CN104429750B - 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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- CN104429750B CN104429750B CN201310429883.3A CN201310429883A CN104429750B CN 104429750 B CN104429750 B CN 104429750B CN 201310429883 A CN201310429883 A CN 201310429883A CN 104429750 B CN104429750 B CN 104429750B
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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 is and in particular to a kind of method of identification 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 water
Rice sown area about 1.4 hundred million hm2, because Global climate change is abnormal, extreme climate occurs frequently, and there are 15,000,000 hm in the whole world2
Above rice makees to be threatened by low temperature., in boot stage to low-temperature sensitive, japonica rice and long-grained nonglutinous rice are respectively at 15 DEG C and less than 18 DEG C for paddy rice
More serious damaging to plants caused by sudden drop in temperature will be produced.Therefore correctly evaluate paddy rice reproductive stage cold resistance at low temperature, be to cultivate paddy rice
Cold-resistant breeding of new varieties is crucial.
The method of boot stage cold-resistant identification includes natural low temperature and processes, long-term cold water irrigation method, constant temperature deep cooling water irrigation method,
The methods such as short-term K cryogenic treatment, its middle or short term K cryogenic treatment refers to before and after paddy pollen meiosis, with 14 DEG C -16 DEG C
30cm deep cooling water or growth cabinet are processed 5-7 days, or with 17 DEG C of cold water treatment 10 days；With 29 DEG C of daytime, 15 DEG C of night
Temperature Treatment 5 days.At present, these methods have larger limitation.Phjytotron processes not only high cost, and be difficult into
The screening of row high-volume material and identification.Additive method is easily affected by outside air temperature.In a word, Identification of Cold Tolerance is easily subject to ring
Border affects, different environment（Weather conditions）The treatment conditions requiring are different, process overweight, then lead to paddy pollen abortion,
Setting percentage is too low, can not screen cold-resistant rice material；Process kicks the beam, then lead to Seed-Setting Percentage in Rice too high, do not have between material
Difference.
Select suitable Rise's boot period Identification of Cold Tolerance method, the boot stage that can correctly evaluate rice material of participating in the experiment is resistance to
Cold energy power, filters out cold resistant paddy rice germplasm, the cold-resistant breeding important in inhibiting to paddy rice.
Content of the invention
The invention provides a kind of method of identification cold resistant paddy rice.
A kind of method of identification cold resistant paddy rice that the present invention provides, methods described comprises the steps：
1）Build pond；
2）Control the temperature of water in pond；
3）Cold treatment is participated in the experiment material.
The long 10m in described pond, wide 2m, the pond that at the bottom of pond is risen progressively in staged, at the bottom of pond bosom is 2m, and the most shallow place is 1m,
Often section ladder rises the height of 20cm..
In described pond, the depth of water is 60-160cm.
In methods described, described pond will carry out antiseepage drain process；In described pond, the temperature of water is specially 18-19 DEG C.
Described antiseepage drain process refers specifically to covered rearing with plastic film basin bottom and 4 sides.
Described plastic sheeting is specially double-plastic.
In methods described, described cold treatment includes for paddy rice to be identified immersing step 1）With 2）7 days in the water in described pond,
The depth standards of described paddy rice to be identified immersion are：The water logging in described pond do not have at the sword-like leave auricle of paddy rice spike of rice to be identified and/
Or the sword-like leave auricle of paddy rice spike of rice to be identified 5cm below the water surface.
In methods described, described paddy rice to be identified refers specifically to be in the paddy rice in boot stage.
In methods described, the described paddy rice being in boot stage refers specifically to plant fringe developmental stage and is in pollen mother cell subtrahend
Anaphase, the described big tripe phase in production, that is, paddy rice visual inspection to Flag Leaf Sheath extract out fall two leaf leaf sheath 0-5cm period.
In methods described, in described control pond, the temperature of water refers specifically to the temperature using water in Groundwater Control pond.
