CN105409708B - A kind of identification method of V. amurensis germ plasm resource rest period winter resistance - Google Patents

A kind of identification method of V. amurensis germ plasm resource rest period winter resistance Download PDF

Info

Publication number
CN105409708B
CN105409708B CN201510814158.7A CN201510814158A CN105409708B CN 105409708 B CN105409708 B CN 105409708B CN 201510814158 A CN201510814158 A CN 201510814158A CN 105409708 B CN105409708 B CN 105409708B
Authority
CN
China
Prior art keywords
amurensis
branch
dyeing
rank
temperature
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN201510814158.7A
Other languages
Chinese (zh)
Other versions
CN105409708A (en
Inventor
赵滢
艾军
杨义明
范书田
王振兴
王春伟
何伟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute Special Animal and Plant Sciences CAAS
Original Assignee
Institute Special Animal and Plant Sciences CAAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute Special Animal and Plant Sciences CAAS filed Critical Institute Special Animal and Plant Sciences CAAS
Priority to CN201510814158.7A priority Critical patent/CN105409708B/en
Publication of CN105409708A publication Critical patent/CN105409708A/en
Application granted granted Critical
Publication of CN105409708B publication Critical patent/CN105409708B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G17/00Cultivation of hops, vines, fruit trees, or like trees
    • A01G17/02Cultivation of hops or vines
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01CPLANTING; SOWING; FERTILISING
    • A01C1/00Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00Botany in general
    • A01G7/06Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Ecology (AREA)
  • Forests & Forestry (AREA)
  • Botany (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Soil Sciences (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The present invention provides a kind of identification methods of V. amurensis germ plasm resource rest period winter resistance, include the following steps:By the V. amurensis branch of rest period under different cryogenic temperatures de-epithelization after freezing processing, be put into chloro triphenyltetrazolium chloride dyeing liquor, be placed in isoperibol and carry out dyeing processing;The dyeing liquor for washing away V. amurensis branch surface determines dyeing rank according to V. amurensis branch xylem stained area, and is come out of retirement and taken up an official post the staining index of grape branch according to dyeing level calculation;Staining index corresponding to the cryogenic temperature of V. amurensis branch determines half lethal low temperature.Half lethal low temperature is lower, shows that the winter resistance of the V. amurensis germplasm is better.It is smaller due to being influenced by extraneous factor in continuous mode using TTC decoration methods, and visual result, it is therefore, simple and easy to do using this method identification V. amurensis germ plasm resource rest period winter resistance, accurately and reliably.

