CN112005657A - Method for screening salt-tolerant germplasm of quinoa by utilizing natural seawater - Google Patents
Method for screening salt-tolerant germplasm of quinoa by utilizing natural seawater Download PDFInfo
- Publication number
- CN112005657A CN112005657A CN202010917060.5A CN202010917060A CN112005657A CN 112005657 A CN112005657 A CN 112005657A CN 202010917060 A CN202010917060 A CN 202010917060A CN 112005657 A CN112005657 A CN 112005657A
- Authority
- CN
- China
- Prior art keywords
- seeds
- natural seawater
- germination
- quinoa
- salt
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Botany (AREA)
- Health & Medical Sciences (AREA)
- Physiology (AREA)
- Soil Sciences (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
The invention relates to a method for screening salt-tolerant germplasm of quinoa by utilizing natural seawater, which comprises the following steps: step 1, collecting seed materials, step 2, selecting seeds, step 3, preparing natural seawater series concentration gradient solution, step 5, testing physicochemical properties of the solution, step 6, analyzing germination characteristics and salt tolerance of the seeds, step 8, determining the concentration of the natural seawater suitable for screening salt-tolerant germplasm, and screening and determining salt-tolerant materials.
Description
Technical Field
The invention belongs to the technical field of crop germplasm screening and evaluation, and particularly relates to a method for screening quinoa salt-tolerant germplasm by utilizing natural seawater stress treatment.
Background
With the continuous reduction of fresh water resources and the increasing of the salinization degree of soil, the planting of halophytes is considered as the most economic mode for improving saline-alkali soil, can increase the organic matters and the microbial biomass of the soil, can prevent the salt accumulation on the surface of the soil, and plays an important role in restoring the agricultural ecological environment. According to statistics, 97.5 percent of water on the earth is salt water, and the water is about 10 hundred million hm all over the world2Salinized land, and warming with global climate, increasing world population and increasing human activitiesThe severity is increasing. The land of salinization of China has abundant resources, and the area reaches 9900 more than ten thousand hm2(ii) a The arable land area is affected by salinization by about 6.62%, and especially in coastal areas, the limitation factor seriously threatens the grain safety. The yellow river delta area of Shandong province is the main distribution area of the Chinese saline-alkali soil and has 40 to ten thousand hm more than the tidal flat wasteland2And the soil and climate conditions are suitable, so that the soil becomes a national important agricultural land reserve resource. However, the salinization severely restricts the land development and utilization in the area, and causes a great amount of resource waste. Therefore, the improvement and utilization of the beach saline-alkali soil is a key problem to be solved urgently in agricultural sustainable development.
Chenopodium quinoa (Chenopodium quinoa # Wild) is an annual facultative halophytic dicotyledonous herbaceous plant of Chenopodiaceae, prefers cold and intense light, and is mainly distributed in Tibetan, Gansu, Qinghai, Shanxi, etc. in China. Quinoa has excellent characteristics of salt and alkali resistance, low temperature resistance, drought resistance and the like, can grow well in arid and salinized marginal agricultural areas, and is a crop with great potential in sustainable development of agricultural ecological systems. Meanwhile, as a green health-care crop, the quinoa seeds are rich in various minerals, vitamins, essential amino acids and fatty acids, wherein the content of unsaturated fatty acids is up to 89.4%, and the content of polyunsaturated fatty acids is 54.2-58.3%, so that the quinoa seeds are very important for human health and food safety. In addition, the quinoa straw is rich in various nutrient substances, the crude protein content reaches 5.42%, and the quinoa straw can be used as a potential backup forage grass resource to relieve the contradiction between livestock in alpine regions and forage grass seasonal imbalance.
At present, researches on quinoa are mostly concentrated on the aspects of introduction and cultivation, resource development and utilization and the like, and the researches on salt tolerance mechanism and salt tolerance variety breeding are less, so that screening of quinoa salt tolerance germplasm resources has very important significance on the salt tolerance mechanism research, new variety cultivation and beach saline-alkali soil improvement. In China, with the aggravation of soil salinization, the deterioration of agricultural ecological environment, the rapid development of ecological animal husbandry and the increase of the demand of human on green healthy food, the chenopodium quinoa has increasingly prominent advantages as a multi-purpose high-quality crop resource of 'grain feeding, appreciation and ecology' in improving the saline-alkali soil, restoring the agricultural ecological environment and improving the dietary nutrition of human. In recent years, the planting area of the Chinese chenopodium quinoa is greatly increased, and the yield of the Chinese chenopodium quinoa is continuously increased. However, effective data for screening salt-tolerant germplasm of quinoa under the seashore beach environment of the yellow river delta region in Shandong do not exist yet. Therefore, the invention researches the quinoa salt-tolerant germplasm material screening method under the stress of natural seawater, finally determines the proper concentration condition and operation procedure of the natural seawater for screening the quinoa salt-tolerant germplasm, and has important significance for screening and applying the salt-tolerant material.