The temperature of water in described control pond, particular by the flow control entered pond and discharge pond of the underground water extracted out
The temperature of water in pond processed.
The temperature of described underground water is specially 16-17 DEG C.
Described underground water is specially well water；Well depth is specially 150 meters of underground.
In methods described, after described cold treatment, also need to take out paddy rice to be identified, plant in land for growing field crops continued growth to maturation
Phase；Then directly observe the setting percentage of paddy rice, for judging the cold resistance of strain；The criterion of described cold resistance is：Work as ginseng
Examination material relative setting rate（The setting percentage of fringe under the setting percentage/normal condition of marked fringe after cold treatment）More than 60% or relatively
Fringe yield（The yield of the yield of marked fringe/this fringe of normal process after cold treatment）More than 40%, or setting percentage or fringe relatively produce
The amount cold sensitive varieties used higher than test, all can determine that rice strain to be identified or kind have cold resistance, relative setting rate
Or fringe production value relatively is higher, illustrates that its cold resistance is stronger；Or, the criterion of described cold resistance is：Will be to be identified
The relative setting rate of paddy rice and/or fringe yield relatively are compared with rice varieties ten and field, if the relative knot of paddy rice to be identified
Real rate and/or fringe yield relatively 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.
It is a further object to provide application in screening cold resistant paddy rice for the described method.
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 boot stage.
In described application, the described paddy rice being in boot stage refers specifically to plant fringe developmental stage and is in pollen mother cell subtrahend
Anaphase, the described big tripe phase in production, that is, paddy rice visual inspection to Flag Leaf Sheath extract out fall two leaf leaf sheath 0-5cm period.
The method is not affected by weather conditions, is not mainly affected by temperature；Can effectively identify that heading stage different
Material of participating in the experiment.Be easy to process bulk materials simultaneously, with low cost, accuracy high it is easy to the cold-resistant breeding of paddy rice reproductive stage
Using with the identification of cold-resistant genetic stocks, assisted Selection boot stage cold-resistant strain etc..The method can directly observe handled rice
The setting percentage of fringe strain, for judging the cold resistance of strain.
Brief description
Fig. 1 is to process, with water cooling pond, material of participating in the experiment.
Fig. 2 is the fringe portion difference of material of participating in the experiment after normal process and cold treatment.
Specific embodiment
Experimental technique used in following embodiments if no special instructions, is conventional method.
Material used, reagent etc. in following embodiments, if no special instructions, all commercially obtain.
Embodiment 1, using water cooling pond method identify strain cold resistance
（One）Participate in the experiment material
Ten and field：For cold sensitivity parent, can be in rice research unit, such as China Agricultural University, Yunnan academy of agricultural sciences, China
Paddy rice Core Germplasms storehouse etc. obtain.
NIL1913：The NIL containing a cold-resistant QTL, i.e. cold-resistant strain；From cold-resistant parent Kunming by force
The height in little Bai paddy and cold sensitivity parent ten and field is for backcross progeny.Concrete Backcrossing methods are from the beginning of hybrid F_2, each generation point
Randomly select 10-20 strain to be returned with ten and field respectively, being differentiated according to the selfing fringe setting percentage on homophyletic during maturation is in material
No reservation cross combination, the higher combination of general setting percentage（Setting percentage is more than 70%）It is accredited as cold-resistant strain, and it is miscellaneous to retain it
Hand over combination, continue backcrossing and cold-resistant identification for of future generation.By this method continuous backcross to BC6.Then, plant each backcrossing combination
BC6F2Colony, filters out cold-resistant strain, then become cold-resistant NIL（NIL）.BC6F2 group to mainly cold-resistant NIL simultaneously
Body carries out cold-resistant identification and cold-resistant QTL positioning, obtains the NIL of each main cold tolerance gene.NIL1913 is exactly to also have No. 4 chromosomes
The NIL of a upper cold-resistant QTL.