Description

A kind of identification method of V. amurensis germ plasm resource rest period winter resistance
Technical field
The present invention relates to seeds cold hardness evaluation fields, anti-in particular to a kind of V. amurensis germ plasm resource rest period Cold identification method.
Background technology
The V. amurensis northeast originating in China (Vitis amurensis Rupr.) and Korea, the Far East Area of the former Soviet Union.China mountain The natural distribution area of grape mainly Jilin Province Changbai Mountain, Wandashan Mountains, Heilongjiang Province, the Xiaoxinanlin Mountains, Liaoning Province the north mountain Area, upper level, the Daqunshan Mountains to the east of Wulanchabu League of Inner Mongolia alliance are also distributed.It is the primary raw material that northeast makes grape wine, Its wine is ruby, bright-coloured transparent, has unique flavor, welcomed by consumers.
V. amurensis is one kind of most cold-resistant in Vitis, and branches and tendrils are resistant to -40 DEG C of severe cold, and root system is resistant to -14~-16 DEG C Low temperature also has stronger resistivity to white rot, powdery mildew, anthracnose and black soya bean disease etc..Therefore, it is current domestic and international Portugal The most important resource of grape cold-tolerance breeding.
The winter resistance of V. amurensis have higher genetic force, its cold tolerance significant difference of different wild grape resources, soon Fast, the reliable relevant index of V. amurensis winter resistance can be provided for the screening of V. amurensis cold hardness evaluation and cold-resistant kind it is important according to According to.But it is still not clear at present suitable for the method for V. amurensis cold hardness evaluation, it is strong to the winter resistance of separate sources V. amurensis germplasm Weak understanding is insufficient, affects the development and utilization of winter resistance germ plasm resource.
The strong and weak most common method of existing identification Vitis winter resistance is conductance method, however, we are in practical measurement It was found that due to CO in temperature, soaking time and air2The factors such as dissolving the electric conductivity value of practical measurement can all be had an impact, Cause measurement result less reproducible, qualification result is inaccurate.
In view of this, special propose the present invention.
Invention content
The purpose of the present invention is to provide a kind of identification methods of V. amurensis germ plasm resource rest period winter resistance.For current Due to CO in temperature, soaking time and air when Means of Electrical Conductivity Vitis winter resistance2The factors such as dissolving cause to measure and tie Fruit is less reproducible, the problem of qualification result inaccuracy, and the present invention identifies V. amurensis germ plasm resource suspend mode using TTC decoration methods The winter resistance of phase.Since TTC decoration methods are by the way that the V. amurensis branch after freezing processing is put into chloro triphenyltetrazolium chloride dye Carry out observing branch xylem staining conditions after dyeing processing in color liquid, in continuous mode by extraneous factor influenced compared with It is small, it is therefore, simple and easy to do using this method measurement V. amurensis germ plasm resource rest period winter resistance, accurately and reliably.
In order to realize that the above-mentioned purpose of the present invention, spy use following technical scheme:
A kind of identification method of V. amurensis germ plasm resource rest period winter resistance, includes the following steps:
(1) by the V. amurensis branch of rest period under different cryogenic temperatures de-epithelization after freezing processing, be put into chloro three In phenyl tetrazole dyeing liquor, it is placed in isoperibol and carries out dyeing processing;
(2) dyeing liquor for washing away V. amurensis branch surface is determined according to the stained area of V. amurensis branch xylem and is dyed Rank, and come out of retirement and taken up an official post the staining index of grape branch according to dyeing level calculation;
(3) staining index corresponding to the cryogenic temperature of V. amurensis branch determines half lethal low temperature.
Chloro triphenyltetrazolium chloride (i.e. TTC) is a kind of color developing agent, can be by the enzyme system institute in active cell Reduction forms red San Ben Ji Jia Za (TTF) not soluble in water.The present invention identifies that V. amurensis germplasm provides using TTC decoration methods The winter resistance of source rest period is peelled off after the V. amurensis branch after freezing processing is carried out freezing processing under different cryogenic temperatures Epidermis is put into TTC dyeing liquors and is dyed.
The red area of V. amurensis branch xylem is bigger after dyeing, then the dyeing for grape branch of going down the hill in the cryogenic temperature Rank is higher, and staining index is bigger.By determining the V. amurensis germplasm after calculating the staining index corresponding to different cryogenic temperatures Half lethal low temperature LT50To weigh the winter resistance of V. amurensis germplasm.LT50It is smaller, show the cold-resistant of the V. amurensis germplasm Property is better.
