CN104303867B - Method for comparing hard-light resistance of plants - Google Patents

Method for comparing hard-light resistance of plants Download PDF

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Publication number
CN104303867B
CN104303867B CN201410549301.XA CN201410549301A CN104303867B CN 104303867 B CN104303867 B CN 104303867B CN 201410549301 A CN201410549301 A CN 201410549301A CN 104303867 B CN104303867 B CN 104303867B
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light
plant
blade
leaf
disjunctor
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CN104303867A (en
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李宏伟
郑琪
李滨
李振声
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Institute of Genetics and Developmental Biology of CAS
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Institute of Genetics and Developmental Biology of CAS
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    • 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
    • A01G7/04Electric or magnetic or acoustic treatment of plants for promoting growth
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/14Measures for saving energy, e.g. in green houses

Abstract

The invention discloses a method for comparing the hard-light resistance of plants. The method for comparing the hard-light resistance of the plants comprises the following steps of flatly paving connected leaf blades of different plants to be measured, which are obtained under the same culture conditions and have the same growing periods, at certain same leaf positions; under the conditions of the same temperature and relative humidity, enabling the connected leaf blades to receive same hard light treatment at the positions perpendicular to the light. The method disclosed by the invention can be used for identifying varieties with the performance of resisting hard-light high temperature or hard-light low temperature, is simple, convenient and quick for operation, is suitable for large-scale identification and has important significance for foundation research on hard-light resistance of the plants and the breeding of the hard-light-resisting plant varieties.

Description

A kind of method comparing plant resistance to high light ability
Technical field
The present invention relates to a kind of method comparing plant resistance to high light ability, belong to biological technical field.
Background technology
Under natural environment, when the light that plant leaf blade absorbs can exceed that the scope that photosynthesis can utilize, these are relative Superfluous luminous energy can promote to produce a large amount of active oxygen, and then causes leaf photosynthesis efficiency to decline and Xanthophyll cycle (Powles occurs et al.,1984).Along with enhancing and the prolongation of high light persistent period of extraneous light intensity, the Xanthophyll cycle that high light causes can be sent out Exhibition is photooxidation, with the reduction of leaf pigment concentration.For annual crops (such as Oryza sativa L., Semen Tritici aestivi etc.), Xanthophyll cycle and photooxidation that high light causes can limit the photosynthate transfer to seed, even can cause blade aging ahead of time And cause the underproduction.Therefore, one of the important channel that the New Crop Varieties of resistance to high light is high-yield breeding of crops is cultivated.
Xanthophyll cycle or photooxidation that high light is caused by plant present species, subspecies and interracial difference.To Different Crop The qualification of kind resistance to high light characteristic helps speed up the New Crop Varieties cultivating resistance to high light.Chlorophyll content and leaf can be used Verdazulene fluorescence parameter such as lightsystemⅡ maximal photochemistry efficiency (Fv/Fm) reflects that the resistance to high light of plant causes Xanthophyll cycle or light The degree of oxidation.It is generally believed that Fv/Fm drops to less than 0.75, show to there occurs Xanthophyll cycle.The kind of resistance to high light After high light processes, chlorophyll content and Fv/Fm fall are little, and to strong light activated kind after high light processes, Chlorophyll content and Fv/Fm fall are big.
Typically manual simulation is utilized to aoxidize bar relative to high light and low concentration methyl viologen (also referred to as MV) simulated light at laboratory Part processes the method for tooth in vitro disk and carries out plant photooxidation resistant research (Iwao et al.1984;Mitsuko et al. 1995).MV is the main component of herbicides paraquat (Paraquat), as artificial electron's receptor of Photosystem I, Produce superoxide anion (O2 -).Because the method is by factors such as artificial light source, operation complexity, do not directly apply to big In the screening varieties of scale.
