CN107389738A - A kind of rapid screening method of tomato Evaluation of Cold Tolerance and application - Google Patents
A kind of rapid screening method of tomato Evaluation of Cold Tolerance and application Download PDFInfo
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- CN107389738A CN107389738A CN201710604796.5A CN201710604796A CN107389738A CN 107389738 A CN107389738 A CN 107389738A CN 201710604796 A CN201710604796 A CN 201710604796A CN 107389738 A CN107389738 A CN 107389738A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
- G01N27/04—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance
- G01N27/041—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance of a solid body
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
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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
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Abstract
The invention provides a kind of method of quickly screening and testing cold resistance of tomatoes.Utilize the different tomato variety of known tolerance to cold, after first carrying out of short duration Osmotic treatment to it, the cold treatment condition to its overall plant is studied again, and determine the electrical conductivity of the aerial part cauline leaf after cold treatment, establish cold-resistant correlation of the degree under the conditions of particular procedure with electrical conductivity size of tomato plant, the number range for the electrical conductivity for distinguishing tomato difference cold-resistant variety is obtained, so as in the short time, tomato varieties different to high-volume（System）The cold resistance that material carries out plant carries out easy, Fast Evaluation, selects and cultivates cold-resistant tomato variety（System）.The cold resistance of whole process of the present invention is screened all under controlled condition, and the selection result is reliable, and method is easy, and screening cost is relatively low.
Description
Technical field
The present invention relates to a kind of rapid screening method of tomato Evaluation of Cold Tolerance and application, available for fruit eggplant class, such as tomato,
The new cold-resistant species such as muskmelon, eggplant（System）Screening and identification.
Background technology
Tomato is a kind of fruit eggplant class for liking temperature.High temperature and low temperature can all produce to it to be had an impact, in Central China area, agricultural
Low temperature cold damage is a kind of serious natural calamity in industry production, and in the serious area of frost, production delay occurs in eggplant class fruits and vegetables,
Yield declines, or even stops growing, gradually withered death, and according to statistics, loss can reach 1000 caused by freeze injury every year in China
More than ten thousand mu.Therefore, the seed selection of cold-resistant crop kind is significant in cultivation Dominant variety in agricultural production.
Tomato to be planted in China rather extensive, in agricultural community, though obtained the eggplant veriety being much possessed of good qualities,
But tolerance is cold and the kind of freeze injury is seldom.In recent years, domestic and international expert is attempted by cell technology engineering and transgenosis
Technology cultivates the new cold-resistant species of eggplant class.With reference to traditional spontaneons screening, occur simultaneously in many new varieties, one simply has
Effect the performance to the new varieties cultivated on identification also into it is very important the problem of.
It has recently been demonstrated that a series of lifes in cell level occur for meeting of the plant in experience cool period, plant
In reason change, the new old generation under low temperature, include the change of plant cell membrane, proline, soluble sugar, antioxidant reductase and some
The regulating and expressing of Cold resistant genes.What is be studied at first in these indexs is the change of cell membrane, and the change of permeability of cell membrane is just
It is plant to one of important indicator of low temperature tolerant, the big I of membrane passage is by the change of plant electrical conductivity come indirectly
Define, relatively other several indexs, it is most simply also most direct detection method that electrical conductivity, which is,.
By the excellent hardy plant of screening of nature cold climate approach, stock number is excessive, and the cycle is oversize, Character instability
Deng.The inaccurate factor of measured quantity again be present in the measure of the screening technique in other room, such as proline and soluble sugar, its
Repeatability is poor.And in the recent period more commonly used Subordinate Function and DSP analysis softwares the methods of, also have that data volume is big, software
The problems such as operation difficulty is big, and time-consuming.Therefore, the present invention provides a kind of rapid screening method of tomato Evaluation of Cold Tolerance with answering
With being substantially shorter seed selection cycle of cold-resistant tomato variety.The conductance measurement of the same type than before adds cold place simultaneously
Osmotic treatment before reason, and convalescence pre- cold period photo-irradiation treatment, the time of convalescence is added, so as to optimize whole plant
Cold treatment condition, reduce because leaf water and illumination are on influence caused by it is metabolized.The whole detection of above-ground plant parts
Reduce the only detection issuable one-sidedness of blade..
The content of the invention
It is an object of the invention to the needs to be resisted cold for existing seed selection in tomato technique improvement, there is provided a kind of easy and effective
Screening and authentication method, reduce the qualification cycle of cold tolerance trait during traditional breeding method and transgenic breeding, improve tomato
The validity of cold-resistant Screening of Strains Against, as a result more reliable, the high-quality resource that reducing may occur in improved seeds qualification process is lost.