It is smaller due to being influenced by extraneous factor in continuous mode using TTC decoration methods, and therefore visual result uses This method identifies that V. amurensis germ plasm resource rest period winter resistance is simple and easy to do, accurately and reliably.
Preferably, in step (2), described the step of determining dyeing rank according to the staining conditions of V. amurensis branch xylem It specifically includes:
By V. amurensis branch it is longitudinal sectional after, according to the stained area of V. amurensis branch vertical section xylem determine dyeing rank.
It is by observing branch cross section clip wood currently, in being studied about TTC Determination Staining shoot tissue vigor Matter portion colours depth degree to be judged.In practical operation, the red TTF of generation is dyed often outside the clip of branch cross section Side is accumulated, and xylem hyperchromasia is caused to generate observation error;And branch cross section thin slice is cut using free-hand slicing method and is difficult Operation;Xylem coloring depth degree range estimation is also difficult to accurate judgement.In the present invention, TTC decoration methods, dyeing observation portion are improved Position is changed to the xylem of branch vertical section, can longitudinally be cut with sharp cutter, easy to operate.
Meanwhile vertical section xylem is in regular rectangular shape substantially, is conducive to stained area estimation so that dyeing gradational boundary is clear Chu can follow, as a result more accurate and reliable to overcome the shortcomings of conventional method.
For the ease of dyeing, coloring is improved, it is preferable that described under different cryogenic temperatures at freezing in step (1) Further include following steps before being put into chloro triphenyltetrazolium chloride dyeing liquor after reason after de-epithelization:
The V. amurensis branch of de-epithelization is cut into the branch section of 0.3~0.5cm.
In order to ensure the accuracy of result, it is further preferred that branch section in the chloro triphenyltetrazolium chloride dyeing liquor Number be 10~15.
Similarly, for the ease of dyeing, coloring is improved, it is further preferred that in step (1), the dyeing processing Temperature be 28~32 DEG C, dyeing time 18~for 24 hours, and being carried out under the conditions of being protected from light.
Preferably, in step (1), a concentration of the 0.5~1.0% of the chloro triphenyltetrazolium chloride dyeing liquor.
The configuration method of chloro triphenyltetrazolium chloride dyeing liquor used is:Weigh the chlorinated triphenyl base four of 0.5~1.0g Nitrogen azoles, with deionized water dissolving and constant volume is in 100mL volumetric flasks, is protected from light after preparing, room temperature preservation.
Dehydrogenase content in Different Crop tissue is different, and activity is different, thus the dyeing condition needed is also different.This hair In bright, suitable TTC dyeing conditions, including dyeing branch segment length, stin of thickness, dyeing temperature, dyeing time have been screened, this A little parameter synergistic effects, improve coloring jointly.
Preferably, described to be specifically included the step of freezing processing under different cryogenic temperatures in step (1):
The V. amurensis branch of rest period is placed at -20~-50 DEG C 12~15h of freezing, after raise the temperature to 20~30 DEG C, and 4-6h is kept at this temperature.
Preferably, it is specifically included the step of freezing processing under different cryogenic temperatures for described every group:
The temperature of V. amurensis branch local environment is down to set temperature according to the cooling rate of 4~5 DEG C/h, in this temperature Degree is lower to keep 12~13h, after be gradually heating to 20~30 DEG C, keep 4~5h at this temperature.
By the V. amurensis branch after above-mentioned freezing processing, when carrying out cold hardness evaluation, result is more accurate.
Preferably, in step (2), the determination method of the dyeing rank is:
When xylem vertical section has 85% or more area to take on a red color, then the dyeing rank is 4 grades;When xylem is longitudinal sectional Face has 46%~85% area to take on a red color, then the dyeing rank is 3 grades;When xylem vertical section has 20%~45% face Product takes on a red color, then the dyeing rank is 2 grades;When xylem vertical section has 20% area below to take on a red color, then the dyeing Rank is 1 grade.
It is further preferred that in step (2), the staining index=Σ (dyeing rank × branch hop count)/(branch section sum × Highest dyes rank).
The staining index determined by the above method can more truly reflect the winter resistance of different V. amurensis kinds, can For distinguishing the winter resistance between different V. amurensis kinds.
Compared with prior art, beneficial effects of the present invention are:
