CN113030037B - Chlorophyll fluorescence kinetics analysis method for Chinese torreya male seedlings and female seedlings under high-temperature stress - Google Patents
Chlorophyll fluorescence kinetics analysis method for Chinese torreya male seedlings and female seedlings under high-temperature stress Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
Abstract
The invention provides a method for analyzing chlorophyll fluorescence kinetics of Chinese torreya seedlings under high-temperature stress, which comprises the following steps: taking a 2-year-old torreya tree original seedling as a stock, respectively harvesting a torreya grandis female seedling and a torreya grandis male Miao Youzhi as scion, grafting and growing for 2 years, and taking the scion as a research object; placing the female grafted seedling and the male grafted seedling in an illumination incubator at 25 ℃,30 ℃,35 ℃ and 40 ℃ respectively, and placing the female grafted seedling and the male grafted seedling in a dark environment at 10 h after illumination of 14 h; after the leaves are subjected to dark adaptation for 30 min, carrying out rapid chlorophyll fluorescence induction kinetic curve measurement, measuring 4 strains of male and female grafted seedlings respectively, and repeatedly measuring each strain for 2 times; the data were processed and analyzed and compared using the least significant difference method. The invention reflects the situation that irreversible damage is caused to seedlings when the torreya grafting seedlings are planted in a large area in the production practice process, and the test conclusion can further provide a theoretical basis for the torreya grafting seedling planting process.
Description
Technical Field
The invention relates to the field of general botanic, in particular to a method for analyzing chlorophyll fluorescence kinetics of Chinese torreya seedlings and female seedlings under high-temperature stress.
Background
Chinese torreyaTorreya grandis cv. 'Merrillii Hu') belonging to the genus Taxaceae (Taxoceae) Torreya @ genus @Torrey) Evergreen arbor is a variety of Chinese torreya tree subjected to asexual propagation. The grafting improvement is used for fruit, oil and other multipurpose excellent economic tree species. The absolute minimum temperature of the suitable life is-8 to-15 ℃, the average annual temperature is 14.5 to 17.5 ℃, and the reference literature shows that: in a continuous sunny and hot high-temperature environment in summer, the production of Chinese torreya is seriously affected, and measures such as no tending operation, shading irrigation and the like are adopted for newly-built Chinese torreya forests to reduce losses. The torreya is sufficient in rainwater and proper in temperature before flowering, the rainwater is moderate in flowering period and falling period, the illumination is sufficient, the rainwater is sufficient in fruit setting period, and the cool climate in summer is favorable for the high yield of the torreya. In the high temperature stress process, the relative membrane permeability, MDA content, SOD, POD, CAT enzyme activity, pro and SS content of the Chinese torreya leaf take 39 ℃ as peak values, and after continuously risingGradually decrease, when the temperature reaches 42 ℃, the torreya leaf damage is serious, and the protective enzyme system is destroyed after short-time stress.
The grafting technology is widely applied in the production process of Chinese torreya, and is also used as one of the measures for improving the yield and the resistance of Chinese torreya. In order to comprehensively reflect the situation that irreversible damage is caused to seedlings when torreya grafting seedlings are planted in a large area in the production practice process, a test conclusion can further provide theoretical basis for the torreya grafting seedling planting process, guide production and reduce loss. Moreover, the sensitivity of the torreya grafted female seedling and the grafted male seedling to the high temperature in summer is not completely the same, so that in order to more accurately guide the cultivation management of the torreya grafted female seedling and the grafted male seedling, the rapid chlorophyll fluorescence induction kinetic parameters and the OJIP curve of the torreya grafted female seedling and the grafted male seedling under high temperature stress are required to be researched and analyzed, and the parameters reflected by partial light systems of the torreya grafted female seedling and the grafted male seedling under high temperature stress are required to be discussed.