Jiao Demao etc. (1991) establish easy extensive photooxidation identification system, and specific practice is by vitro for Oryza sativa L. merit Energy blade (sword-like leave or two leaves), fills in the ceramic whiteware dish of 2.5 centimetres of water layers, the outer warp of holding chamber with clear glass press-in Fine day daylight continuous processing 5-7 days, keeps green degree to carry out classification to identify that Oryza sativa L. is resistance to by force according to processing rear blade Light kind.They utilize the method that the rice varieties of 165 parts of separate sources has been carried out photooxidation qualification, it was found that one Kind (the Jiao et al.1992 of a little resistance to high lights;Gu Hangying etc. 1992;Jiao Demao etc. 1994).Fang Xianwen etc. (1998) Use the method that the photooxidation resistant characteristic of 10 wheat breeds is detected.Jiang Hua etc. (2008) and Sun little Xia etc. (2009) utilize the method that Oryza sativa L. photooxidation resistant gene has been carried out QTL location.Owing to the method requires continuous 5-7 It fine day just can be identified, therefore its qualification result is easily affected by weather, and its accuracy and repeatability are wayward. Li etc. (2010) improve on the basis of the method.Mainly by the Ou Silang metal halide lamp of 5 × 5 (HQI-TS 150W/D) is as artificial light source, it is provided that light-receiving area is 1 square metre, and light intensity is about 1000 μm ol m-2 s-1Stable state homogenizing light source, specific practice is that the leaf section of in the middle part of wheat flag leaf about 5 centimetres is sandwiched in two sheet glass Between, it is placed in the aquarium of the 1m × 1m filling 5 centimetres of water layers, through 1000 μm ol m-2 s-1Artificial light Source continuous processing 3 days, changes the resistance to of reflection Wheat Cultivars by comparison process chlorophyll content and Fv/Fm High light characteristic.Li etc. (2010) utilize the method that one DH colony is carried out photooxidation process, located Semen Tritici aestivi resistance to High light QTL.
Above photooxidation authentication method is carried out with excised leaf for material, although the method that excised leaf high light processes The plant toleration to high light can be reflected to a certain extent, but this method cannot reflect plant self disjunctor leaf completely The sheet response process to high light.Typically directly plant can be carried out under artificial light source photo-irradiation treatment, then than Relatively functional leaf photosynthetic efficiency after high light processes, carries out Xanthophyll cycle or photooxidation research.Shared by whole plant Space relatively big, Different Leaf-position Leaf Blades is blocked mutually, blade angle, the factor such as blocks mutually between plant, is studied spy Fixed blade light is not consistent, and therefore, the method is not suitable for that more plant carries out accurate resistance to high light and identifies. The method of the disjunctor blade resistance to high light process setting up scale is the research of plant resistance to high light and the crop breed breeding of resistance to high light Important foundation.Therefore, it is contemplated that set up a kind of plant disjunctor blade authentication method of resistance to high light.
Summary of the invention
It is an object of the invention to provide a kind of method comparing plant resistance to high light ability.
The present invention provides a kind of method comparing plant resistance to high light ability, is identical by obtain under same culture conditions The difference of trophophase treats the disjunctor blade tiling of certain same leaf position of measuring plants, and at mutually synthermal and relative humidities Under, described disjunctor blade is processed at the high light that the position acceptance vertical with light is identical so that the company of this same leaf position The lightsystemⅡ maximal photochemistry efficiency Fv/Fm of body blade is respectively less than 0.75, obtains difference after the process of measuring plants This disjunctor blade;Measure the lightsystemⅡ maximal photochemistry efficiency that difference treats this disjunctor blade before treatment of measuring plants Fv/Fm, is denoted as A;Measure the lightsystemⅡ maximal photochemistry efficiency of different this disjunctor blade after the process of measuring plants Fv/Fm, is denoted as B;The value that the same A treating measuring plants deducts B gained is denoted as C;If treating the C value of measuring plants X It is less than the C value treating measuring plants Y, then treats that the resistance to high light ability of measuring plants X is better than and treat measuring plants Y;
Described disjunctor blade is the Non in vitro blade being connected on whole strain plant to be measured, can be by plant to be measured for whole strain during operation Put in high light processing equipment and operate;
Described plant is same plant.