(1) due to CO in temperature, soaking time and air when being directed to current Means of Electrical Conductivity Vitis winter resistance2It is molten The factors such as solution cause measurement result less reproducible, the problem of qualification result inaccuracy, and the present invention uses chloro triphenyltetrazol Azoles decoration method identifies the winter resistance of V. amurensis germ plasm resource rest period.Since TTC decoration methods are by will be after freezing processing V. amurensis branch is put into chloro triphenyltetrazolium chloride dyeing liquor carry out dyeing processing after observation branch xylem staining conditions i.e. Can, it is influenced by extraneous factor in continuous mode smaller, therefore, V. amurensis germ plasm resource rest period cold-resistant is measured using this method Property is simple and easy to do, accurately and reliably.
(2) currently, in being studied about TTC Determination Staining shoot tissue vigor, cut by observing branch cross section Mouth xylem colours depth degree to be judged.In practical operation, the red San Ben Ji Jia Za (TTF) for dyeing generation often exists Accumulation on the outside of the clip of branch cross section, causes xylem hyperchromasia to generate observation error;And it is cut using unarmed cross section method Thin slice is difficult operation;Xylem coloring depth degree range estimation is also difficult to accurate judgement.In the present invention, TTC decoration methods are improved, are contaminated Color look-out station is changed to the xylem of branch vertical section, can longitudinally be cut with sharp cutter, easy to operate.Meanwhile vertical section Xylem is in regular rectangular shape substantially, is conducive to stained area estimation so that dyeing gradational boundary understands and can follow, to overcome tradition side The deficiency of method, it is as a result more accurate and reliable.
(3) in the present invention, suitable TTC dyeing conditions, including dyeing branch segment length, stin of thickness, dyeing have been screened Temperature, dyeing time, these parameters synergistic effect, improve coloring jointly.
(4) operating condition of the invention is not harsh, and TTC concentration is in 0.5~1.0% (W (g)/V (mL)) in operating process Can, be not necessarily to particularly accurate preparation, stained area is easy to estimate, and each step operation is simple, to instrument, reagent etc. require compared with It is low, it can be promoted and applied in V. amurensis germplasm rest period cold hardness evaluation.
(5) mirror the present invention provides staining index and half lethal low temperature as V. amurensis germ plasm resource winter resistance Determine index, the winter resistance of different V. amurensis kinds can be accurately distinguished.
Description of the drawings
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below There is attached drawing needed in technology description to be briefly described.
Fig. 1 is the schematic diagram that V. amurensis branch difference dyes rank;
Fig. 2 is the matched curve of " double rich " and " northern ice is red " that are obtained according to assay method of the present invention;
Fig. 3 correlations between the obtained half lethal low temperature of assay method of the present invention and cold- resisting index average membership Analysis result figure.
Specific implementation mode
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is The conventional products that can be obtained by commercially available purchase.
Embodiment 1
The assay method of V. amurensis germ plasm resource rest period winter resistance provided in this embodiment, includes the following steps:
The freezing processing of V. amurensis branch:
The V. amurensis branch of " double rich " this germ plasm resource is selected to carry out freezing processing.By the mountain of the germ plasm resource rest period Grape branch carries out freezing processing at -25 DEG C, -30 DEG C, -35 DEG C, -40 DEG C, -45 DEG C, -50 DEG C respectively.By V. amurensis branch Place 10h under above-mentioned 6 cryogenic temperatures, after be warming up to 20 DEG C after place 3h again.
The V. amurensis branch of rest period used is to acquire V. amurensis germplasm annual, that maturity is consistent mid-January to stop Dormancy phase branch is cut into the branch section of long 20~25cm and is packed into the valve bag of sealing after atoleine sealing, is protected in -15 DEG C of refrigerators It deposits.
Freeze the dyeing processing of branch:
By the V. amurensis branch de-epithelization Jing Guo above-mentioned freezing processing, the V. amurensis branch of de-epithelization is put into concentration For in 1.5% chloro triphenyltetrazolium chloride dyeing liquor, 15h is dyed in the light protected environment at 20 DEG C.
V. amurensis shoot tissue vitality test:
By above-mentioned dyeing, treated that V. amurensis branch is rinsed with deionized water, with blade that branch is crosscutting, and observation is crosscutting The stained area of face xylem determines dyeing rank, goes out staining index according to dyeing level calculation.
According to every group of cryogenic temperature and its corresponding staining index, improved Logistic is combined with nonlinear regression analysis Equation model temperature inflection point analyzes the half lethal low temperature for determining each germ plasm resource branch using DPS9.0 softwares.
