CN104429750A - Method used for identifying cold tolerance of rice at booting stage - Google Patents
Method used for identifying cold tolerance of rice at booting stage Download PDFInfo
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- CN104429750A CN104429750A CN201310429883.3A CN201310429883A CN104429750A CN 104429750 A CN104429750 A CN 104429750A CN 201310429883 A CN201310429883 A CN 201310429883A CN 104429750 A CN104429750 A CN 104429750A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/20—Cereals
- A01G22/22—Rice
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Abstract
The invention provides a method used for identifying cold tolerance of rice at a reproductive stage. The method provided by the invention comprises the following steps: 1) building a pool; 2) controlling the temperature of water in the pool; and 3) carrying out cold treatment on experimental materials. With the method, cold tolerance of different rice strains are identified, and identification results show that the method provided by the invention can effectively identify cold tolerance of different experimental materials at a booting stage. The method provided by the invention has the advantages of insusceptibility to weather conditions, easy treatment of large quantities of experimental materials at the same time, low cost and high accuracy and facilitates utilization of cold tolerance breeding and identification of cold-tolerant genetic materials at the reproductive stage of rice and assistant selection of varieties with cold tolerance at the booting stage.
Description
Technical field
The invention belongs to Crop Genetic Breeding, be specifically related to a kind of method identifying Rise's boot period cold resistance.
Background technology
Paddy rice is one of China and staple food crop in the world, is the staple food grain of China more than 60% population.Whole world rice growing area about 1.4 hundred million hm
2, because Global climate change is abnormal, extreme climate occurs frequent, and there are 1,500 ten thousand hm in the whole world
2above rice is subject to low temperature and threatens.Paddy rice is in booting stage to low-temperature sensitive, and japonica rice and long-grained nonglutinous rice will produce comparatively serious damaging to plants caused by sudden drop in temperature respectively below 15 DEG C and 18 DEG C.Therefore correctly evaluating paddy rice reproductive stage cold resistance at low temperatures, is cultivate rice cold tolerance breeding of new varieties key.
The method of cold-resistant qualification in booting stage comprises natural low temperature process, long-term cold water irrigation method, constant temperature deep cooling water irrigation method, the methods such as short-term low temperature treatment, its, low temperature treatment referred to before and after paddy pollen reduction division a middle or short term, with the 30cm deep cooling water of 14 DEG C-16 DEG C or climatic cabinate process 5-7 days, or by 17 DEG C of cold water treatment 10 days; With daytime 29 DEG C, night the Temperature Treatment of 15 DEG C 5 days.At present, these methods have larger limitation.Phytotron process not only cost is high, and not easily carries out the screening and identification of material in enormous quantities.Additive method is easily by the impact of outside air temperature.In a word, Identification of Cold Tolerance is easily affected by environment, and the treatment conditions that different environment (weather conditions) requires are different, and process overweight, then cause paddy pollen abortion, ripening rate is too low, can not screen cold-resistant rice material; Process kicks the beam, then cause Seed-Setting Percentage in Rice too high, do not have difference between material.
Select suitable Rise's boot period Identification of Cold Tolerance method, correctly can evaluate the cold-resistant ability in booting stage of rice material of participating in the experiment, filter out cold resistant paddy rice kind matter, to the cold-resistant breeding important in inhibiting of paddy rice.
Summary of the invention
The invention provides a kind of method identifying cold resistant paddy rice.
A kind of method identifying cold resistant paddy rice provided by the invention, described method comprises the steps:
1) pond is built;
2) temperature of water in pond is controlled;
3) cold treatment is participated in the experiment material.
The long 10m in described pond, wide 2m, the pond of rising progressively in staged at the bottom of pond, bosom at the bottom of pond is 2m, and the most shallow place is 1m, often saves the height of ladder rising 20cm.。
In described pond, the degree of depth of water is 60-160cm.
In described method, antiseep process will be carried out in described pond; In described pond, the temperature of water is specially 18-19 DEG C.
Described antiseep process specifically refers to covered rearing with plastic film basin bottom and 4 sides.
Described plastic film is specially double-plastic.
In described method, described cold treatment comprises immerses step 1) and 2 by paddy rice to be identified) in the water in described pond 7 days, the depth standards that described paddy rice to be identified is immersed is: the water logging in described pond does not have sword-like leave auricle 5cm below the water surface of the sword-like leave auricle place of paddy rice spike of rice to be identified and/or paddy rice spike of rice to be identified.
In described method, described paddy rice to be identified specifically refers to the paddy rice being in booting stage.