Disclosure of Invention
The invention aims to provide a method for analyzing chlorophyll fluorescence kinetics of Chinese torreya seedlings and female seedlings under high-temperature stress.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a method for analyzing chlorophyll fluorescence kinetics of Chinese torreya seedlings and female seedlings under high temperature stress comprises the following steps:
step 1): taking a 2-year-old torreya tree original seedling as a stock, respectively harvesting a torreya grandis female seedling and a torreya grandis male Miao Youzhi as scion, grafting and growing for 2 years, and taking the scion as a research object;
step 2): placing the female grafted seedling and the male grafted seedling in illumination incubator at 24-26deg.C, 29-31deg.C, 34-36 deg.C and 39-41 deg.C respectively, controlling the rest variables, illuminating 13-15 and h, reducing temperature by 1-3deg.C, and placing in dark environment 9-11 h;
step 3): after the leaves are subjected to dark adaptation for 25-35 min, selecting the leaves at the 3 rd-4 th leaf positions under the top leaves of the new shoots of the plants to carry out rapid chlorophyll fluorescence induction kinetic curve measurement, measuring 4 plants of the male and female grafted seedlings respectively, repeatedly measuring each plant for 2 times, and measuring the fluorescence parameters as follows:
F O : initial fluorescence intensity (50 [ mu ] s) after dark adaptation; f (F) M : maximum fluorescence intensity (0.3-2 s); f (F) J : fluorescence intensity when 2 ms was illuminated; f (F) I : fluorescence intensity at 30 ms under illumination;V t ≡(F t -F O )/(F M -F O ): relatively variable fluorescence at time t;V J =(F J -F O )/(F M -F O ): point J relative variable fluorescence;Ψ O ≡ET O /TR O =(1-V J ): the trapped excitons transfer electrons into the electron transfer chain beyondQ A - Other electron acceptor probabilities of (c) in the electron acceptor;φ Eo ≡ET O /ABS=[1-(F O /F M )]Ψ O : the absorbed energy is used for the quantum yield of electron transfer;φ Do ≡1-φ Po =(F O /F M ): quantum ratio for heat dissipation;δ Ro ≡REo/ETo=(F M -F I )/(F M -F J ): the captured energy can be transferred to the quantum yield at the end of the electron chain; ABS/CSm approximatelyF M : light energy absorbed per unit area; TRo/CSm =φ Po (ABS/CSm): light energy captured per unit area; ETo/csm=φ Eo (ABS/CSm): quantum yield per unit area of electron transfer; DIo/csm= (ABS/CSm) - (TRo/CSm): heat dissipation per unit area; RC/csm=φ Po (V J Mo) (ABS/CSm): the number of reaction centers per unit area; PI (proportional integral) ABS ≡(RC/ABS)[φ Po /(1-φ Po )][Ψ O /(1-Ψ O )]: performance index based on absorbed light energy; PI (proportional integral) total ≡PI ABS (δ Ro /1-δ Ro ): and (5) comprehensive performance index.
Step 4): all data were processed and analyzed and multiple comparisons were made using the least significant difference method.
Preferably, in the step 2), the grafted female seedling and the grafted male seedling are placed in an illumination incubator at a temperature of 25 ℃,30 ℃,35 ℃ and 40 ℃ respectively, and after illumination of 14 and h, the temperature is reduced by 2 ℃ and placed in a dark environment of 10 h.
Preferably, in step 3), the rapid chlorophyll fluorescence induction kinetics is determined using a multifunctional plant efficiency analyzer (M-PEA, hansatech, norfolk, UK).
The technical scheme has the following beneficial effects:
according to the invention, research and analysis are carried out on quick chlorophyll fluorescence induction kinetic parameters and OJIP curves of torreya grandis grafted female seedlings and grafted male seedlings under high temperature stress, and the parameters reflected by partial light systems of the torreya grandis grafted female seedlings and grafted male seedlings under the high temperature stress environment are discussed, so that irreversible damage to seedlings can be caused under the conditions when torreya grandis grafted seedlings are planted in a large area in the production practice process, and a test conclusion can further provide theoretical basis for the torreya grandis grafted seedling planting process, guide production and reduce losses.
Drawings
The invention is further described below with reference to the accompanying drawings:
FIG. 1 is a comparison of rapid fluorescence induction curves of torreya grandis grafted male and female seedling leaves under different temperature stresses.
FIG. 2 shows a torreya grandis grafted male and female seedling leaf F under different temperature stresses V /F M And (5) comparing the values.
FIG. 3 shows the torreya grandis grafted male and female seedling leaf PI under different temperature stresses ABS And (5) comparing the values.
FIG. 4 shows a torreya grandis grafted male and female seedling leaf PI under different temperature stresses total And (5) comparing the values.
FIG. 5 is a comparison of net photosynthetic rate values of torreya grandis grafted male and female seedling leaves under different temperature stresses.
FIG. 6 is a comparison of the stomatal conductance values of torreya grandis grafted male and female seedling leaves under different temperature stresses.