In said method, described relative humidity is 40%-60%.
In any of the above-described described method, described trophophase was two leaf one heart stages;
Described same leaf position is the second leaf.
In any of the above-described described method, the light intensity that described high light processes is 700-1000 μm ol m-2 s-1, it is specially 700μmol m-2 s-1、800μmol m-2 s-1Or 1000 μm ol m-2 s-1
In any of the above-described described method, the process time that described high light processes is 10min-4h, specially 10min, 20min, 30min, 1h, 1.5h, 3h or 4h.
In any of the above-described described method, described in obtain difference this disjunctor blade after the process of measuring plants and described survey Determine also to wrap between the lightsystemⅡ maximal photochemistry efficiency Fv/Fm of the difference this disjunctor blade after the process of measuring plants Include the step of dark processing.
In any of the above-described described method, the time of described dark processing is 15-20min, specially 15min;
When described lightsystemⅡ maximal photochemistry efficiency Fv/Fm measures, saturated light intensity is set as 3000 μm ol m-2 s-1, Saturated flash time is 1s;
Described temperature is 20-42 DEG C, specially 23-26 DEG C, more specially 25-26 DEG C, more specifically 20 DEG C, 25 DEG C, 31 DEG C, 37 DEG C and 42 DEG C.
The lightsystemⅡ maximal photochemistry efficiency Fv/Fm of a kind of disjunctor blade the making plant method less than 0.75 also belongs to In protection scope of the present invention, it is that the disjunctor blade of plant is tiled, by described disjunctor blade in the position vertical with light Put and accept high light process;
The light intensity that described high light processes is specially 700-1000 μm ol m-2 s-1, specially 700 μm ol m-2 s-1、 800μmol m-2 s-1Or 1000 μm ol m-2 s-1
Described disjunctor blade is the Non in vitro blade being connected on whole strain plant, whole strain plant can be put into high light during operation Processing equipment operates.
In any of the above-described described method, described plant is grass, specially Semen Tritici aestivi or Oryza sativa L..
The application in plant selection-breeding of any of the above-described described method falls within protection scope of the present invention;
Described plant is grass, specially Semen Tritici aestivi or Oryza sativa L..
The method that the present invention provides can be used for the high temperature of resistance to high light or the low temperature of resistance to high light cultivar identification, and the method is easy and simple to handle, Quickly, being suitable for Large scale identification, the basic research of high light resistance to plant and the selection-breeding of resistance to high light plant variety have important Meaning.
Accompanying drawing explanation
Fig. 1 is that the resistance to high light of blade to be identified identifies schematic diagram.
Fig. 2 is wheat leaf blade Fv/Fm change after high light processes different time.
Fig. 3 is wheat leaf blade Fv/Fm change at different temperatures after high light processes.
Fig. 4 is the Fv/Fm change after Semen Tritici aestivi various position leaves disjunctor vanes high light processes.
Fig. 5 is the Fv/Fm change before and after rice leaf high light processes.
Detailed description of the invention
Experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, the most commercially obtain.
Little lay down 54 (Triticumaestivum) passes through the authorization of Crop breed audit committee of Shaanxi Province for 2000.
Entrusted in capital 411 (Triticumaestivum L.) for 1991,1992 respectively by Crops in Beijing authorization Member can be with the authorization of Crop breed audit committee of country.
Little lay down 54 and capital 411 be all grass family Triticum common wheat winter wheat variety.
Two excellent 2163 (Oryza sativa) are authorized by Fujian province crops variety validation board in April, 2000.
