CN103558141B - A kind of method utilizing tooth in vitro disk to identify Fructus Capsici thermostability - Google Patents
A kind of method utilizing tooth in vitro disk to identify Fructus Capsici thermostability Download PDFInfo
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Abstract
The invention discloses a kind of method utilizing tooth in vitro disk to identify Fructus Capsici thermostability, the method mainly includes that with Fructus Capsici heat-resistant strain B35 be heat-resisting comparison, compare using thermo-labile strain B6 of Fructus Capsici as temperature-sensitive, the Heat tolerance identification of Fructus Capsici tooth in vitro disk, tooth in vitro disk heat evil grade scale etc..The feature of the method is, at specified temp (high temperature and room temperature), specific salts solution concentration and process Fructus Capsici tooth in vitro disk under certain time, in leaf disk phenotype symptom, notable difference is there is between different heat-resistant strains, with this by adding up, calculate the heat injury index of a certain strain colony to be checked, and according to the colony's heat evil grade scale formulated, Fructus Capsici thermostability strain and thermal sensitivity strain can be identified.This method is particularly suitable for those the precious rare and qualification of transgenic line Fructus Capsici material thermal resistance and genetic analyses, is a kind of method of qualification hot pepper germ plasm resource thermostability simple to operate, quick, effective, can greatly accelerate Fructus Capsici heat enduring vegetables breeding process.
Description
Technical field
The invention belongs to vegetable breeding field, particularly relate to Fructus Capsici tooth in vitro disk and distinguish different heat-resisting
A kind of method of property hot pepper germ plasm resource, particularly relates to Fructus Capsici tooth in vitro disk Heat tolerance identification, and
The method that Fructus Capsici tooth in vitro disk thermostability staging hierarchy is set up.The method is ensureing the survival of thermo-labile plant
On the premise of it is carried out Heat tolerance identification, it is simple to thermo-labile strain is reserved seed for planting and descendant inheritting analysis, especially
It is suitable for strain colony little, and the Heat tolerance identification of the precious pepper breeding strain such as transgenic line and something lost
Pass and analyze, be the method for a kind of qualification simple to operate, quick, effective a large amount of hot pepper germ plasm resource thermostability.
Background technology
Fructus Capsici (Capsicum annuum L.) belongs to typical thermophily plant, and high temperature is unfavorable for that it grows
Grow.During more than 35 DEG C, then growth promoter is obstructed, and falls leaves, fallen flowers phenomenon, fruit-setting rate be down to 10%～
20%, current global climate has warming trend significantly, particularly China the Changjiang river areas to the south extremely hot for summer,
Early red pepper is difficult to the more summer, in, late variety yield fall sharply, affect economic benefit.
The selection-breeding of the heat-resisting new varieties of Fructus Capsici is the effective way preventing or alleviating the harm of its heat stress.Accurately mirror
The thermostability determining Fructus Capsici is research Fructus Capsici heat stress and the important foundation improving Fructus Capsici thermostability.At present, mirror
The common method determining Fructus Capsici thermostability has field test, manual simulation adverse circumstance identification method, and stress physiology
Identification method etc., wherein:
Field test is very big by environmental influence, and result repeatability is poor, and required time is long, and workload is big；
Manual simulation adverse circumstance identification method needs certain facility condition, it is difficult to high-volume is carried out；
Stress physiology identifies it is a kind of indirect identification process, and its reliability is poor.
Above-mentioned these authentication methods existing are wasted time and energy, inefficient, are available for identifying especially for some
The less strain of sample and transgenic line, have significant limitation especially.
The following is relevant list of references:
1、Cloning and Expression under Heat Stress of A Small Heat Shock Protein
Gene CaHSP24 from Pepper.Zhu Wei, Lu Minghui, Gong Zhenhui and Chen
Rugang.African Journal of Biotechnology, 2011,10 (25): 4968-4976.