In described method, described in be in the paddy rice in booting stage and specifically refer to that plant fringe developmental stage is in the pollen mother cells later stage, the described large tripe phase in production, namely paddy rice visual inspection extracts the period of two leaf leaf sheath 0-5cm out to Flag Leaf Sheath.
In described method, in described control pond, the temperature of water specifically refers to the temperature utilizing water in Groundwater Control pond.
The temperature of water in described control pond, particular by the temperature entering water in the flow-control pond in pond and discharge pond of the underground water extracted out.
The temperature of described underground water is specially 16-17 DEG C.
Described underground water is specially well water; The well degree of depth is specially 150 meters, underground.
In described method, after described cold treatment, also need paddy rice to be identified to take out, plant in land for growing field crops continued growth to the maturing stage; Then the ripening rate of paddy rice is directly observed, for judging the cold resistance of strain; The criterion of described cold resistance by: when material relative setting rate of participating in the experiment (after cold treatment mark fringe ripening rate/normal condition under the ripening rate of fringe) more than 60% or relatively the fringe output output of output/this fringe of normal process of mark fringe (after the cold treatment) is more than 40%, or ripening rate or relatively fringe output are higher than testing cold sensitive varieties used, all can judge that rice strain to be identified or kind have cold resistance, relative setting rate or relatively fringe production value are higher, illustrate that its cold resistance is stronger; Or, the criterion of described cold resistance is: by the relative setting rate of paddy rice to be identified and/or relatively fringe output and rice varieties ten and field compare, if the relative setting rate of paddy rice to be identified and/or relatively fringe output are significantly higher than ten and field, then judge that the cold resistance of described paddy rice to be identified is higher than ten and field.
Another object of the present invention is to provide the application of described method in screening cold resistant paddy rice.
In described application, described paddy rice is the paddy rice being in reproductive stage.
The paddy rice of described reproductive stage is specially the paddy rice in booting stage.
In described application, described in be in the paddy rice in booting stage and specifically refer to that plant fringe developmental stage is in the pollen mother cells later stage, the described large tripe phase in production, namely paddy rice visual inspection extracts the period of two leaf leaf sheath 0-5cm out to Flag Leaf Sheath.
The method, not by the impact of weather conditions, is not mainly subject to the impact of temperature; Effectively can identify the material of participating in the experiment that heading stage is different.Be easy to process bulk materials, with low cost, accuracy is high simultaneously, is easy to the qualification of the cold-resistant breeding utilization of paddy rice reproductive stage and cold-resistant genetic stocks, assisted Selection cold-resistant strain in booting stage etc.The method directly can observe the ripening rate of handled spike of rice strain, for judging the cold resistance of strain.
Accompanying drawing explanation
Fig. 1 is material of participating in the experiment with water cooling pond process.
Fig. 2 is the fringe portion difference of material of participating in the experiment after normal process and cold treatment.
Embodiment
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Embodiment 1, water cooling pond method is utilized to identify the cold resistance of strain
(1) to participate in the experiment material
Ten and field: be cold responsive parent, can in rice research unit, as China Agricultural University, Yunnan academy of agricultural sciences, rice in China the acquisition such as Core Germplasms storehouse.
NIL1913: be the near-isogenic line containing a cold-resistant QTL, i.e. cold-resistant strain; Derive from the height in strong cold-resistant parent's Kunmingxiaobaigu and cold responsive parent ten and field for backcross progeny.Concrete Backcrossing methods is from hybrid F_2, in each generation parting material, random selecting 10-20 strain backcrosses with ten and field respectively, differentiate whether retain hybrid combination according to the selfing fringe ripening rate on homophyletic during maturation, the combination (ripening rate is greater than 70%) that general ripening rate is higher is accredited as cold-resistant strain, and retain its hybrid combination, continue to backcross and cold-resistant qualification for the next generation.By this method continuous backcross to BC
6.Then, each combination BC that backcrosses is planted
6f
2colony, filters out cold-resistant strain, then become cold-resistant near-isogenic line (NIL).Cold-resistant qualification is carried out to the BC6F2 colony of main cold-resistant NIL and cold-resistant QTL locates simultaneously, obtain the NIL of each main cold tolerance gene.NIL1913 is exactly the near-isogenic line also having a cold-resistant QTL on No. 4 chromosomes.
Kunmingxiaobaigu can obtain in China national germplasm resource bank.
T-1, T-5, T-12: from NIL1913, cloned cold tolerance gene qCTB4-1-1 and proceed to parent ten and Tanaka, the separate transformation events of the overexpression cold tolerance gene obtained; Need through carrier construction and carry out conversion obtain transgenic line.