Bright extensive 63 (Oryza sativa) document " thank to Huaan etc. indica hybrid rice Shanyou 63 and restorer thereof are bright The election effects of extensive 63. Fujian Academy journal .1987. " disclosed in mistake, the public can be from Fujian Agricultural science Institute obtains, and in December, 1997 is by Fujian Province's appraisal of scientific and technological achievements, is the restorer of China's currently used area maximum.
Two excellent 2163 is grass family paddy rice cross breeding rice varieties, and bright extensive 63 is grass family paddy rice cross breeding rice restorer.
Following embodiment lightsystemⅡ maximal photochemistry efficiency (Fv/Fm) reflect the resistance to high light of plant cause Xanthophyll cycle or The degree of photooxidation.It is generally believed that Fv/Fm drops to less than 0.75, show to there occurs Xanthophyll cycle.The product of resistance to high light Planting after high light processes, Fv/Fm fall is little, and to strong light activated kind after high light processes, Fv/Fm declines Amplitude is big.
Embodiment 1, resistance to high light test experience
One, the cultivation of wheat seedling
By little lay down 54 and the seed in capital 411 through 30%H2O2(10-30%H2O2) seed soaking sterilization 10min (10-20min After), it is transferred to be placed with in the culture dish of moistening filter paper and is placed in 22 DEG C of (20-25 DEG C) calorstats germination, The seed showed money or valuables one carries unintentionally put in second day in fill tap water nylon Dropbox (30cm × 20cm × 8.5cm (long × wide × In deeply)), at intelligence growth cabinet, (model is HP1000GS, public purchased from the auspicious China in Wuhan instrument and equipment Limited Liability Department) middle growth.Condition of culture is: 14h photoperiod (12-14h photoperiod), light intensity 150 μm ol m-2 s-1 (LED light source carried by instrument provides, 100-300 μm ol m-2 s-1);Day temperature is 20 DEG C (15-22 DEG C ), nocturnal temperature is 15 DEG C (10-15 DEG C), relative humidity 40%-60%.Cultivate that (7-10 days equal in 7 days Can) after, seedling is moved into 30cm × 30cm × 15cm (long × wide × deep) and fills pancebrin (such as table 1 institute Show) plastic casing in, continue intelligence growth cabinet in cultivate (condition of culture is identical) open up completely to second leaf Open (be in for two leaf one heart stages, be about germinate after 20-24 days) time carry out high light process.
Table 1 Semen Tritici aestivi pancebrin formula (1000 times, surplus is deionized water)
Note: the Semen Tritici aestivi pancebrin of working concentration is the Semen Tritici aestivi pancebrin deionized water dilution 1000 of above-mentioned table 1 Times.
Two, resistance to high light is identified
Two leaf one heart stages that step one is obtained little lay down 54 and second leaf (disjunctor blade) in capital 411 carry out at high light Reason.
First with double faced adhesive tape, blade is fixed on the flat board of 30cm × 16cm, makes blade to be identified be arranged in order It is maintained at same level, keeps vertical (as shown in Figure 1) with top-down light.Utilize growth chamber (E36HO Type, purchased from Percival company of the U.S.) carry out high light process, largest light intensity is 1000 μm ol m-2 s-1.By adjusting Whole process blade adjusts light intensity with the distance of light, and relative humidity controls, at 40%-60%, before optical processing, first to open Lamp preheating 30min, starts to process after isocandela is stable.
(1) different light intensity process wheat seedling disjunctor blade
At 25-26 DEG C, with 600-1000 μm ol m-2 s-1As relative high light, lay down 54 and 411 liang of capital leaf to little One heart stage seedling the second leaf (disjunctor blade) carries out the process of 1h, and relative humidity controls at 40%-60%.After processing Semen Tritici aestivi disjunctor vanes dark adaptation 15min (15-20min) after, with Handy-pea chlorophyll fluorescence instrument (purchased from Hansatech company of Britain) measures lightsystemⅡ maximal photochemistry efficiency (Fv/Fm), saturated light during mensuration It is set as by force 3000 μm ol m-2 s-1, saturated flash time is 1s.