In the present embodiment, the determination method for dyeing rank is:When xylem vertical section has 85% or more area in red Color, then the dyeing rank is 4 grades;When xylem vertical section has the area of 46%-85% to take on a red color, then the dyeing rank is 3 grades;When xylem vertical section has the area of 20%-45% to take on a red color, then the dyeing rank is 2 grades;When xylem vertical section There is 20% area below to take on a red color, then the dyeing rank is 1 grade.
The calculation formula of staining index is:Staining index=Σ (dyeing rank × branch hop count)/(branch section sum × highest dye Color rank).
Embodiment 2
In the assay method of V. amurensis germ plasm resource rest period winter resistance provided in this embodiment, the freezing of V. amurensis branch Processing, the dyeing processing step of freezing branch are identical with embodiment 1.
Unlike the V. amurensis shoot tissue vitality test the step of in, in the present embodiment,
By above-mentioned dyeing, treated that V. amurensis branch is rinsed with water, with blade that branch is longitudinal sectional, and observation vertical section is wooden The stained area in portion determines dyeing rank, goes out staining index according to dyeing level calculation.
Embodiment 3
The identification method of V. amurensis germ plasm resource rest period winter resistance provided in this embodiment, includes the following steps:
The freezing processing of V. amurensis branch:
The V. amurensis branch of " double rich " this germ plasm resource is selected to carry out freezing processing.By the mountain of the germ plasm resource rest period The grape branch freezing processing at -20 DEG C, -25 DEG C, -30 DEG C, -35 DEG C, -40 DEG C, -45 DEG C, -50 DEG C respectively.By V. amurensis branch Local environment is down to the cryogenic temperature of setting according to 4 DEG C/h cooling rates, and places 12h under corresponding cryogenic temperature, after 4h is placed again after being warming up to 20 DEG C.
The V. amurensis branch of rest period used is to acquire V. amurensis germplasm annual, that maturity is consistent mid-January to stop Dormancy phase branch is cut into the branch section of long 20~25cm and is packed into the valve bag of sealing after atoleine sealing, is protected in -15 DEG C of refrigerators It deposits.
Freeze the dyeing processing of branch:
By the V. amurensis branch de-epithelization Jing Guo above-mentioned freezing processing, the V. amurensis branch of de-epithelization is cut into 0.3~ The branch section of 0.5cm, every group of freezing processing are cut into 10 branch sections, after be put into a concentration of 0.5% tetrazole dyeing liquor, 18h is dyed in light protected environment at 28 DEG C.
V. amurensis shoot tissue vitality test:
By above-mentioned dyeing, treated that V. amurensis branch section is rinsed with water, with blade that branch section is longitudinal sectional, observes vertical section The stained area of xylem determines dyeing rank, goes out staining index according to dyeing level calculation.
According to every group of cryogenic temperature and its corresponding staining index, improved Logistic is combined with nonlinear regression analysis Equation model temperature inflection point analyzes the half lethal low temperature for determining each germ plasm resource branch using DPS9.0 softwares.
In the present embodiment, the determination method for dyeing rank is:When xylem vertical section has 85% or more area in red Color, then the dyeing rank is 4 grades;When xylem vertical section has the area of 46%-85% to take on a red color, then the dyeing rank is 3 grades;When xylem vertical section has the area of 20%-45% to take on a red color, then the dyeing rank is 2 grades;When xylem vertical section There is 20% area below to take on a red color, then the dyeing rank is 1 grade.
The calculation formula of staining index is:Staining index=Σ (dyeing rank × branch hop count)/(branch section sum × highest dye Color rank).
In embodiment 3, the staining index corresponding to every group of cryogenic temperature is as shown in table 1.And in order to verify each freezing temperature Can the lower staining index obtained of degree accurately reflect the cryogenic temperature and go down the hill the activity of grape branch, and the application has detected often respectively It organizes under cryogenic temperature, the branch germination rate of " double rich " V. amurensis branch, TTC reduction rates, succinate dehydrogenase activity, result is equal It is shown in Table 1.
The corresponding staining index of 1 cryogenic temperature of table, germination rate, TTC reduction rates and succinate dehydrogenase activity
As the reduction of cryogenic temperature, the staining index of V. amurensis branch continuously decrease it can be seen from the result of table 1, This point also complies with common sense:When V. amurensis branch freezes at a lower temperature, the number of competent cell is then fewer.
And branch germination rate, TTC reduction rates and the succinate dehydrogenase activity and staining index detected is in extremely notable positive It closes (as shown in table 2), shows that cold hardness evaluation method provided by the invention being capable of freeze injury change of the real simulation plant after enduring cold Change process.
Correlation analysis figure between 2 staining index of table and germination rate, TTC reduction rates, succinate dehydrogenase activity