(2) water cooling pond process is utilized to participate in the experiment the process of material
1, water cooling pond is built
Select a place to irrigate field easily, dig a long 10m, wide 2m, the pond of rising progressively in staged at the bottom of pond, bosom at the bottom of pond is 2m, and the most shallow place is 1m, often saves the height of ladder rising 20cm.Bottom and 4 sides are covered, with antiseep with the double-plastic of long 15m × wide 8m.Extract the underground water of 16-17 DEG C from deep well (150m), inject water cooling pond, make the depth of water 60-160cm in water cooling pond, utilize water inlet and drain discharge to control the temperature of cold water pool inner water, pool inner water temperature is remained between 18-19 DEG C round the clock.In pond, laying temperature meter is monitored in real time to water temperature.
2, to participate in the experiment the screening of material and mark
Select tiller number, plant height, booting stage almost consistent ten and field, NIL1913, T-1, T-5 and T-12 strain plant as material of participating in the experiment, in addition, 3 main spikes of rice in each plant, to be had to be in booting stage.The described plant being in booting stage specifically refers to that plant fringe developmental stage is in the pollen mother cells later stage, the described large tripe phase in production, and namely paddy rice visual inspection extracts the period of two leaf leaf sheath 0-5cm out to Flag Leaf Sheath.Utilize to hang label or make to paint modes such as spot printing of drawing a circle and carry out marking the main fringe and tillering tassel that are in this period, for investigating the ripening rate after process.
3, water cooling pond process is utilized to participate in the experiment material
As shown in Figure 1, the plant that above-mentioned steps 2 is screened and marked loads according to different strains in the plastic casing of respective experimental group, immerse in the water cooling pond of step 1 construction, the standard of the degree of depth that plant is immersed is: the water logging in described pond does not have the sword-like leave auricle place of spike of rice to be identified or sword-like leave auricle 5cm below the water surface of spike of rice to be identified.
Cold water treatment 7 days, material of participating in the experiment takes out, and conveniently implantation methods is planted in land for growing field crops continued growth to the maturing stage, then, investigates its ripening rate.
Each kind or strain set up 3 repetitions, and one is repeated to investigate 3 strains respectively, and the spike of rice (marking before process) that 3 are in optimization process period is investigated in every strain; Altogether mark 27 fringes carry out the investigation of later stage ripening rate.
(3) result of water cooling pond method qualification strain cold resistance
Qualification result is in table 1 and Fig. 2.When material relative setting rate of participating in the experiment (marking the ripening rate of the ripening rate/normal condition of fringe under fringe after cold treatment) more than 60% or relatively fringe output (marking the output of the output/normal process fringe of fringe after cold treatment) more than 40%, all can judge that material has certain Cold Tolerance at Booting Stage.
Parent ten and field, NIL1913 and transgenic line T-1, T-5, T-12 have larger difference in the output of relative setting rate, relatively fringe.Described cold treatment refers to the process carried out according to above-mentioned (two) described method; Described normal process refers to the process of conveniently implantation methods plantation.
Table 1
The result display of table 1 and Fig. 2, the relative setting rate in cold responsive parent ten and field and relative fringe output are all low than near-isogenic line NIL1913, transgenic line T-1, T-5, T-12.Cold responsive parent ten and field every economical character after cold treatment all shows the injury being subject to low temperature, have reduction in various degree, and every Agronomic of cold-resistant near-isogenic line NIL1913 is all better than ten and field; Transgenic line T-1, T-5, T-12 of overexpression cold tolerance gene in relative setting rate all than ten and field high.
The qualification result of material of participating in the experiment is illustrated, utilizes water cooling pond method process material of participating in the experiment respectively to participate in the experiment the cold-resistant ability of material by precise Identification.Method of the present invention accurately processes for fringe portion, and remains the selection of natural environment to plant to greatest extent, and this method is applicable to the cold resistance identifying rice germplasm.
Claims (10)
1. identify a method for cold resistant paddy rice, described method comprises the steps:
1) pond is built;
2) temperature of water in pond is controlled;
3) cold treatment is participated in the experiment material.
2. method according to claim 1, is characterized in that: antiseep process will be carried out in described pond; In described pond, the temperature of water is specially 18-19 DEG C.
3. method according to claim 1 and 2, it is characterized in that: described cold treatment comprises immerses step 1) and 2 by paddy rice to be identified) in the water in described pond 7 days, the standard of the degree of depth that described paddy rice to be identified is immersed is: the water logging in described pond does not have sword-like leave auricle 5cm below the water surface of the sword-like leave auricle place of paddy rice spike of rice to be identified and/or paddy rice spike of rice to be identified.