The invention researches and determines specific parameters for screening the salt-tolerant germplasm of quinoa under the stress of natural seawater. On the technical application program, a set of standardized operation system consisting of seed material collection, seed selection, natural seawater series concentration gradient solution preparation, solution physicochemical property determination, seed germination test, seed germination characteristic and salt tolerance analysis, natural seawater concentration determination suitable for screening salt-tolerant germplasm, and salt-tolerant material screening and determination is formed, and errors caused by different laboratories or technicians in the technical application process are avoided. The invention provides a standardized operation method for screening the salt-tolerant germplasm of the quinoa by natural seawater stress treatment, and the technology is more complete and convenient to apply.
Disclosure of Invention
In order to achieve the aim, the invention provides a method for screening salt-tolerant seeds of quinoa by utilizing natural seawater stress treatment, which comprises the following steps:
step 1, collecting seed material
The method comprises the steps of selecting seed materials of 'Long Cheng No. 1, Long Cheng No. 3, Long Cheng No. 4 and YL (non-species name)', judging the vitality level of the seeds by using a method of calculating the germination rate of the seeds by taking the standard that the radicle breaks through at least 2mm of the seed coat, and taking Chenopodium quinoa seeds with high vitality level (standard germination rate is 100%) as the materials of the technology. The basic information of the seed material is shown in table 1.
TABLE 1 Chenopodium quinoa seed basic information
After the seed material was collected, the seeds that were undamaged, of consistent size, mature and full were manually screened in the laboratory and stored in a-20 ℃ freezer for testing.
Step 3, preparing natural seawater series concentration gradient solution
The natural seawater is taken from the red island region (N36 degrees 13 '7' and E120 degrees 8 '9') of Qingdao city, Shandong province, a series of concentration gradients are set, and the concentration gradients are respectively 0%, 20%, 40%, 60%, 70%, 80%, 90% and 100%, and the solutions with corresponding concentrations are prepared. The concentration percentage of the solution is the volume percentage of the natural seawater in the total volume of the solution.
Step 4, measuring physicochemical properties of the solution #
And (3) measuring the pH value and the conductivity value of the natural seawater series concentration gradient solution.
Step 5, seed Germination test #
The method comprises the steps of taking selected quinoa seeds as materials, taking natural seawater series concentration gradient solutions as germination media, and measuring the salt tolerance of different quinoa seeds in the germination process. #
Judging the seed germination by taking the condition that the radicle breaks through at least 2mm of the seed coat as a standard, counting the number of the germinated seeds, and calculating the indexes related to the seed germination characteristics and the indexes related to salt tolerance.
Step 7, determining the concentration of the natural seawater suitable for screening the salt-tolerant germplasm
According to the change of germination characteristic related indexes and salt tolerance related indexes of different quinoa seeds under the stress of natural seawater, the proper concentration condition of the natural seawater for the quinoa seeds to germinate is determined by combining statistical analysis.
Step 8, screening and determining salt-resistant materials
And screening and determining the quinoa salt-tolerant material by comparing the salt tolerance differences of different quinoa seeds under the stress of natural seawater.
Wherein, step 1, collecting seed material
This procedure requires the collection of different quinoa seed materials. Seeds with high initial activity level are selected as test samples, and the germination characteristics and the change of salt tolerance indexes of the seeds under the stress of natural seawater are compared to determine the proper concentration of natural seawater and salt-tolerant materials screened by the quinoa salt-tolerant germplasm.
Wherein, step 3, preparing natural seawater series concentration gradient solution
The natural seawater stress concentration is an important factor influencing the germination characteristics of seeds and the change of salt tolerance indexes.