The identification method provided by embodiment 3 detects to obtain dye of the V. amurensis kind " double rich " under different cryogenic temperatures After colour index, improved Logistic equation models temperature inflection point is combined using nonlinear regression analysis, utilizes DPS9.0 softwares Analysis determines that the half lethal low temperature of " double rich " is -38.63 DEG C, and fit equation is y=0.9887/ (1+e(-11.5911-0.3002x)), degree of fitting 0.9987.
Experimental example 1
It can be by the V. amurensis winter resistance area of different germ plasm resources in order to verify winter resistance detection method provided by the invention It separates, the assay method provided using embodiment 3 also has detected the mountain of 39 germplasm in addition to " double rich " this kind respectively Grape, including " Zuo Shanyi ", " double red ", " northern ice is red ", " Zuo Youhong ", " 75047 ", " 73099 ", " 84005 " etc. be at -20 DEG C, - 25 DEG C, -30 DEG C, -35 DEG C, -40 DEG C, -45 DEG C, -50 DEG C of staining index, and its low temperature semilethal temperature is determined according to its result Degree.The results are shown in Table 3 for it.
The Measurement of Cold Resistance result of the V. amurensis of the different germ plasm resources of table 3
As seen from Figure 3, for 40 parts of V. amurensis germplasm LT of examination50Value has differences, can be accordingly by different V. amurensis Germ plasm resource winter resistance power distinguishes.Wherein " 75047 ", " 74003-1 ", " 73099 ", " 73068 ", " 84005 " are height Cold-resistant germplasm, " 73134 ", " Zuo Youhong ", " northern ice is red " are low cold-resistant germplasm, remaining is medium cold-resistant germplasm.
By taking " double rich " and " northern ice is red " as an example, the winter resistance of the two can not be distinguished, and led to according to traditional conductance method Assay method provided by the invention is crossed, the fit equation of the two and matched curve are significantly different (being shown in Table 3 and Fig. 2), can be by two The winter resistance of person distinguishes.
In addition, cold- resisting index average membership can also be used as V. amurensis winter resistance comprehensive evaluation index to V. amurensis germplasm Resource winter resistance carries out overall merit.It is whether accurate and reliable in order to further verify identification method provided by the invention, the present invention In, 40 parts of germplasm LT of V. amurensis for being acquired with improved TTC decoration methods50Value is with its cold- resisting index average membership in extremely significantly just Correlation, related coefficient square value are 0.7794 (see Fig. 3), illustrate that the two cold hardness evaluation result is consistent.Therefore, improved TTC Decoration method can be as a kind of short-cut method of V. amurensis germ plasm resource rest period cold hardness evaluation, and qualification result is accurate and reliable.
Experimental example 2
The winter resistance of V. amurensis germ plasm resource rest period is identified using conductance method.Its method is as follows:
Relative conductivity measures:It takes the wild grape resource branch of each low-temperature treatment, removes epidermis, avoid bud eye, it is cut into 2~ The thin slice of 3mm is uniformly mixed, accurately weighs 0.50g and be put into 10ml graduated centrifuge tubes, adds 8ml deionized waters, is evacuated 40min Afterwards, it shakes up, its first electric conductivity value R is measured with DDS-308A types conductivity gauge (thunder magnetic)1, then test tube is placed in boiling water water-bath, it boils Water-bath 1h after standing 8h at room temperature later, then surveys its whole electric conductivity value R2, calculate relative conductivity.Each processing is repeated 3 times.Phase To Conductivity Calculation formula:Relative conductivity R (%)=(R1/R2) × 100%.
Half lethal low temperature (LT50) determine:The staining index of each low-temperature treatment branch is in " S " with the decline for the treatment of temperature Type curvilinear motion combines improved Logistic equation models temperature inflection point with nonlinear regression analysis, utilizes DPS9.0 softwares Analysis determines the LT of each germplasm branch50
By the data of Means of Electrical Conductivity compared with the data that the present invention measures, the results are shown in Table 4.
The Comparative result of 4 two kinds of assay methods of table
Using the V. amurensis branch LT of Means of Electrical Conductivity it can be seen from the testing result of table 450Average value be -28.86 DEG C, the LT that the present invention measures50Average value be -36.80 DEG C.And illustrated by the measurement result of table 2, the mountain under -35 DEG C of processing Grape branch remains able to rudiment, the LT that conductance method determines50It is only -28.86 DEG C to be worth average value, so using Means of Electrical Conductivity mountain When the winter resistance of Grape Germplasm resource, measurement result is inaccurate.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from the present invention's Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims Including belonging to all such changes and modifications in the scope of the invention.

