CN111279834A - Method for improving seed vitality - Google Patents
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- CN111279834A CN111279834A CN202010072334.5A CN202010072334A CN111279834A CN 111279834 A CN111279834 A CN 111279834A CN 202010072334 A CN202010072334 A CN 202010072334A CN 111279834 A CN111279834 A CN 111279834A
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- 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
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Abstract
The invention discloses a method for improving seed vigor, which comprises the step of soaking corn seeds in a wet state for 3-48h for 5-90min by using helium normal-pressure room-temperature plasma, so that the germination vigor of the corn seeds is improved under the condition of at least ensuring the germination rate, the vitality of the seeds is improved, and the high uniformity of the seedling emergence is ensured.
Description
Technical Field
The invention belongs to the technical field of agriculture, relates to a method for improving seed vigor, and particularly relates to a method for improving seed vigor by irradiating soaked seeds with normal-pressure room-temperature plasma.
Background
Corn is an important feed and grain crop in China, the planting area and the total yield of the corn are the first in China, and the corn is used for providing guarantee for grain safety and agricultural product supply in China. The germination rate of the corn seeds is reduced, the vitality of the seeds is low, the emergence of seedlings is irregular, the utilization efficiency of the seeds is seriously influenced, and a large amount of corn seeds are wasted.
ARTP is a short name for Atmospheric pressure Room Temperature Plasma (Atmospheric and from Temperature Plasma) capable of generating a Plasma jet having a high concentration of active particles (including helium atoms, oxygen atoms, nitrogen atoms, OH radicals, etc. in an excited state) at a Temperature of 25-40 ℃ under Atmospheric pressure.
ARTP has attracted increasing attention in recent years in the fields of microbial breeding and biomedicine, and has become a currently quite active cross-discipline research field. In addition, it has been found that plasma can carry out complex and controllable biochemical processes, such as catalysis, diagnosis, excitation reaction, etc., with biological tissues or cells. The chemical properties of plasma can also be applied in the biomedical field by treating the surface of the material and changing the properties of the material. Plasma surface treatment is increasingly used due to its excellent properties, such as high purity, sterility, and surface diversity.
No report on the application or research of ARTP in improving the vitality of seeds, particularly corn seeds, is found.
Disclosure of Invention
In order to solve the defects in the prior art, the invention provides a method for improving the seed viability, which comprises the following steps: the wet seeds were treated with atmospheric room temperature plasma.
In some embodiments, the wet state of the seed is obtained by soaking the seed in water for 3-48h (e.g., 5h, 10h, 15h, 20h, 25h, 30h, 35h, 39h, 43h, 45h, 47 h).
In some embodiments, the seeds are soaked with water for 12-36h to obtain the seeds in the wet state.
In some embodiments, the seed is a corn seed.
In some embodiments, the seed is a 4W, 6WC, Chang 7-2 or PH4CV corn seed.
In some embodiments, the seed is treated with the atmospheric room temperature plasma for 5-90min (e.g., 7min, 10min, 20min, 40min, 50min, 58min, 70min, 80min, 88 min).
In some embodiments, the seed is treated with the atmospheric room temperature plasma for 15-30min (e.g., 17min, 19min, 22min, 25min, 28 min).
In some embodiments, the embryo of the seed is subjected to the treatment towards an atmospheric room temperature plasma.
In some embodiments, the distance between the plasma emission source and the seed of the normal-pressure room-temperature plasma generating device is 2-5 mm.
In some embodiments, the working gas of the atmospheric-pressure room-temperature plasma generating device is helium.
In some embodiments, the helium gas has a purity of 95-99.99% (e.g., 96%, 97%, 98%, 99%).
In some embodiments, the helium gas has a purity of 99.99%.
In some embodiments, the atmospheric room temperature plasma generating device operates at a temperature of 20-30 ℃ (e.g., 21 ℃, 23 ℃, 24 ℃, 27 ℃, 29 ℃).
In some embodiments, the power supply power of the normal-pressure room-temperature plasma generation device is 240-400W (e.g., 250W, 280W, 310W, 340W, 390W).
In some embodiments, the working gas flow rate of the atmospheric room temperature plasma generating device is 10-20L/min (e.g., 11L/min, 13L/min, 15L/min, 17L/min, 19L/min).
