CN110907417A - Method for rapidly evaluating heat resistance of alum roots - Google Patents
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Abstract
The invention discloses a method for rapidly evaluating heat resistance of alum roots, which comprises the following steps: subjecting the variety of alum root to be evaluated to heat stress treatment, and measuring the value of the chlorophyll fluorescence kinetic parameter Fv/Fm, wherein Fv/Fm is more than or equal to 0.70, the variety has strong heat resistance, and 0.55<Fv/Fm<0.70 is a medium heat-resistant variety, and Fv/Fm is less than or equal to 0.55 is a weak heat-resistant variety. According to the invention, the heat stress treatment at 45 ℃ for 3h and 6h is carried out on 37 varieties, and the corresponding heat damage index and chlorophyll fluorescence kinetic parameters are determined. Through the correlation analysis of the chlorophyll fluorescence kinetic parameter indexes and F after high-temperature stress treatmentv/FmThe correlation analysis of the value and the heat injury index shows that the Fv/Fm value after 3h treatment is the optimal index for the heat resistance analysis of different alum root varieties. Comprehensively evaluating the heat resistance of the alum root according to the thermal injury index and the chlorophyll fluorescence kinetic parameters to establish a method capable of quickly evaluating the heat resistance of the alum root variety。
Description
Technical Field
The invention belongs to the field of plant tolerance heat evaluation, and particularly relates to a method for quickly evaluating heat resistance of alum roots.
Background
Alum root (Heuchera spp.) is a general name of plants of the genus alum in the family saxifragaceae, is native to north america, and is a perennial ground cover plant with colored leaves which can be enjoyed in all seasons (xie ying et al, 2018). It has been widely used in europe and america gardens, with a reputation of "garden palette" (invaliqing et al, 2018). In recent years, the number of alum root varieties introduced into China is increased year by year, and about 100 alum root varieties are accumulated. The flower pot has the outstanding advantages of various varieties, rich colors, long ornamental period, strong adaptability and the like, is applied to flower beds, family gardening and three-dimensional greening in various forms, is deeply favored by the market and has wide development prospect (Lichuaiqi and the like, 2017). The alum roots are cold-resistant, and are distributed in 4-9 areas according to the division standard of vegetation cold-resistant areas of the United states department of agriculture, so that the alum roots can successfully overwinter in open field in most areas of China. However, in introduction and cultivation of alum roots and garden application in various places in China, it is found that the difficulty in plant growth and even death during the over-summer process are key problems limiting the cultivation and popularization of part of alum root varieties.
In recent years, with the rising global temperature, plant growth and development face more serious challenges. Under high temperature stress, the phenotype and physiological properties of cell membrane systems, transpiration, photosynthesis, etc. of many ornamental plants are affected to varying degrees (xu lan xin and tian, e.g. men, 2019).
The heat damage index is the most direct phenotypic parameter for evaluating the heat resistance of the plant, and can more intuitively reflect the damage degree of the plant under high temperature stress. The smaller the value of the thermal injury index, the stronger the heat resistance of the plant. However, when a significant change in phenotype occurs, the heat stress time required is long and plant damage is already severe, often difficult to recover.
The chlorophyll fluorescence parameters can directly reflect the photochemical activity of the plant, and the operation is simple, the speed is high, and the chlorophyll fluorescence parameters are widely applied to the research of the ornamental plant responding to the high-temperature stress (Wangzhong et al, 2012; Zhoushui et al, 2016; Zhang Fang et al, 2019). Fv/Fm in chlorophyll fluorescence parameters belongs to a thermosensitive index and is considered as an important index for evaluating plant heat resistance (Wujizi36191et al, 2019). Currently, studies on the evaluation of the heat resistance of alum roots are not deep, and only certain studies on the photosynthetic properties of a few varieties at high temperatures have been conducted (qiden et al, 2014).
Disclosure of Invention
The method is to perform heat stress treatment on different alum root varieties, record the plant states after treatment and calculate the heat damage index according to a formula; the chlorophyll fluorescence parameters are measured and analyzed by using a continuous excitation type fluorometer, and the heat resistance of different alum root varieties is comprehensively evaluated from the phenotype and physiological levels, so that a method capable of quickly evaluating the heat resistance of the alum root varieties is established.
The invention provides a method for rapidly evaluating the heat resistance of alum root, which comprises the following steps: carrying out heat stress treatment on the alum root variety to be evaluated, and measuring the value of chlorophyll fluorescence kinetic parameter Fv/Fm, wherein Fv/Fm is more than or equal to 0.70 and is a variety with stronger heat resistance, Fv/Fm is more than 0.55 and less than 0.70 and is a variety with middle heat resistance, and Fv/Fm is less than or equal to 0.55 and is a variety with weaker heat resistance.
