CN112930760B - Tetrazole determination method for sweet pepper seed viability - Google Patents
Tetrazole determination method for sweet pepper seed viability Download PDFInfo
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- CN112930760B CN112930760B CN202110364433.5A CN202110364433A CN112930760B CN 112930760 B CN112930760 B CN 112930760B CN 202110364433 A CN202110364433 A CN 202110364433A CN 112930760 B CN112930760 B CN 112930760B
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- 240000004160 Capsicum annuum Species 0.000 title claims abstract description 42
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 title claims abstract description 41
- 230000035899 viability Effects 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 22
- 150000003536 tetrazoles Chemical class 0.000 title claims abstract description 9
- 238000004043 dyeing Methods 0.000 claims abstract description 42
- KJUGUADJHNHALS-UHFFFAOYSA-N 1H-tetrazole Chemical compound C=1N=NNN=1 KJUGUADJHNHALS-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000008055 phosphate buffer solution Substances 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 210000001161 mammalian embryo Anatomy 0.000 claims abstract description 10
- 239000008399 tap water Substances 0.000 claims abstract description 8
- 235000020679 tap water Nutrition 0.000 claims abstract description 8
- 238000005406 washing Methods 0.000 claims abstract description 8
- 238000002791 soaking Methods 0.000 claims abstract description 7
- 238000004040 coloring Methods 0.000 claims abstract description 5
- 238000012360 testing method Methods 0.000 claims description 33
- 238000005520 cutting process Methods 0.000 claims description 16
- 239000000243 solution Substances 0.000 claims description 11
- 230000001338 necrotic effect Effects 0.000 claims description 9
- 239000007853 buffer solution Substances 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 4
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 3
- YZSUTKMFUHJPNB-UHFFFAOYSA-M 2,3,5-triphenyltetrazol-2-ium;bromide Chemical compound [Br-].C1=CC=CC=C1C(N=[N+]1C=2C=CC=CC=2)=NN1C1=CC=CC=C1 YZSUTKMFUHJPNB-UHFFFAOYSA-M 0.000 claims description 2
- PKDBCJSWQUOKDO-UHFFFAOYSA-M 2,3,5-triphenyltetrazolium chloride Chemical compound [Cl-].C1=CC=CC=C1C(N=[N+]1C=2C=CC=CC=2)=NN1C1=CC=CC=C1 PKDBCJSWQUOKDO-UHFFFAOYSA-M 0.000 claims description 2
- KDQPSPMLNJTZAL-UHFFFAOYSA-L disodium hydrogenphosphate dihydrate Chemical compound O.O.[Na+].[Na+].OP([O-])([O-])=O KDQPSPMLNJTZAL-UHFFFAOYSA-L 0.000 claims description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- 238000007689 inspection Methods 0.000 abstract description 9
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 230000000007 visual effect Effects 0.000 abstract description 2
- 230000035784 germination Effects 0.000 description 18
- 244000025254 Cannabis sativa Species 0.000 description 10
- 239000000463 material Substances 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000003026 viability measurement method Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 210000002257 embryonic structure Anatomy 0.000 description 3
- 231100000747 viability assay Toxicity 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 241000208292 Solanaceae Species 0.000 description 2
- 241001520823 Zoysia Species 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 229910000397 disodium phosphate Inorganic materials 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 125000003831 tetrazolyl group Chemical group 0.000 description 2
- 235000002566 Capsicum Nutrition 0.000 description 1
- 241000208293 Capsicum Species 0.000 description 1
- 235000002567 Capsicum annuum Nutrition 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000001511 capsicum annuum Substances 0.000 description 1
- 239000001390 capsicum minimum Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000005059 dormancy Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/02—Germinating apparatus; Determining germination capacity of seeds or the like
- A01C1/025—Testing seeds for determining their viability or germination capacity
Abstract
The invention discloses a tetrazole determination method for sweet pepper seed viability, which belongs to the technical field of crop seed quality inspection and comprises the following steps: soaking the cleaned seeds of sweet peppers in clear water for 12-18 hours, and then taking out the seeds; one third of the seed is slit longitudinally along the flat plane toward the radicle and cotyledon tip while the seed coat near the base of the seed is excised without damaging the radicle, thereby opening the embryo cavity of the seed. Placing the treated seeds into a tetrazole solution with the mass concentration of 1.0% prepared by a phosphate buffer solution, and dyeing for 6-12 hours at the constant temperature of 30-35 ℃ in a dark place; after dyeing is finished, washing the seeds for 2-3 times by using tap water, extruding the flat surfaces of the seeds by using tweezers to extrude radicles from the embryo cavities along the cuts of the base parts, and judging whether the seeds have viability or not according to the coloring condition of the radicles. The invention provides a simple and easy method for the quality inspection of the sweet pepper seeds, the identification standard is visual, the result is reliable, and the seed quality identification in the production and circulation of the sweet pepper seeds is convenient.
