CN115144546A - Identification and evaluation method for low-temperature resistance of sesame in germination stage - Google Patents
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- 235000003434 Sesamum indicum Nutrition 0.000 title claims abstract description 89
- 241000207961 Sesamum Species 0.000 title claims abstract description 87
- 230000035784 germination Effects 0.000 title claims abstract description 32
- 238000011156 evaluation Methods 0.000 title claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 26
- 238000012258 culturing Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 206010057040 Temperature intolerance Diseases 0.000 claims 1
- 239000000463 material Substances 0.000 description 27
- 230000005200 bud stage Effects 0.000 description 5
- 238000003556 assay Methods 0.000 description 4
- 238000007821 culture assay Methods 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
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Abstract
The invention relates to the technical field of crop identification methods, and particularly provides an identification and evaluation method for low-temperature resistance characteristics of sesame in a germination stage, which comprises the following steps: s1, a step: germinating sesame seeds until the sesame seeds are exposed to the white; and S2, a step: determining the bud length, the root length and the fresh weight of seedlings of the exposed and white sesame seeds after dark culture for the same time at a first temperature and a second temperature respectively, wherein the first temperature is 25-30 ℃, and the second temperature is 16-18 ℃; calculating relative bud length, relative root length and relative fresh seedling weight according to a formula of the specification; and S3, a step: and determining the low temperature resistance level according to the relative bud length, the relative root length and the relative fresh weight of the seedling. The method is simple and easy to operate, strong in controllability and good in repeatability, can accurately reflect the characteristic of the reaction of the sesame bud period to low-temperature stress, and is reliable in result.
Description
Technical Field
The invention relates to the technical field of crop identification methods, in particular to an identification and evaluation method for low-temperature resistance characteristics of sesame in a germination stage.
Background
Sesame (Sesamum indicum L.) is a traditional characteristic oil crop in China and is a thermophilic crop extremely sensitive to low-temperature reaction.
The influence of low temperature on sesame is mainly reflected in that the low temperature directly influences the germination and the bud stage growth of spring sesame seeds in the north, and the cold exposure influences the late growth and the seed development of autumn sesame seeds in the south. Therefore, the method for identifying and evaluating the low-temperature resistance of the sesame in the bud stage is established, and technical support and material sources are provided for low-temperature resistance breeding. The application of the low temperature resistant variety can enhance the temperature stress resistance capability in the bud stage and improve the stable yield of the sesame on the one hand, and can properly advance the sesame sowing period or the sesame production area to realize north expansion on the other hand, thereby improving the sesame yield and the sesame planting area.
The identification and evaluation of the low temperature resistance of the sesame are developed, sesame germplasm resources with strong low temperature resistance are excavated, and low temperature resistant excellent varieties are cultivated, so that the method has important significance for improving the stable yield of the sesame and 'north expansion' of sesame planting in China. However, no method for identifying and evaluating the low temperature resistance characteristics of sesame has been reported so far.
Disclosure of Invention
Therefore, the invention aims to provide the method for identifying and evaluating the low-temperature resistance of the sesame in the germination stage, the method is simple and easy to implement, low in cost, not influenced by the growing season, accurate and reliable in identification result, and can provide technical and material support for identification and development of low-temperature resistant germplasm of sesame and cultivation of new low-temperature resistant varieties.
The invention provides a method for identifying and evaluating low-temperature resistance of sesame in a germination stage, which comprises the following steps:
s1, a step: germinating the sesame seeds until the sesame seeds are exposed to the white;
and S2, a step: determining the bud length, the root length and the fresh weight of seedlings of the exposed and white sesame seeds after dark culture for the same time at a first temperature and a second temperature respectively, wherein the first temperature is 25-30 ℃, and the second temperature is 16-18 ℃; calculating relative bud length, relative root length and relative fresh seedling weight according to the following formula;
relative shoot length = shoot length cultured at second temperature/shoot length cultured at first temperature × 100%;
relative root length = root length cultured at second temperature/root length cultured at first temperature × 100%;
relative shoot fresh weight = shoot fresh weight cultured at second temperature/shoot fresh weight cultured at first temperature × 100%;
and S3, a step: and determining the low temperature resistance level according to the relative bud length, the relative root length and the relative fresh weight of the seedling.