FIG. 7 shows the intercellular CO of the leaves of torreya grandis grafted male and female seedlings under different temperature stresses 2 Concentration values are compared.
Fig. 8 is a comparison of transpiration rate values of torreya grandis grafted male and female seedling leaves under different temperature stresses.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and examples.
The invention provides a method for analyzing chlorophyll fluorescence kinetics of Chinese torreya seedlings under high-temperature stress, which comprises the following steps:
step 1): taking a 2-year-old torreya tree original seedling as a stock, respectively harvesting a torreya grandis female seedling and a torreya grandis male Miao Youzhi as scion, grafting and growing for 2 years, and taking the scion as a research object;
step 2): placing the female grafted seedling and the male grafted seedling in illumination incubator at 24-26deg.C, 29-31deg.C, 34-36 deg.C and 39-41 deg.C respectively, controlling the rest variables, illuminating 13-15 and h, reducing temperature by 1-3deg.C, and placing in dark environment 9-11 h;
in the embodiment, the grafting female seedling and the grafting male seedling are respectively placed in an illumination incubator with the temperature of 25 ℃,30 ℃,35 ℃ and 40 ℃, and after illumination of 14 and h, the temperature is reduced by 2 ℃ and placed in a dark environment of 10 h;
step 3): after the leaves are subjected to dark adaptation for 25-30 min, rapid chlorophyll fluorescence induction kinetic curve determination is carried out, 4 plants are respectively measured for male and female grafted seedlings, each plant is repeatedly determined for 2 times, and the determined fluorescence parameters are as follows: fo: initial fluorescence intensity (50 [ mu ] s) after dark adaptation; f (F) M : maximum fluorescence intensity (0.3-2 s); f (F) J : fluorescence intensity when 2 ms was illuminated; f (F) I : fluorescence intensity at 30 ms under illumination;V t ≡(F t –Fo)/(F M -Fo): relatively variable fluorescence at illumination time t;V J =(F J –Fo)/(F M -Fo): point J relative variable fluorescence;Ψo≡ETo/TRo=(1-V J ): the trapped excitons transfer electrons into the electron transfer chain beyondQ A - Other electron acceptor probabilities of (c) in the electron acceptor; phi (phi) Eo ≡ETo/ABS=[1-(Fo/F M )]Ψ O : the absorbed energy is used for the quantum yield of electron transfer;φ Do ≡1-φ Po ＝(Fo/F M ): quantum ratio for heat dissipation;δ Ro ≡REo/ETo=(F M -F I )/(F M -F J ): the captured energy can be transferred to the quantum yield at the end of the electron chain; ABS/CSm approximately equal to F M : light energy absorbed per unit area; TRo/CSm =φ Po (ABS/CSm): light energy captured per unit area; ETo/csm=φ Eo (ABS/CSm): quantum yield per unit area of electron transfer; DIo/csm= (ABS/CSm) - (TRo/CSm): heat dissipation per unit area; RC/csm=φ Po (V J Mo) (ABS/CSm): the number of reaction centers per unit area; PI (proportional integral) ABS ≡(RC/ABS)[φ Po /(1–φ Po )][Ψ O /(1–Ψ O )]: performance index based on absorbed light energy; PI (proportional integral) total ≡PI ABS (δ Ro /1–δ Ro ): and (5) comprehensive performance index.
In this example, a rapid chlorophyll fluorescence induction kinetics profile (OJIP) was determined using a multifunctional plant efficiency analyzer (M-PEA, hansatech, norfolk, UK). All leaves were measured after 30 min dark adaptation and JIP-test analysis was performed on the OJIP curve.
Step 4): all data were processed and analyzed and multiple comparisons were made using the least significant difference method.
In FIG. 1, after high temperature stress, the rapid chlorophyll fluorescence induction kinetics curve of the torreya grandis grafted female seedlings and male seedlings leaves is obviously changed. Within 35 ℃, the initial fluorescence of the fluorescent dye and the fluorescent dye is not larger than the change amplitude of normal temperature, and the initial fluorescence of the fluorescent dye and the fluorescent dye is greatly increased under the high-temperature stress of 40 ℃. The J-I phase inflection points of both began to disappear under high temperature stress at 40 ℃. The maximum fluorescence intensity of the two is reduced compared with the normal temperature, the high temperature reduction at 40 ℃ is particularly obvious, the J-I phase in the curve almost tends to be gentle, but is not completely disappeared, and the fluorescence output limit of the point I and the point P is obviously reduced. The results show that the damage degree of the photosynthetic mechanism of the torreya grandis grafted female seedling and the torreya grandis grafted male seedling is basically the same under the high temperature stress.