Wherein, step 4, the physicochemical property of the solution is measured #
The basic data of physicochemical properties of natural seawater solutions with different concentrations is very important for screening and researching the salt-tolerant germplasm of quinoa.
According to the method, chenopodium quinoa seeds of 'chenopodium quinoa No. 1, chenopodium quinoa No. 3, chenopodium quinoa No. 4 and YL' with high activity level are selected, natural seawater series concentration gradient solution preparation, solution physicochemical property determination and seed germination tests are carried out according to a design scheme, and natural seawater suitable concentration conditions and salt-tolerant materials screened from chenopodium quinoa salt-tolerant germplasm are determined by determining seed germination characteristics and salt-tolerant index changes under natural seawater stress and combining statistical analysis.
Preferably, the method of the invention comprises the following steps:
step 1, collecting seed material
In order to ensure the practicability of screening research and application of the salt-tolerant seeds of the chenopodium quinoa, the activity level of the seeds is judged by a method of calculating the germination rate by taking the embryo root breaking through at least 2mm as a standard, and the chenopodium quinoa seeds with the germination rate of 100% are selected as test materials.
In order to ensure the accuracy of research results and eliminate errors caused by uneven seed quality, the invention selects seeds which are not damaged, have consistent sizes and are mature and plump for testing.
Step 3, preparing natural seawater series concentration gradient solution
According to the series of concentration gradients of 0%, 20%, 40%, 60%, 70%, 80%, 90% and 100%, the volumes of the natural seawater and the distilled water corresponding to the prepared solutions with various concentrations are respectively calculated. The natural seawater and the distilled water with corresponding volumes are measured by using a measuring cylinder, and are uniformly mixed to prepare solutions with different concentrations.
Step 4, measuring the physical and chemical properties of the solution
The pH value and the conductivity value of the solution with each concentration are measured by a pH meter and a conductivity meter respectively. And (3) cleaning the electrode, drying by using filter paper, inserting into the solution to be measured, and reading and recording when the number displayed by the pH meter or the conductivity meter is stable. When the conductivity is measured, if the reading exceeds the maximum measuring range of the conductivity meter, the solution needs to be diluted by a certain multiple, and then the measurement is carried out again.
Step 5, seed germination test
Carefully selecting 50 mature and plump chenopodium quinoa seeds without damage and with consistent size, placing the seeds into a culture dish with a cover of 110mm multiplied by 110mm and containing 3 layers of filter paper, wetting the filter paper with 10mL of natural seawater with various concentrations, and wetting the filter paper with distilled water as a control. Each treatment was repeated 3 times. Placing the culture dish in an illumination incubator (GXZ-380B-LED) to culture under the constant temperature and dark condition of 25 ℃, observing the germination condition of the seeds every 12h, judging the germination of the seeds by taking the criterion that the radicle breaks through at least 2mm of the seed coat, and counting the number of the germinated seeds until the seeds germinate for 7 d.
And respectively calculating the average germination time (MGT), the germination vigor (GE), the germination rate (GP), the Germination Index (GI), the Salt Tolerance Index (STI) and the Relative Salt Damage Rate (RSDR) of the chenopodium quinoa seeds according to a formula.
MGT (days), wherein n is the number of newly germinated seeds at time t, t is the number of days from seed arrangement to counting, and Σ n is the total number of all germinated seeds;
GE (%) ═ number of seeds germinated on day 3/total number of seeds tested) × 100%;
GP (%) (N/N) x 100%, N being the total number of all germinated seeds in the first 7 days, N being the total number of test seeds;
GI ═ Σ (Gt/Dt), Gt is the number of germination in t days, Dt is the corresponding number of germination days;
STI ═ Σ (GP + GE + GI)/mn, m is the number of calculation index terms, n is the number of salt concentration gradients;
RSDR (%) - (control germination rate-treatment germination rate)/control germination rate × 100%.
The experimental data were statistically analyzed for significance using Excel 2010 and SPSS17.0 software and plotted using sigmaplot10.0 software. Results are expressed as mean ± sem.
Step 7, determining the concentration of the natural seawater suitable for screening the salt-tolerant germplasm
And determining the stress concentration corresponding to the inflection point with obvious difference as the proper concentration condition of the natural seawater for salt-tolerant germination of the quinoa seeds according to the result change of the seed germination characteristic related index and the salt-tolerant related index.