Description
Technical Field
The invention relates to the field of crop seed quality inspection, in particular to a method for determining the viability of sweet pepper seeds.
Background
Seed viability, which means the potential ability of a seed to germinate or the vitality possessed by the embryo, is expected to have the potential to grow into normal seedlings. Many plant seeds have a dormant character and cannot germinate or have a low germination rate after harvesting even when given suitable germination conditions. In practice, however, most non-germinating seeds may be viable, and the seeds in a dormant state still have normal physiological and biochemical metabolic characteristics. Therefore, the method for measuring the vitality of the seeds can be utilized to show the life phenomenon and the vitality of the dormant seeds and judge the potential ability of the dormant seeds to grow normal seedlings and plants so as to indicate the physiological quality of the dormant seeds. The determination of the seed viability can not only know the physiological quality of the dormant seeds, but also quickly predict the seed quality and analyze the reasons of non-germination and abnormal germination of the seeds. In production practice, the determination of seed viability is an indispensable important method in the work of crop breeding, seed production, processing, purchasing, storage, transportation, inspection, research and the like. The tetrazole determination is a practical, convenient, time-saving, rapid and reliable seed viability determination method, and is widely applied to crop seed quality inspection and detection.
Sweet pepper (sweet pepper)Capsicum annuum) Is a subspecies of capsicum of Solanaceae (Solanaceae), and is named sweet pepper because it can produce sweet fruit. Because sweet pepper is rich in nutrient components such as organic acid and vitamins, the sweet pepper is a widely cultivated vegetable variety in recent years. Planting sweet peppers in ChinaThe region is wide, the seed consumption is large, but the sweet pepper seeds belong to the species which is not storage-resistant and are easy to naturally age; and because partial seeds may have dormancy, the existing germination test technology cannot ensure that all viable seeds germinate completely, and cannot accurately evaluate the viability of the sweet pepper seeds and predict the maximum germination potential. Therefore, in the current scientific research and practice, how to research a simple, convenient and reliable-result dyeing method for determining the viability of sweet pepper seeds, determine the maximum germination potential of the seeds, evaluate the quality of the seeds and provide a reference basis for seed production and sale becomes a main problem to be solved urgently at present.
Disclosure of Invention
In view of this, the invention aims to provide a tetrazole determination method for sweet pepper seed viability, and the method has accurate detection result and good repeatability for sweet pepper seed viability. The invention provides a simple and easy method for the quality inspection of the sweet pepper seeds, the identification standard is visual, the result is reliable, and the seed quality identification in the production and circulation of the sweet pepper seeds is convenient.
The technical scheme of the invention is as follows:
1. soaking the cleaned seeds of sweet peppers in clear water for 12-18 hours, and then taking out the seeds;
2. the prewetted seed longitudinally cuts one third of the seed along the flat surface towards the radicle and cotyledon tip direction, and simultaneously cuts off the seed coat close to the base of the seed without damaging the radicle, thereby opening the embryo cavity of the seed;
3. placing the treated seeds into a tetrazole solution with the mass concentration of 1.0% prepared by a phosphate buffer solution, and dyeing for 6-12 hours at the constant temperature of 30-35 ℃ in a dark place;
4. after dyeing is finished, washing the seeds for 2-3 times by using tap water, extruding a flat surface of the seeds by using tweezers to extrude radicles from a germ cavity along the incision of the base part, judging whether the seeds have viability or not by using the coloring condition of the radicles, wherein the judgment standard is as follows:
the viable seeds are identified as those in which the radicles are fully stained, or less than one third of the radicles are not stained, weakly stained, or necrotic; the radicles are not stained, weakly stained or necrotic in their entirety, or more than one third of the radicles are not stained, weakly stained or necrotic and are identified as non-viable or dead seeds. Percent of viable seeds is (N/N). times.100%, wherein: n represents the number of viable seeds and N represents the number of seeds under test.