2、Effects of Antitranspirant Spray and Potassium:Calcium:Magnesium
Ratio on Photosynthesis, Nutrient and Water Uptake, Growth, and Yield of Sweet
Pepper.Francisco M.del Amor and Jose S.Rubio.Journal of Plant Nutrition,
2009,32:97 111.
Summary of the invention
In place of the deficiencies in the prior art, it is an object of the invention to, it is provided that a kind of utilize from
Body leaf disk identifies the fast method of Fructus Capsici thermostability, the method step simple and fast, reliable and stable, mirror
Other result is consistent with Natural infection result, reproducible, is particularly well-suited to strain colony little, and
The Heat tolerance identification of the precious pepper breeding strain such as transgenic line and genetic analysis, it is possible to improve well
Fructus Capsici thermostability screening efficiency, accelerates breeding process.
In order to realize above-mentioned task, the present invention uses following technical solution:
A kind of method utilizing tooth in vitro disk to differentiate Fructus Capsici thermostability, it is characterised in that the method utilizes
Fructus Capsici 6～7 leaf Seedling Stage tooth in vitro disk, carries out Heat tolerance identification in growth cabinet, by system
Count, calculate the heat injury index of a certain strain colony to be checked, and according to colony's heat evil grade scale of initiative,
The thermostability of Fructus Capsici can be identified.Specifically comprise the following steps that
1) selecting Fructus Capsici heat-resistant strain B35 is heat-resisting comparison, and thermo-labile strain B6 of Fructus Capsici is as temperature-sensitive
Comparison；
2) by the Fructus Capsici material seed to be checked that thermostability is unknown, and the kind that heat-resisting comparison and temperature-sensitive compare
Son, respectively through 55 DEG C of water seed soaking 15min, and constantly agitation makes water temperature be down to 35 DEG C of 8h that soak seed again,
Pull out and clean with clear water, load and clean wet cloth bag is placed in 28 DEG C of illumination boxs, completely black dark situation
Carry out accelerating germination, show money or valuables one carries unintentionally through 4d seed, be sowed in hole tray；Then hole tray is placed in temperature photoperiod bar round the clock
The growth cabinet of part is cultivated, and the described photoperiod condition of temperature round the clock is: relative humidity 50%～
70%, illumination 3000～4000lx, day temperature 25 DEG C～30 DEG C, 16h, nocturnal temperature 18 DEG C, 8h,
Hoagland ' s nutritional solution is watered during cultivation；
3) collection of functional leaf and the making of tooth in vitro disk.When seedling length to 6～7 slice true leaf, choosing
Take the 5th, 6 normal band handle functional leafs of growth above Shoot Organogenesis from Cotyledon of Pepper, utilize the punching of diameter 0.9cm
Device gathers the leaf disk sample as its thermostability of qualification, and each Fructus Capsici strain to be checked gathers diameter 0.9cm
Tooth in vitro wafer sample 20, is repeated 3 times；
4) by the Fructus Capsici tooth in vitro disk that gathers by Fructus Capsici line numbers to be checked, immerse equipped with 30mL,
Concentration is in the culture dish with cover of 300mmol/L NaCl, a diameter of 9.0cm of culture dish, each cultivation
Ware immerses leaf disk 10, indicates Fructus Capsici line numbers to be checked with marking pen after adding a cover on culture dish；
Then, the saline solution culture dish that will be equipped with testing sample leaf disk is respectively placed in 38 DEG C (high in duplicate
Temperature) and the growth cabinet of 20 DEG C (room temperature) in, illumination 20000lx, continuous illumination process 24h
After, add up heat injury index；During identifying, check culture dish middle water level every 12h and add in time
Distilled water is to 30mL water level；
5) Fructus Capsici strain blade to be checked is added up according to following Fructus Capsici excised leaf Heat tolerance identification grade scale
Progression:
0 grade: leaf disk is dark green, without heat evil symptom；
1 grade: chlorosis that leaf disk edge is slight；
3 grades: chlorosis that leaf disk edge is slight, leaf disk center is water stain shape；
5 grades: lamellar chlorosis symptom occurs in the middle part of leaf disk；
7 grades: in the middle part of leaf disk, light green colour circle occur in the aggravation of lamellar chlorosis, only leaf disk surrounding；
9 grades: the equal chlorosis in the middle part of leaf disk and leaf disk edge.