Fv/Fm reflects an important parameter of the light energy conversion efficiency of the PS II active center. In the figure, different capital/lowercase letters respectively represent that the difference between different varieties is obvious (P is less than 0.05) at different temperatures of the same variety and at the same temperature, and each numerical value is the average value plus or minus standard deviation and is the same as the following. From FIGS. 2 andtable 1 shows that F of torreya grafting seedlings V /F M The value gradually decreases with increasing stress (P < 0.05). Under normal conditions, F of grafting male seedlings and female seedlings of torreya grandis at the temperature of above 35 ℃ without obvious change in 30℃ treatment V /F M The values are obviously reduced by 37.3 percent and 25.7 percent compared with the control; the temperature of 40 ℃ is reduced to the minimum, the temperature is respectively reduced by 83 percent and 66.7 percent, and the male seedling is obviously lower than the female seedling, which indicates that electrons are directed from the PS II reaction centerQ A 、Q B And the transfer of the PQ library is blocked, the Fv/Fm value is obviously reduced, and the electron transfer inhibition in the Chinese torreya grafted male seedling leaves is higher than that of the grafted female seedling.
PI ABS Is a performance index based on the absorption of light energy. From fig. 3 and table 1, the photosynthetic performance index of the torreya grandis grafted seedlings is remarkably reduced under the high temperature stress (40 ℃). Starting from the environment culture at 25 ℃, the photosynthetic performance indexes of the Chinese torreya and the male seedlings show a trend of decreasing after rising, the performance index of the Chinese torreya grafted male seedlings reaches the highest value at about 30 ℃ and is always lower than that of female seedlings, and the difference is obvious. Under 40 ℃ high temperature stress, the photosynthesis performance index of the two is obviously reduced to reflect PI ABS Sensitivity to high temperature stress.
Combination Performance index PI total The electronic transfer activity of the torreya grafting Miao Guang system is mainly researched to reflect the influence of high temperature stress on torreya grafting seedling leaves PS I. As can be seen from fig. 8 and table 1, torreya grafting female and male seedlings PI under high temperature environmental stress total Significantly reduced. Both of them show a tendency to decrease after increasing from 25℃environmental culture, and reach the highest value at about 30 ℃. The comprehensive performance index of the torreya grafting male seedling is always higher than that of the female seedling, and the difference is obvious.
TABLE 1 maximum photochemical efficiency, photosynthetic Performance index and comprehensive Performance index differential analysis of leaves under the stress of torreya grandis grafting female and male Miao Gaowen
As can be seen from table 2, at normal temperature, the torreya grafted female seedling always absorbs higher light energy per unit area than the male seedling, the absorption amount is more, and the difference is obvious. Under the high-temperature stress at 30 ℃,35 ℃ and 40 ℃, the light energy (ABS/CSm) absorbed per unit area, the captured light energy (TRo/CSm) and the transmitted light energy (ETo/CSm) all rise and fall firstly, and the share of the absorption, capture and transmission electrons of the grafted female seedlings is obviously higher than that of the grafted male seedlings. The heat dissipation per unit area (DIo/CSm) increases significantly in a progressively higher temperature environment and the data in table 2 shows that the grafted female shoots dissipate a lower proportion of energy than the male shoots. The energy share of the torreya grandis leaves for electron transfer is reduced, the electron transfer is blocked, the energy share of heat dissipation of the torreya grandis leaves and the torreya grandis leaves is increased, and the electron blocking degree of the grafting female seedling leaves is lower than that of the grafting male seedling leaves. In general, the quantum efficiency parameters of the torreya grafting female seedlings and the male seedlings in unit area have large variation with the comparison temperature ratio under high temperature stress, and the difference is obvious, wherein the energy flow parameters of the torreya grafting female seedlings in unit area have small variation after the high temperature stress.
TABLE 2 energy flow parameters per unit cross-sectional area and differences in reactive center density under high temperature stress of torreya grandis grafted female and male seedlings leaves
V J Reflecting the closing degree of the active reaction center of the light system II, the torreya grafting female seedling and male seedling is more normal temperature under the high temperature stress of 40 DEG CV J Up by 123.76% and 324.55%, respectively.