Among them, the heat stress temperature is preferably 45 ℃.
Among them, the heat stress treatment time is preferably 3 hours.
Wherein, chlorophyll fluorescence kinetic parameter determination is carried out on mature leaves of the middle wheel of the alum root.
The method is established by taking 37 alum root varieties as test materials, performing heat stress treatment on different alum root varieties, and comprehensively evaluating the heat resistance of the alum root varieties according to the heat damage index and the chlorophyll fluorescence kinetic parameters, and can quickly evaluate the heat resistance of the alum root. Accordingly, 12 varieties with strong heat resistance, such as 'sweet tea', 'orange jam', 'royal wine', and the like, 17 varieties with moderate heat resistance, such as 'peach flame', 'berry fruit', and the like, are screened out; 8 varieties with weak heat resistance, such as 'fire alarm' and 'lemon yellow' are adopted. The research can lay a good foundation for the deep research of the heat-resisting mechanism of the alum root, and simultaneously can provide important references for the popularization of alum root varieties with strong adaptability in summer and the breeding work of new heat-resisting alum root varieties by using resistant parents.
The invention combines the visual thermal injury index with F capable of responding to heat stress earlyv/FmThe method is combined, and comprehensive evaluation of heat resistance of different alum root varieties is carried out from phenotype and physiological level through mutual evidence between the two, so that a method for rapidly evaluating alum root heat resistance is established, and the operation is simpler and more convenient on the basis of science and effectiveness.
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FIG. 1 shows different alum root varieties F after high temperature stress treatment at 45 DEG Cv/FmThe variation of the value with time.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Example 1
1.1 test materials
The test germplasm resources of the research come from a alum root germplasm resource garden of academy of agricultural sciences of Shanghai city, and the names of 37 alum root varieties are shown in Table 1. Annual potted plants were used as test material for subsequent experimental treatments, with 10 individual plants per variety, and 3 biological replicates were set.
1.2 test methods
The tests were conducted in the institute of supple, shanghai academy of agricultural sciences during the period of 2019, 4 months to 2019, 5 months. The test material was moved into a climatic chamber (Shanghai Haozhuang apparatus Co., Ltd., model HZLD-1000C) for pre-cultivation at 25 ℃ for 3h, and then subjected to high temperature stress treatment at 45 ℃ for 0h, 3h and 6 h. The relative humidity of air in the test process is 50% +/-5%, and the illumination intensity is 22000 Lx. And (4) after the high-temperature stress treatment is finished, putting the tested plant into a resource garden for recovery cultivation.
1.2.1 plant State Observation
And photographing in the test process, recording the plant states after 0h, 3h and 6h of treatment, calculating the proportion according to the conditions of leaf burn, curling and the like, and counting the heat-resisting grade. The heat resistance grade is divided into 5 grades: grade 0, normal growth of plants; grade 1, less than 25% of the leaves exhibited curling; grade 2, 25% -50% of the leaves are curled, and the leaf edges are burnt; grade 3, 50% -75% of the leaves appear curled and burnt; grade 4, more than 75% of the leaves are curled or wilted; and 5, stage: the plants died. And calculating the heat damage index through a formula. The thermal injury index (%) is Σ (number of grades × number of grades)/maximum number of grades × number of total treated plants × 100 (field and country, etc., 2011).
1.2.2 determination of chlorophyll fluorescence parameters
The kinetic parameters of chlorophyll fluorescence of the vanadium leaves were determined using a continuous excitation fluorometer (Pocket Plant Efficiency Analyzer, Hansatech, UK). And (3) carrying out chlorophyll fluorescence kinetic parameter determination on the mature leaves of the middle wheel of different plants after high-temperature stress treatment for 0h, 3h and 6h respectively. Dark adaptation was carried out for 20min before measurement, with fluorescence at 3500. mu. mol. m-2·s-1For each variety, 6 individuals were assayed and repeated 3 times.
1.2.3 data processing and analysis
Data obtained by the test are subjected to data entry by using EXCEL software, and the mean value and the standard deviation are calculated. The data were analyzed for differential significance using analysis of variance (ANOVA) and multiple comparison analysis (Duncan) with SPSS17.0 software, at a significance level of 0.05(P < 0.05); correlation analysis was performed using bivariates of the SPSS17.0 software.
