阅读说明：本技术 一种构建植物种子活力梯度材料的方法 (Method for constructing plant seed vigor gradient material ) 是由 麻浩 程浩 王鑫慧 叶晨惠 钱芳 何军言 刘永祥 谢磊 于 2021-01-26 设计创作，主要内容包括：本发明公开了一种构建植物种子活力梯度材料的方法。针对同一植物品种或遗传型种子,称取等量的7份样品,采用56℃-68℃水浴加热处理,分别处理0、2、4、6、8、10和12min,从而形成一套具有7个种子活力等级的材料；同时针对这套材料,可直接做标准发芽实验,或采用人工加速老化法处理再做标准发芽实验,测定并明确不同种子活力等级材料的具体活力指标。本发明方法简单,耗时短,可操作性强,成本低,无环境污染,不仅适合高等院校和科研院所,而且特别适合于基层单位应用。(The invention discloses a method for constructing a plant seed vigor gradient material. Weighing 7 equal samples of the same plant variety or genetic type seeds, and respectively treating the samples for 0, 2, 4, 6, 8, 10 and 12min by adopting water bath heating treatment at the temperature of 56-68 ℃ so as to form a set of materials with the activity grades of 7 seeds; meanwhile, the standard germination experiment can be directly carried out on the set of materials, or the standard germination experiment is carried out after the artificial accelerated aging method is adopted for treatment, and the specific activity indexes of the materials with different seed activity grades are determined and determined. The method is simple, short in time consumption, strong in operability, low in cost and free of environmental pollution, and is not only suitable for universities and scientific research institutions, but also particularly suitable for basic units.)

1. A method for constructing a plant seed vigor gradient material is characterized in that 7 equal samples are weighed aiming at the same plant variety or genetic type seeds, and are respectively treated for 0, 2, 4, 6, 8, 10 and 12min by adopting 56-68 ℃ water bath heating treatment, so that a set of material with 7 seed vigor grades is formed; meanwhile, the standard germination experiment can be directly carried out on the set of materials, or the standard germination experiment is carried out after the materials are treated by adopting an artificial accelerated aging method, the specific activity indexes of the materials with different seed activity grades are determined and determined, and then a seed activity index standard curve is drawn.

2. The method of claim 1, wherein the conditions of the artificially accelerated aging process are 40-45 ℃ and 70% -100% relative humidity for 3 days.

3. The method of claim 1, wherein the indicators of germination comprise: counting the percentage of the number of germinated seeds in the total number of the tested seeds from the beginning of the seed germination experiment to the 7 th day, and recording as the germination rate; counting the percentage of the number of the germinated seeds in the total number of the tested seeds from the time of putting the seeds to the 4 th day, and recording the percentage as the germination vigor; on the 7 th day of germination, 50 whole seedlings are taken and washed clean by tap water, and the length of the measured seedlings is accurate to cm; after surface moisture is sucked dry, the mixture is dried at 80 ℃ to constant weight and weighed, the weight is measured to be g, and the average value is taken and recorded as dry weight.

4. The method according to claim 3, wherein the germination index is GI ∑ (Gt/Dt), where Gt is the number of germination on day t and Dt is the number of germination days.

5. The method of claim 3, wherein the seed vigor index VI is SxGI, wherein VI is the vigor index and S is the length (cm) or dry weight (g) of the individual seedling.

6. The method of claim 1, wherein the seed vigor index curve: and drawing 7 seed vigor index curve graphs with different vigor grades by taking the treatment time as an abscissa and the seed vigor index as an ordinate.

7. The method according to claim 1, wherein the water bath heating treatment is performed in a constant temperature water bath; after each batch of samples is processed, the samples are taken out from the water bath quickly, the surface moisture of the seeds is absorbed by filter paper, and the samples are dried again at 25 ℃ until the original moisture content is reached.

8. The method as claimed in claim 1, wherein before the artificial accelerated aging treatment, the plant seed sample is soaked in 1% sodium hypochlorite solution for 15min for surface disinfection, then washed with sterile water for 3 times, and dried at 25 ℃ until the original water content is reached.

Technical Field

The invention relates to a construction method of a plant seed vigor gradient material, belonging to the field of research of seedlogy.

