CN101695278A - Method for quickly screening and testing cold resistance of tomatoes - Google Patents
Method for quickly screening and testing cold resistance of tomatoes Download PDFInfo
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- CN101695278A CN101695278A CN200910184966A CN200910184966A CN101695278A CN 101695278 A CN101695278 A CN 101695278A CN 200910184966 A CN200910184966 A CN 200910184966A CN 200910184966 A CN200910184966 A CN 200910184966A CN 101695278 A CN101695278 A CN 101695278A
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Abstract
The invention discloses a method for quickly screening and testing the cold resistance of tomatoes, which comprises: firstly, culturing tomato seedlings growing for 3 to 5 weeks, and processing stem apex leaves at 3 to 5 DEG C for 1 to 3 hours and at 3 DEG C below zero to 5 DEG C below zero for 7 to 9 hours; secondly, restoring the stem apex leaves at 3 to 5 DEG C for 10 to 14 hours and placing the stem apex leaves in light at 20 to 22 DEG C for 14 to 18 hours; and finally, measuring the electrical conductivity of the cold-treated and restored leaves, placing the leaves in distilled water, performing vacuumization and volume fixing, shaking the leaves at 21 to 23 DEG C under a sealed condition for 1 to 3 hours, measuring an electrical conductivity C1, sealing the leaves, boiling the leaves for 10 to 20 minutes, cooking the leaves to room temperature, measuring an electrical conductivity C2 and calculating a relative conductivity according to a formula of C=(C1-C0)/(C2-C0)x100, wherein when the value domain of the relative conductivity C is less than or equal to 42 percent, the tomato variety is a cold-resistance variety. In the invention, the cold resistance screening of the whole system is performed under controllable conditions, the result is reliable, the method is simple, convenient and easy to master, the breeding period is shortened greatly and the method can be used for test the cold resistance of an introduced variety at the same time.
Description
One, technical field
The present invention relates to a kind of discrimination method of fruit trees and vegetables cold resistance kind, exactly is the method for a quickly screening and testing cold resistance of tomatoes.
Two, background technology
Low temperature freezing-disaster is a kind of serious natural calamity during agriculture and forestry are produced, and according to statistics, the annual loss that therefore causes in the world is up to 2,000 hundred million dollars.Therefore, the seed selection of cold-resistant crop kind has become key issue that needs to be resolved hurrily in the agricultural production.
Tomato is that a kind of economic benefit is than higher vegetable crop.Its fruits nutrition is abundant, contains multivitamin, is one of modern's best nutritional food, and tomato also is the cosmetics of health food, functional food and the ladies that like to be beautiful favor simultaneously.Tomato is former to originate from South America, China's common cultivated, and grow seedlings in protection ground in general Winter-Spring, and spring cultivation is main, and winter greenhouse is cultivated.Though tomato in China has obtained the kind that much is possessed of good qualities, but the kind of tolerance cold and freeze injury seldom, tomato in China is done vegetables for cultivation throughout the year in addition, and the expert attempts to cultivate the tomato new cold-resistant species by technique for gene engineering and cell technology engineering both at home and abroad in recent years.Therefore, seek a kind of rapid screening and identify that the method for cold-resistant tomato variety seems particularly necessary.
The method amount of the tomato germ plasm resource of the good cold-resistant proterties of the tradition natural cold climate approach Screening and Identification of dependence is big, and the cycle is long, shortcomings such as proterties instability.Therefore, the invention provides the method that a kind of rapid screening is identified cold-resistant tomato variety (being), can shorten the seed selection cycle of cold-resistant tomato variety greatly.
Three, summary of the invention
The present invention is directed to the defective of existing tomato cold-resistant variety process for screening and identifying, aim to provide a kind of quick simple and effective process for screening and identifying to accelerate the process of the cold-resistant breeding of tomato, technical problem to be solved is the reliability of process for screening and identifying, avoids losing of fine germplasm resources simultaneously.
Thinking of the present invention is to establish histoorgan in the cold-resistant plant of tomato and the correlation between certain physical quantity under given conditions, thereby the Screening and Identification work of tomato variety cold resistance is converted into the detection of this physical quantity.Research by experiment, the present invention utilizes the cold treatment under certain cold stress conditions, reflect the plant degree that resists cold according to the numerical value of tomato excised leaf relative conductivity, can make fast and effectively the power of the tolerance to cold of plant and estimating, accelerate the process of the cold-resistant breeding of tomato.