2 results and analysis
2.1 Effect of high temperature stress treatment on Heat injury index of different vanadium root species
After the different alum root varieties are subjected to high-temperature stress treatment at 45 ℃, the heat resistance indexes of the alum root varieties are counted according to the plant states. The results show (Table 1) that all the leaves of the alum root varieties tested did not show any burning or curling after the high temperature stress treatment for 3 hours, and the heat damage index was 0%. After 6 hours of treatment, the heat resistance of different alum root varieties is obviously different. In 37 alum root varieties, 13 plant leaves do not have any burning and curling conditions, and the heat damage index is 0 percent, namely, the heat damage is not shown, which indicates that the heat resistance of the varieties is stronger. The leaves of the remaining 24 varieties appeared to burn or curl to different degrees, indicating that they were relatively weak in heat resistance. The variety with the largest heat damage index is 'fire alarm' and is 40 percent. According to the test results, the heat resistance of different alum root varieties can be obviously different after 6 hours of high-temperature stress treatment at 45 ℃ at the phenotype level. According to the heat damage index measurement result, the alum root varieties can be divided into 2 types, namely, the alum root variety with stronger heat resistance (the heat damage index is 0%) and the alum root variety with poorer heat resistance (the heat damage index is more than 0%).
Heat damage index of different alum root varieties subjected to high-temperature stress treatment at 145 ℃ in table and F of different alum root varietiesv/FmHigh temperature response behavior of values
Note: different lower case letters after the same column of data indicate significant difference (P <0.05)
2.2 correlation between the Alum chlorophyll fluorescence kinetics parameter indexes after high temperature stress
The chlorophyll fluorescence kinetic parameters of the alum root leaves subjected to the high-temperature stress for 3h and 6h are measured by adopting a continuous excitation type fluorometer, 15 parameter indexes are subjected to correlation analysis respectively, and the results are shown in tables 2 and 3. Among the kinetic parameters of chlorophyll fluorescence, Fv/FmIs a main parameter of high temperature stress resistance of plants. As can be seen from Table 2, F was observed after 3h of high temperature stress treatmentv/FmThe correlation with the rest 14 indexes is a very significant relation; wherein Fv/FmAnd Fm、Fv/FO、ETO/RC、ΦEO、ψEO、PIABS、RCQAShows very significant positive correlation with FO、dVG/dtO、dV/dtO、ABS/RC、DIO/RC、TRO/RC、DIO/CSOShows extremely obvious negative correlation. After 6h high temperature stress treatment (Table 3), Fv/FmAnd TROthe/RC is in positive correlation, but the correlation is not obvious. Except for TROOutside of/RC, Fv/FmThe correlation condition with the other 13 indexes is consistent with that after the high-temperature stress treatment for 3 h. Thus, it is considered that Fv/FmThe value can be used as a representative index of the fluorescence kinetic parameters of the alum root chlorophyll to be used for evaluating the heat resistance of different alum root varieties.
TABLE 245 ℃ correlation of chlorophyll fluorescence parameter index after 3h high temperature stress treatment
Note:＊＊indicating significant correlation at the 0.01 level (bilateral).
Relevance of chlorophyll fluorescence parameter index after high-temperature stress treatment for 6h at 345 ℃ in table
Note:＊＊indicating significant correlation at the 0.01 level (bilateral).
2.3 chlorophyll fluorescence kinetics parameters F of different vanadium root varietiesv/FmResponse to high temperature stress
Leaf chlorophyll fluorescence kinetic parameter F after being treated for 0h, 3h and 6h by high temperature stress at 45 DEG Cv/FmAnd (4) measuring values, and researching the response conditions of different alum root varieties to high-temperature stress. The results show (Table 1) that, during the initial stage of high temperature stress (0 h treatment), all the leaves F of the alum root varieties testedv/FmThe values are all larger than 0.8, which indicates that the chloroplast is not affected by stress and can normally function. After 3h and 6h of 45 ℃ high-temperature stress treatment, the leaves F of 37 alum root varieties to be testedv/FmAll values are obviously reducedAnd F of most varietiesv/FmThe value decreased with increasing high temperature stress time (fig. 1). It shows that high temperature stress can cause the function of chloroplast to be reduced, and the longer the stress is, the more obvious the function is reduced. Although the phenotype of the plants subjected to the 3h heat stress treatment is not obviously different, the phenotype shows that the heat injury indexes of all varieties after the 3h treatment are all 0%, the Fv/Fm value of the plants is obviously reduced, and the fact that the alum root chlorophyll can rapidly respond to the high temperature stress is shown. At this time, the more heat-resistant variety and the less heat-resistant variety are already separated to some extent and expressed as Fv/FmThe degree of value reduction varies. Such as sweet tea of high heat resistance variety Fv/FmThe value can be maintained at 0.7613, and the product with weak heat resistance is 'lemon yellow', and F thereofv/FmThe value rapidly dropped to 0.3429. Varieties with heat injury index of 0% after 6h treatment, such as 'autumn maple', 'golden zebra', 'flower blanket', etc., and F thereofv/FmThe values do not differ significantly between 3h and 6 h; varieties with higher heat injury indexes after 6h treatment, such as 'fire alarm', 'lemon yellow', 'egg tart' and the like, wherein F is the number of the varietiesv/FmThe value was also not significantly different between 3h and 6 h; and the varieties with intermediate heat injury indexes after 6h treatment, such as 'Shanghai', 'peach flame', and the like, have Fv/FmThe values show a tendency to gradually decrease with increasing heat treatment time. According to the test results, before the vitriol root chloroplast F can show high-temperature stress on the phenotype, the vitriol root chloroplast Fv/FmThe value can reflect the rapid response of the alum root plant to high temperature stress. F after 3h treatment of alum root variety with weak heat resistancev/FmThe values already exhibit lower levels; and F after 6h treatment of alum root variety with strong heat resistancev/FmThe value can still be maintained at a higher level. The variety of alum can be divided into 3 types according to Fv/Fm value, namely F after 6h treatmentv/FmThe varieties with still higher values are mostly expressed as varieties with stronger heat resistance and lower Fv/Fm values after 3h treatment, and the varieties with poor heat resistance, and Fv/FmThe varieties with gradually decreasing values are mostly characterized by moderate heat resistance.