Compared with the prior art, the invention has the following beneficial effects:
1. in the current research on the viability of sweet pepper seeds measured by the tetrazole method, in the preparation stage before dyeing, the crop seed test protocol (GB/T3543.7-1995) stipulates that the seed coat and endosperm are punctured in the center of the seed or the end of the seed is cut off, and the method of cutting off the seed coat near the base of the seed is adopted in the International handbook of tetrazolium test protocol (International society for seed testing, 2003). The two treatments are not beneficial to the rapid and full penetration of the tetrazole solution into the seeds in the actual operation, so that the dyeing time is prolonged, generally 18-24 hours, and the dyeing effect is not ideal. In the invention, the prewetted sweet pepper seeds are longitudinally cut, and are dyed after the embryo cavities are opened, so that the dyeing effect of the seeds is enhanced, the dyeing time is reduced, and the time is generally shortened to 6-12 hours.
2. For the identification after dyeing, the sweet pepper seeds need to be longitudinally cut into two halves, and the dyeing condition of the cut surfaces is observed, so that the seed coats of the sweet pepper seeds are smooth and are not easy to cut in the actual operation. According to the invention, the seeds are squeezed by the tweezers to enable the radicles to be extruded out of the embryo cavities along the incision of the base part, the operation is convenient to implement, the working intensity is low in efficiency and high in efficiency, the working time is greatly shortened, the dyeing effect of the radicles can be favorably observed under a stereoscopic microscope, and the existence of the vitality of the seeds can be judged.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a schematic view of the cutting process of pimento seeds before dyeing in the present invention.
Detailed Description
In order to make the objects and advantages of the present invention more apparent, the technical solutions of the present invention will be further described in detail below with reference to the accompanying drawings in the embodiments of the present invention.
A tetrazole determination method for sweet pepper seed viability comprises the following steps:
A. soaking the cleaned seeds of sweet peppers in clear water for 12-18 hours, and then taking out the seeds;
B. longitudinally cutting the soaked and pre-wetted seeds towards the direction of radicles and cotyledon tips, simultaneously cutting off seed coats close to the bases of the seeds, not damaging the radicles, and opening embryo cavities of the seeds;
C. placing the treated seeds into a tetrazole solution with the mass concentration of 1.0% prepared by a phosphate buffer solution, and dyeing for 6-12 hours at the constant temperature of 30-35 ℃ in a dark place;
D. after dyeing, washing the seeds for 2-3 times by tap water, extruding a seed longitudinal cutting plane by using tweezers to extrude radicles from a germ cavity along the incision of the base part, judging whether the seeds have viability or not by the coloring condition of the radicles, wherein the judgment standard is as follows:
the viable seeds are identified as those in which the radicles are fully stained, or less than one third of the radicles are not stained, weakly stained, or necrotic; the radicles are not stained, weakly stained or necrotic in their entirety, or more than one third of the radicles are not stained, weakly stained or necrotic and are identified as non-viable or dead seeds. Percent of viable seeds is (N/N) × 100%, wherein: n represents the number of viable seeds and N represents the number of seeds under test.
As shown in FIG. 1, A is the longitudinal cutting direction of the soaked and pre-wetted seed towards the radicle and the cotyledon tip, and B is the cutting direction of the seed coat close to the base of the seed.
And C, longitudinally cutting one third of the seeds soaked in the pre-wet seeds in the step B along the flat surface towards the direction of the radicle and the cotyledon tip.
Placing the treated seeds into a tetrazole solution with the mass concentration of 1.0% and prepared by phosphate buffer solution, and dissolving 9.078 g of monopotassium phosphate KH in 1000 mL of distilled water2PO4A solution I was prepared and 9 was dissolved in 1000 mL of distilled water.472 g disodium hydrogen phosphate Na2HPO4Or 11.876 g of disodium hydrogenphosphate dihydrate Na2HPO4·2H2And preparing a solution II by using O, mixing two parts of the solution I by weight and three parts of the solution II by weight to obtain a buffer solution, weighing 1.0 g of 2,3, 5-triphenyltetrazolium chloride or triphenyltetrazolium bromide, dissolving the buffer solution by using a phosphate buffer solution to a constant volume of 100 mL, and preparing a tetrazolium solution with a mass percentage concentration of 1.0%.