Background

The seeds are the basic unit of agricultural production, and the sowing and seedling establishment are the primary links related to the high yield and high quality production success or failure of plants. For a long time, people mostly measure the sowing quality of seeds by using the germination rate, but the condition that the germination rate does not accord with the field emergence rate is often encountered in production. Therefore, people try to find an index capable of accurately predicting the emergence situation of seeds in the field, and a new concept, namely seed vigor, is provided.

The influence factors of the seed vitality mainly include genetic factors and external environment factors. The genetic factor of the seed vitality determines the vitality characteristic of the seed, and the external condition determines the vitality expression degree of the seed. For a long time, people have more researches on the comparison of seed vigor and field emergence characteristics among different plant varieties, but the seed vigor characteristics among different varieties are controlled by genetic factors, so that the comparison cannot truly and accurately reflect the activity performance of the seeds due to the influence of genetic factor difference; for the same seed batch of a certain kind (the seed batch belongs to the concept in the field of seedscience, and refers to seeds with the same source, the same variety, the same year, the same period, harvesting and quality basically consistent and within the specified quantity), the former generally adopts natural aging for 1-2 years or an artificial aging box to accelerate aging for 3-12 days to form seed materials with different activities, but the methods (1) have the disadvantages of long processing time and inconvenience; (2) the equipment of the aging box is expensive, each equipment is more than ten thousand yuan, the popularization of part of units (particularly basic units) is difficult, and the operation is complex; (3) during the course of our research, it was found that artificial ageing treatments could not be applied to the treatment of seeds of certain plant species, such as mildew-prone seed material. In the practical research and production practice processes, materials with different seed vigor grade gradients are often and extremely required to be constructed for the same plant variety or different plant varieties, and are used for (1) researching the seedling emergence characteristics and stress resistance of seeds with different vigor gradient under different adverse conditions, determining the correlation between the vigor gradient and the stress resistance, and predicting the actual seedling emergence and growth conditions in the field of the variety; (2) evaluating a seed vigor detection method, and determining the optimal vigor determination method of different crops; (3) the method is combined with a non-destructive detection technology of seeds, and can be used for establishing and (every year) correcting a non-destructive identification model of the seed vitality, accelerating the detection speed of the seed vitality, rapidly screening high-vitality seeds and the like. Therefore, in view of the above scientific problems and the urgent need for solving the problems, it is urgently needed to establish a method for constructing a gradient material of different seed vigor of plants so as to prepare a plant seed material with different vigor.

Disclosure of Invention

Aiming at the problems, the invention provides a method for constructing different plant seed vigor gradient materials, and a set of plant seed materials with vigor gradients is formed, and the specific embodiment is as follows:

a method for constructing a plant seed vigor gradient material is characterized in that 7 equal samples are weighed aiming at the same plant variety or genetic type seeds, and are respectively treated for 0, 2, 4, 6, 8, 10 and 12min by adopting 56-68 ℃ water bath heating treatment, so that a set of material with 7 seed vigor grades is formed; meanwhile, the standard germination experiment can be directly carried out on the set of materials, or the standard germination experiment is carried out after the materials are treated by adopting an artificial accelerated aging method, the specific activity indexes of the materials with different seed activity grades are determined and determined, and then a seed activity index standard curve is drawn.

The seed vitality index calculation and standard curve drawing method comprises the following steps:

(1) the germination indexes are as follows: counting the percentage of the number of germinated seeds in the total number of the tested seeds from the beginning of the seed germination experiment to the 7 th day, and recording as the germination rate; counting the percentage of the number of the germinated seeds in the total number of the tested seeds from the time of putting the seeds to the 4 th day, and recording the percentage as the germination vigor; on the 7 th day of germination, 50 whole seedlings are taken and washed clean by tap water, and the length of the measured seedlings is accurate to cm; after surface moisture is sucked dry, drying at 80 ℃ to constant weight, weighing until the weight is accurate to g, and taking an average value to be recorded as dry weight;

(2) the germination index GI ═ sigma (Gt/Dt), wherein Gt is the germination number on the t day and Dt is the germination number on the t day;

(3) the seed vigor index VI is S × GI, wherein VI is the vigor index and S is the length (cm) or dry weight (g) of a seedling individual;

(4) seed vigor index curve: and drawing 7 seed vigor index curve graphs with different vigor grades by taking the treatment time as an abscissa and the seed vigor index as an ordinate.

The condition of the artificial accelerated aging method is 40-45 ℃, 70% -100% relative humidity, and the treatment is carried out for 3 days.