Kunmingxiaobaigu can obtain in China national germplasm resource bank.
T-1、T-5、T-12：Clone cold tolerance gene qCTB4-1-1 and proceeded to parent ten and Tanaka from NIL1913, obtained
The separate transformation events of the overexpression cold tolerance gene arriving；Through carrier construction and conversion acquisition transgenic line need to be carried out.
（Two）Process the process of material of participating in the experiment using water cooling pond
1st, build water cooling pond
Select to irrigate easily field at one, dig a long 10m, wide 2m, the pond that at the bottom of pond is risen progressively in staged, at the bottom of pond is
Depths is 2m, and the most shallow place is 1m, and often section ladder rises the height of 20cm.Cover bottom with the double-plastic of long 15m × wide 8m
Portion and 4 sides, with antiseep.From deep well（150m）Extract 16-17 DEG C of underground water, inject water cooling pond, make in water cooling pond
Depth of water 60-160cm, using water inlet and drain discharge control cold water pool inner water temperature, make pool inner water temperature be maintained at 18- round the clock
Between 19 DEG C.Place thermometer in pond and monitor in real time is carried out to water temperature.
2nd, participate in the experiment the screening of material and mark
Select tiller number, plant height, boot stage almost consistent ten and field, the plant of NIL1913, T-1, T-5 and T-12 strain
Strain as participating in the experiment material, additionally, 3 main spikes of rice will be had in each plant to be in boot stage.The described plant being in boot stage
Refer specifically to plant fringe developmental stage and be in the pollen mother cellses later stage, the described big tripe phase in production, i.e. paddy rice outward appearance sight
Observe the period that Flag Leaf Sheath extracts two leaf leaf sheath 0-5cm out.Using hanging label or carried out using paint modes such as spot printing of drawing a circle
Mark is in main fringe and the tillering tassel in this period, the setting percentage after processing for investigation.
3rd, process, using water cooling pond, material of participating in the experiment
As shown in figure 1, above-mentioned steps 2 are screened and the plant that marks loads respective experimental group according to different strains
In plastic casing, in the water cooling pond that immersion step 1 is built, the standard of the depth of plant immersion is：The water logging in described pond is not waited to reflect
Determine at the sword-like leave auricle of spike of rice or spike of rice to be identified sword-like leave auricle below the water surface 5cm.
Cold water treatment 7 days, material of participating in the experiment takes out, and plants in land for growing field crops continued growth to maturation according to general planting method
Phase, then, investigate its setting percentage.
Each kind or strain set up 3 repetitions, and one is repeated to investigate respectively 3 plants, investigate 3 for every plant and are in optimization process
The spike of rice in period（Before processing is labeled）；Have 27 fringes of mark altogether and carry out the investigation of later stage setting percentage.
（Three）Water cooling pond method identifies the result of strain cold resistance
Qualification result is shown in Table 1 and Fig. 2.When material relative setting rate of participating in the experiment（The setting percentage of marked fringe after cold treatment/normal
Under the conditions of fringe setting percentage）More than 60% or with respect to fringe yield（The yield of the yield of marked fringe/normal process fringe after cold treatment）
More than 40%, all can determine that material has certain Cold Tolerance at Booting Stage.
Parent ten and field, NIL1913 and transgenic line T-1, T-5, T-12 are in the yield side of relative setting rate, relative fringe
There is larger difference in face.Described cold treatment refers to according to above-mentioned（Two）The process that methods described is carried out；Described normal process is
Refer to the process according to general planting method plantation.
Table 1
The result of table 1 and Fig. 2 shows, cold sensitivity parent ten all waits base than near with the relative setting rate in field with relative fringe yield
Because being that NIL1913, transgenic line T-1, T-5, T-12 are low.Cold sensitivity parent ten and field every economical character after cold treatment is equal
Show and injured by low temperature, have different degrees of reduction, and every economical character of cold-resistant NIL NIL1913
Performance is superior to ten and field；Transgenic line T-1, T-5, T-12 of overexpression cold tolerance gene equal ratio ten in terms of relative setting rate
High with field.