Be kind cold resistance of establishment tomato and the correlation between the relative conductivity size, at first in existing tomato variety respectively to cold-resistant, mediumly cold-resistant carry out the relative conductivity assay determination with frigolabile three kinds of kind excised leafs, cooperate the restorative test of blade, observe its mode of appearance variation characteristic, this method is optimized, by optimizing the cold treatment condition of determining blade, the relative conductivity testing conditions, obtain the corresponding relation between tomato variety cold resistance and the relative conductivity numerical value simultaneously, in view of the above the tomato variety of cold resistance the unknown is experimentized then and differentiate checking, establish technical scheme of the present invention at last.
Cold-resistant, medium cold-resistant being meant by tradition nature cold climate approach with frigolabile filter out the tomato germplasm of different cold-resistant proterties in the existing tomato variety according to the difference of its survival rate and output size.
Technical scheme of the present invention comprises the cultivation of tomato seedling, cold treatment and recovery, the mensuration of electrical conductivity and the calculating of relative conductivity of excised leaf, described cold treatment and recovery are exactly to get growth 3～5 all tomato seedling stem end blades to handle 1～3 hour under 3～5 ℃ of conditions, under-3～-5 ℃ of conditions, handled 7～9 hours again, under 3～5 ℃ of conditions, recover 10～14h then, at last 20～22 ℃ of following illumination 14～18 hours; The mensuration of described electrical conductivity is exactly that the blade that will recover after the cold treatment vacuumized in distilled water 15～25 minutes, then with the distilled water constant volume, seal, under 21～23 ℃, vibration 1～3 hour, survey electrical conductivity C
1, the 10～20min that seals, seethes with excitement is cooled to room temperature and surveys electrical conductivity C
2Relative conductivity C=(C
1-C
0)/(C
2-C
0) * 100%, C in the formula
0Be the distilled water electrical conductivity.When the codomain of relative conductivity C≤42% then is the cold-resistant variety tomato.
Compare with the conventional screen selecting technology, the inventive method is easy, is easy to grasp, and has shortened breeding cycle greatly.The cold resistance Screening and Identification of whole system is all carried out under controlled condition, make The selection result more reliable, utilize the relative conductivity of excised leaf can reflect the destructiveness of low temperature cell membrane, simultaneously in conjunction with the recovery growth experiment after damaging to plants caused by sudden drop in temperature, both identified the direct tolerance of cell, identified the growth recovery ability of material after removing low temperature again injury from low temperature.In addition, the present invention also can be used for introducing a fine variety cold resistance and identifies, can accelerate new varieties and identify speed, puts into production as early as possible, can improve the validity of the cold-resistant screening mutant of tomato, avoids losing of contingent in process of production improved seeds resource.
Four, description of drawings
Fig. 1 is the seedling phenotype after around A, B, three kinds of development growths of C.
Fig. 2 is that the blade phenotype after A, B, three kinds of excised leaf cold treatments of C recover changes.The A kind blade degree minimum of crispaturaing is green, and the C kind blade degree maximum of crispaturaing is aterrimus, and the B kind falls between.
Fig. 3 is A, B, the diagram of three kinds of relative conductivity data of C.
Fig. 4 is in the demonstration test 1
#, 2
#, 3
#Recover upgrowth situation figure after three kinds of seedling cold treatments.
Five, embodiment
Experiment is provided by Academy of Agri-Science and Technology Anhui Province gardening research institute with tomato variety.Wherein (1) cold-resistant tomato variety (being) is named: A
1, A
2, A
3(2) medium cold-resistant tomato variety, name: B
1, B
2, B
3(3) not cold-resistant tomato variety, name: C
1, C
2, C
3Every group of three parallel tests.
1, the cultivation of tomato seedling:
With A
1, A
2, A
3, B
1, B
2, B
3And C
1, C
2, C
3Tomato seeds is seeded in the earth culture flowerpot, and soil is configured to vermiculite: black earth: perlite=9: 3: 0.5, the black earth 20min that need sterilize before using.Sufficient plant soil medium nutrient solution is watered in sowing first, waters once weekly later on.At 21 ℃, after the 16h illumination condition cultivated for 4 weeks down, its upgrowth situation was seen Fig. 1.