6) it is calculated as follows heat injury index:
Heat injury index=∑ is (each under heat injury index leaf disk numbers at different levels × this heat evil class value-room temperature under high temperature
Level heat injury index leaf disk number × this heat evil class value) ÷ (investigating total leaf disk number × superlative degree value) × 100；
When heat injury index calculates, as between repetition, difference is notable, take the meansigma methods repeated 3 times；Such as difference
Significantly, repeat 3 times, take its 3 inapparent meansigma methodss of difference as these detection Fructus Capsici thermostability product
The heat injury index of system；Finally, Fructus Capsici strain (or strain) to be measured is determined according to following colony grade scale
Thermostability:
Heat injury index≤35.0 are heat-resistant strain；
Heat injury index is relatively heat-resistant strain 35.1～54.9；
Heat injury index >=55.0 are temperature-sensitive strain.
The method utilizing tooth in vitro disk qualification Fructus Capsici thermostability of present invention initiative, is mainly planted by collection
The blade that strain is a small amount of, utilizes card punch to gather tooth in vitro disk and carries out high-temperature process, by statistics, calculate
Heat injury index identifies the thermostability of Fructus Capsici strain.This method is reliable and stable, and qualification result is natural with field
Qualification result is consistent, reproducible, and speed is fast, and time saving and energy saving, the wound causing plant is little, especially
It is to solve in Fructus Capsici preciousness germ plasm resource, and during the Heat tolerance identification of transfer-gen plant, is available for
The few problem of plant selected, can carry out again identifying that strain is reserved seed for planting and descendant inheritting analysis, thus significantly carry
The high process of Fructus Capsici Heat-tolerance Breeding.
Accompanying drawing explanation
Fig. 1 is that high temperature (38 DEG C), 300mmol/L NaCl coerce lower different stage Fructus Capsici tooth in vitro disk
Symptom.Figure is from left to right followed successively by 0 grade, 1 grade, 3 grades, 5 grades, 7 grades and 9 grades.
Below in conjunction with drawings and Examples, the present invention is described in further detail.
Detailed description of the invention
It should be noted that following example are merely illustrative, the invention is not restricted to these embodiments,
Proving through the test of many times of applicant, the method for the present invention is suitable for current existing all capsicum varieties
(being).
Embodiment 1:
The present embodiment utilizes Fructus Capsici 6～7 leaf Seedling Stage tooth in vitro disk, carries out resistance in growth cabinet
Hot qualification, by adding up, calculate the heat injury index of a certain strain colony to be checked, and according to the group of initiative
Body heat evil grade scale, can identify the thermostability of Fructus Capsici.Specifically comprise the following steps that
1, it is heat-resisting comparison with Fructus Capsici heat-resistant strain B35, using thermo-labile strain B6 of Fructus Capsici as temperature-sensitive
Comparison (Zhu Wei, Lu Minghui, Gong Zhenhui, and Chen Rugang.Cloning and
Expression under Heat Stress of A Small Heat Shock Protein Gene CaHSP24
From Pepper [J] .African Journal of Biotechnology, 2011,10(25): 4968-4976),
All it is known in the art for detection Fructus Capsici strain respectively PP97-7195-1, Mito Lee Seln-1(
Fructus Capsici strain, by applicant seminar provide).