Step 8, screening and determining salt-resistant materials
Screening the quinoa salt-tolerant materials according to the comparison of the salt tolerance differences of different quinoa seed materials under the stress of natural seawater.
Through experimental research and result analysis, the inventor discovers that: the technology can be used as an effective method for screening the quinoa salt-tolerant germplasm materials. On one hand, the technology determines that the proper concentration condition of the natural seawater for screening the salt-tolerant germplasm of the quinoa is 90-100%, and is the result of analyzing and comparing the change of the seed germination index and the salt-tolerant index under the stress of a series of concentrations in the range of 0-100%; on the other hand, the technology screens the Chenopodium quinoa No. 4 seed material with strong salt tolerance by comprehensively comparing the index changes of the 4 Chenopodium quinoa seeds under the natural seawater stress. Therefore, the quinoa seed material with strong salt tolerance can be screened by utilizing the technology; meanwhile, the standardized operation procedure of the technology is established to provide theoretical guidance for solving the problem that quinoa seeds germinate in the seaside beach environment. #
Compared with the prior art, the technology of the invention has the following advantages:
(1) the technology of the invention determines the concentration parameter of natural seawater for screening the salt-tolerant seeds of quinoa, and the concentration of 90-100% can be used for identifying the salt tolerance of different quinoa seed materials; meanwhile, the technology screens the seeds with stronger salt tolerance, namely the Chenopodium longum No. 4;
(2) aiming at the relatively ideal condition that the traditional quinoa salt-tolerant germplasm screening method mostly adopts NaCl single salt stress treatment with different concentration gradients and the limitation of manual control, the technology adopts natural seawater stress to determine the quinoa salt-tolerant germplasm screening method, and has important significance for simulating the quinoa salt-tolerant germplasm in the seaside beach environment; #
(3) The technology of the invention provides a feasible and easy-to-operate method capable of screening the salt-tolerant seeds of quinoa in a short time;
(4) the technical method is simple and convenient, low in cost, high in screening efficiency, standardized and normalized in operation procedure and easy to popularize and use.
According to the invention, the natural seawater concentration parameters for screening the salt-tolerant germplasm of the quinoa are determined, and meanwhile, the quinoa No. 4 screened by the method is a quinoa seed material with stronger salt tolerance, and then the quinoa seed material is used for breeding, planting, cultivating and managing the quinoa in saline-alkali soil of yellow river delta, and the further work is as follows:
(1) the quinoa cultivation technology comprises the following steps: the screened Chenopodium quinoa No. 4 with strong salt tolerance is planted in saline-alkali soil of the yellow river delta, the technical points of Chenopodium quinoa planting and management suitable for the yellow river delta area are explored, and a high-quality Chenopodium quinoa cultivation technology is developed;
(2) chenopodium quinoa breeding work: taking the natural seawater concentration and the Chenopodium quinoa No. 4 Chenopodium quinoa seeds determined in the invention as materials, excavating salt-tolerant related genes of the Chenopodium quinoa through the combined analysis of transcriptome, metabolome and other multiomics, and carrying out salt-tolerant germplasm creation by utilizing biotechnology to obtain a new salt-tolerant Chenopodium quinoa material;
(3) and (3) development and utilization of quinoa: exploring a new mode for improving the saline-alkali mud flat of the yellow river delta through quinoa breeding; meanwhile, the quinoa straws are used as high-quality coarse feed for livestock and poultry, and the processing and utilization technology of the quinoa straws is developed to serve the production and development of modern animal husbandry.
Through the work, the conditions suitable for the growth of the quinoa seeds are obtained, and a foundation is laid for the popularization and the application of the technology disclosed by the invention.
Description of the drawings:
FIG. 1 is the influence of natural seawater of different concentrations on the germination potential of quinoa seeds
Note: different capital letters indicate that the difference between the natural seawater treatments with different concentrations of the same seed material is obvious, and different lowercase letters indicate that the difference between the natural seawater treatments with the same concentration and the different seed materials is obvious (P is less than 0.05).
FIG. 2 is the influence of natural seawater of different concentrations on the germination rate of quinoa seeds
Note: different capital letters indicate that the difference between the natural seawater treatments with different concentrations of the same seed material is obvious, and different lowercase letters indicate that the difference between the natural seawater treatments with the same concentration and the different seed materials is obvious (P is less than 0.05).
