CN109729870A - The identification method of cold-resistant sex differernce between a kind of wheat breed - Google Patents
The identification method of cold-resistant sex differernce between a kind of wheat breed Download PDFInfo
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- CN109729870A CN109729870A CN201910159989.3A CN201910159989A CN109729870A CN 109729870 A CN109729870 A CN 109729870A CN 201910159989 A CN201910159989 A CN 201910159989A CN 109729870 A CN109729870 A CN 109729870A
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Abstract
The present invention relates to a kind of identification methods of cold-resistant sex differernce between wheat breed, belong to wheat and combat a natural disaster mitigation technical field.The present invention passes through simulation low temperature stress, it obtains by low temperature stress treated wheat leaf blade, and determine the conductivity of wheat leaf blade leachate, it is calculated from the formula wheat leaf blade relative electrolyte leaching rate, to judge that wheat leaf blade cell leakage changes, and the size changed according to cell leakage, the winter resistance to judge wheat breed are strong and weak.The present invention is applied widely, using laboratory routine equipment, simple and easy, method is reliable, it identifies that the cold resistance of many plant varieties and winter resistance cultivate effect and provides effective means for early stage or optimum period, avoids the disadvantages of growth cycle that crop field is identified is long, condition is not easy to control.
Description
Technical field
The invention belongs to wheats to combat a natural disaster mitigation technical field, and in particular to the identification of cold-resistant sex differernce between a kind of wheat breed
Method.
Background technique
Wheat winter freeze calamity, Early spring freezing injury (cold spell in later spring) and chilling injury have become the master for threatening China's Wheat Production safety
Want natural calamity.Since the power of wheat breed tolerance to cold is to determine the main internal factor of freeze injury occurrence degree, it is quasi-
The really cold-resistant degree of identification wheat breed (being), cultivates new variety of wheat and its popularization and application are of great significance.
Currently, the identification method of wheat breed winter resistance, main includes the conventional winter resistance mirror of investigation crop field dead seedling situation
It is fixed, and Determination of Physiological And Biochemical Indices relevant to cold resistance etc..Wheat breed cold hardness evaluation is sent out in the application of these methods
Important function has been waved, but still has been come with some shortcomings.Such as conventional cold hardness evaluation method, it is affected, is added by ambient temperature
Usually occur warm winter weather in recent years, not only seriously affected the accuracy of cold hardness evaluation, but also cannot definitely identify its cold-resistant
Degree；Determination of Physiological And Biochemical Indices method, operating procedure is many and diverse, and chemical reagent used easily causes environmental pollution, and wheat breed
(being) slightly low temperature stress can cause biggish physiological acoustic signals, and strong low temperature stress easily causes wheat plant dead, nothing
Method accurately determines its cold-resistant degree according to the related index of measurement.
The cold-resistant wheat breed of breeding is to solve wheat by one of the main means of cold damage, and the key of breeding cold-resistant variety is
To the precise Identification of cold-resistant variety.According to the identification method of current existing wheat breed winter resistance, rely primarily on after enduring cold
Phenotypic characteristic carry out cold-resistant classification, but the cold spell in later spring that or spring low for winter cooling degree occurs, since it is by cold damage
Phenotypic characteristic is unobvious, then cannot carry out accurate judgement to the winter resistance of kind.
Summary of the invention
The purpose of the present invention is provide a kind of identification of cold-resistant sex differernce between wheat breed to solve above-mentioned technical problem
Method goes out the cold-resistant sex differernce between wheat breed with precise Identification.