2, by the Fructus Capsici material seed to be checked that thermostability is unknown, and the kind that heat-resisting comparison and temperature-sensitive compare
Son, respectively through 55 DEG C of water seed soaking 15min, and constantly agitation makes water temperature be down to 35 DEG C of 8h that soak seed again,
Pull out and clean with clear water, load and clean wet cloth bag is placed in 28 DEG C of illumination boxs, completely black dark situation
Carry out accelerating germination, show money or valuables one carries unintentionally through 4d seed, be sowed in hole tray；Then hole tray is placed in temperature photoperiod bar round the clock
The growth cabinet of part is cultivated, and the described photoperiod condition of temperature round the clock is: relative humidity 50%～
70%, illumination 3000～4000lx, day temperature 25 DEG C～30 DEG C, 16h, nocturnal temperature 18 DEG C, 8h,
Hoagland ' s nutritional solution (Francisco M.del Amor and Jose S.Rubio. is watered during cultivation
Effects of Antitranspirant Spray and Potassium:Calcium:Magnesium Ratio on
Photosynthesis, Nutrient and Water Uptake, Growth, and Yield of Sweet Pepper.
Journal of Plant Nutrition, 2009,32:97 111)；
3, the collection of functional leaf and the making of tooth in vitro disk.
When seedling length to 6～7 slice true leaf, choose the 5th, 6 normal bands of growth above Shoot Organogenesis from Cotyledon of Pepper
Handle functional leaf, utilizes the card punch of diameter 0.9cm to gather the leaf disk sample as its thermostability of qualification,
Each Fructus Capsici strain to be checked gathers diameter 0.9cm tooth in vitro wafer sample 20, is repeated 3 times.
4, by the Fructus Capsici tooth in vitro disk that gathers by Fructus Capsici line numbers to be checked, immerse equipped with 30mL,
Concentration is in the culture dish with cover of 300mmol/L NaCl, a diameter of 9.0cm of culture dish, each cultivation
Ware immerses leaf disk 10, indicates Fructus Capsici line numbers to be checked with marking pen after adding a cover on culture dish；
Then, the saline solution culture dish that will be equipped with testing sample leaf disk is respectively placed in 38 DEG C (high in duplicate
Temperature) and the growth cabinet of 20 DEG C (room temperature) in, illumination 20000lx, continuous illumination process 24h after,
Statistics heat injury index；During identifying, check culture dish middle water level every 12h and add distilled water in time
To 30mL water level；
5. add up Fructus Capsici strain leaf to be checked according to following Fructus Capsici tooth in vitro disk Heat tolerance identification grade scale
Sheet progression:
0 grade: leaf disk is dark green, without heat evil symptom；
1 grade: chlorosis that leaf disk edge is slight；
3 grades: chlorosis that leaf disk edge is slight, leaf disk center is water stain shape；
5 grades: lamellar chlorosis symptom occurs in the middle part of leaf disk；
7 grades: in the middle part of leaf disk, light green colour circle occur in the aggravation of lamellar chlorosis, only leaf disk surrounding；
9 grades: the equal chlorosis in the middle part of leaf disk and leaf disk edge.
6. it is calculated as follows heat injury index:
Heat injury index=∑ is (each under heat injury index leaf disk numbers at different levels × this heat evil class value-room temperature under high temperature
Level heat injury index leaf disk number × this heat evil class value) ÷ (investigating total leaf disk number × superlative degree value) × 100；
When heat injury index calculates, as between repetition, difference is notable, take the meansigma methods repeated 3 times；Such as difference
Significantly, repeat 3 times, take its 3 inapparent meansigma methodss of difference as these detection Fructus Capsici thermostability product
The heat injury index of system；Finally, Fructus Capsici strain (or strain) to be measured is determined according to following colony grade scale
Thermostability:
Heat injury index≤35.0 are heat-resistant strain；
Heat injury index is relatively heat-resistant strain 35.1～54.9；
Heat injury index >=55.0 are temperature-sensitive strain.