FIG. 3 is the influence of natural seawater of different concentrations on germination index of quinoa seeds
Note: different capital letters indicate that the difference between the natural seawater treatments with different concentrations of the same seed material is obvious, and different lowercase letters indicate that the difference between the natural seawater treatments with the same concentration and the different seed materials is obvious (P is less than 0.05).
FIG. 4 influence of natural seawater of different concentrations on salt tolerance index of quinoa seeds
Note: different capital letters indicate that the difference between the natural seawater treatments with different concentrations of the same seed material is obvious, and different lowercase letters indicate that the difference between the natural seawater treatments with the same concentration and the different seed materials is obvious (P is less than 0.05).
FIG. 5 the effect of natural seawater of different concentrations on the relative salt damage rate of quinoa seeds
Note: different capital letters indicate that the difference between the natural seawater treatments with different concentrations of the same seed material is obvious, and different lowercase letters indicate that the difference between the natural seawater treatments with the same concentration and the different seed materials is obvious (P is less than 0.05).
Detailed Description
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto.
Example 1
Step 1, collecting seed material
The standard germination rates of seeds of Long Cheng No. 1, Long Cheng No. 3, Long Cheng No. 4 and YL selected in the present invention are shown in Table 2.
TABLE 2 quinoa seed Standard Germination Rate
The quinoa seeds are selected one by means of a microscope and tweezers, and mature and plump seeds without damage and with consistent sizes are selected for later use.
Step 3, preparing natural seawater series concentration gradient solution
According to the series of concentration gradients of 0%, 20%, 40%, 60%, 70%, 80%, 90% and 100%, the volumes of the natural seawater and the distilled water corresponding to the prepared solutions with various concentrations are respectively calculated. The natural seawater and the distilled water with corresponding volumes are measured by using a measuring cylinder, and are uniformly mixed to prepare solutions with different concentrations.
Step 4, measuring the physical and chemical properties of the solution
The pH value and the conductivity value of the solution with each concentration are measured by a pH meter and a conductivity meter respectively. And (3) cleaning the electrode, drying by using filter paper, inserting into the solution to be measured, and reading and recording when the number displayed by the pH meter or the conductivity meter is stable. When the conductivity is measured, if the reading exceeds the maximum measuring range of the conductivity meter, the solution needs to be diluted by a certain multiple, and then the measurement is carried out again.
In the present invention, the pH and conductivity values of the natural seawater at each concentration gradient are shown in table 3.
TABLE 3 physicochemical Properties of Natural seawater at various concentrations
Step 5, seed germination test
Carefully selecting 50 mature and plump chenopodium quinoa seeds without damage and with consistent size, placing the seeds into a culture dish with a cover of 110mm multiplied by 110mm and containing 3 layers of filter paper, wetting the filter paper with 10mL of natural seawater with various concentrations, and wetting the filter paper with distilled water as a control. Each treatment was repeated 3 times. Placing the culture dish in an illumination incubator (GXZ-380B-LED) to culture under the constant temperature and dark condition of 25 ℃, observing the germination condition of the seeds every 12h, judging the germination of the seeds by taking the criterion that the radicle breaks through at least 2mm of the seed coat, and counting the number of the germinated seeds until the seeds germinate for 7 d.
1. Influence of natural seawater with different concentrations on average germination time of quinoa seeds
MGT was prolonged for different quinoa seeds as the concentration of natural seawater increased (table 4). Compared with the reference concentration (0%), the 20-60% of natural seawater has no significant influence difference (P is more than or equal to 0.05) on MGT of the Chenopodium album Gaertn seed No. 1, and the influence difference is respectively 0.77, 0.89 and 1.22 d; 70-100% of natural seawater obviously (P is less than 0.05) prolongs the seed MGT, and the MGT is respectively 1.56, 2.62, 2.80 and 3.00 d; although the differences between the 80%, 90% and 100% concentrations were not significant (P.gtoreq.0.05), they were all significant (P < 0.05) over the MGT of each of the other concentration treatments. For the seeds of Long Cheng 3 and Long Cheng 4, the difference between the MGT effect of 20-40% natural seawater on the seeds is not significant (P is more than or equal to 0.05), and the MGT effect of 60-100% concentration is significant (P is less than 0.05). Except that 20-60% of natural seawater has no obvious influence on the MGT of the YL seed (P is more than or equal to 0.05), the MGT of the YL seed is prolonged obviously at 70-100% concentration (P is less than 0.05).