Result is as shown in table 2.
Table 2 and the different light intensity of blade before treatment process the Fv/Fm change after 1h
Table 2 shows, through 600 μm ol m-2 s-1Light intensity processes after 1h, little lays down 54 and the Fv/Fm in capital 411 from by force Before optical processing 0.807 drops to 0.754 and 0.749 respectively, close to 0.75, shows obvious Xanthophyll cycle. And through 700 μm ol m-2 s-1Above light intensity processes 1h, and little laying down 54 and the Fv/Fm in capital 411 is remarkably decreased to 0.75 Hereinafter, 700 μm ol m are shown-2 s-1Above light intensity processes 1h, and the two wheat breed can be caused to occur substantially Xanthophyll cycle, wherein 800-1000 μm ol m-2 s-1Light intensity processes after 1h, little lay down 54 Fv/Fm be significantly higher than capital 411, this is consistent with the conclusion of little 54 high lights more resistance to than capital 411 of laying down reported before this.
Under the conditions of above-mentioned, wheat leaf blade is through 1000 μm ol m-2 s-1Light intensity processes Fv/Fm change such as figure after different time Shown in 2.
Fig. 2 shows, through 1000 μm ol m-2 s-1After light intensity process 10min, the Fv/Fm of wheat leaf blade is from before treatment 0.802 (little lay down 54) and 0.803 (capital 411) are remarkably decreased to about 0.75, show that the two wheat breed is sent out Give birth to Xanthophyll cycle.Hereafter, processing the prolongation of time with high light, Fv/Fm is gradually lowered, and high light processes 4h and drops to 0.67 (little lay down 54) and 0.62 (capital 411).High light process front capital 411 with little lay down 54 Fv/Fm identical, by force After optical processing, the Fv/Fm in capital 411 lays down 54 less than little, it is demonstrated experimentally that little lay down 54 resistance to high light ability more than capital 411.
(2) under different temperatures, high light processes wheat seedling disjunctor blade
E36HO type growth chamber temperature controllable is 0-40 DEG C of change.Due to leaf temperature and growth case detection temperature There are differences, with ZDR20 humiture monitor (Zhejiang University's Electrical utility firms produces) real time record blade, institute is in place The temperature put.When controlling leaf temperature and being 20 DEG C, 25 DEG C, 31 DEG C, 37 DEG C and 42 DEG C with 800 μm ol m-2 s-1 Light intensity processes little lays down 54 and 411 liang of capital leaf one heart stage seedling the second leaf (disjunctor blade) 1h, and relative humidity controls 40%-60%.
Wheat leaf blade after processing is after dark 15min (15-20min), glimmering with Handy-pea chlorophyll Light instrument (purchased from Hansatech company of Britain) measures lightsystemⅡ maximal photochemistry efficiency (Fv/Fm), full during mensuration It is set as 3000 μm ol m with light intensity-2 s-1, saturated flash time is 1s.
Wheat leaf blade is at different temperatures through 800 μm ol m-2 s-1High light processes the Fv/Fm after 1h to be changed such as Fig. 3 institute Show.
Before processing, little laying down 54 and the Fv/Fm of 411 liang of leaf one heart stage seedling second leaves in capital is 0.803 and 0.805.
Fig. 3 shows, after wheat seedling high light processes 1h, Fv/Fm is different because processing the difference of leaf temperature.Little Lay down and 54 and the Fv/Fm in capital 411 process slightly below higher temperature (31-37 DEG C) at lower temperature (20-25 DEG C) high light High light processes, and the Fv/Fm that high temperature (42 DEG C) high light processes blade is then strong significantly lower than higher temperature (31-37 DEG C) Fv/Fm after optical processing.