According to the method for the present embodiment, testing result is as follows:
Heat injury index=33.9 of PP97-7195-1, for heat-resistant strain；The heat evil of Mito Lee Seln-1 refers to
Number=32.9, for heat-resistant strain.This result and the field test result (heat injury index of PP97-7195-1
=18.2, for heat-resistant strain；Heat injury index=32.6 of Mito Lee Seln-1, for heat-resistant strain) consistent,
Illustrate that the tooth in vitro disk that the present invention formulates identifies that the method for Fructus Capsici thermostability can truly reflect Fructus Capsici field
Thermostability, it is adaptable to the Rapid identification of Fructus Capsici thermostability.
Embodiment 2:
The present embodiment utilizes Fructus Capsici 6～7 leaf Seedling Stage tooth in vitro disk, carries out resistance in growth cabinet
Hot qualification, by adding up, calculate the heat injury index of a certain strain colony to be checked, and according to the group of initiative
Body heat evil grade scale, can identify the thermostability of Fructus Capsici.Specifically comprise the following steps that
1, it is heat-resisting comparison with Fructus Capsici heat-resistant strain B35, using thermo-labile strain B6 of Fructus Capsici as temperature-sensitive
Comparison (Zhu Wei, Lu Minghui, Gong Zhenhui and Chen Rugang.Cloning and
Expression under Heat Stress of A Small Heat Shock Protein Gene CaHSP24
From Pepper [J] .African Journal of Biotechnology, 2011,10 (25): 4968-4976),
For detection Fructus Capsici strain respectively Huang marshal No. 2 choosing system, beautiful jade choosing is, satisfy sub-green pepper and Early
Calwonder(is Fructus Capsici strain well known in the art, the seminar of applicant provide).
2, by the Fructus Capsici material seed to be checked that thermostability is unknown, and the kind that heat-resisting comparison and temperature-sensitive compare
Son, respectively through 55 DEG C of water seed soaking 15min, and constantly agitation makes water temperature be down to 35 DEG C of 8h that soak seed again,
Pull out and clean with clear water, load and clean wet cloth bag is placed in 28 DEG C of illumination boxs, completely black dark situation
Carry out accelerating germination, show money or valuables one carries unintentionally through 4d seed, be sowed in hole tray；Then hole tray is placed in temperature photoperiod bar round the clock
The growth cabinet of part is cultivated, and the described photoperiod condition of temperature round the clock is: relative humidity 50%～
70%, illumination 3000～4000lx, day temperature 25 DEG C～30 DEG C, 16h, nocturnal temperature 18 DEG C, 8h,
Hoagland ' s nutritional solution (Francisco M.del Amor and Jose S.Rubio. is watered during cultivation
Effects of Antitranspirant Spray and Potassium:Calcium:Magnesium Ratio on
Photosynthesis,Nutrient and Water Uptake,Growth,and Yield of Sweet Pepper.
Journal of Plant Nutrition, 2009,32:97 111)；
3, the collection of functional leaf and the making of tooth in vitro disk:
When seedling length to 6～7 slice true leaf, choose the 5th, 6 normal bands of growth above Shoot Organogenesis from Cotyledon of Pepper
Handle functional leaf, utilizes the card punch of diameter 0.9cm to gather the leaf disk sample as its thermostability of qualification,
Each Fructus Capsici strain to be checked gathers diameter 0.9cm tooth in vitro wafer sample 20, is repeated 3 times.