After high temperature stress, torreya grandis is grafted with female seedlings and male seedling leavesΨo andφeo gradually decreases, and the difference between the decreasing amplitude and normal temperature is remarkable. Grafting female seedling leafΨo andφeo decreased to a lesser extent than male seedlings, but the difference was not significant. The quantum ratio of heat dissipation of the grafted female seedlings and the grafted male seedlings is respectively increased by 257.95 percent and 279.09 percent, and the increase value of the female seedlings is lower than that of the male seedlings, but the difference is not obvious.
TABLE 3 difference in Vj and Quantum yield of torreya grandis grafted female and male seedlings under high temperature stress
Measuring gas exchange parameters of torreya leaf by using portable photosynthetic system, wherein the gas flow during measurement is 500 mu mol s -1 Under the measurement condition of CO 2 Concentration of 400 + -5 mu mol -1 Measuring net photosynthetic rate (Pn), stomatal conductance (Gs) and intercellular CO of Chinese torreya grafted female and male seedling leaves 2 The measurement time of the parameters such as concentration (Ci), transpiration rate (Tr) and the like is 11:00-13:00 in noon on 3 rd day of continuous stress at different temperatures.
TABLE 4 gas exchange parameter differences under high temperature stress of torreya grandis grafted female and male seedlings
The data of table 4 and fig. 8 and the comparative graph show: the net photosynthetic rate of the leaf blade of the torreya grandis grafted female seedling and the leaf blade of the torreya grandis grafted male seedling are firstly increased and then decreased under different temperature stresses, the net photosynthetic rate of the female seedling is higher than that of the male seedling, the net photosynthetic rate of the female seedling and the net photosynthetic rate of the male seedling reach the highest value at about 30 ℃, and the difference is obvious from 35 ℃. The air pore conductivity value is continuously and obviously reduced from 25 ℃, and the air pore conductivity of female seedlings is higher than that of male Miao Yaolve. Intercellular CO 2 The concentration is higher than 30-35 ℃ in the room temperature environment, and the intercellular CO is in the high temperature environment 2 The concentration is obviously increased, and the intercellular CO of the male and female grafted seedling leaves at each temperature 2 The concentration difference is significant. The transpiration rate of the two leaves at different temperatures gradually decreases along with the temperature rise, and the transpiration rate of the female seedling leaves is higher than that of the male Miao Yaolve. The gas exchange parameters of the subject blades indicate: the gas exchange capacity of the leaves of the torreya grafting female seedling is stronger than that of the grafting male seedling under the high-temperature environment.
PS ii is the most sensitive region of the plant photosynthetic system to high temperature stress. The PS II inhibition condition of the Chinese torreya seedlings under continuous high-temperature treatment is studied, and the adaptability of the Chinese torreya seedling leaves to the high-temperature environment can be reflected. The change of the plant thylakoid membrane can be rapidly captured by chlorophyll fluorescence, and the main reason for the K point of the rapid induction curve of the chlorophyll fluorescence is thatIs the cracking process of water and partial electron primary quinone acceptor in the electron transfer processQ A ) Is inhibited, indicating that the oxygen-releasing complex is damaged to some extent. In the study, the OJIP curve of the high-temperature stress torreya grafting female seedling and male seedling has great change compared with room temperature, the maximum fluorescence intensity (P) of the female seedling is Gao Yuxiong seedling, the O-J-I curve of the female seedling and the O-J-I curve of the male seedling tend to be horizontal straight lines at the temperature of 40 ℃, the O-J-I curve reaches the highest value at the temperature of 35 ℃ and the O-J-I curve is in a trend of rising first and then reducing with the gradual rising of the temperature, wherein the O-J-I curve is obviously reduced in the environment of 40 ℃ high-temperature stress. The initial fluorescence of female seedlings is obvious Gao Yuxiong seedlings under the high-temperature stress at 40 ℃, the curve is more gentle compared with male seedlings, and the J-I inflection point of the female seedlings disappears later than the male seedlings. The decrease of the fluorescence yields of the points I and P of female seedlings in each temperature environment is smaller than that of male seedlings, and the female seedlings are more obvious at 40 ℃. Further indicates that the damage of high temperature stress to the torreya grafting female seedlings is smaller than that of the male seedlings.
