the prior art, the improved rice of the embodiments 1, 2 and 3 has obvious rice blast resistance, can greatly reduce the damage of rice blast, has a wider gene range for resisting rice blast, can stabilize the rice blast resistance and has a wider resistance range, and meanwhile, the rice seeds are planted by adopting a method of artificial vernalization treatment, so that the growth of the rice is not limited by seasons and geographical positions, the improvement period of the rice is shortened, and the north-south running of improvement personnel can be avoided.
Claims (7)
1. A method for improving rice blast resistance, which is characterized by comprising the following specific steps:
step one: screening parents: respectively selecting three rice breeding materials carrying Pia, pikm and (Pizt, pi9 and Pigm) genes as donors, and selecting rice breeding materials needing improvement as recurrent parents;
step two: hybridization: hybridizing the three donors in the first step with recurrent parent respectively to obtain first filial generation (F) 1 -a、F 1 -km and F 1 -（9、zt、gm）；
Step three: first backcross: planting the first filial generation obtained in the second step, backcrossing the flowering period of the first filial generation with the recurrent parent in the first step to obtain a first-generation backcross hybrid, planting the first-generation backcross hybrid, selecting the first-generation backcross hybrid carrying Pia, pikm and (Pizt, pi9 and Pigm) genes by utilizing a pyrosequencing quantitative genetic analysis system, selecting plants with the same agronomic characters as the recurrent parent in the selected first-generation backcross hybrid, and marking the plants as BC respectively 1 F 1 -a、BC 1 F 1 -km and BC 1 F 1 -（9、zt、gm）；
Step four: multiple backcross screening: at BC 1 F 1 -a、BC 1 F 1 -km and BC 1 F 1 The flowering period of the- (9, zt, gm) plant is back crossed with the recurrent parent again, the three back crossed steps are repeated for m times to obtain an mth generation back crossed hybrid, the mth generation back crossed hybrid is planted, the pyrosequencing quantitative genetic analysis system is again utilized to select a homozygous plant carrying Pia, pikm and (Pizt, pi9, pigm) genes, and the homozygous plant with the same agronomic characters as the recurrent parent is selected from the homozygous plants and marked as BC m F 1 -a、BC m F 1 -km and BC m F 1 - (9, zt, gm), wherein m > 2;
step five: crossbreeding: in BC m F 1 -a and BC m F 1 The Km is taken as parent, hybridization is carried out to obtain a hybrid generation, the hybrid generation is planted, a pyrosequencing quantitative genetic analysis system is utilized to select the hybrid generation carrying Pia and Pikm genes simultaneously as a second generation parent, and the second generation parent is combined with BC m F 1 -（9、zt、gm) the homozygous plants are hybridized to obtain hybrid second generation, the hybrid second generation is planted, the hybrid second generation is taken as an inbred parent, the pedigree is bred to a seventh generation, and the homozygous plants with the same agronomic characters as the recurrent parent are selected from the stable strain plants, so that the pre-improved rice is obtained;
step six: infection test: planting the pre-improved rice, and then carrying out resistance identification on each stage of the pre-improved rice by using rice blast pathogenic bacteria with strong pathogenicity;
step seven: post-treatment: harvesting the qualified mature rice in the step six, and airing and storing the rice to obtain the improved rice.
2. The method for improving rice blast resistance according to claim 1, wherein said planting in step one to step six is carried out by using a seedling artificial vernalization technique, comprising the steps of:
s1, enabling the abdominal furrows of rice seeds to face downwards and inoculating the rice seeds into a culture dish, enabling the rice seeds to be induced artificially to obtain young rice plants, and transplanting the young rice plants into an incubator when the plant height of the young rice plants is 4-6 cm;
s2, placing the incubator in a manual climate chamber, simulating the vernalization process of the rice under the natural condition of the field, and performing manual vernalization.
3. The method for improving rice blast resistance according to claim 2, wherein said young rice plant in step S1 is transplanted to a depth of 3 to 4cm.
4. The method according to claim 2, wherein the parameter of the artificial climate chamber in step S2 is 16-26℃in the presence of light, 8-12℃in the absence of light, 50-70% in the presence of humidity, and 8-10 hours of light per 24 hours.
5. The method for improving rice blast resistance according to claim 1, wherein said rice breeding material carrying the Pia gene in step one is Aichi Asahi variety.
6. The method for improving rice blast resistance according to claim 1, wherein said rice breeding material carrying the Pikm gene in the first step is a Tsuyuake variety.
7. The method for improving rice blast resistance according to claim 1, wherein said rice breeding material carrying (pizz, pi9, pigm) genes in step one is obtained by cross breeding of Toride1, 75-1-127 and Chimonanthus praecox 4 varieties.
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| WO2016101859A1 (en) * | 2014-12-22 | 2016-06-30 | 广东省农业科学院植物保护研究所 | Rice blast resistant gene pi50, preparation methods therefor, and applications thereof |
| CN104969855A (en) * | 2015-07-30 | 2015-10-14 | 江苏里下河地区农业科学研究所 | Method for cultivating rice breeding material with broad spectrum and lasting spike blast resistance |
| CN106544396A (en) * | 2016-10-18 | 2017-03-29 | 湖南杂交水稻研究中心 | Indoor rice blast sick seedling pestilence identification and the method for quick breeding disease resisting rice |
| CN106566888A (en) * | 2016-11-08 | 2017-04-19 | 淮阴师范学院 | Method for selecting and breeding variety with diversified resistance by molecular marker-assisted selection |
| CN109924121A (en) * | 2018-12-11 | 2019-06-25 | 华智水稻生物技术有限公司 | A method of improvement Rice Resistance To Rice Blast |
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