Compared with the prior art, the invention has the advantages and beneficial effects that: (1) the normal-pressure room-temperature plasma technology adopted by the invention ensures that the plasma jet temperature is less than or equal to 40 ℃, the active particles are uniformly distributed, and the method is harmless to human bodies and the environment; (2) the germination potential of the soaked seeds is increased after the seeds are treated by the normal-pressure room-temperature plasma jet generating device, the germination rate is not reduced at least, the seed utilization rate is improved, and the seedlings are ensured to be tidy and have strong vitality.
Drawings
FIG. 1 is a bar graph of the change in seed germination potential under different treatment conditions as in example 2.
FIG. 2 is a bar graph of the germination rate of seeds for different treatments in example 2.
FIG. 3 is a bar graph of the change in germination index of seeds for different treatments in example 2.
FIG. 4 is a bar graph of the change in seed viability index for the different treatment cases in example 2.
Wherein, G-CK: dry seeds are not subjected to ARTP treatment; G-CL 1: treating dry seeds with ARTP for 15 min; G-CL 2: treating dry seeds with ARTP for 30 min; J-CK: the dry seeds are not subjected to ARTP treatment after being soaked for 12 hours; J-CL 1: soaking the dry seeds for 12h, and then performing ARTP treatment for 15 min; J-CL2 Dry seeds were soaked for 12h and then treated with ARTP for 30 min.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in detail with reference to the accompanying drawings.
The instrument comprises the following steps: an ARTP generating device, namely a normal-pressure low-temperature plasma generating device, is a normal-pressure low-temperature plasma generating device produced by Luoyang Huaqingtianmu biotechnology limited. The model is as follows: ARTP-P.
Corn seeds 4W, 6WC, Chang 7-2 and PH4CV were all commercially available varieties from: the Qingdao city is the center of the germplasm resources of main crops.
Example 1: corn seed ARTP treatment test for different varieties
(1) And (4) selecting seeds. 300 complete seeds with full seeds and the humidity of about 14 percent are respectively selected from 4W, 6WC and Chang 7-2. It is group-numbered every fifty pieces.
(2) Sterilizing the corn seeds screened in the step (1) with 1% sodium hypochlorite aqueous solution for 10min, washing with redistilled water for 3-5 times, and soaking for 0-48 h.
(3) And (6) processing the groups. The treated seeds of (2) were designed as 6 treatments, each of which was repeated three times. When the ARTP generating device is used for processing, the corn seeds are neatly placed in a carrying disc of the plant type ARTP generating device, wherein the carrying disc is suitable for larger plant seeds, each seed is preferably ensured to be embryo upwards (which is beneficial to improving the ATRP processing efficiency), 99.99 percent helium is taken as working gas, the working pressure is ambient atmospheric pressure, the working temperature is 30 ℃, the power supply is 380W, the flow velocity of working gas flow is 15L/min, and the distance between a plasma emission source and the corn seeds is 2-5 mm.
Treatment 1: 4W does not carry out any treatment;
and (3) treatment 2: treating 4W dry seeds with ARTP for 30 min;
and (3) treatment: treating the Chang 7-2 dry seeds with ARTP for 90 min;
and (4) treatment: soaking Chang 7-2 for 12h, and performing ARTP treatment for 90 min;
and (4) treatment 5: soaking in 6WC for 36h, and performing ARTP treatment for 0 min;
and (6) treatment: soaking 6WC for 36h, and performing ARTP treatment for 60 min;
(4) and (4) germination test. The seeds treated with ARTP in (3) were germinated in an incubator at a constant temperature in dark at 28 ℃ for seven days according to the germination test method for corn seeds in 1996 International seed test protocol, in which the seeds were placed on a wet filter paper and covered with another layer of paper. Spraying distilled water 4 times daily, and maintaining incubator humidity above 75%. Recording the number of the germinated seeds day by day, counting for the first time at the 4 th day, measuring the bud length and the root length of the normal seedlings, calculating the germination index, counting for the last time at the 7 th day and calculating the vitality index.
When measuring the dry weight, the seedlings with the measured fresh weight are placed in a clean glass culture dish, dried for 8 hours in a constant-temperature drying oven at the temperature of 103 +/-2 ℃, and when the temperature is reduced to the room temperature, the dry weight of the seedlings is measured by a balance with the precision of 0.01. The germination vigor, germination rate, germination index and vigor index of the seeds are calculated according to the following formulas.