When the natural seawater concentration is 0-60% and 80%, the MGT of the longli No. 3 seed is significantly (P < 0.05) higher than that of the longli No. 1, 4 and YL, but the MGT difference between the longli No. 4 and YL seeds is not significant (P is more than or equal to 0.05), and is the minimum value among 4 materials. When the natural seawater concentration is 70% and 90%, the MGT of the longli No. 1 and longli No. 3 seeds is significantly higher (P < 0.05) than that of the longli No. 4 and YL seeds, and the difference between the longli No. 1 and longli No. 3 and the difference between the longli No. 4 and YL are not significant (P is more than or equal to 0.05). When the natural seawater concentration is 100%, the MGT difference of the 4 quinoa seeds is not significant (P is more than or equal to 0.05).
TABLE 4 influence of Natural seawater of different concentrations on average Germination time of Chenopodium quinoa seeds
Note: the difference between different natural seawater treatments with different concentrations is obvious when different capital letters in the same row represent the same variety, and the difference between different varieties when different capital letters in the same column represent the same concentration natural seawater treatments (P is less than 0.05).
2. Influence of natural seawater with different concentrations on germination potential of quinoa seeds
As the concentration of the natural seawater increases, the germination potential GE of the 4 chenopodium quinoa seeds shows a decreasing trend (figure 1). Compared with the control (0%), the difference between the 20-60% natural seawater and the GE of the seeds of Chenopodium longum No. 1 and Chenopodium longum No. 3 is not significant (P is more than or equal to 0.05), the GE of the seeds is significantly reduced at 70-100% (P is less than 0.05), and the GE is reduced to the lowest value at 100%, which is 1%. For the seeds of Chenopodium longum No. 4, the GE of 20-80% of natural seawater could not be significantly reduced (P is greater than or equal to 0.05) compared with the control, the GE was significantly reduced (P is less than 0.05) only when the concentration was 90-100%, and the concentration was still 31% when the concentration was 100%. Except that 20% of natural seawater can not obviously reduce YL GE (P is more than or equal to 0.05), 40% -100% of natural seawater can obviously reduce GE (P is less than 0.05), and the content is reduced to the minimum value of 27% when the concentration is 100%.
When the concentration of the natural seawater is 0-20%, the GE difference of the 4 quinoa seeds is not significant (P is more than or equal to 0.05); when the natural seawater concentration is 40%, the GE of seeds of Long Cheng No. 1 and Long Cheng No. 4 is significantly (P < 0.05) higher than that of YL; when the natural seawater concentration is 60%, the GE of the Long Cheng No. 1 seeds is obviously (P is less than 0.05) higher than that of the Long Cheng No. 3 and YL seeds; when the natural seawater concentration is increased to 70-80%, the GE of the Long Cheng No. 4 seeds is obviously higher (P is less than 0.05) than that of the other 3 seeds; when the natural seawater concentration continues to rise to 90% -100%, the langli No. 4 and YL seeds GE are significantly (P < 0.05) higher than the langli No. 1 and langli No. 3 seeds GE, but the difference between langli No. 4 and YL, and between langli No. 1 and langli No. 3 is not significant (P is more than or equal to 0.05).
3. Influence of natural seawater with different concentrations on germination rate of quinoa seeds
The germination rates GP of the 4 chenopodium quinoa seeds all show a decreasing trend with the increasing concentration of the natural seawater (figure 2). Compared with the control (0%), the difference of the influence of 20-60% of natural seawater on the longeron No. 1, the longeron No. 3 and the YL seed GP is not significant (P is more than or equal to 0.05); the seed GP is obviously reduced by 70-100 percent of natural seawater (P is less than 0.05), the seed GP is reduced to the lowest value at the concentration of 100 percent, the value is respectively 3 percent, 1 percent and 37 percent, and the difference between the treatment of each concentration is obvious (P is less than 0.05). For Chenopodium longum seed No. 4, GP was not significantly reduced at 20-80% (P is greater than or equal to 0.05), but was significantly reduced at 90-100% (P is less than 0.05), and remained at 45% at 100%. As can be seen from fig. 2, the concentrations of the natural seawater at the key nodes where the longus No. 1, longus No. 3, longus No. 4 and YL seeds GP significantly decreased are 70%, 90% and 70%, respectively, and the corresponding GP is 89%, 85%, 77% and 88%, respectively.