The object of the present invention is achieved like this: the identification method of cold-resistant sex differernce between a kind of wheat breed comprising with
Lower step:
(1) simulation low temperature stress processing: take wheat breed to be measured without disease pest plant as processing group, take its leaf, clean afterwards it is clean
Net filter paper exhaustion surface moisture, is put into the refrigerator for setting treatment temperature, stands 3 h, while setting one group and not making freezing processing
Control group；
(2) measure blade leachate conductivity: the processing group blade after taking out freezing mixes after being broken into fritter, is added
Detection liquid is made in deionized water；Then above-mentioned detection liquid is vacuumized, to extract space between cells air out, is then slowly put into sky
Gas, water penetrate into space between cells, and blade becomes transparence；Then the detection liquid concussion Jing Guo above-mentioned processing is mixed into 0.5-1h,
Obtain blade leachate；Same processing is done to control group blade, finally with the blade of conductivity gauge measurement processing group and control group
The conductivity of leachate, is denoted as C respectively1And C0；
(3) the leachate conductivity after measurement blade cell is entirely killed: the blade that conductivity has been measured in step (2) is soaked
Liquid progress boiling water bath is cooled to room temperature after taking-up out with killing leaf tissue, shakes up the sample in test tube, measurement processing again
The conductivity for organizing the leachate after blade cell is entirely killed, is denoted as C2；
(4) it calculates relative electrolyte leaching rate: calculating the relative electrolyte leaching rate of processing group blade, phase according to following equation
To electrolyte osmotic rate (%)=(C1-C0)/(C2-C0) × 100%；
(5) it concludes that with the size of the relative electrolyte leaching rate of blade, the winter resistance to judge wheat breed is strong and weak, phase
Bigger to electrolyte osmotic rate, the winter resistance of wheat breed is weaker, conversely, relative electrolyte leaching rate is smaller, wheat breed
Winter resistance is stronger.
Further, the treatment temperature in the step (1) can be set as the present invention: -2 DEG C, -4 DEG C, -6 DEG C, -8 DEG C and -10
DEG C, while carrying out 5 processing groups.
The blade that freeze post-processing group further, in the step (2), can be cut into 1mm segment by the present invention, or by leaf
Piece stacks, and is beaten with stainless steel punch and takes disk.
The present invention further, in the step (2), is added 100 mL deionized waters, detection is made in every gram of blade
Liquid.
The present invention further, in the step (2), the detection liquid is put into vacuum desiccator, vacuum pump is started,
Take its pumping.
Further, in the step (2), the number for starting vacuum pump pumping is 2 times to the present invention, each 10-15min.
Further, in the step (3), the time for carrying out boiling water bath is 10-30min to the present invention.
Further, every group of processing group or control group can set 3-4 and repeat to test the present invention, as a result take its average value.
Further, container used must be cleaned thoroughly the present invention, then be washed down with deionized water, be inverted on clean filter paper
It is spare.
Theoretical basis of the invention:
The selection permeability of cell membrane is one of the most important condition that it maintains physiological function.Various adverse circumstance injuries can all cause matter
Film selects the change or forfeiture of permeability, and plant tissue is by various unfavorable environmental conditions (such as low temperature, high temperature and atmosphere dirts
Dye) when endangering, the structure and function of cell membrane is first subjected to injury, shows as cell leakage and increases.The journey that membrane permeability increases
Degree is related with environment stress intensity, and coercive intensity is bigger, and injury is heavier, exosmoses more, the increase of conductivity is also bigger.Simultaneously
Related with the power of stress resistance of plant, resistance is stronger, and injury is lighter, exosmoses fewer, the increase of conductivity is also smaller.So logical
Cross the variation for measuring outer sepage conductivity, so that it may reflect the extent of injury of cell membrane and measure and monitor the growth of standing timber and expect the size of resistance.
Beneficial effects of the present invention:
1, it is first that the membrane permeability variation that the present invention is measured by electrolyte leakage method significantly appears in formalness variation, therefore
The present invention identifies that the cold resistance of many plant varieties and winter resistance cultivate effect and provides effective means for early stage or optimum period；
2, low temperature-controlled temperature ranges of the present invention are wide, and minimum temperature can be easy to operate down to -35 DEG C, have certain degree
Sensitivity and accuracy, the cold-resistant degree of wheat can be accurately determined, and efficiently avoid crop field identification growth week
The disadvantages of phase is long, condition is not easy to control；
3, the present invention is applied widely, can be applied to cold hardness evaluation, the wheat hybridizing parent of Wheat Germplasm Resources and its sibling species
This Screening For Cold, the cold hardness evaluation of wheat breeding offspring and new varieties (being) and the winter resistance of wheat breed (being) is ground
Study carefully；
4, the present invention uses laboratory routine equipment, and simple and easy, method is reliable.