Claims (7)

1. a kind of identification method of V. amurensis germ plasm resource rest period winter resistance, which is characterized in that include the following steps:
(1) by the V. amurensis branch of rest period under different cryogenic temperatures de-epithelization after freezing processing, be put into chloro triphenyl In tetrazole dyeing liquor, it is placed in isoperibol and carries out dyeing processing;
(2) dyeing liquor for washing away V. amurensis branch surface determines dyeing rank according to the stained area of V. amurensis branch xylem, And the staining index for grape branch of being come out of retirement and taken up an official post according to dyeing level calculation;
(3) staining index corresponding to the cryogenic temperature of V. amurensis branch determines half lethal low temperature;
In step (2), described the step of determining dyeing rank according to the stained area of V. amurensis branch xylem, specifically includes:
By V. amurensis branch it is longitudinal sectional after, according to the stained area of V. amurensis branch vertical section xylem determine dyeing rank;
In step (2), the determination method of the dyeing rank is:
When xylem vertical section has the area more than 85% to take on a red color, then the dyeing rank is 4 grades;When xylem vertical section has 46%~85% area takes on a red color, then the dyeing rank is 3 grades;It is in when xylem vertical section has 20%~45% area Red, then the dyeing rank is 2 grades;When xylem vertical section has 20% area below to take on a red color, then the dyeing rank It is 1 grade;
In step (2), staining index=Σ (dyeing rank × branch hop count)/(branch section sum × highest dyes rank).
2. identification method according to claim 1, which is characterized in that described cold under different cryogenic temperatures in step (1) Further include following steps before being put into chloro triphenyltetrazolium chloride dyeing liquor after jelly processing after de-epithelization:
The V. amurensis branch of de-epithelization is cut into the branch section of 0.3~0.5cm.
3. identification method according to claim 2, which is characterized in that the number of the branch section is 10~15.
4. identification method according to claim 2, which is characterized in that in step (1), the temperature of the dyeing processing is 28 ~32 DEG C, dyeing time 18~for 24 hours, and carried out under the conditions of being protected from light.
5. identification method according to claim 2, which is characterized in that in step (1), the chloro triphenyltetrazolium chloride dye A concentration of the 0.5~1.0% of color liquid.
6. identification method according to claim 1, which is characterized in that described cold under different cryogenic temperatures in step (1) The step of freezing processing specifically includes:
The V. amurensis branch of rest period is placed at -20~-50 DEG C 12~15h of freezing, after raise the temperature to 20~30 DEG C, And 4~6h is kept at this temperature.
7. identification method according to claim 6, which is characterized in that the step of the freezing processing under different cryogenic temperatures Suddenly it specifically includes:
The temperature of V. amurensis branch local environment is down to set temperature according to the cooling rate of 4~5 DEG C/h, at this temperature Keep 12~13h, after be gradually heating to 20~30 DEG C, at this temperature keep 4~5h.
CN201510814158.7A 2015-11-20 2015-11-20 A kind of identification method of V. amurensis germ plasm resource rest period winter resistance Expired - Fee Related CN105409708B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510814158.7A CN105409708B (en) 2015-11-20 2015-11-20 A kind of identification method of V. amurensis germ plasm resource rest period winter resistance