2.4 analysis and evaluation of Heat resistance of different types of Kalanchoe roots
In view of Fv/FmThe value is an important index for evaluating the heat resistance of the alum root, so that F is subjected to 45 ℃ high-temperature stress treatment according to different alum root varietiesv/FmThe correlation of the values with the thermal injury index was analyzed (Table 4). The results show a thermal hazard index after 6h treatment and F after 0h treatmentv/FmThe values are not related, but to F after 3h and 6h treatmentv/FmThe values are extremely obviously and negatively related, which indicates that F is measured after the high-temperature stress treatment at 45 ℃ for 3h and 6hv/FmThe values can be used as important parameters for evaluating the heat resistance of the vanadium root variety. And the correlation coefficient comparison condition is | -0.893| > | -0.888|, which shows that the correlation is compared with the thermal injury index after 6h treatment, and F after 3h treatmentv/FmThe value is slightly stronger than 6 h. And after 3h treatment, the plants do not show obvious heat damage and have small damage to the plants. From the viewpoint of efficiency and simplicity, F after 3h treatment was usedv/FmThe value is more suitable as an effective index for analyzing the heat resistance of different vanadium root varieties.
TABLE 4 different alum root varieties after 45 ℃ high temperature stress treatment Fv/FmCorrelation of value with Heat hazard index
Note:＊＊indicating significant correlation at the 0.01 level (bilateral).
F of alum root variety after 3h of heat stress treatmentv/FmThe values and the 6h heat damage index conditions are comprehensively analyzed, and the 37 alum root varieties to be tested are divided into 3 types, which are shown in the table 5. The heat damage indexes after 6 hours of treatment are correspondingly arranged, and the distribution rule is that the heat damage index of the variety with stronger heat resistance is 0 percent, the heat damage index of the variety with medium heat resistance is 0 to 30 percent, and the heat damage index of the variety with stronger heat resistance is more than or equal to 30 percent. Heat stress corresponding theretoF of alum root variety after 3h treatmentv/FmThe values are divided into three types of more than or equal to 0.70, (0.55, 0.70) and less than or equal to 0.55, which respectively correspond to the types of the varieties with stronger heat resistance (type 1), moderate heat resistance (type 2) and weaker heat resistance (type 3) and are used as the standard for evaluating the heat resistance of the alum roots. Of the 37 alum-root varieties, the varieties with stronger heat resistance, moderate heat resistance and weaker heat resistance were 12, 17 and 8, respectively. The research result can provide theoretical reference for the alum root heat-resistant variety breeding work.
TABLE 5 Heat resistance status classification chart for different alum root varieties
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the technical principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (4)
1. A method for rapidly evaluating the heat resistance of alum root comprises the following steps: carrying out heat stress treatment on the alum root variety to be evaluated, and measuring the value of chlorophyll fluorescence kinetic parameter Fv/Fm, wherein Fv/Fm is more than or equal to 0.70 and is a variety with stronger heat resistance, Fv/Fm is more than 0.55 and less than 0.70 and is a variety with middle heat resistance, and Fv/Fm is less than or equal to 0.55 and is a variety with weaker heat resistance.
2. The method for rapidly evaluating the heat resistance of alum root according to claim 1, wherein the heat stress temperature is 45 ℃.
3. The method for rapidly evaluating the heat resistance of alum root according to claim 1, wherein the heat stress treatment is performed for 3 hours.
4. The method for rapidly evaluating the heat resistance of the alum root according to claim 1, wherein chlorophyll fluorescence kinetic parameters of mature leaves of the alum root medium wheel are measured.
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| CN113176235A (en) * | 2021-03-25 | 2021-07-27 | 浙江农林大学暨阳学院 | Hickory nut high temperature resistance evaluation method based on rapid fluorescence induction kinetics |
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