By with process before little lay down 54 and 411 liang of leaf one heart stage seedling second leaves in capital Fv/Fm compared with, little 54 Hes of laying down The fall of the Fv/Fm that capital 411 high light at different temperatures processes all there are differences, and this Differences is low Become apparent under temperature (20-25 DEG C) and high temperature (42 DEG C), and to little lay down 54 and capital 411 at variant temperature The fall of Fv/Fm that processes of high light compare, it is also possible to proves, little lay down 54 resistance to high light ability be more than capital 411, the qualification result that this conclusion is reported with known references is consistent.
Embodiment 2, the high light of different times Semen Tritici aestivi the second leaf process
At 23-26 DEG C, with 1000 μm ol m-2 s-1As relative high light, step one in embodiment 1 is cultivated After germination, little 54 seedling disjunctor the second leaves of laying down after 18-26 days carry out 1h high light process, and relative humidity controls 40%-60%.After Semen Tritici aestivi disjunctor vanes dark adaptation 15-20min after processing, glimmering with Handy-pea chlorophyll Light instrument (purchased from Hansatech company of Britain) measures lightsystemⅡ maximal photochemistry efficiency (Fv/Fm), full during mensuration It is set as 3000 μm ol m with light intensity-2 s-1, saturated flash time is 1s.
Result is as shown in table 3.
Semen Tritici aestivi disjunctor second leaf of different number of days Fv/Fm after high light processes after table 3 germination
Natural law after germination 18 20 22 24 26
Fv/Fm 0.683 0.693 0.708 0.705 0.686
Table 3 shows, wheat seedling the second leaf germinateed latter 18 days and about 26 days is through 1000 μm ol m-2 s-1G is strong After optical processing 1h, its Fv/Fm is more lower slightly than the Fv/Fm of 20-24 days wheat seedling the second leaves after germinateing.This experiment is said Bright, for wheat seedling the second leaf, within after germination 20-24 days, relatively stable testing result can be obtained.
Embodiment 3, Semen Tritici aestivi Different Leaf-position Leaf Blades high light process
One, wheat seedling earth culture
Little 54 seeds of laying down directly are seeded in and fill 0.8-1.0 kilogram of Vermiculitum: in the plastic flowerpot of soil (2:1), mould Material flowerpot size is upper bore 17 centimetres, and lower bore 12 centimetres is high 15 centimetres.It is placed in glasshouse growth.Temperature Room condition is: temperature 18-25 DEG C (daytime)/10-15 DEG C (night), photoperiod 10-12h, humidity 30-50%, Light intensity is 200-600 μm ol m-2 s-1.Until its long to one heart stage of leaf time, water weekly Semen Tritici aestivi pancebrin (such as table 1 Shown in), until long to 5-6 leaf phase (the most after planting 50-60 days), carefully dig out whole strain seedling, with washing from the beginning After root, plant disjunctor 1-5 blade is carried out high light process.
Two, the high light of Different Leaf-position Leaf Blades processes
Length step one obtained is to wheat seedling during the 5-6 leaf phase, respectively to its disjunctor 1-5 leaf (from bottom to top) At 23-26 DEG C, with 1000 μm ol m-2 s-1Carry out high light as relative high light and process 30min.Relative humidity controls At 40%-60%.After Semen Tritici aestivi disjunctor vanes dark adaptation 15-20min after processing, green with Handy-pea leaf Element luminoscope (purchased from Hansatech company of Britain) measures lightsystemⅡ maximal photochemistry efficiency (Fv/Fm), measures Time saturated light intensity be set as 3000 μm ol m-2 s-1, saturated flash time is 1s.
Semen Tritici aestivi various position leaves disjunctor vanes 1000 μm ol m-2 s-1High light processes Fv/Fm change such as figure after 30min Shown in 4.
In Fig. 4, L1-L5 represents 1-5 blade respectively.