To the explanation of the qualification result of material of participating in the experiment, being processed material of participating in the experiment can precise Identification respectively be participated in the experiment material using water cooling pond method
The cold-resistant ability of material.The method of the present invention is accurately processed for fringe portion, and remains natural environment to greatest extent to plant
Selection, this method be applied to identification rice germplasm cold resistance.
Claims (5)
1. a kind of method of identification cold resistant paddy rice, methods described comprises the steps：
1) build pond；
2) control the temperature of water in pond；
3) cold treatment paddy rice to be identified；
Described pond will carry out antiseepage drain process；In described pond, the temperature of water is 18-19 DEG C；
Described cold treatment includes for paddy rice to be identified immersing step 1) and 2) 7 days in the water in described pond, described paddy rice to be identified
The standard of depth of immersion is：The water logging in described pond does not have at the sword-like leave auricle of paddy rice spike of rice to be identified and/or paddy rice to be identified
The sword-like leave auricle of spike of rice 5cm below the water surface；
Described paddy rice to be identified refers specifically to be in the paddy rice in boot stage；
In described control pond, the temperature of water refers to the temperature using water in Groundwater Control pond.
2. method according to claim 1 it is characterised in that：Described it is in boot stage and refers specifically at plant fringe developmental stage
In the pollen mother cellses later stage, the big tripe phase of indication in production, that is, paddy rice visual inspection to Flag Leaf Sheath bag extract out fall
Period at two leaf leaf sheath 0-5cm.
3. the method according to claim 1 or 2 it is characterised in that：After described cold treatment, also need to take out paddy rice to be identified, plant
Plant in land for growing field crops continued growth to the maturity period；Then directly observe the setting percentage of paddy rice, for judging the cold resistance of strain；
The criterion of described cold resistance is：When paddy rice relative setting rate to be identified is more than 60%, or fringe yield relatively exceedes
40%, all can determine that paddy rice to be identified has cold resistance, relative setting rate or fringe production value relatively are higher, and its cold resistance is described
Stronger；
Or, the criterion of described cold resistance is：By the relative setting rate of paddy rice to be identified and/or relative fringe yield and paddy rice product
Kind ten and field are compared, if the relative setting rate of paddy rice to be identified and/or fringe yield relatively are significantly higher than ten and field, judge
The cold resistance of described paddy rice to be identified is higher than ten and field.
4. application in screening cold resistant paddy rice for the arbitrary described method of claim 1-3, described paddy rice is to be in boot stage
Paddy rice.
5. according to claim 4 application it is characterised in that：Described it is in boot stage and refers specifically at plant fringe developmental stage
In the pollen mother cellses later stage, the big tripe phase of indication in production, that is, paddy rice visual inspection to Flag Leaf Sheath bag extract out fall
Period at two leaf leaf sheath 0-5cm.
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| CN105794508B (en) * | 2016-03-11 | 2019-11-15 | 江西农业大学 | A kind of simple and easy method of quick screening cold resistance rice varieties |
| 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 |
| CN107006366B (en) * | 2017-02-27 | 2019-11-08 | 湖北省农业科学院粮食作物研究所 | A kind of method that big mass screening cultivates rice resistant to high temperatures |
| 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 |
| 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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| CN1220425C (en) * | 2002-11-21 | 2005-09-28 | 江西省农业科学院水稻研究所 | Screening method for breeding strong cold-resisting new rice variety |
| CN100350829C (en) * | 2005-01-27 | 2007-11-28 | 中国科学院等离子体物理研究所 | Breeding method of continuously and directly seeding carly and late rice breed |
| CN101129123A (en) * | 2007-09-26 | 2008-02-27 | 中国科学院东北地理与农业生态研究所 | Cold resistant high-production synthetic method for planting rice |
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