Example 1
Selecting sweet pepper seeds with batch number PV05.91/GWD-05 which are sent to a grass and lawn grass seed quality supervision and inspection test center (Lanzhou) of agricultural rural department from Shanxi Xiaozhuang trading Limited company of 10-11 months in 2020, soaking the clean sweet pepper seeds in clear water for 15 hours, taking out the seeds, randomly counting 200 seeds, dividing the seeds into 4 repetitions, and repeating the 50 seeds every time; the prewetted seed longitudinally cuts one third of the seed along the flat surface towards the radicle and the cotyledon tip, and simultaneously cuts off the seed coat close to the base of the seed without damaging the radicle, thereby opening the embryo cavity of the seed; placing the treated seeds into a tetrazole solution with the concentration of 1.0 percent prepared by a phosphate buffer solution, and dyeing for 12 hours in a dark place at the constant temperature of 30 ℃; and after dyeing is finished, washing the seeds for 3 times by using tap water, extruding the flat surfaces of the seeds by using tweezers to extrude radicles from the embryo cavity along the incision of the base part, judging whether the seeds have viability or not according to the coloring condition of the radicles, and calculating the viability of the seeds.
Performing germination test on the batch of seeds, randomly counting 4 times from clean seeds, and respectively placing 50 seeds in a culture dish with 2 layers of wet filter paper; placing the culture dish in a variable-temperature illumination incubator at 20-30 ℃, keeping the low temperature for 16 hours and the high temperature for 8 hours, and illuminating for 8 hours every day; examining the culture dish every day, and keeping the filter paper wet; the germination test time is 14 days, and the germination results are counted according to normal seedlings, fresh ungerminated seeds, abnormal seedlings and dead seeds.
Example 2
Example 1 was repeated for viability assay with the following differences: the dyeing temperature was 35 ℃ and the dyeing time was 8 hours.
Comparative example 1
According to the method of the crop seed inspection regulation, sweet pepper seeds with the batch number of PV05.91/GWD-05 are soaked in clear water for 15 hours and then taken out, 200 seeds are randomly counted and divided into 4 repetitions, and 50 seeds are repeated each; the pre-wetted seeds pierce the seed coat and endosperm at the center; placing the treated seeds into a tetrazole solution with the concentration of 1.0% prepared by a phosphate buffer solution, and dyeing in a dark place at the constant temperature of 30 ℃; after dyeing, washing the seeds with tap water for 3 times, longitudinally cutting the seeds into two halves, judging whether the seeds have viability according to the dyeing conditions of the embryos and the endosperms, and calculating the viability of the seeds.
Comparative example 2
According to the method of the crop seed inspection regulation, sweet pepper seeds with the batch number of PV05.91/GWD-05 are soaked in clear water for 15 hours and then taken out, 200 seeds are randomly counted and divided into 4 repetitions, and 50 seeds are repeated each; cutting off the tail end of the seed by the pre-wetted seed; placing the treated seeds into a tetrazole solution with the concentration of 1.0 percent prepared by a phosphate buffer solution, and dyeing in a dark place at the constant temperature of 30 ℃; after dyeing, washing the seeds with tap water for 3 times, longitudinally cutting the seeds into two halves, judging whether the seeds have viability according to the dyeing conditions of the embryos and the endosperms, and calculating the viability of the seeds.
Comparative example 3
Soaking sweet pepper seeds with the batch number of PV05.91/GWD-05 in clear water for 15 hours according to the regulation of an International Manual for testing seed tetrazole, taking out the seeds, randomly counting 200 seeds, dividing the seeds into 4 times of repetition, and repeating 50 seeds every time; cutting off seed coats close to the base parts of the seeds in the prewetted seeds; placing the treated seeds into a tetrazole solution with the concentration of 1.0 percent prepared by a phosphate buffer solution, and dyeing in a dark place at the constant temperature of 30 ℃; after dyeing, washing the seeds with tap water for 3 times, longitudinally cutting the seeds into two halves, judging whether the seeds have viability according to the dyeing conditions of the embryos and the endosperms, and calculating the viability of the seeds.
Statistics were made on the staining time, the pretreatment time for identification, and the viability and germination test results of examples 1-2 and comparative examples 1-3, as shown in table 1.
As can be seen from Table 1, compared with the existing method, the method provided by the invention has the advantages that the dyeing time and the pretreatment time for identification are both greatly shortened, and the detection efficiency is effectively improved.