Fig. 4 shows, the 1st and the 2nd leaf high light Fv/Fm before treatment from bottom to top is that 0.77-0.78 is less than 0.80, Show that it there occurs a certain degree of aging, and the Fv/Fm of remaining 3-5 leaf is all near 0.8, shows its physiology State is normal.Semen Tritici aestivi Different Leaf-position Leaf Blades is through 1000 μm ol m-2 s-1After g light intensity processes 30min, its Fv/Fm is equal Significantly reducing before processing than high light, wherein the 1st leaf is reduced by up to, and is down to about 0.67, and 2-4 leaf is down to 0.72 Left and right, the 5th leaf is about 0.75.This embodiment illustrates, utilizes the method to compare the resistance to high light of Wheat Cultivars Ability is not limited to the second leaf, and same the method can be used for the different times resistance to high light of various position leaves Semen Tritici aestivi and identifies.
Embodiment 4, Oryza sativa L. disjunctor blade high light process
One, rice seedling is cultivated
By excellent for Oryza sativa L. two 2163 and the bright seed of extensive 63 through 30%H2O2(10-30%) seed soaking sterilization 10min After (10-20min), it is transferred to be placed with in the culture dish of moistening filter paper that to be placed in 30 DEG C (28-31 DEG C) permanent Incubator germinates, the seed showed money or valuables one carries unintentionally was put in the nylon Dropbox (30cm × 20cm × 8.5cm filling tap water in second day (long × wide × deep)) in, at intelligence growth cabinet, (model is HP1000GS, purchased from the auspicious China in Wuhan instrument and equipment Co., Ltd) middle growth.Condition of culture is: 14h photoperiod (12-14h), light intensity 150-200 μ mol m-2 s-1(100-300μmol m-2 s-1) LED light source that carried by instrument provides;Day temperature is 28 DEG C (26-28 DEG C), nocturnal temperature is 25 DEG C (23-26 DEG C), relative humidity 40%-60%.After one week, will Seedling moves into 30cm × 30cm × 15cm (long × wide × deep) and fills the plastics of pancebrin (as shown in table 4) In box, continue to cultivate (condition of culture is identical) in intelligence growth cabinet and open up completely to two leaf one heart stage that is second leaves When opening standby.
Table 4 Oryza sativa L. pancebrin formula (1000 times, surplus is deionized water)
Note: the Oryza sativa L. pancebrin of working concentration is the Oryza sativa L. pancebrin deionized water dilution 1000 of above-mentioned table 4 Again (adjusting pH is 5.5).
Two, the resistance to high light of rice seedling disjunctor blade is identified
Second leaf of the two leaf one heart stage rice seedlings obtaining step one carries out high light process.Will first with double faced adhesive tape Blade is fixed on the flat board of 30cm × 16cm, makes blade to be identified be arranged in order and is maintained at same level, with oneself Upper and lower light keeps vertically (as shown in Figure 1).At 25-26 DEG C, through 800 μm ol m-2 s-1Light intensity processes The process of 1h, relative humidity controls at 40%-60%.Rice leaf after processing is through dark adaptation 15min (15-20min After), measure lightsystemⅡ maximal photochemistry efficiency (Fv/Fm) with Handy-pea chlorophyll fluorescence instrument, measure Time saturated light intensity be set as 3000 μm ol m-2 s-1, saturated flash time is 1s.
Rice leaf is through 800 μm ol m-2 s-1Light intensity processes the Fv/Fm after 1h and changes as shown in Figure 5.
Fig. 5 shows, high light processes front Oryza sativa L. two excellent 2163 and the bright Fv/Fm of extensive 63 is respectively 0.810 and 0.805; Through 800 μm ol m-2 s-1After light intensity processes 1h, two excellent 2163 and the bright Fv/Fm of extensive 63 reduce to 0.744 He respectively 0.711.After high light process, the Fv/Fm of two excellent 2163 is slightly above bright extensive 63, shows the resistance to high light characteristic of two excellent 2163 It is better than bright extensive 63.This embodiment illustrates, the method can be used for the resistance to high light of other grasses in addition to Semen Tritici aestivi and identifies.