4, by the Fructus Capsici tooth in vitro disk that gathers by Fructus Capsici line numbers to be checked, immerse equipped with 30mL,
Concentration is in the culture dish with cover of 300mmol/L NaCl, a diameter of 9.0cm of culture dish, each cultivation
Ware immerses leaf disk 10, indicates Fructus Capsici line numbers to be checked with marking pen after adding a cover on culture dish；
Then, the saline solution culture dish that will be equipped with testing sample leaf disk is respectively placed in 38 DEG C (high in duplicate
Temperature) and the growth cabinet of 20 DEG C (room temperature) in, illumination 20000lx, continuous illumination process 24h after,
Statistics heat injury index；During identifying, check culture dish middle water level every 12h and add distilled water in time
To 30mL water level；
5, Fructus Capsici strain leaf to be checked is added up according to following Fructus Capsici tooth in vitro disk Heat tolerance identification grade scale
Sheet progression:
0 grade: leaf disk is dark green, without heat evil symptom；
1 grade: chlorosis that leaf disk edge is slight；
3 grades: chlorosis that leaf disk edge is slight, leaf disk center is water stain shape；
5 grades: lamellar chlorosis symptom occurs in the middle part of leaf disk；
7 grades: in the middle part of leaf disk, light green colour circle occur in the aggravation of lamellar chlorosis, only leaf disk surrounding；
9 grades: the equal chlorosis in the middle part of leaf disk and leaf disk edge
6. it is calculated as follows heat injury index:
Heat injury index=∑ is (each under heat injury index leaf disk numbers at different levels × this heat evil class value-room temperature under high temperature
Level heat injury index leaf disk number × this heat evil class value) ÷ (investigating total leaf disk number × superlative degree value) × 100；
When heat injury index calculates, as between repetition, difference is notable, take the meansigma methods repeated 3 times；Such as difference
Significantly, repeat 3 times, take its 3 inapparent meansigma methodss of difference as these detection Fructus Capsici thermostability product
The heat injury index of system；Finally, Fructus Capsici strain (or strain) to be measured is determined according to following colony grade scale
Thermostability:
Heat injury index≤35.0 are heat-resistant strain；
Heat injury index is relatively heat-resistant strain 35.1～54.9；
Heat injury index >=55.0 are temperature-sensitive strain.
According to the method for the present embodiment, testing result is as follows:
Heat injury index=55.3 that Huang marshal No. 2 choosing is, for temperature-sensitive strain；The heat injury index that beautiful jade choosing is
=36.1, for relatively heat-resistant strain；Heat injury index=37.2 of full sub-green pepper, for relatively heat-resistant strain；Early
Heat injury index=36.9 of Calwonder, for relatively heat-resistant strain.This result is (yellow with field test result
Heat injury index=78.0 that marshal No. 2 choosing is, for temperature-sensitive strain；Heat injury index=52.8 that beautiful jade choosing is,
For relatively heat-resistant strain；Heat injury index=37.6 of full sub-green pepper, for relatively heat-resistant strain；Early Calwonder
Heat injury index=36.6, for relatively heat-resistant strain) consistent, illustrate that the tooth in vitro disk that the present invention formulates reflects
The method determining Fructus Capsici thermostability can truly reflect the thermostability in Fructus Capsici field, it is adaptable to the thermostability mirror of Fructus Capsici
Fixed.
Claims (1)
1. one kind utilizes the method that tooth in vitro disk identifies Fructus Capsici thermostability, it is characterized in that, the method utilizes Fructus Capsici 6～7 leaf Seedling Stage tooth in vitro disk, Heat tolerance identification is carried out in growth cabinet, colony's heat injury index of a certain strain to be checked is added up, calculated to Fructus Capsici tooth in vitro disk heat evil grade scale according to initiative, and according to the colony's heat evil grade scale formulated, determine the thermostability of Fructus Capsici strain to be identified, specifically comprise the following steps that
1) selecting Fructus Capsici heat-resistant strain B35 is heat-resisting comparison, and thermo-labile strain B6 of Fructus Capsici compares as temperature-sensitive；
