CN109618921A - A kind of method and application of the stress critical-temperature of identification zasiokaurin high temperature resistant - Google Patents
A kind of method and application of the stress critical-temperature of identification zasiokaurin high temperature resistant Download PDFInfo
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- CN109618921A CN109618921A CN201811488806.4A CN201811488806A CN109618921A CN 109618921 A CN109618921 A CN 109618921A CN 201811488806 A CN201811488806 A CN 201811488806A CN 109618921 A CN109618921 A CN 109618921A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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Abstract
The invention discloses a kind of methods of identification zasiokaurin high temperature resistant stress critical-temperature, comprise the following steps: A, normal culture；B, fixed temperature culture；C, pollen activity measures；D, establish analysis model: with different temperatures gradient for x value, the zasiokaurin vigor to cultivate at each temperature constructs y=f (x) regression analysis model, is up to standard with the coefficient of determination of model and determines final regression model for y value；E, it determines critical-temperature: with y >=50% for critical value, substituting into equation y=f (x) and solve x value, obtain the critical-temperature of zasiokaurin high temperature resistant stress.Method of the invention not only compensates for the blank of the prior art, has specific theory significance and good practical value, while being with a wide range of applications in terms of the breeding of corn high temperature resistant, corn high temperature resistant resistance screening.
Description
Technical field
The present invention relates to field of agricultural production technologies, and in particular to the breeding of corn high temperature resistant, the screening of high temperature resistant varietal resistance
Correlation subdivision technical field.
Background technique
One of an important factor for zasiokaurin vigor is influence corn grain number per spike, high temperature can make pollen development abnormal or lose
It educates, pollen quantity is caused to reduce, on the other hand it is also possible that pollen grain division is abnormal, form deformity.Therefore, high temperature causes to spend
The quantity and vigor of powder decline, and influence fertilization, eventually lead to setting percentage reduction and the underproduction, even No kernels or seeds are gathered, as in a year of scarcity when serious.
At present in research and production, the method mould of the excessively in vitro zasiokaurin high-temperature process of the measurement multi-pass of zasiokaurin vigor
Harm of the high temperature to pollen under quasi- natural conditions, using iodine-potassium iodide method, triphenyltetrazolium chloride (Triphenyl
Tetrazolium chloride, TTC) method and germination in vitro method be measured pollen activity.It is generally acknowledged that TTC method is surveyed
The vigor for determining pollen is the simplest accurate.
But lacking in research and production can accurately determine different corn varieties to the research side of high temperature tolerance range at present
Method and high temperature critical indicator especially influence the critical-temperature of pollen activity.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of methods of identification zasiokaurin high temperature resistant stress critical-temperature
And its application in terms of the breeding of corn high temperature resistant, corn high temperature resistant resistance screening.
In order to solve the above technical problems, the technical solution used in the present invention is as follows.
A method of identification zasiokaurin high temperature resistant coerces critical-temperature, comprises the following steps:
A, normal culture: corn variety to be measured is cultivated under non-stress condition, until the tasseling stage of corn, obtaining can be just
Often take out plant that is male and generating pollen；
B, fixed temperature culture: being transferred to rapidly phjytotron/climate box after corn takes out male loose powder, the temperature ladder manually set
Degree culture；The setting of artificial climate room temperature includes at least five temperature gradients, and it is beautiful that a collection of tasseling stage is cultivated under the conditions of each temperature
Rice；
C, pollen activity measures: measuring pollen activity after daily high-temperature process with TTC method, replication is averaged；
D, establish analysis model: with different temperatures gradient for x value, the zasiokaurin vigor to cultivate at each temperature constructs for y value
Y=f (x) regression analysis model is up to standard with the coefficient of determination of model and determines final regression model；
E, it determines critical-temperature: with y >=50% for critical value, substituting into equation y=f (x) and solve x value, obtain zasiokaurin high temperature resistant
The critical-temperature of stress.
As a preferred technical solution of the present invention, non-stress condition described in step A refers in preference temperature, light
According to carrying out corn culture under, nutrient and moisture condition.
As a preferred technical solution of the present invention: in step A, at interval of a collection of plant of culture in 5 days, add up to
Culture 5 batches, as subsequent processing and the basis for establishing analysis model.