The germination vigor (%) of the seeds was equal to the number of germinated seeds for 4 days of germination x 100/number of test seeds.
The germination rate (%) of the seeds was equal to the number of germinated seeds in 7 days of germination x 100/number of test seeds.
Germination index Ig ═ Σ (Gt/Dt) of seeds in which: gt is the germination percentage at different times (t days), and Dt is the germination test days.
The vigor index Iv of the seed is sxig sxΣ (Gt/Dt), where S is the dry weight of the germination vigor count seed.
Results and analysis
The differences between the three replicates of each treatment were not significant by statistical analysis. Referring to table 1, it can be seen from table 1 that the germination potential, germination rate, germination index and vitality index are all decreased after ARTP treatment without soaking the seeds, and the germination potential, germination rate, germination index and vitality index are all increased after ARTP treatment with the soaked seeds.
TABLE 1 comparison of different treatment modalities for the same species
| Processing number | Variety-soak time-treatment time | Vigor of germination | Germination rate | Index of germination | Vitality index |
| 1 | 4W--0h--0min | 0.88 | 0.97 | 1.07 | 5.42 |
| 2 | 4W--0h--30min | 0.79 | 0.83 | 0.89 | 3.82 |
| 3 | Chang 7-2-0 h-90 min | 0.24 | 0.57 | 0.06 | 1.22 |
| 4 | Chang 7-2-12 h-90 min | 0.54 | 0.70 | 0.67 | 1.95 |
| 5 | 6WC--36h--0min | 0.80 | 0.92 | 1.14 | 5.90 |
| 6 | 6WC--36h--60min | 0.94 | 0.94 | 1.52 | 8.99 |
Example 2: PH4CV corn seed ARTP treatment test under different soaking conditions
(1) And (4) selecting seeds. At least 900 full and complete PH4CV corn seeds were taken, and the test was designed by random grouping, with three replicates per treatment group.
(2) ① Dry seeds, without any treatment, 50 seeds are repeated, ② wet seeds are soaked in clean water for 12h at 25 ℃, and 50 seeds are repeated.
(3) And (4) ARTP treatment. And (3) treating the two kinds of seeds in the step (2) by using an ARTP generating device, taking 99.99% helium as working gas, taking the working pressure as the ambient atmospheric pressure, taking the working temperature as 30 ℃, and taking the treatment time as three levels of 0, 15 and 30min, wherein the power of a power supply is 380W, the flow rate of working gas flow is 15L/min, the distance between a plasma emission source and the corn seeds is 2-5 mm.
(4) According to the germination test method of the corn seeds in 1996 International seed test protocol, the PH4CV corn seeds treated in the step (3) are paved with three layers of filter paper in a culture dish, 20mL of sterilized water is added, and the seeds germinate in the culture dish (embryo faces upwards) in a dark environment at a constant temperature of 28 ℃ for seven days. The humidity of the incubator is kept above 75%. The number of germinated seeds, the number of the germinated seeds counted at the 4 th day, the number of the germinated seeds counted at the 7 th day, the root length and the bud length are measured, and the germination index and the vitality index are calculated. Wherein, the standard of germination is that the length of the bud reaches half of the length of the seed. The germination vigor, germination rate, germination index and vigor index of the seeds are calculated according to the following formulas. When measuring the dry weight, the seedlings with the measured fresh weight are placed in a clean glass culture dish, dried for 8 hours in a constant-temperature drying oven at the temperature of 103 +/-2 ℃, and when the temperature is reduced to the room temperature, the dry weight of the seedlings is measured by a balance with the precision of 0.01.
The germination vigor (%) of the seeds was equal to the number of germinated seeds for 4 days of germination x 100/number of test seeds.
The germination rate (%) of the seeds was equal to the number of germinated seeds in 7 days of germination x 100/number of test seeds.
Germination index Ig ═ Σ (Gt/Dt) of seeds in which: gt is the germination percentage at different times (t days), and Dt is the germination test days.
The vigor index Iv of the seed is sxig sxΣ (Gt/Dt), where S is the dry weight of the germination vigor count seed.