Further, in the step S1, plump and healthy sesame seeds are selected for germination, and preferably, the plump and healthy sesame seeds are placed in a sealed germination box paved with filter paper, water is added, and the seeds are cultured until the seeds are white.
Further, the culture temperature in the S1 step was 25 to 30 ℃.
For example, double-layer or three-layer filter paper can be laid in the sealed germination box, and 20-40mL (e.g. 30 mL) of distilled water can be added into 50-100 seeds.
Further, the culture time from germination of the sesame seeds to exposure to white is 12-24h.
Further, the standard of seed blush is that the germinating radicle exposes the seed coat and does not exceed half the length of the seed itself.
Further, the culture time in the S2 step is 5 to 7 days, preferably 6 days.
Further, in the step S2, two parts of the white exposed seeds with the same grain number are selected, and each part of the white exposed seeds is cultured at 50-100 grains respectively at different temperatures.
Further, the low temperature resistance level is divided into the following according to the relative bud length, the relative root length and the relative fresh weight of seedlings:
high resistance: relative bud length of more than or equal to 70% in 100%, relative root length of more than or equal to 80% in 100%, and relative seedling fresh weight of more than or equal to 85% in 100%;
resisting: 70 percent more than 50 percent of relative bud length, 80 percent more than 60 percent of relative root length and 85 percent more than 65 percent of relative fresh weight of the seedling;
medium tolerance: 50 percent more than 40 percent of relative bud length, 60 percent more than 55 percent of relative root length and 65 percent more than 60 percent of relative seedling fresh weight;
intolerance of: 40% more than 35% of relative bud length, 55% more than 45% of relative root length and 60% more than 55% of relative fresh seedling weight;
extremely weak: 35 percent more than the relative bud length more than or equal to 0 percent, 45 percent more than the relative root length more than or equal to 0 percent, and 55 percent more than the relative fresh weight of the seedling more than or equal to 0 percent.
Further, if the results of low temperature resistance level classification are inconsistent in terms of relative bud length, relative root length, and relative fresh seedling weight, the result with the lowest evaluation grade is taken as the standard.
Further, the first temperature is 28 ℃ and the second temperature is 18 ℃.
The technical scheme of the invention has the following advantages:
1. the invention provides a method for identifying and evaluating low temperature resistance of sesame in a germination period, which comprises the following steps of S1: germinating sesame seeds until the sesame seeds are exposed to the white; and S2, a step: determining the bud length, the root length and the fresh weight of seedlings of the exposed and white sesame seeds after dark culture for the same time at a first temperature and a second temperature respectively, wherein the first temperature is 25-30 ℃, and the second temperature is 16-18 ℃; calculating relative bud length, relative root length and relative fresh seedling weight according to the following formula; relative shoot length = shoot length cultured at second temperature/shoot length cultured at first temperature × 100%; relative root length = root length cultured at second temperature/root length cultured at first temperature × 100%; relative shoot fresh weight = shoot fresh weight cultured at second temperature/shoot fresh weight cultured at first temperature × 100%; and S3, a step: the method can be carried out in a laboratory, is not influenced by growth seasons, is simple and easy to operate, has strong controllability and good repeatability, can accurately reflect the characteristic of the reaction of the sesame bud stage to low-temperature stress, has accurate and reliable results, can accelerate the development process of low-temperature resistant germplasm identification of sesame, and provides technical and material support for breeding of new low-temperature resistant varieties of sesame.
2. According to the method for identifying and evaluating the low temperature resistance of the sesame in the germination period, disclosed by the invention, in order to more accurately identify the low temperature resistance of a sesame sample, normal temperature (28 ℃) is selected for exposing white (exposed white seeds are viable seeds), and then the sesame is moved to a low temperature (18 ℃) condition for treatment, so that the low temperature resistance of the material can be more objectively reflected.