Claims (16)

1. the method comparing plant resistance to high light ability, it it is the disjunctor blade tiling of certain same leaf position that the difference of the identical trophophase obtained under same culture conditions is treated measuring plants, and under mutually synthermal and relative humidities, described disjunctor blade is processed at the high light that the position acceptance vertical with light is identical, the lightsystemⅡ maximal photochemistry efficiency Fv/Fm making the disjunctor blade of this same leaf position is respectively less than 0.75, obtains difference this disjunctor blade after the process of measuring plants;Measure the lightsystemⅡ maximal photochemistry efficiency Fv/Fm that difference treats this disjunctor blade before treatment of measuring plants, be denoted as A;Measure the lightsystemⅡ maximal photochemistry efficiency Fv/Fm of different this disjunctor blade after the process of measuring plants, be denoted as B;The value that the same A treating measuring plants deducts B gained is denoted as C;If treating that the C value of measuring plants X is less than the C value treating measuring plants Y, then treat that the resistance to high light ability of measuring plants X is better than and treat measuring plants Y;
Described disjunctor blade is the Non in vitro blade being connected on whole strain plant to be measured;
Described plant is same plant;
Described disjunctor blade is the Non in vitro blade being connected on whole strain plant to be measured, is put in high light processing equipment by plant to be measured for whole strain and operate during operation;Before described high light processes, will treat that measuring plants is cultivated a period of time in pancebrin;
Described high light processing equipment is intelligence growth cabinet.
Method the most according to claim 1, it is characterised in that: described relative humidity is 40%-60%.
Method the most according to claim 1 and 2, it is characterised in that: described trophophase was two leaf one heart stages;
Described same leaf position is the second leaf.
Method the most according to claim 1 and 2, it is characterised in that: the light intensity that described high light processes is 700-1000 μm ol m-2 s-1
Method the most according to claim 4, it is characterised in that: the light intensity that described high light processes is 700 μm ol m-2 s-1、800μmol m-2 s-1Or 1000 μm ol m-2 s-1
Method the most according to claim 1 and 2, it is characterised in that: the process time that described high light processes is 10min-4 h.
Method the most according to claim 6, it is characterised in that: the process time that described high light processes is 10min, 20min, 30min, 1h, 1.5h, 3h or 4h.
Method the most according to claim 1 and 2, it is characterised in that obtain difference this disjunctor blade after the process of measuring plants described in: and also include the step of dark processing between the lightsystemⅡ maximal photochemistry efficiency Fv/Fm of the described mensuration difference this disjunctor blade after the process of measuring plants.
Method the most according to claim 8, it is characterised in that: the time of described dark processing is 15-20min.
Method the most according to claim 9, it is characterised in that: the time of described dark processing is 15min.
11. methods according to claim 1 and 2, it is characterised in that: described plant is grass.
12. methods according to claim 1 and 2, it is characterised in that: described plant is Semen Tritici aestivi or Oryza sativa L..
The lightsystemⅡ maximal photochemistry efficiency Fv/Fm of 13. 1 kinds of disjunctor blades the making plant method less than 0.75, is to be tiled by the disjunctor blade of plant, described disjunctor blade accepts in the position vertical with light high light and processes;
Described disjunctor blade is the Non in vitro blade being connected on whole strain plant.
14. methods according to claim 13, it is characterised in that: described plant is grass.
15. methods according to claim 13, it is characterised in that: described plant is Semen Tritici aestivi or Oryza sativa L..
The application in plant selection-breeding of the 16. claim 1-15 arbitrary described method.
CN201410549301.XA 2014-10-16 2014-10-16 Method for comparing hard-light resistance of plants Expired - Fee Related CN104303867B (en)

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