Further strictly controlling the time of hot water treatment, adding distilled water into a constant-temperature water bath kettle to keep the temperature constant, putting the prepared plant seeds into the water bath kettle, completely immersing and starting timing; after each batch of samples is processed, the samples are taken out from the water bath quickly, the surface moisture of the seeds is absorbed by filter paper, and the samples are dried again at 25 ℃ until the original moisture content is reached.

Meanwhile, before the artificial accelerated aging treatment, a plant seed sample is soaked in a 1% sodium hypochlorite solution for 15min for surface disinfection, then washed for 3 times by using sterile water, and dried at 25 ℃ until the original water content is reached.

Advantageous effects

The invention provides a method for constructing a plant seed vigor gradient material, which can form a set of plant seed materials with different vigor gradients. The beneficial effects are as follows: (1) the same seed of the same plant variety forms different activity gradients in batch, so that the influence of genetic background difference on the activity of the seeds in research is avoided; (2) by constructing the plant seed vigor gradient material, the method can be used for researching the seedling emergence characteristics and the stress resistance of seeds with different vigor gradients under different stress conditions, determining the correlation between the vigor gradients and the stress resistance and predicting the actual seedling emergence and growth conditions of the variety in the field; (3) applying the constructed plant seed vigor gradient material to evaluation of a seed vigor detection method, and determining the optimal vigor determination methods of different crops; (4) the method is combined with a non-destructive detection technology of seeds, and can be used for establishing and correcting a non-destructive identification model of seed vitality, accelerating the vitality detection speed of the seeds and quickly screening high-vitality seeds.

Drawings

FIG. 1 is a standard curve of activity index of different activity gradient materials of soybean seinnong 36 seed

FIG. 2 is an index standard curve of different activity gradient materials of Anhui bean 22337 seed

FIG. 3 is a standard curve of activity index of different gradient materials of haloxylon ammodendron seeds

FIG. 4 is the standard curve of vitality index of Xiangyan No. 5 seeds with different vitality gradient materials

Detailed Description

The present invention will be described in detail with reference to specific examples.

Example 1:

(1) selecting the following crops: the Suiyong 36 soybean seeds are harvested in 2019 and stored in a refrigerator at 4 ℃.

(2) Hot water treatment is carried out on the seinnong 36 at the temperature of 58 +/-1 ℃, and the treatment is carried out for 0, 2, 4, 6, 8, 10 and 12min respectively to form 7 vitality grades.

(3) Experimental treatment

And hot water treatment is started in 2019, 10 and 20 days, and the materials with different activities can be formed by airing the materials to constant weight at room temperature after the treatment is finished. Further, the seed viability of these materials can be determined by an artificial aging test. During measurement, the material to be measured is put into an aging box and aged for 3 days at 40 ℃ and 100 percent of humidity; and then performing a standard germination test, sterilizing the seeds, distributing the seeds on 2019, 11/1, putting wet filter paper in each germination box, uniformly putting 100 soybean seeds on the filter paper, putting the soybean seeds into an illumination incubator with the temperature of 25 ℃ for illumination for 16 hours and the temperature of 20 ℃ for darkness for 8 hours, recording the putting time, and recording the germination number day by day on the next day. And in 2019, 11, 8 days, taking out the germination box to measure indexes such as dry weight, bud length and the like, then calculating vitality indexes, and drawing a vitality index curve.

(4) The experimental results are as follows:

TABLE 1 vitality indexes of different vitality gradient materials of the Suiyong 36 seeds

Note: different letters in the table indicate that the difference reached a significant level (P <0.05), the same below.

And (4) analyzing results: materials of the Suiyongong 36 seeds with different activity gradients are constructed by the method, activity indexes of the Suiyongong 36 seeds with different activity gradients have significant differences, and the processing time and the activity index of the seeds form a very significant linear relation; with the reduction of the activity grade, the germination rate and the activity index are reduced continuously. Therefore, by adopting the method, a set of good materials with different seed vigor gradients can be constructed for the seinhong 36.

Example 2:

(1) selecting the following crops: the Anhui bean 22337 soybean seeds were harvested in 2019 and stored in a refrigerator at 4 ℃.

(2) Processing Anhui beans 22337 with hot water at 58 + -1 deg.C for 0, 2, 4, 6, 8, 10, and 12min to obtain 7 activity grades.