2) by the Fructus Capsici material seed to be checked of thermostability the unknown, and heat-resisting comparison and the seed of temperature-sensitive comparison, respectively through 55 DEG C of water seed soaking 15min, and constantly agitation makes water temperature be down to 35 DEG C of 8h that soak seed again, pulling out and clean with clear water, load and be placed in 28 DEG C of illumination boxs in clean wet cloth bag, completely black dark situation carries out accelerating germination, show money or valuables one carries unintentionally through 4d seed, be sowed in hole tray；The growth cabinet that then hole tray is placed in temperature photoperiod condition round the clock is cultivated, and the described photoperiod condition of temperature round the clock is: relative humidity 50%～70%, illumination 3000～4000lx, day temperature 25 DEG C～30 DEG C, 16h, nocturnal temperature 18 DEG C, 8h, waters Hoagland ' s nutritional solution during cultivation；
3) collection of functional leaf and the making of tooth in vitro disk:
When seedling length to 6～7 slice true leaf, choose the 5th, 6 normal band handle functional leafs of growth above Shoot Organogenesis from Cotyledon of Pepper, the card punch utilizing diameter 0.9cm gathers the leaf disk sample as its thermostability of qualification, and each Fructus Capsici strain to be checked gathers diameter 0.9cm tooth in vitro wafer sample 20, is repeated 3 times；
4) by the Fructus Capsici tooth in vitro disk that gathers by Fructus Capsici line numbers to be checked, immerse equipped with 30mL, concentration be 300mmol/L NaCl culture dish with cover in, on culture dish, indicate Fructus Capsici line numbers to be checked with marking pen after adding a cover；Then, the saline solution culture dish that will be equipped with testing sample leaf disk is respectively placed in the growth cabinet of 38 DEG C and 20 DEG C in duplicate, illumination 20000lx, after continuous illumination processes 24h, adds up heat injury index；During identifying, check culture dish middle water level every 12h and add distilled water in time to 30mL water level；
5) according to following Fructus Capsici tooth in vitro disk Heat tolerance identification grade scale statistics Fructus Capsici strain blade progression to be checked:
0 grade: leaf disk is dark green, without heat evil symptom；
1 grade: chlorosis that leaf disk edge is slight；
3 grades: chlorosis that leaf disk edge is slight, leaf disk center is water stain shape；
5 grades: lamellar chlorosis symptom occurs in the middle part of leaf disk；
7 grades: in the middle part of leaf disk, light green colour circle occur in the aggravation of lamellar chlorosis, only leaf disk surrounding；
9 grades: the equal chlorosis in the middle part of leaf disk and leaf disk edge；
6) it is calculated as follows heat injury index:
Heat injury index=∑ (heat injury index leaf disk numbers at different levels × this heat evil class value under heat injury index leaf disk numbers at different levels × this heat evil class value-room temperature under high temperature) ÷ (investigating total leaf disk number × superlative degree value) × 100；
When heat injury index calculates, as between repetition, difference is notable, take the meansigma methods repeated 3 times；Such as significant difference, repeat 3 times, take its 3 inapparent meansigma methodss of difference heat injury index as this detection Fructus Capsici thermostability strain；Finally, Fructus Capsici strain to be measured or the thermostability of strain are determined according to following colony grade scale:
Heat injury index≤35.0 are heat-resistant strain；
Heat injury index is relatively heat-resistant strain 35.1～54.9；
Heat injury index >=55.0 are temperature-sensitive strain.
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| CN109164212A (en) * | 2018-09-08 | 2019-01-08 | 华中农业大学 | A kind of measuring method using excised leaf identification Chinese rose germplasm heat resistance |
| CN109041876A (en) * | 2018-09-12 | 2018-12-21 | 中国农业科学院麻类研究所 | A kind of identification method of Heat Tolerance for Radish |
| CN109374843B (en) * | 2018-09-19 | 2021-03-23 | 西北农林科技大学 | Method for manually and rapidly identifying cracking resistance of hot pepper |
| CN114830866A (en) * | 2022-04-13 | 2022-08-02 | 徐州院博辣椒产业技术研究院有限公司 | Method for identifying authenticity of pepper variety |
| CN115299299A (en) * | 2022-08-12 | 2022-11-08 | 武汉市农业科学院 | Method for identifying heat resistance of pakchoi |
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| CN102911945A (en) * | 2011-08-05 | 2013-02-06 | 中国科学院上海生命科学研究院 | High temperature resistant gene of plants and application thereof |
| CN102805010A (en) * | 2012-08-28 | 2012-12-05 | 西北农林科技大学 | Method for identifying chilli heat resistance by using detached leaves |
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