Example 3
The viability test and germination test of example 1 were repeated using sweet pepper seeds of lot number PV51/HF-12 as the test material from Shanxi Xiaozhuang Jiangzhuang trade company, 10-11 months 2020 to the center for quality supervision and test of grass and turf grass seeds, Lanzhou, in the rural areas of agriculture, with the following differences: the prewetting time was 12 hours, the dyeing temperature was 30 ℃ and the dyeing time was 8 hours.
Example 4
The viability test and germination test of example 1 were repeated using sweet pepper seeds of lot number PV51/HF-13 as the test material, sent to the center for quality supervision and test of grass and turf grass seeds from Shanxi Xiaozhuang Zhuang trade company, 10-11 months, 2020, as the test material, with the following differences: the prewetting time was 12 hours, the dyeing temperature was 30 ℃ and the dyeing time was 10 hours.
Example 5
The viability measurement and germination test of example 1 were repeated using, as the test material, sweet pepper seeds of lot number CH0050/HF-01, which were sent from Shanxi Xiaozhuang Jiangzhuang trade company, No. 10-11 months 2020 to the center for quality supervision and test of grass and turf grass seeds, Lanzhou, of the agricultural rural area, with the following differences: the prewetting time was 18 hours, the dyeing temperature was 35 ℃ and the dyeing time was 6 hours.
Example 6
The viability assay and germination test of example 1 were repeated using sweet pepper seeds of lot number CH0050/HF-04, sent to the center for quality supervision and test of grass and turf grass seeds from Shanxi Xiaozhuang Jiangzhuang trade company, 10-11 months, 2020 as test material, with the following differences: the prewetting time was 18 hours, the dyeing temperature was 35 ℃ and the dyeing time was 9 hours.
Example 7
The viability and germination tests of example 1 were repeated using, as test materials, sweet pepper seeds of lot number CH0050/HF-05, which were sent from Shanxi Xiaozhuang trading Co., Ltd to the center for quality supervision and test of grass and lawn grass seeds, located in rural areas in 2020, as test materials, with the following differences: the prewetting time was 15 hours, the dyeing temperature was 35 ℃ and the dyeing time was 12 hours.
The results of viability assay and germination test of examples 3-7 are shown in Table 2
As can be seen from Table 2, the method provided by the invention has obvious positive correlation with the germination test result of seeds, which shows that the method has reliable results for testing the viability of the seeds of the sweet peppers.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (1)
1. The tetrazole determination method for the viability of sweet pepper seeds is characterized by comprising the following steps of:
A. soaking the cleaned seeds of sweet pepper in clear water for 12-18 hours, and taking out;
B. soaking the pre-wetted seeds, longitudinally cutting one third of the seeds along the flat surface in the directions of radicles and cotyledon tips, simultaneously cutting off seed coats close to the base parts of the seeds, not damaging the radicles, and opening embryo cavities of the seeds;
C. placing the treated seeds into a tetrazole solution with the mass concentration of 1.0% prepared by a phosphate buffer solution, and dyeing for 6-12 hours at the constant temperature of 30-35 ℃ in a dark place; dissolving 9.078 g of monopotassium phosphate in 1000 mL of distilled water to prepare a solution I, dissolving 9.472 g of disodium hydrogen phosphate or 11.876 g of disodium hydrogen phosphate dihydrate in 1000 mL of distilled water to prepare a solution II, mixing two parts by weight of the solution I and three parts by weight of the solution II to obtain a buffer solution, wherein the pH value of the buffer solution is 6.5-7.5, weighing 1.0 g of 2,3, 5-triphenyltetrazolium chloride or triphenyltetrazolium bromide, dissolving the buffer solution with a phosphate buffer solution to a constant volume of 100 mL, and preparing a tetrazole solution with the mass percentage concentration of 1.0%;
D. after dyeing is finished, washing the seeds for 2-3 times by tap water, extruding a seed longitudinal cutting plane by using tweezers, extruding radicles from a germ cavity along the incision of the base part, judging whether the seeds have viability or not according to the coloring condition of the radicles, wherein the judgment standard is as follows:
the viable seeds are identified as those in which the radicles are fully stained, or less than one third of the radicles are not stained, weakly stained, or necrotic; the non-viable or dead seeds are identified as either none, weak or necrotic radicles, or more than one third of the radicles are not stained, weak or necrotic, the percentage of viable seeds being (N/N) × 100%, wherein: n represents the number of viable seeds and N represents the number of seeds under test.
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