3. According to the identification and evaluation method for the low temperature resistance of the sesame in the germination period, researches find that the low temperature has small influence on seed germination and mainly influences seedling growth, so that the low temperature resistance level of a sesame sample is determined by selecting the relative bud length, the relative root length and the relative fresh weight of seedlings as indexes, and the three indexes can very reflect the low temperature resistance of different varieties. In addition, the material which is only 5-7 days in the determination treatment (the culture time in the S2 step) has the advantage of short culture time, preferably 6 days, and is not suitable for identification and evaluation due to the short period, small biomass accumulation, no obvious difference in indexes such as fresh weight, dry weight and the like of the root system.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a photograph of sesame seedlings after 6 days of cultivation at different temperatures in Experimental example 1;
FIG. 2 is the results of the effect of low temperature stress treatment on the relative shoot length (A), the relative root length (B) and the relative shoot fresh weight (C) of sesame in Experimental example 1.
Detailed Description
The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
Experimental example 1 determination of suitable temperature for Low temperature stress treatment in sesame sprout stage
1. Material
The sesame materials used in the experiment are 5 sesame varieties of 'shangshang cap', 'Shancai white sesame', 'Xiaozihuang', 'Yiyesan' and 'Shayang sesame', which are all provided by crop research institute of agricultural academy of sciences in Jiangxi province, and the sources are shown in Table 1.
2. Method of producing a composite material
(1) Selecting 1500 full seeds from sesame materials of each variety, respectively placing the seeds in 15 germination boxes (10 cm multiplied by 5 cm) paved with double-layer wet filter paper in advance, respectively placing 100 seeds in each box, adding 30mL of distilled water into each box, and carrying out dark culture at the constant temperature of 28 ℃ for 16h until the seeds germinate and reveal white, wherein the standard of the seed reveal white is as follows: the germinated radicles are exposed from the seed coat and are no longer than half of the length of the seeds.
(2) After the seeds germinate and are exposed to white, respectively placing 15 boxes of each sesame material in a constant-temperature incubator with 5 temperature gradients of 28 ℃, 25 ℃, 21 ℃, 18 ℃ and 15 ℃ for continuous culture, namely 3 boxes for continuous 28 ℃ constant-temperature dark culture, 3 boxes for constant-temperature dark culture at 25 ℃,3 boxes for constant-temperature dark culture at 21 ℃,3 boxes for constant-temperature dark culture at 18 ℃, and 3 boxes for constant-temperature dark culture at 15 ℃; continuing to culture for 6d;1 box was considered as 1 replicate, 3 boxes per temperature, considered as three replicates;
(3) After culturing for 6 days, randomly selecting 10 seedlings in each box, and then measuring the bud length, the root length and the fresh weight of the seedlings; calculating the average value of the three repetitions; then, the relative values were calculated by dividing the average values of the 28 ℃ culture assay, 25 ℃ assay, 21 ℃ assay, 18 ℃ assay, and 15 ℃ assay by the average values of the 28 ℃ culture assay, respectively.
As shown in FIG. 1, the temperature greatly affects the growth of sesame seedlings, and the seedling bud length and root length become shorter as the temperature is lowered. When the temperature reaches 18 ℃, the seedlings grow slowly, the main roots are basically difficult to grow, and abnormal seedlings are formed. When the temperature reaches 15 ℃, sesame seeds hardly grow and normal buds cannot be formed. Therefore, 15 ℃ can be regarded as the critical temperature for the growth of sesame sprouts.
As shown in fig. 2, the relative value of the measured 3 personality indexes is constantly reduced along with the reduction of the temperature, the relative bud length is slowly reduced at 25-21 ℃, and the relative bud length is suddenly reduced at 18-15 ℃ (fig. 2A); the relative root length and the relative fresh weight of the seedlings are sharply reduced at 21-15 ℃ (FIG. 2B, C).
By combining the indexes, the relative bud length, the root length and the fresh seedling weight of the sesame are all greatly reduced when the dark culture temperature is 16-18 ℃, particularly the relative bud length, the root length and the fresh seedling weight of the sesame at 18 ℃ and 28 ℃ reach extremely obvious levels, namely the temperature condition can be regarded as the turning temperature influencing the normal growth of the sesame. Therefore, 16-18 ℃ can be used as the reference stress temperature for low-temperature identification of the sesame bud stage, and the relative bud length, the relative root length and the relative fresh weight of the seedling can be used as indexes for low-temperature resistance evaluation, and the temperature is preferably 18 ℃.