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510814158.7A CN105409708B (en) 2015-11-20 2015-11-20 A kind of identification method of V. amurensis germ plasm resource rest period winter resistance

Publications (2)

Publication Number Publication Date
CN105409708A CN105409708A (en) 2016-03-23
CN105409708B true CN105409708B (en) 2018-10-23

Family

ID=55488636

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510814158.7A Expired - Fee Related CN105409708B (en) 2015-11-20 2015-11-20 A kind of identification method of V. amurensis germ plasm resource rest period winter resistance

Country Status (1)

Country Link
CN (1) CN105409708B (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109521128A (en) * 2019-01-15 2019-03-26 中国农业科学院特产研究所 A kind of method of the overwintering winter resistance index of correlation analysis of V. amurensis
CN114062436B (en) * 2021-10-28 2023-12-22 甘肃省农业科学院林果花卉研究所 Cold resistance comparison method for dormant branches of fruit trees
CN117999999B (en) * 2024-04-09 2024-06-14 平利县八仙云雾茶业有限公司 Antifreezing method and system for tea tree in winter

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103712921A (en) * 2013-12-12 2014-04-09 安徽农业大学 Method for authenticating and evaluating cold resistance of tea tree

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103712921A (en) * 2013-12-12 2014-04-09 安徽农业大学 Method for authenticating and evaluating cold resistance of tea tree

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
有核葡萄品种的抗寒性测定;苏来曼 ·艾则孜 等;《北京农业》;20140625(第18期);第73~75页 *
杜仲15个初选优良无性系抗寒性研究;张博勇等;《西北林学院学报》;20030625;第18卷(第2期);第4~5页 *
核桃属植物休眠期的抗寒性鉴定;田景花等;《园艺学报》;20130625;第40卷(第6期);第1051~1060页 *
橡胶芽条人工模拟低温鉴定抗寒性的研究;郭祁源等;《热带作物研究》;19840828(第4期);第35~41页 *
温185核桃一年生枝条的抗寒力测定分析;潘俨等;《新疆农业科学》;20120330;第49卷(第3期);第454~460页 *
葡萄品种及砧木抗寒性评价方法研究;何伟等;《果树学报》;20150907;第32卷(第6期);第1135~1142页 *
葡萄根系抗寒性鉴定方法研究;郭修武;《葡萄栽培与酿酒》;19941230(第4期);第26~29页 *

Also Published As

Publication number Publication date
CN105409708A (en) 2016-03-23

Similar Documents

Publication Publication Date Title
CN106248739B (en) Method for identifying and evaluating cold resistance of winter rape
Armstrong Rhizosphere oxidation in rice: An analysis of intervarietal differences in oxygen flux from the roots
CN105409708B (en) A kind of identification method of V. amurensis germ plasm resource rest period winter resistance
CN106446596B (en) To the significant relevant molecular labeling of waterlogging tolerance of chrysanthemum and its identification method and application
Meiqing et al. Early diagnosis and monitoring of nitrogen nutrition stress in tomato leaves using electrical impedance spectroscopy
CN107941989A (en) A kind of Kiwi berry cold hardness evaluation and the method for evaluation
CN107966434A (en) A kind of method of salt tolerance of eggplant Rapid identification
De Pascale et al. Water and fertilization management of vegetables: State of art and future challenges
CN104805080A (en) Rapeseed pod number major QTL molecular marker and application thereof
CN102965443A (en) Method for identifying purity of tobacco variety Zhongyan 90 by using specific molecular marker method
CN106950262B (en) A method of prediction crop salt stress is horizontal
CN109105091A (en) A kind of rapid assay methods of the tree species drought resistance based on stomata margin of safety index
Paligi et al. Accuracy of the pneumatic method for estimating xylem vulnerability to embolism in temperate diffuse-porous tree species
Reynolds et al. Defining Niagara terroir by chemical and sensory analysis of Chardonnay wines from various soil textures and vine sizes
Sihombing et al. Detection of air temperature, humidity and soil pH by using DHT22 and pH sensor based Arduino nano microcontroller
Winkler et al. Xylem, phloem and transpiration flows in developing strawberries
CN102577852A (en) Method for identifying salt tolerance of cotton
CN109596502A (en) A kind of method and its application of Rapid identification sweet potato Genome Size
Van Leeuwen et al. Comparison of three operational tools for the assessment of vine water status: stem water potential, carbon isotope discrimination measured on grape sugar and water balance
GB2571684A (en) Nutrient Solution Management Technique Based on Length of Leaf of Greenhouse Tomato
CN109430052A (en) A kind of rapid identification method of B. campestris L.ssp. Chinensis cold resistance
JP2013128479A (en) Method for judging spontaneous dormancy awakening period of low temperature requiring deciduous tree
Fukuzawa et al. Fine root dynamics and root respiration
Lee et al. Can net photosynthesis and water relations provide a clue on the forest decline of Quercus suber in North Tunisia?
CN103088148A (en) Soybean lodging-resistant major gene locus and application

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20181023