As a preferred technical solution of the present invention, in step B, five temperature gradients include 30 degree, 32 degree, 35
Degree, 38 degree, 40 degree.
As a preferred technical solution of the present invention, in step B, is handled to the manual temperature of every plant of corn daily the time
10:00--15:00.
As a preferred technical solution of the present invention, in step B, every group of corn is connected respectively according to set temperature gradient
Continuous processing 3 days.
As a preferred technical solution of the present invention, in step C, each processing, which is repeated 3 times, to be averaged.
It is living with the zasiokaurin for cultivating the 3rd day at each temperature in step D as a preferred technical solution of the present invention
Power is y value.
As a preferred technical solution of the present invention, in step D, pollen of the different corn varieties under temperature gradient is living
Power variation tendency is different, and regression model is linear, quadratic form, power model or other models, in the final regression model of determination
When standard is up to the coefficient of determination of model.
As a preferred technical solution of the present invention, in step E, with the pollen activity after high-temperature cultivation for three days on end >=
50% is used as evaluation criterion, substitutes into equation y=f (x) and solves x value.
Application of the above method in terms of the breeding of corn high temperature resistant, corn high temperature resistant resistance screening.
The beneficial effects of adopting the technical scheme are that
There is genetic variety, the degree of different cultivars tolerance high temperature stress disaster to have obviously for the heat-resistant quality of zasiokaurin
Difference determines different corn varieties to the tolerance degree of high temperature and to high temperature resistant screening varieties, high temperature resistant corn breeding and height
Cultivation etc. is produced to have a very important significance.The present invention is for shortage can accurately determine different jade in scientific research, production at present
Rice kind gives different corn varieties to high temperature tolerance by the research and test of systematization to the status of high temperature tolerance range
Research method, the high temperature tolerance critical indicator for obtaining different corn varieties is given especially by the foundation of analysis model,
So as to directly obtain the critical-temperature for influencing pollen activity.
This original technology scheme not only compensates for the blank of the prior art, has specific theory significance and good
Practical value, while being with a wide range of applications in terms of the breeding of corn high temperature resistant, corn high temperature resistant resistance screening.
Detailed description of the invention
Fig. 1 is that the pollen activity of three different genotype corn varieties at different temperatures measures numerical value.
Specific embodiment
The present invention is described in detail in following embodiment.Various raw materials used in the present invention and items of equipment are conventional city
Product is sold, can be bought and be directly obtained by market.
Embodiment 1 cultivates corn to tasseling stage under the conditions of being suitble to temperature.
Corn potting culture is carried out under preference temperature, illumination, nutrient and moisture condition.It is spaced a batch of culture in 5 days, is trained altogether
Support 5 batches.This purpose is to cultivate related corn variety to can normally take out male and generation pollen.
Embodiment 2, phjytotron high-temperature cultivation zasiokaurin.
It is transferred to different temperatures phjytotron (or climate box) rapidly after corn takes out male loose powder and carries out artificial set temperature
Culture.Artificial climate room temperature is set as 30 degree, 32 degree, 35 degree, 38 degree, 40 degree, cultivates a batch under the conditions of each temperature and takes out hero
Phase corn, 5 temperature gradients totally 5 batches.The daily Temperature Treatment time is 10:00--15:00, continuous processing 3 days.
Embodiment 3, measurement zasiokaurin vigor.
(15:00) measures pollen activity with TTC method after daily high-temperature process, and each processing is repeated 3 times.
Embodiment 4 establishes analysis model.
With different temperatures gradient for x value, the zasiokaurin vigor to cultivate the 3rd day at each temperature constructs y=f (x) for y value
Regression analysis model.Since pollen activity variation tendency of the different corn varieties under temperature gradient is different, regression model can
It can be linear, quadratic form, power model etc., therefore mark is up to the coefficient of determination of model in determination final regression model
It is quasi-.
Embodiment 5 determines critical-temperature.
It is critical value with y >=50%, i.e., pollen activity >=50% after high-temperature cultivation for three days on end is used as evaluation criterion, band
Enter equation y=f (x) solution, then x value is the critical-temperature of zasiokaurin high temperature resistant stress.
The test result of 6, three groups of maize genotypes of embodiment.
It is that the pollen activity of three different genotype corn varieties at different temperatures measures numerical value referring to attached drawing 1.