Detailed description of the invention
Relative electrolyte leaching rate of Fig. 1 Wheat Cultivars under Different hypothermia.
Specific embodiment
The present invention will be further explained with reference to the examples below.
The identification method of cold-resistant sex differernce between a kind of wheat breed exists in the present embodiment aobvious with winter resistance in production practices
The wheat breed for writing difference is expert evidence, tested variety 10, respectively 1 grade: mountain agriculture 20, Jimai 22；2 grades: Henan wheat 49,
AK58；3 grades: Zhou Mai 18, hundred agricultures 207；4 grades: middle burnt No. 3, Zheng Mai 9023；5 grades: river agriculture 16, E Mai 15.
Specifically includes the following steps:
(1) simulation low temperature stress processing: above-mentioned each variety selection, as processing group, takes its leaf without disease pest wheat plant, cleans
Clean filter paper exhausts surface moisture afterwards, is put into the refrigerator for setting treatment temperature, each kind totally 5 processing groups, 5 processing
The refrigerator temperature of group is successively set as -2 DEG C, -4 DEG C, -6 DEG C, -8 DEG C, -10 DEG C, and each processing group sets 3 parallel tests, stands 3
H, while setting one group of control group for not making freezing processing；
(2) measure blade leachate conductivity: the processing group blade after taking out freezing wipes that be cut into 1 mm after dry surface moisture small
Section, or blade is stacked, it is beaten with stainless steel punch and takes disk, blade is mixed, accurately weighed 0.2 g and mix sample, be placed in
In 20mL plug test tube, 20 mL deionized waters are added, detection liquid is made；Then above-mentioned detection liquid vacuum is put into do
In dry device, vacuum pump is started, to extract space between cells air out, is evacuated 2 times, then each 10-15min is slowly put into air, water
Space between cells is penetrated into, blade becomes transparence；Then the detection liquid concussion Jing Guo above-mentioned processing is mixed into 0.5-1h, obtains leaf
Piece leachate；Same processing is done to control group blade, finally with the blade leachate of conductivity gauge measurement processing group and control group
Conductivity, be denoted as C respectively1And C0；
(3) the leachate conductivity after measurement blade cell is entirely killed: will be with sample to measured conductivity in step (2)
Test tube, which is put into water-bath, carries out boiling water bath, time 10-30min, to kill leaf tissue, is cooled to room temperature after taking-up, then
The secondary sample shaken up in test tube measures the conductivity of the leachate after processing group blade cell is entirely killed, is denoted as C2；
(4) it calculates relative electrolyte leaching rate: calculating the relative electrolyte leaching rate of processing group blade, phase according to following equation
To electrolyte osmotic rate (%)=(C1-C0)/(C2-C0) × 100%, when cold hardiness is variant between unknown kind, with this formula meter
It calculates, error caused by Electrolyte Leakage amount is different between kind can be overcome, achieve the purpose that precise Identification；
(5) it concludes that with the size of the relative electrolyte leaching rate of blade, the winter resistance to judge wheat breed is strong and weak, phase
Bigger to electrolyte osmotic rate, the winter resistance of wheat breed is weaker, conversely, relative electrolyte leaching rate is smaller, wheat breed
Winter resistance is stronger；
(6) result verification: with small in " variety of crops regional testing technical regulation wheat " (NY/T 1301-2007) appendix A
Wheat winter resistance rank is as control, as a result such as Fig. 1.It will be seen from figure 1 that using the method for the present invention and conventional cold hardness evaluation knot
Fruit is almost the same, illustrates that the method for the present invention is accurately reliable.
Specific embodiment is to further explanation of the invention rather than limits, and is existed for those of ordinary skills
Further transformation is done in the case where not departing from substantive content of the present invention, and all these transformation all should belong to right appended by the present invention
It is required that protection scope.