For purpose as realization, the present invention establishes tomato plant on ground using the specific cold treatment condition after optimization
The corresponding linear relation of the numerical values recited of relative conductivity and the cold-resistant degree of plant, so as to reach the tolerance to cold of plant and spy
Fixed physical quantity is associated, and cold-resistant can make of tomato plant is fast and effectively judged according to the data of gained, so as to
To accelerate the seed selection process of the cold-resistant breeding of tomato.
In technical scheme, the key problem in technology in each period includes：
(1) correlation established between relative conductivity and the cold-resistant degree of the whole strain of tomato：
1. the culture of tomato seedling：By the different tomato of known cold-resistant degree (cold-resistant strain, general strain, can not resist cold strain) kind
Son, it is seeded into by 5 per basin in small-sized earth culture flowerpot（Soil is configured to vermiculite：Black earth：Perlite=9:3:0.5, black earth needs 121
DEG C sterilizing 30min）If 3 groups, every group 10 parallel, after planting irrigates soil first, weekly waters later, is placed on temperature
21 DEG C of degree, humidity 65%, after the culturing room under 16h illumination conditions cultivates 5 weeks, observe its upgrowth situation.
2. the Osmotic treatment before cold treatment：Cold treatment the last week stops watering, shadow of the moisture to cold treatment in reduction plant
Ring.
3. cold treatment：The tomato seedling taking-up for growing 5-6 weeks will be cultivated, choose growth conditions in 3 groups, size height phase
Seemingly, germinate seedling number identical difference cold-resistant variety, and setting 6 is parallel.Seedling after selection is put into 4 DEG C of refrigerators, illumination bar
Precooling treatment 4h under part, dark processing 8h in -4 DEG C of refrigerating chambers is placed into, 0 DEG C and 4 DEG C of illumination cool place cabinets are sequentially placed into after taking-up
Middle recovery 2h and 12h.
4. renewal cultivation：After recovery terminates, continue to be placed on 16h illumination cultivations room（21 DEG C of room temperature, humidity 65%）In,
The metamorphosis of its blade is observed after 24h and cuts the relative conductivity that every group of 6 parallel plant survey its ground plant.
5. the measure of relative conductivity：The material handled well is taken out from Culture basin, is put into 100ml syringes, adds
20min is vacuumized after water 20ml, is transferred in 250ml conical flasks, adds distilled water 80ml to be put into shaking table, 21 DEG C, 180r.min-1 shakes
Bed culture 2h, takes out and surveys electrical conductivity C1；Detection seals conical flask with aluminium-foil paper after terminating for the first time, boils 15min, cools down
To room temperature, C2 is surveyed after constant volume to 100ml；Statistics relative conductivity C=（C1—C0）/（C2-C0）, wherein C0 is distilled water conductance
Rate.
6. ground will be obtained after the different tomato of cold-resistant degree (cold-resistant strain, general strain, can not resist cold strain) cold treatment
The relative conductivity of upper overall plant carries out correlation analysis, it is established that the cold-resistant degree of tomato plant and relative conductivity size
Correlation, obtain distinguishing the value range of the electrical conductivity of tomato difference cold-resistant variety.
The tomato plants relative conductivity of three kinds of cold treatments see the table below 1：
After cold treatment method using this kind optimization, the relative conductivity value scope of strain plant of the same race reduces in 10%-13%
The discrete type of data, repeatability increase.The relative conductivity model 20.75%-33.67% of cold-resistant germline, normal germline it is relative
Conductivity range is 50.27%-62.34%, and the relative conductivity scope that can not resist cold germline is 78.88%-89.63%.
Obtained by changing data analysis：The relative conductivity that cold-resistant germline tomato plant obtains after treatment is minimum, no
The tomato plant relative conductivity of cold-resistant germline is maximum, and tomato plant relative conductivity circle of normal germline is between the two.Cause
This, relative conductivity carries out correlation analysis and obtained：The cold-resistant germline that it is tomato variety less than 33% that relative conductivity, which is,.
Brief description of the drawings
Fig. 1 is K2, the production restoration state diagram after P2 kind cold treatments.
Fig. 2 is K3, the production restoration state diagram after P3 kind cold treatments.
Embodiment：
The tomato breeding lines of experiment are that Chinese University of Science and Technology's school of life and health sciences provides, the skill by specific example to the present invention
Art scheme is further described：
The kind participated in the experiment is：The different tomato variety of three groups of cold-resistant degree, cold-resistant variety K1, K2, K3；Kind P1, P2 are can not resist cold,
P3.Take K1 and P1 to do under specified conditions the measure of the relative conductivity after cold treatment, experimental method with the content of the invention technical scheme,
K1 is measured, P1 conductivity value see the table below 2, K2, K3 and P2, continue to cultivate one week in normal culturing room after P3 cold treatments, growth
Situation is shown in Fig. 1, Fig. 2.