Experimental example 2 evaluation of low temperature resistance characteristics in germination stage of 188 parts of sesame seed material
1. Material
188 sesame seed materials with inconsistent geographical sources and rich genetic bases are selected.
TABLE 1 germplasm information and identification results of sesame seeds tested
2. Method of producing a composite material
(1) Selecting 600 plump seeds from sesame materials of each variety, respectively placing the seeds in 6 germination boxes (10 cm multiplied by 5 cm) paved with double-layer wet filter paper in advance, respectively placing 100 seeds in each box, adding 30mL of distilled water into each box, and performing dark culture at the constant temperature of 28 ℃ for about 16-20h until the seeds germinate and reveal white, wherein the standard of the seed reveal white is as follows: the germinated radicles are exposed from the seed coat and are no longer than half of the length of the seeds.
(2) After the seeds germinate and reveal white, continuously culturing 3 boxes of each sesame material at the constant temperature of 28 ℃ in dark, placing the 3 boxes at the constant temperature of 18 ℃ in dark, and continuously culturing for 6 days; 1 box was considered as 1 replicate, 3 boxes per temperature, considered as three replicates;
(3) After culturing for 6 days, selecting 10 seedlings with consistent growth vigor from each box, and then measuring the bud length, the root length and the fresh weight of the seedlings; calculating the average value of the three repetitions; relative values were then calculated as the average of the 18 ℃ culture assay divided by the average of the 28 ℃ culture assay, respectively.
(4) Determination of the Low temperature resistance level: and comprehensively determining the low temperature resistance level of the sample material according to three indexes of relative bud length, relative root length and relative fresh seedling weight, and referring to table 2.
The result shows that 5 parts of the high and low temperature resistant resource material accounts for 2.65 percent of the tested material; 49 parts of low-temperature-resistant resource material, which accounts for 26.06 percent of the tested material; 51 parts of medium and low temperature resistant resource material, which accounts for 27.13 percent of the test material; 44 parts of low-temperature-intolerant resource material, which accounts for 23.40 percent of the test material; 39 parts of extremely low temperature-resistant resource material accounts for 20.74 percent of the tested material.
From the evaluation result of the low temperature resistant identification of the 188 parts of sesame resources in the germination period, various materials with low temperature resistant levels are very consistent with sesame thermophily, the high low temperature resistant type and the low temperature resistant type are less in resources, the percentage of the high low temperature resistant type and the low temperature resistant type is respectively 2.65% and 26.06%, and the resistance of other data materials to low temperature is poor. Wherein, 5 parts of high-low temperature resistant materials of Zhi 19, zhi 1, zhi 16, shayang sesame and Luo Zhi 16 have stronger resistance to low temperature in practice, which shows that the method of the invention has reliable identification result and can better reflect the characteristic of the sesame bud period to low temperature.
TABLE 2 Low temperature resistant grade Standard and resource distribution
Wherein, if the results of low temperature resistance level division are inconsistent according to relative bud length, relative root length and relative fresh seedling weight, the result with the lowest evaluation grade is taken as the standard.
Experimental example 3 repeatability test
100 sesame seeds with 19 full sesame seeds are taken and randomly divided into two parts, the two parts are respectively placed in a germination box (10 cm multiplied by 5 cm) paved with double-layer wet filter paper in advance, 30mL of distilled water is added into each box, and the dark culture is carried out at the constant temperature of 28 ℃ for 20h until the seeds germinate and are exposed to the white.
And after the seeds germinate and reveal white, continuously culturing 1 box in dark at a constant temperature of 28 ℃, placing 1 box in dark at a constant temperature of 18 ℃, continuously culturing for 6d, selecting 10 seedlings with consistent growth vigor from each box, measuring the bud length, the root length and the fresh weight of each seedling, taking an average value, dividing the average value measured by the culture at 18 ℃ by the average value measured by the culture at 28 ℃ respectively, and calculating a relative value. And determining the low temperature resistance level of the sample material according to the grade division standard of the table 2 according to three indexes of relative bud length, relative root length and relative fresh seedling weight.