The result shows that the damage degree of the photosynthetic mechanism of the torreya grandis grafted female seedling is lower than that of the torreya grandis grafted male seedling under high temperature stress. The OJIP curve can intuitively reflect the degree of the adverse injury of plants, and the high-temperature stress resistance of the torreya grandis grafted female seedlings is stronger than that of the male seedlings.
In this experiment, high temperature not only damages the electron transfer on the donor side of the leaf, but also suppresses and reduces the rate of electron transfer on the acceptor side of the PS ii. After the experimental result is analyzed by JIP-test, after the torreya grafting female seedlings and male seedlings are subjected to high-temperature stress,V J all rise significantly, indicating that electron transfer is affected. Subjected to both high temperature stress at 40 DEG CΨo significantly decreases, resulting inφEo drops, the subject spends too many electrons activated,φdo increases significantly. From test data, after 40 ℃ high temperature stress, torreya grafting female seedlingsV J The ascending amplitude is small, the energy consumption is low, more electrons enter a transmission chain, the electron-measuring transmission capacity of a receptor in the leaf light system II is larger than that of a torreya grafting male seedling, and the capacity of a photosynthetic mechanism of the leaf light system II for coping with high-temperature stress environment is stronger than that of the grafting male seedling.
In the research, fv/Fm of torreya grafted female seedlings and male seedlings is reduced due to temperature stress, which accords with the research result of plants after high temperature stress, and the research result shows that the efficiency of converting light energy of PS II after high temperature stress into electron transfer is reduced. Fv/Fm values of the Chinese torreya grafted female seedlings are less reduced than those of the male seedlings after high-temperature stress, so that the light energy conversion rate and the electron transfer efficiency of the Chinese torreya grafted female seedlings PS II in a high-temperature environment are higher than those of the grafted male seedlings under the same conditions. Under the stress of temperature, the energy flow distribution of the reaction centers of the two leaves is obviously changed, and the descending amplitude of the ABS/CSm, TRo/CSm and ETo/CSm of the torreya grafted female seedlings is obviously higher than that of the torreya grafted male seedlings.
PI ABS Mainly reflecting the efficiency of the PS II reaction center. The study considers PI ABS The operation state of a plant photosynthetic mechanism can be accurately reflected, and the plant photosynthetic mechanism is more sensitive to adversity stress than Fv/Fm. In the present study, torreya grandis was grafted with female and male seedling leaves PI ABS Are sensitive to temperature changes as are Fv/Fm, but the photosynthetic performance index is much more sensitive to this. Simultaneously PIabs and PItotal appear simultaneously, solve the problem that can not reflect optical system I, reflect the transmission ability between optical system I and optical system II and other relevant performances. Compared with the prior art, the method has the advantages that the conditions of rapid measurement and the like after 2 hours of high-temperature stress on plants are different, the continuous high-temperature stress environment is adopted to carry out simulation experiments on plants, and the temperature is 30 ℃ higher than the normal-temperature PI ABS And PI (proportional integral) total Rise to PI ABS The increase is significant, both of which show a gradual decrease trend with increasing temperature, which is mainly related to the gradual destruction of photosynthetic mechanisms, consistent with the trend of Fv/Fm. From the data, the torreya grafting female seedling is in PI under high temperature environmental stress than the grafting male seedling ABS And PI (proportional integral) total The upper part of the product has better performance.
In the experimental process, the temperature change causes the gas exchange parameters of the Chinese torreya grafted male and female seedling leaves, and as the temperature rises, the net photosynthetic rate (Pn), the stomatal conductance (Gs) and the transpiration rate (Tr) gradually decrease to meet the research results after the high-temperature stress of plants, which shows that the photosynthesis of the plants is inhibited. Intercellular CO 2 The concentration (Ci) is caused by the damage of photosynthetic mechanisms of torreya grandis leaves along with the rise of temperature, the respiration is continuous, the photosynthesis is weakened, and the intercellular CO is caused 2 The concentration increases. Experimental data indicate that: ring at room temperatureIn the environment, the Pn, gs and Tr values of the torreya grandis grafted male and female seedling leaves are not obvious, and the difference is kept at a certain level. In the process of increasing the temperature, except that Pn slightly increases at about 30 ℃, each numerical value is obviously reduced in a high-temperature environment, and three numerical values of the torreya grafted female seedlings are higher than those of the grafted male seedlings in each temperature environment. CO between two cells 2 The concentration is lowest within the range of 30-35 ℃, which shows that in the temperature range, the photosynthesis and respiration difference are closest, and the intercellular CO of the torreya grafted female seedlings is the same in the whole temperature change process 2 The concentration is obviously lower than that of the grafted male seedlings.