(3) Experimental treatment

Starting hot water treatment in 7-10 months in 2020, and drying at room temperature to constant weight after the treatment is finished to obtain materials with different activities. Standard germination experiments can be directly carried out on the materials, and specific values of the vitality of the materials are determined. Starting to distribute seeds at 20 days 7 months in 2020, putting wet filter paper in each germination box, then uniformly putting 100 soybean seeds on the filter paper, then putting the box into an illumination incubator with illumination at 25 ℃ for 16 hours and darkness at 20 ℃ for 8 hours, recording the putting time, and starting to record the germination number day by day the next day. And 7, 27 days in 2020, taking out indexes such as measured dry weight, bud length and the like from a germination box, then calculating vitality indexes, and drawing a vitality index curve.

The experimental results are as follows:

TABLE 2 Activity indexes of different activity gradient materials of Anhui bean 22337 seed

And (4) analyzing results: materials with different activity gradients of the Anhui bean 22337 seeds are constructed by the method, activity indexes of the Anhui bean 22337 seeds with different activity gradients have significant differences, and the treatment time and the activity indexes of the seeds have a very significant linear relationship; as the viability grade decreases significantly, the germination rate and viability index decrease continuously. Therefore, by using this method, a good set of materials with different seed vigor gradients can be constructed for Wan beans 22337.

Example 3:

(1) the selected plants were: the seeds of the haloxylon ammodendron are harvested in 2019 and stored in a refrigerator at 4 ℃.

(2) The seeds of the haloxylon ammodendron are treated with hot water at the temperature of 56 +/-1 ℃ for 0, 2, 4, 6, 8, 10 and 12min respectively to form 7 vitality grades.

(3) Experimental treatment

Starting hot water treatment in 9/10/2020, and air drying at room temperature to constant weight after the treatment is finished to obtain materials with different vitality. For these materials, standard germination experiments can be directly carried out to clarify specific values of the vitality. Starting to distribute seeds at 20 days 9 and 9 in 2020, putting wet filter paper in each germination box, then uniformly putting 100 haloxylon ammodendron seeds on the filter paper, then putting the haloxylon ammodendron seeds into an illumination incubator which is illuminated for 16 hours at 25 ℃ and dark for 8 hours at 20 ℃, recording the putting time, and starting to record the germination number day by day on the next day. And taking out the germination box for measuring indexes such as dry weight, bud length and the like in 28 days 9 and 2020, then calculating the vitality index and drawing a vitality index curve.

TABLE 3 vitality indexes of different vitality gradient materials of haloxylon ammodendron seeds

And (4) analyzing results: the method constructs materials of the haloxylon ammodendron seeds with different activity gradients, activity indexes of the haloxylon ammodendron seeds with different activity gradients have significant difference, the processing time and the activity indexes of the seeds form a very significant linear relation, and the germination rate and the activity index are continuously reduced along with the reduction of the significance of the activity grades. Therefore, by adopting the method, a set of materials with different vigor gradients of the haloxylon ammodendron seeds can be constructed.

Example 4:

(1) the selected plants were: xiangyan No. 5 tobacco seeds are harvested in 2019, and stored in a refrigerator at 4 ℃.

(2) Xiangyan No. 5 was treated with hot water at 58 + -1 deg.C for 0, 2, 4, 6, 8, 10, and 12min to obtain 7 vitality grades.

(3) Experimental treatment

Starting hot water treatment at 5-10 months in 2020, drying at room temperature to constant weight after treatment, directly performing standard germination experiments, starting seed distribution at 5-20 months in 2020, putting wet filter paper into each germination box, uniformly putting 100 haloxylon ammodendron seeds on the filter paper, putting into a light incubator with the temperature of 25 ℃ being 16 hours and the temperature of 20 ℃ being 8 hours in darkness, recording the putting time, and starting to record the germination number day by day on the next day. And taking out the germination box for measuring indexes such as dry weight, bud length and the like in 6-3 months in 2020, then calculating the vitality index and drawing a vitality index curve.

TABLE 4 vitality indexes of different vitality gradient materials of Xiangyan No. 5 seeds

And (4) analyzing results: the method constructs the materials of Xiangyan No. 5 seeds with different activity gradients, the activity indexes of the Xiangyan No. 5 seeds have significant difference, and the processing time and the activity indexes of the seeds have extremely significant linear relation; along with the reduction of the significance of the activity grade, the germination rate and the activity index are also reduced continuously, and the difference between the activity grades is significant. Therefore, by adopting the method, a set of Xiangyan No. 5 seeds with different activity gradients can be constructed.