Standard is up to the coefficient of determination of model in determination final regression model.Obtain the optimum regression mould of kind 1
Type are as follows:
y=-0.7475x2+42.46x-501.8 (R2=0.9995)。
The best regression model of kind 2 are as follows:
y=0.869x2-71.407x+1463 (R2=0.9922)。
The best regression model of kind 3 are as follows:
y=-10.221x+406.32 (R2=0.9978)。
The critical-temperature that x value of the solution as pollen activity y=50% can obtain kind 1 is: 36.7 DEG C；Kind 2 it is critical
Temperature is: 33.1 DEG C；The critical-temperature of kind 3 is: 34.9 DEG C.To obtain the final product.
Foregoing description is only proposed as the enforceable technical solution of the present invention, not as to the single of its technical solution itself
Restrictive condition.
Claims (10)
1. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature, it is characterised in that: this method comprises the following steps:
A, normal culture: corn variety to be measured is cultivated under non-stress condition, until the tasseling stage of corn, obtaining can be just
Often take out plant that is male and generating pollen；
B, fixed temperature culture: being transferred to rapidly phjytotron/climate box after corn takes out male loose powder, the temperature ladder manually set
Degree culture；The setting of artificial climate room temperature includes at least five temperature gradients, and it is beautiful that a collection of tasseling stage is cultivated under the conditions of each temperature
Rice；
C, pollen activity measures: measuring pollen activity after daily high-temperature process with triphenyl tetrazolium chloride method, repeats
Measurement is averaged；
D, establish analysis model: with different temperatures gradient for x value, the zasiokaurin vigor to cultivate at each temperature constructs for y value
Y=f (x) regression analysis model is up to standard with the coefficient of determination of model and determines final regression model；
E, it determines critical-temperature: with y >=50% for critical value, substituting into equation y=f (x) and solve x value, obtain zasiokaurin high temperature resistant
The critical-temperature of stress.
2. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature according to claim 1, it is characterised in that:
Non-stress condition described in step A refers to the progress corn culture under preference temperature, illumination, nutrient and moisture condition.
3. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature according to claim 1, it is characterised in that:
In step A, at interval of a collection of plant of culture in 5 days, add up to culture 5 batches, as subsequent processing and the base for establishing analysis model
Plinth.
4. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature according to claim 1, it is characterised in that:
In step B, five temperature gradients include 30 degree, 32 degree, 35 degree, 38 degree, 40 degree.
5. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature according to claim 1, it is characterised in that:
In step B, time daily 10:00--15:00 is handled to the manual temperature of every plant of corn.
6. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature according to claim 1, it is characterised in that:
In step B, every group of corn is distinguished continuous processing 3 days according to set temperature gradient.
7. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature according to claim 1, it is characterised in that:
In step D, to cultivate the 3rd day zasiokaurin vigor at each temperature as y value.
8. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature according to claim 1, it is characterised in that:
In step D, pollen activity variation tendency of the different corn varieties under temperature gradient is different, and regression model is linear, secondary
Type, power model or other models are up to standard in the final regression model of determination with the coefficient of determination of model.
9. a kind of method of identification zasiokaurin high temperature resistant stress critical-temperature according to claim 1, it is characterised in that:
In step E, using pollen activity >=50% after high-temperature cultivation for three days on end as evaluation criterion, substitutes into equation y=f (x) and solve x value.
10. application of the method for any one of claim 1-9 in terms of the breeding of corn high temperature resistant, corn high temperature resistant resistance screening.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110999655A (en) * | 2019-12-04 | 2020-04-14 | 河南省农业科学院粮食作物研究所 | Preparation method and application of standardized sample for high-throughput determination of maize pollen viability under high-temperature stress |
| CN112772410A (en) * | 2021-01-18 | 2021-05-11 | 重庆市农业科学院 | Method for screening high-temperature-resistant corn material based on flowering characteristics |
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Cited By (2)
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| CN110999655A (en) * | 2019-12-04 | 2020-04-14 | 河南省农业科学院粮食作物研究所 | Preparation method and application of standardized sample for high-throughput determination of maize pollen viability under high-temperature stress |
| CN112772410A (en) * | 2021-01-18 | 2021-05-11 | 重庆市农业科学院 | Method for screening high-temperature-resistant corn material based on flowering characteristics |
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Application publication date: 20190416 |