In conclusion, the photosynthetic performance of the leaves of the torreya grandis grafted seedlings is the most stable under the environment of about 30 ℃; through experimental data and discussion and a rapid chlorophyll fluorescence induction kinetic curve, the adaptability of torreya grandis grafted female seedlings to high temperature is stronger than that of grafted male seedlings, and the female grafted seedlings have wider seed adaptation range than that of the male grafted seedlings in the production practice process.
In addition to the above preferred embodiments, the present invention has other embodiments, and various changes and modifications may be made by those skilled in the art without departing from the spirit of the invention, which is defined in the appended claims.
Claims (3)
1. The method for analyzing chlorophyll fluorescence kinetics of Chinese torreya seedlings under high-temperature stress is characterized by comprising the following steps:
step 1): taking a 2-year-old torreya tree original seedling as a stock, respectively harvesting a torreya grandis female seedling and a torreya grandis male Miao Youzhi as scion, grafting and growing for 2 years, and taking the scion as a research object;
step 2): placing the female grafted seedling and the male grafted seedling in illumination incubator at 24-26deg.C, 29-31deg.C, 34-36 deg.C and 39-41 deg.C respectively, controlling the rest variables, illuminating 13-15 and h, reducing temperature by 1-3deg.C, and placing in dark environment 9-11 h;
step 3): after the leaves are subjected to dark adaptation for 25-35 min, selecting the leaves at the 3 rd-4 th leaf positions under the top leaves of the new shoots of the plants to carry out rapid chlorophyll fluorescence induction kinetic curve measurement, measuring 4 plants of the male and female grafted seedlings respectively, repeatedly measuring each plant for 2 times, and measuring the fluorescence parameters as follows:
F O : initial fluorescence intensity after dark adaptation; f (F) M : maximum fluorescence intensity; f (F) J : fluorescence intensity when 2 ms was illuminated; f (F) I : fluorescence intensity at 30 ms under illumination;V t ≡(F t -F O )/(F M -Fo): relatively variable fluorescence at time t;V J =(F J -F O )/(F M -F O ): point J relative variable fluorescence;Ψ O ≡ET O /TR O =(1-V J ): the trapped excitons transfer electrons into the electron transfer chain beyondQ A - Other electron acceptor probabilities of (c) in the electron acceptor;φ Eo ≡ET O /ABS=[1-(F O /F M )]Ψ O : the absorbed energy is used for the quantum yield of electron transfer;φ Do ≡1-φ Po =(F O /F M ): quantum ratio for heat dissipation;δ Ro ≡REo/ETo=(F M -F I )/(F M -F J ): the captured energy can be transferred to the quantum yield at the end of the electron chain; ABS/CSm approximatelyF M : light energy absorbed per unit area; TRo/CSm =φ Po (ABS/CSm): light energy captured per unit area; ETo/csm=φ Eo (ABS/CSm): quantum yield per unit area of electron transfer; DIo/csm= (ABS/CSm) - (TRo/CSm): heat dissipation per unit area; RC/csm=φ Po (V J Mo) (ABS/CSm): the number of reaction centers per unit area; PI (proportional integral) ABS ≡(RC/ABS)[φ Po /(1-φ Po )][Ψ O /(1-Ψ O )]: performance index based on absorbed light energy; PI (proportional integral) total ≡PI ABS (δ Ro /1-δ Ro ): a comprehensive performance index;
step 4): all data were processed and analyzed and multiple comparisons were made using the least significant difference method.
2. The method for analyzing chlorophyll fluorescence kinetics of Chinese torreya seedlings and female seedlings under high temperature stress according to claim 1, wherein in the step 2), the grafted female seedlings and the grafted male seedlings are respectively placed in an illumination incubator at the temperature of 25 ℃,30 ℃,35 ℃ and 40 ℃, and after illumination of 14 and h, the temperature is reduced by 2 ℃ and placed in a dark environment of 10 h.
3. The method for analyzing chlorophyll fluorescence kinetics of Chinese torreya seedlings under high temperature stress according to claim 1, wherein in the step 3), a multifunctional plant efficiency analyzer is used for measuring a rapid chlorophyll fluorescence induction kinetics curve.
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