When the concentration of the natural seawater is 0-20%, the GP difference of the 4 quinoa seeds is not significant (P is more than or equal to 0.05); when the natural seawater concentration is 40%, the seeds GP of Long Cheng No. 1 and Long Cheng No. 4 are significantly higher (P < 0.05) than those of YL seeds GP; when the natural seawater concentration is 60%, the GP of the Long Cheng No. 1 seed is obviously higher (P is less than 0.05) than that of the other 3 seeds GP; when the natural seawater concentration is increased to 70-80%, the GP of the Long Cheng No. 4 seed is obviously higher (P is less than 0.05) than that of the other 3 seeds GP; when the natural seawater concentration continues to rise to 90-100%, the longli No. 4 and YL seeds GP are significantly (P < 0.05) higher than the longli No. 1 and longli No. 3 seeds GP, but the difference between longli No. 4 and YL, and between longli No. 1 and longli No. 3 is not significant (P is more than or equal to 0.05).
4. Influence of natural seawater with different concentrations on germination index of quinoa seeds
The germination index GI of the 4 chenopodium quinoa seeds shows a decreasing trend along with the increase of the concentration of the natural seawater (figure 3). Compared with the control, 20-100% of natural seawater remarkably (P < 0.05) reduces GI of seeds of Long Cheng No. 1 and Long Cheng No. 3, and the difference between the concentration treatments is remarkable (P < 0.05). For Chenopodium longum seed No. 4, GI reduction was not significant at 20-40% (P is greater than or equal to 0.05), but was significant at 60-100% (P is less than 0.05), and the difference between the treatments was significant (P is less than 0.05). Except that 20% of natural seawater can not obviously reduce YL GI (P is more than or equal to 0.05), 40% -100% of natural seawater can obviously reduce GI (P is less than 0.05), and the difference between treatment of each concentration is obvious (P is less than 0.05).
When the concentration of the natural seawater is 0-100%, the GI of the Long Cheng No. 4 and YL seeds is significantly (P < 0.05) higher than that of the Long Cheng No. 1 and No. 3 seeds. For the seeds GI of Chenopodium Long. No. 1 and Chenopodium Long. 3, the GI of Chenopodium Long. No. 1 is significantly (P < 0.05) higher than that of Chenopodium Long. 3 when the natural seawater concentration is 0-60%, and the difference between them is not significant (P is greater than or equal to 0.05) when the natural seawater concentration is 70-100%. For GI seeds of Chenopodium longum No. 4 and YL seeds, when the natural seawater concentration is 0% -20% and 60% -70%, the difference between the two is not significant (P is more than or equal to 0.05); when the natural seawater concentration is 40%, GI of the Long Cheng No. 4 seeds is obviously (P is less than 0.05) higher than GI of YL seeds; when the natural seawater concentration is further increased to 80% -100%, the YL seed GI is significantly (P < 0.05) higher than the GI of Chenopodium album Gaertn seed No. 4.
5. Influence of natural seawater with different concentrations on salt tolerance index of quinoa seeds
As the concentration of the natural seawater rises, the salt tolerance index STI of the 4 chenopodium quinoa seeds shows a reduced variation trend (figure 4). Compared with the control, the 20-100% natural seawater has the advantages that the STI of the seeds of the Long Cheng No. 1 and the Long Cheng No. 3 are remarkably reduced (P is less than 0.05), and the difference between the concentration treatments is remarkable (P is less than 0.05). For Chenopodium longum seed No. 4, STI can not be significantly reduced at 20-40% (P is greater than or equal to 0.05), but can be significantly reduced only at 60-100% (P is less than 0.05), and the difference between the treatments of each concentration is significant (P is less than 0.05). Besides STI with YL reduced by 20% natural seawater (P is more than or equal to 0.05), STI is reduced remarkably by 40% -100% natural seawater (P is less than 0.05), and difference between concentration treatments is remarkable (P is less than 0.05).