Results and analysis
Fig. 1 shows the variation of the germination vigor of seeds under different treatments, fig. 2 shows the variation of the germination rate of seeds under different treatments, the germination index of seeds under different treatments is shown in fig. 3, and the vigor index is shown in fig. 4. The letters on the bar graph indicate the significance of the difference between the different treatment groups.
The differences between the three replicates of each treatment were not significant by statistical analysis.
Therefore, the germination rate, the germination potential, the germination index and the vigor index of the PH4CV corn dry seeds are remarkably reduced along with the prolongation of the ARTP treatment time, while the germination potential, the germination index and the vigor index of the PH4CV corn dry seeds soaked for 12 hours are remarkably increased along with the prolongation of the ARTP treatment time, and the germination rate is close to 1. This shows that the combination of soaking and ARTP treatment helps to increase the germination potential of the PH4CV corn seeds, increase the vitality and vitality of the seeds and ensure the uniform emergence of the seedlings.
In conclusion, the embodiments show that the seed soaked by ARTP treatment can ensure the vitality increase of the corn seed, obviously improve the germination vigor, the germination index and the vitality index of the corn seed, has no obvious damage to plants, and is beneficial to improving the utilization efficiency of the corn seed.
It will be appreciated by those skilled in the art that the invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The embodiments disclosed above are therefore to be considered in all respects as illustrative and not restrictive. All changes which come within the scope of or equivalence to the invention are intended to be embraced therein.
Claims (10)
1. A method of increasing seed vigor, the method comprising: the wet seeds were treated with atmospheric room temperature plasma.
2. The method of claim 1, wherein said seeds are soaked in water for 3-48 hours to obtain said seeds in a wet state;
preferably, the seeds are soaked in water for 12-36h to obtain the seeds in the wet state.
3. The method of claim 1, wherein the seed is corn seed;
preferably, the seeds are 4W, 6WC, Chang 7-2 or PH4CV corn seeds.
4. The method of claim 1, wherein the seed is treated with the atmospheric room temperature plasma for 5-90 min;
preferably, the seeds are treated with the normal-pressure room-temperature plasma for 15-30 min.
5. The method of claim 1, wherein said treating of said embryo of said seed is performed towards an atmospheric room temperature plasma.
6. The method according to claim 1, wherein the distance between the plasma emission source and the seed of the normal-pressure room-temperature plasma generating device is 2-5 mm.
7. The method of claim 1, wherein the working gas of the atmospheric room temperature plasma generating device is helium.
8. The method of claim 7 wherein said helium gas has a purity of 95-99.99%;
preferably, the purity of the helium gas is 99.99%.
9. The method of claim 1, wherein the normal pressure room temperature plasma generating device has a working temperature of 20-30 ℃;
preferably, the power supply power of the normal-pressure room-temperature plasma generating device is 240-400W.
10. The method of claim 1, wherein the flow rate of the working gas stream of the atmospheric room temperature plasma generating device is 10-20L/min.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109463062A (en) * | 2017-09-07 | 2019-03-15 | 洛阳华清天木生物科技有限公司 | A method of utilizing atmospheric pressure at room corona treatment crop seeds |
| CN109496839A (en) * | 2018-12-27 | 2019-03-22 | 河南师范大学 | A kind of pair of dichromatism golden wave carries out the breeding method of physical mutagenesis |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109463062A (en) * | 2017-09-07 | 2019-03-15 | 洛阳华清天木生物科技有限公司 | A method of utilizing atmospheric pressure at room corona treatment crop seeds |
| CN109496839A (en) * | 2018-12-27 | 2019-03-22 | 河南师范大学 | A kind of pair of dichromatism golden wave carries out the breeding method of physical mutagenesis |
Non-Patent Citations (4)
| Title |
|---|
| ANNA ZAHORANOVÁ等: "Effect of Cold Atmospheric Pressure Plasma on Maize Seeds: Enhancement of Seedlings Growth and Surface Microorganisms Inactivation", 《PLASMA CHEM PLASMA PROCESS》 * |
| 清华大学无锡应用技术研究院生物育种研究中心: "ARTP（常压室温等离子体）诱变育种仪", 《百度快照》 * |
| 许言: "常压室温等离子体对怀牛膝和昆仑雪菊辐照的生物学效应", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 赵喜亭等: "ARTP辐照对两色金鸡菊种子诱变的生物学效应", 《河南师范大学学报（自然科学版）》 * |
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