4., according to the arbitrary described method of claim 1-3, it is characterized in that: described paddy rice to be identified specifically refers to be in the paddy rice in booting stage.
5., according to the arbitrary described method of claim 1-4, it is characterized in that: described in be in the paddy rice in booting stage and specifically refer to that plant fringe developmental stage is in the pollen mother cells later stage, the described large tripe phase in production.
6., according to the arbitrary described method of claim 1-5, it is characterized in that: in described control pond, the temperature of water specifically refers to utilize the temperature of water in Groundwater Control pond.
7., according to the arbitrary described method of claim 1-6, it is characterized in that: after described cold treatment, also need paddy rice to be identified to take out, plant in land for growing field crops continued growth to the maturing stage; Then the ripening rate of paddy rice is directly observed, for judging the cold resistance of strain;
The criterion of described cold resistance is: when material relative setting rate of participating in the experiment is more than 60%, or relative fringe output is more than 40%, or ripening rate or relatively fringe output are higher than testing cold sensitive varieties used, all can judge that paddy rice to be identified has cold resistance, relative setting rate or relatively fringe production value are higher, illustrate that its cold resistance is stronger;
Or, the criterion of described cold resistance is: by the relative setting rate of paddy rice to be identified and/or relatively fringe output and rice varieties ten and field compare, if the relative setting rate of paddy rice to be identified and/or relatively fringe output are significantly higher than ten and field, then judge that the cold resistance of described paddy rice to be identified is higher than ten and field.
8. the application of the arbitrary described method of claim 1-7 in screening cold resistant paddy rice.
9. application according to claim 8, is characterized in that: described paddy rice is the paddy rice being in booting stage.
10. application according to claim 8 or claim 9, is characterized in that: described in be in the paddy rice in booting stage and specifically refer to that plant fringe developmental stage is in the pollen mother cells later stage, the described large tripe phase in production.
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Cited By (7)
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| CN105794508A (en) * | 2016-03-11 | 2016-07-27 | 江西农业大学 | Simple method for quickly screening out cold-tolerance rice varieties |
| CN107006366A (en) * | 2017-02-27 | 2017-08-04 | 湖北省农业科学院粮食作物研究所 | A kind of method that big mass screening cultivates high temperature resistance paddy rice |
| CN107085005A (en) * | 2017-04-05 | 2017-08-22 | 吉林省农业科学院 | A kind of authentication method of rice cold tolerance |
| CN107432222A (en) * | 2016-12-29 | 2017-12-05 | 中国科学院植物研究所 | A kind of authentication method of the cold-resistant phenotype of rice seedling |
| CN110128514A (en) * | 2018-02-08 | 2019-08-16 | 中国农业大学 | Rise's boot period cold resistance GAP-associated protein GAP CTB4b and encoding gene and application |
| CN111896686A (en) * | 2020-08-03 | 2020-11-06 | 黑龙江省农业科学院水稻研究所 | Device for detecting cold resistance of rice in booting stage |
| CN113728828A (en) * | 2021-06-16 | 2021-12-03 | 袁隆平农业高科技股份有限公司 | Method for identifying heat resistance and cold resistance of rice |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105794508A (en) * | 2016-03-11 | 2016-07-27 | 江西农业大学 | Simple method for quickly screening out cold-tolerance rice varieties |
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| CN107432222A (en) * | 2016-12-29 | 2017-12-05 | 中国科学院植物研究所 | A kind of authentication method of the cold-resistant phenotype of rice seedling |
| CN107006366A (en) * | 2017-02-27 | 2017-08-04 | 湖北省农业科学院粮食作物研究所 | A kind of method that big mass screening cultivates high temperature resistance paddy rice |
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| CN107085005A (en) * | 2017-04-05 | 2017-08-22 | 吉林省农业科学院 | A kind of authentication method of rice cold tolerance |
| CN110128514A (en) * | 2018-02-08 | 2019-08-16 | 中国农业大学 | Rise's boot period cold resistance GAP-associated protein GAP CTB4b and encoding gene and application |
| CN110128514B (en) * | 2018-02-08 | 2020-12-29 | 中国农业大学 | Rice cold tolerance associated protein CTB4b in booting stage, coding gene and application |
| CN111896686A (en) * | 2020-08-03 | 2020-11-06 | 黑龙江省农业科学院水稻研究所 | Device for detecting cold resistance of rice in booting stage |
| CN113728828A (en) * | 2021-06-16 | 2021-12-03 | 袁隆平农业高科技股份有限公司 | Method for identifying heat resistance and cold resistance of rice |
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