As a result show：Under above-mentioned condition cold treatment, the electrical conductivity of the tomato K systems of cold-resistant variety is
23.44%, less than 33%, cold-resistant variety is accredited as, during the cold treatment in the later stage recovers experiment, survival rate
It is very high.All death after cold treatment without cold-resistant variety.
Experiment shows, can be reflected entirely by determining the numerical value of relative conductivity of plant after above-mentioned ad hoc approach cold treatment
The cold-resistant sex differernce of plant, it is consistent that the evaluation result and tomato plant of this kind of cold-resistant identification damage to plants caused by sudden drop in temperature rear restoration ecosystem phenotype, table
The bright data are feasible..
Claims (3)
1. rapid screening method and the application of a kind of tomato Evaluation of Cold Tolerance, it is characterised in that carry out according to the following steps：Cultivate bar
The control of part, the Osmotic treatment before cold treatment, the control of cold treatment condition, the measure of relative conductivity, establish relative conductivity
Correlation between the degree that resisted cold with the whole strain of tomato：
（1）The culture of tomato seedling：Soil is after planting irrigated first, is weekly watered later, condition of culture is 21 DEG C of room temperature,
Humidity 65%, 16h illumination.
2. after culture 5 weeks, its upgrowth situation is observed；
（2）Osmotic treatment before cold treatment：Cold treatment the last week stops watering；
（3）Cold treatment：The tomato seedling that growth 5-6 weeks will be cultivated is taken out, and selection growth conditions are similar, and different cold-resistant varieties are each
3 groups, every group 6 parallel.
3. seedling is put into 4 DEG C of refrigerators into precooling treatment 4h under illumination condition, dark processing 8h in -4 DEG C of refrigerating chambers is placed into, is taken
Recovery 2h and 12h in 0 DEG C and 4 DEG C of illumination cool place cabinets is sequentially placed into after going out；
（4）After recovery terminates, continue to be placed on 16h illumination cultivations room（21 DEG C of room temperature, humidity 65%）In, its blade is observed after 24h
Metamorphosis and survey the relative conductivity of its ground plant；
（5）The measure of in vitro plant electrical conductivity after tomato cold treatment：The material handled well is taken out from Culture basin, vacuumized
20min；Add water and be put into shaking table 2h（21 DEG C, 180r.min-1）Take out afterwards and survey electrical conductivity C1；15min is boiled, is cooled to room
Temperature（Constant volume）, survey C2；Statistics relative conductivity C=（C1—C0）/（C2-C0）, wherein C0 is distillation water conductivity；
（6）The correlation established between relative conductivity and the cold-resistant degree of the whole strain of tomato, obtains the electrical conductivity of cold-resistant tomato variety
Scope be 0-35%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575600A (en) * | 2018-04-24 | 2018-09-28 | 江苏省农业科学院 | A method of quick and precisely comparing tomato variety lower temperature resistance |
| CN110100593A (en) * | 2019-05-10 | 2019-08-09 | 浙江大学 | The identification method of tea tree breed cold-hartliness |
| CN111587745A (en) * | 2020-06-05 | 2020-08-28 | 浙江农林大学 | Cherry tomato variety screening method suitable for overwintering seedling raising |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101695278A (en) * | 2009-10-23 | 2010-04-21 | 合肥工业大学 | Method for quickly screening and testing cold resistance of tomatoes |
| CN102415253A (en) * | 2011-08-25 | 2012-04-18 | 刘慧民 | Method for screening cold-resistant variety of steeplebush |
-
2017
- 2017-07-24 CN CN201710604796.5A patent/CN107389738A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101695278A (en) * | 2009-10-23 | 2010-04-21 | 合肥工业大学 | Method for quickly screening and testing cold resistance of tomatoes |
| CN102415253A (en) * | 2011-08-25 | 2012-04-18 | 刘慧民 | Method for screening cold-resistant variety of steeplebush |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575600A (en) * | 2018-04-24 | 2018-09-28 | 江苏省农业科学院 | A method of quick and precisely comparing tomato variety lower temperature resistance |
| CN110100593A (en) * | 2019-05-10 | 2019-08-09 | 浙江大学 | The identification method of tea tree breed cold-hartliness |
| CN110100593B (en) * | 2019-05-10 | 2021-04-23 | 浙江大学 | Method for identifying cold resistance of tea tree variety |
| CN111587745A (en) * | 2020-06-05 | 2020-08-28 | 浙江农林大学 | Cherry tomato variety screening method suitable for overwintering seedling raising |
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