2, the cold treatment of tomato excised leaf:
Tomato seedling around the incubation growth is taken out, and its stem end blade of clip keeps the blade size of institute's clip consistent, with A as far as possible
1, B
1, C
1Be one group, put into plate, count three groups, three every group parallel.Plate is put into 4 ℃ of refrigerator 2h, put into-4 ℃ of refrigerating chambers again and handle 8h, put into 4 ℃ of refrigerators after the taking-up and recover 12h, at last plate is placed in the 16h illumination cultivation chamber (about 21 ℃ of room temperatures), the metamorphosis of its blade is seen Fig. 2 behind the 24h.
3, the mensuration of excised leaf electrical conductivity after the tomato cold treatment:
The conductance measurement method: the material after the cold treatment recovery is taken out from plate, and every kind of 3 of material is parallel.1. vacuumize 20min with 50ml syringe adding distil water 10ml; 2. change over to again in the 50ml triangular flask, adding distil water 40ml (constant volume 50ml), the aluminium film seals, and puts into 22 ℃ of constant temperature gas/shaking baths, 180r.min
-1Rotation, regularly 2h; 3. take out, survey electrical conductivity C
14. institute is surveyed solution aluminium film and seal, boil 15min, be cooled to room temperature (constant volume 50ml), survey C
25. calculate relative conductivity C=(C
1-C
0)/(C
2-C
0) * 100%, wherein C
0Be the distilled water electrical conductivity.Three groups of relative conductivity magnitude range see Table 1, and Fig. 3 is seen in data analysis.
Table 1
Hence one can see that, and the relative conductivity of the excised leaf of cold-resistant variety is less than not cold-resistant variety, and the relative conductivity of medium kind is between resisting cold and not resisting cold.Therefore, relative conductivity size after cold-resistant degree of the whole strain of tomato and the excised leaf cold treatment is set up correlative relationship is: relative conductivity is at the cold-resistant variety that is below 42.62%, between the 42.62%-62.67% is medium cold-resistant variety, must be cold-resistant variety not greater than 62.67%.
Statistical analysis obtains: behind-4 ℃ of cold treatment growths tomato excised leaf all around, the relative conductivity of cold-resistant variety more than 90% is below 42.62%, the electrical conductivity of cold-resistant variety more than 90% be not more than 62.67%: the electrical conductivity of medium cold-resistant variety more than 90% is (table 1, Fig. 3) between the 47.62%-62.67% near 52.05%.
4, demonstration test
Get 3 tomato varieties of cold-resistant proterties the unknown, (provided by Anhui Province's academy of agricultural sciences horticulture, cold-resistant character pair applicant is unknown) called after 1
#, 2
#, 3
#The identical stem apex blade (three parallel) of clip size is put into plate, 4 ℃ of refrigerator and cooled are handled 2h, put into-4 ℃ of refrigerating chambers again and handle 8h, put into 4 ℃ of refrigerators after the taking-up and recover 12h, at last plate is placed in the 16h illumination cultivation chamber (about 21 ℃ of room temperatures), surveys its conductivity value (table 2) behind the 24h.
Table 2
With 1 of checking gained
#Cold-resistant, 2
#Medium cold-resistant, 3
#Seedling plant-6 ℃ cold treatment 8h around cold-resistant three kinds are cultivated, behind 4 ℃ of recovery 12h, 16h illumination cultivation chamber (about 21 ℃ of room temperatures) recovered to cultivate a week, recovers upgrowth situation and sees Fig. 4.Among the figure as seen, 1
#Cold-resistant degree is best, and 2
#Take second place 3
#The poorest.
Claims (1)
1. the method for a quickly screening and testing cold resistance of tomatoes, comprise the cultivation of tomato seedling, cold treatment and recovery, the mensuration of electrical conductivity and the calculating of relative conductivity of excised leaf, it is characterized in that: described cold treatment and recovery are to get growth 3～5 all tomato seedling stem end blades to handle 1～3 hour under 3～5 ℃ of conditions, under-3～-5 ℃ of conditions, handled 7～9 hours again, under 3～5 ℃ of conditions, recovered 10～14 hours illumination 14～18 hours under 20～22 ℃ of conditions more then; The mensuration of described electrical conductivity is exactly that the blade that will recover after the cold treatment vacuumized in distilled water 15～25 minutes, then with the distilled water constant volume, seal, in 21～23 ℃ of vibrations 1～3 hour, survey electrical conductivity C
1, seal, seethe with excitement 10～20 minutes again, be cooled to room temperature and survey electrical conductivity C
2Relative conductivity C=(C
1-C
0)/(C
2-C
0) * 100%, C in the formula
0Be the distilled water electrical conductivity; When the codomain of relative conductivity C≤42%, be cold-resistant tomato variety.