The above operations are repeated for 4 times, and the results are shown in the following table, and the results show that the identification results of 4 times of measurement are consistent, the RSD values are all less than 3%, and the repeatability is good.
TABLE 3 germplasm information and identification results of sesame for test
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. The method for identifying and evaluating the low temperature resistance of the sesame at the germination stage is characterized by comprising the following steps of:
s1, a step: germinating sesame seeds until the sesame seeds are exposed to the white;
and S2, a step: determining the bud length, the root length and the fresh weight of seedlings of the exposed and white sesame seeds after dark culture for the same time at a first temperature and a second temperature respectively, wherein the first temperature is 25-30 ℃, and the second temperature is 16-18 ℃; calculating relative bud length, relative root length and relative fresh seedling weight according to the following formula;
relative shoot length = shoot length cultured at second temperature/shoot length cultured at first temperature × 100%;
relative root length = root length cultured at second temperature/root length cultured at first temperature × 100%;
relative shoot fresh weight = shoot fresh weight cultured at second temperature/shoot fresh weight cultured at first temperature × 100%;
and S3, a step: and determining the low temperature resistance level according to the relative bud length, the relative root length and the relative fresh weight of the seedling.
2. The method for evaluating the cold resistance of sesame at the germination stage according to claim 1,
and S1, selecting full and healthy sesame seeds for germination, preferably, putting the full and healthy sesame seeds into a sealed germination box paved with filter paper, adding water, and culturing until the seeds are exposed to white.
3. The method for identifying and evaluating low temperature resistance of sesame at a germination stage according to claim 1 or 2, wherein the culture temperature in the step S1 is 25 to 30 ℃.
4. The method for identifying and evaluating low temperature resistance of sesame at a germination stage according to any one of claims 1 to 3, wherein a culture time from germination to white emergence of sesame seeds is 12 to 24 hours.
5. The method for evaluating the cold resistance of sesame at the germination stage according to any one of claims 1 to 4, wherein the standard of exposure of the seed to white is that the germinated radicle exposes the seed coat and the length of the germinated radicle is not more than half of the length of the seed.
6. The method for identifying and evaluating low temperature resistance characteristics of sesame at the germination stage according to any one of claims 1 to 5, wherein the culture time of the S2 step is 5 to 7 days, preferably 6 days.
7. The method for identifying and evaluating low temperature resistance characteristics of sesame at the germination stage according to any one of claims 1 to 6, wherein in the step S2, two exposed white seeds with the same grain number are selected, and each 50-100 exposed white seeds are respectively cultured at different temperatures.
8. The method for evaluating the cold resistance of sesame at the germination stage according to any one of claims 1 to 7, wherein the cold resistance level is divided into:
high and low temperature resistant: relative bud length of more than or equal to 70% in 100%, relative root length of more than or equal to 80% in 100%, and relative fresh seedling weight of more than or equal to 85% in 100%;
low temperature resistance: 70 percent more than 50 percent of relative bud length, 80 percent more than 60 percent of relative root length and 85 percent more than 65 percent of relative fresh weight of the seedling;
medium and low temperature resistant type: 50 percent more than 40 percent of relative bud length, 60 percent more than 55 percent of relative root length and 65 percent more than 60 percent of relative fresh weight of the seedling;
low temperature intolerance type: 40% more than 35% of relative bud length, 55% more than 45% of relative root length and 60% more than 55% of relative fresh seedling weight;
extremely low temperature resistant type: 35 percent more than the relative bud length more than or equal to 0 percent, 45 percent more than the relative root length more than or equal to 0 percent, and 55 percent more than the relative fresh weight of the seedling more than or equal to 0 percent.
9. The method for evaluating cold resistance of sesame at a germination stage according to claim 8, wherein if the results obtained by classifying the cold resistance levels according to the relative bud length, the relative root length, and the relative fresh weight of the seedling are not uniform, the result with the lowest evaluation level is used as the criterion.
10. The method for evaluating the cold resistance of sesame at the germination stage according to any one of claims 1 to 9, wherein the first temperature is 28 ℃ and the second temperature is 18 ℃.
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