Claims (9)
1. the identification method of cold-resistant sex differernce between a kind of wheat breed, which comprises the following steps:
(1) simulation low temperature stress processing: take wheat breed to be measured without disease pest plant as processing group, take its leaf, clean afterwards it is clean
Net filter paper exhaustion surface moisture, is put into the refrigerator for setting treatment temperature, stands 3 h, while setting one group and not making freezing processing
Control group；
(2) measure blade leachate conductivity: the processing group blade after taking out freezing mixes after being broken into fritter, is added
Detection liquid is made in deionized water；Then above-mentioned detection liquid is vacuumized, to extract space between cells air out, is then slowly put into sky
Gas, water penetrate into space between cells, and blade becomes transparence；Then the detection liquid concussion Jing Guo above-mentioned processing is mixed into 0.5-1h,
Obtain blade leachate；Same processing is done to control group blade, finally with the blade of conductivity gauge measurement processing group and control group
The conductivity of leachate, is denoted as C respectively1And C0；
(3) the leachate conductivity after measurement blade cell is entirely killed: the blade that conductivity has been measured in step (2) is soaked
Liquid progress boiling water bath is cooled to room temperature after taking-up out with killing leaf tissue, shakes up the sample in test tube, measurement processing again
The conductivity for organizing the leachate after blade cell is entirely killed, is denoted as C2；
(4) it calculates relative electrolyte leaching rate: calculating the relative electrolyte leaching rate of processing group blade, phase according to following equation
To electrolyte osmotic rate (%)=(C1-C0)/(C2-C0) × 100%；
(5) it concludes that with the size of the relative electrolyte leaching rate of blade, the winter resistance to judge wheat breed is strong and weak, phase
Bigger to electrolyte osmotic rate, the winter resistance of wheat breed is weaker, conversely, relative electrolyte leaching rate is smaller, wheat breed
Winter resistance is stronger.
2. the identification method of cold-resistant sex differernce between a kind of wheat breed according to claim 1, which is characterized in that the step
Suddenly the treatment temperature in (1) can be set as: -2 DEG C, -4 DEG C, -6 DEG C, -8 DEG C and -10 DEG C, while carrying out 5 processing groups.
3. the identification method of cold-resistant sex differernce between a kind of wheat breed according to claim 1, which is characterized in that the step
Suddenly in (2), the blade for freezing post-processing group can be cut into 1mm segment, or blade is stacked, is beaten with stainless steel punch and takes circle
Piece.
4. the identification method of cold-resistant sex differernce between a kind of wheat breed according to claim 1, which is characterized in that the step
Suddenly in (2), 100 mL deionized waters is added in every gram of blade, detection liquid is made.
5. the identification method of cold-resistant sex differernce between a kind of wheat breed according to claim 1, which is characterized in that the step
Suddenly in (2), the detection liquid is put into vacuum desiccator, vacuum pump is started, it is evacuated.
6. the identification method of cold-resistant sex differernce between a kind of wheat breed according to claim 5, which is characterized in that the step
Suddenly in (2), the number for starting vacuum pump pumping is 2 times, each 10-15min.
7. the identification method of cold-resistant sex differernce between a kind of wheat breed according to claim 1, which is characterized in that the step
Suddenly in (3), the time for carrying out boiling water bath is 10-30min.
8. the identification method of cold-resistant sex differernce between a kind of wheat breed according to claim 1, which is characterized in that at every group
Reason group or control group can set 3-4 and repeat to test, and as a result take its average value.
9. the identification method of cold-resistant sex differernce between a kind of wheat breed according to claim 1, which is characterized in that appearance used
Device must be cleaned thoroughly, then be washed down with deionized water, be inverted in spare on clean filter paper.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111665324A (en) * | 2020-06-02 | 2020-09-15 | 西藏自治区农牧科学院农业研究所 | Method for identifying cold resistance difference between wheat varieties |
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2019
- 2019-03-04 CN CN201910159989.3A patent/CN109729870A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 曹文昕等: ""小麦不同生育期抗寒性研究"", 《安徽农学通报》 * |
| 杨敏生等: "《杨树抗逆性研究》", 30 June 2008, 天津科学技术出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111665324A (en) * | 2020-06-02 | 2020-09-15 | 西藏自治区农牧科学院农业研究所 | Method for identifying cold resistance difference between wheat varieties |
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Application publication date: 20190510 |