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| CN2009101849664A CN101695278B (en) | 2009-10-23 | 2009-10-23 | Method for quickly screening and testing cold resistance of tomatoes |
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|---|---|---|---|
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| CN101695278B CN101695278B (en) | 2011-11-23 |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102415253A (en) * | 2011-08-25 | 2012-04-18 | 刘慧民 | Method for screening cold-resistant variety of steeplebush |
| CN102726186A (en) * | 2012-06-11 | 2012-10-17 | 西北农林科技大学 | Seedling identification method for cold-resistant breeding of watermelon |
| CN106804280A (en) * | 2017-04-01 | 2017-06-09 | 中国农业科学院油料作物研究所 | A kind of method of Rapid identification rape cold resistance |
| CN106896128A (en) * | 2017-04-20 | 2017-06-27 | 黑龙江省农业科学院耕作栽培研究所 | A kind of method of red bean cold hardness evaluation with evaluating |
| CN107389738A (en) * | 2017-07-24 | 2017-11-24 | 安徽普元生物科技股份有限公司 | A kind of rapid screening method of tomato Evaluation of Cold Tolerance and application |
| CN109618896A (en) * | 2019-01-22 | 2019-04-16 | 温州科技职业学院 | A kind of rapid screening method of tomato Evaluation of Cold Tolerance and application |
| CN110073974A (en) * | 2019-06-04 | 2019-08-02 | 中国农业科学院棉花研究所 | It is a kind of to screen the method and its application for resisting cold cotton 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 |
| CN111855586A (en) * | 2020-06-04 | 2020-10-30 | 广东省农业科学院果树研究所 | Method for rapidly screening pineapple variety cold resistance |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BG51851A1 (en) * | 1993-03-08 | 1993-10-15 | Bg Inst Genetika Doncho Kostov | Method for creating genotype plants tolerant to low temperatures |
| MD3638G2 (en) * | 2008-02-11 | 2009-02-28 | Институт Генетики, Физиологии И Защиты Растений Академии Наук Молдовы | Method of selecting tomato cold resistant genotypes |
-
2009
- 2009-10-23 CN CN2009101849664A patent/CN101695278B/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102415253A (en) * | 2011-08-25 | 2012-04-18 | 刘慧民 | Method for screening cold-resistant variety of steeplebush |
| CN102726186A (en) * | 2012-06-11 | 2012-10-17 | 西北农林科技大学 | Seedling identification method for cold-resistant breeding of watermelon |
| CN106804280A (en) * | 2017-04-01 | 2017-06-09 | 中国农业科学院油料作物研究所 | A kind of method of Rapid identification rape cold resistance |
| CN106896128A (en) * | 2017-04-20 | 2017-06-27 | 黑龙江省农业科学院耕作栽培研究所 | A kind of method of red bean cold hardness evaluation with evaluating |
| CN106896128B (en) * | 2017-04-20 | 2020-02-21 | 黑龙江省农业科学院耕作栽培研究所 | Method for identifying and evaluating cold resistance of small red beans |
| CN107389738A (en) * | 2017-07-24 | 2017-11-24 | 安徽普元生物科技股份有限公司 | A kind of rapid screening method of tomato Evaluation of Cold Tolerance and application |
| CN109618896A (en) * | 2019-01-22 | 2019-04-16 | 温州科技职业学院 | A kind of rapid screening method of tomato Evaluation of Cold Tolerance and application |
| CN110073974A (en) * | 2019-06-04 | 2019-08-02 | 中国农业科学院棉花研究所 | It is a kind of to screen the method and its application for resisting cold cotton variety |
| CN111855586A (en) * | 2020-06-04 | 2020-10-30 | 广东省农业科学院果树研究所 | Method for rapidly screening pineapple variety cold resistance |
| 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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|---|---|
| CN101695278B (en) | 2011-11-23 |
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