When the concentration of the natural seawater is 20-100%, the STI of the Long Cheng No. 4 and YL seeds is obviously (P < 0.05) higher than that of the Long Cheng No. 1 and No. 3 seeds. For the seeds STI of longli No. 1 and longli No. 3, when the natural seawater concentration is 0% -60% and 90%, the STI of longli No. 1 is significantly (P < 0.05) higher than that of longli No. 3, and when the natural seawater concentration is 70% -80% and 100%, the difference between them is not significant (P is greater than or equal to 0.05). For the Chenopodium Long Li No. 4 and YL seed STI, except that the STI of the Chenopodium Long Li No. 4 seed is more significant (P < 0.05) than the STI of the YL seed when the natural seawater concentration is 40%, the STI difference between the two is not significant (P is more than or equal to 0.05) when the natural seawater concentration is treated.
6. Influence of natural seawater with different concentrations on relative salt damage rate of quinoa seeds
The relative salt damage rate RSDR of the 4 chenopodium quinoa seeds shows a rising trend along with the rising of the concentration of the natural seawater (figure 5). Compared with the control, 20-60% of natural seawater has no significant influence on the RSDR of the Long Cheng No. 1, the Long Cheng No. 3 and the YL seeds (P is more than or equal to 0.05), the RSDR of the seeds is significantly increased (P is less than 0.05) by 70-100% of concentration, and the difference between the concentration treatments is significant (P is less than 0.05). For Chenopodium longum seed No. 4, the RSDR cannot be significantly affected by the concentration of 20-80% (P is greater than or equal to 0.05), and the RSDR can be significantly increased only when the natural seawater concentration is increased to 90-100% (P is less than 0.05).
When the concentration of the natural seawater is 0-40%, the RSDR difference of the 4 quinoa seeds is not significant (P is more than or equal to 0.05); when the natural seawater concentration is 70-80%, the RSDR of the Long Cheng No. 4 seed is obviously lower (P is less than 0.05) than that of the other 3 seeds; when the natural seawater concentration is 90-100%, the RSDR of longli No. 4 and YL seeds is significantly lower (P < 0.05) than that of longli No. 1 and longli No. 3, and the difference between longli No. 4 and YL and between longli No. 1 and longli No. 3 is not significant (P is more than or equal to 0.05).
In summary, regarding the seeds of longli No. 1, under 20% -60% of natural seawater concentration stress, the differences of MGT, GE, GP and RSDR from the control are not significant; stress of 70-100% concentration causes all the measured indexes to be changed obviously. For the Chenopodium longum seed No. 3, when the concentration of 20-40% natural seawater is stressed, the other measurement indexes except GI and STI are not obviously different from the control indexes; the 60% concentration stress obviously increases the MGT of the seeds and reduces GI and STI; when the natural seawater concentration increased to 70% or more, all measurement indexes were significantly changed. For the seeds of Chenopodium longum No. 4, all the determination indexes have no significant difference from the control when 20-40% of the natural seawater concentration is stressed; the 60% -80% concentration stress obviously increases the MGT of the seeds and reduces GI and STI; when the natural seawater concentration is increased to 90% -100%, all measurement indexes are changed remarkably. Regarding YL seeds, all assay indexes were not significantly different from controls when 20% natural seawater concentration was stressed; stress of 40-60% concentration obviously reduces GE, GI and STI of seeds; when the natural seawater concentration is increased to 70% -100%, all measurement indexes are changed remarkably. Therefore, 90-100% of natural seawater can be used as a proper concentration condition for screening the salt-tolerant germplasm of quinoa.
In addition, according to the germination index and salt tolerance index changes of the 4 chenopodium quinoa seed materials under the stress of natural seawater with different concentrations, the salt tolerance of longli No. 4 and YL is obviously superior to that of longli No. 1 and longli No. 3. Under the stress of 90-100% high-concentration natural seawater, although the salt tolerance differences between Long Cheng No. 4 and YL seeds are not significant, and the salt tolerance differences between Long Cheng No. 1 and Long Cheng No. 3 seeds are also not significant, most of the indexes for determining the Long Cheng No. 4 seeds show that the salt tolerance is slightly better than that of the YL seeds, and most of the indexes for determining the Long Cheng No. 1 seeds show that the salt tolerance is slightly better than that of the Long Cheng No. 3 seeds. Therefore, the salt tolerance of the 4 kinds of quinoa seed materials is longli 4 # YL > longli 1 # longli 3, and longli 4 # can be used as the seed material with stronger salt tolerance.
Claims (5)
1. A method for screening salt-tolerant germplasm of quinoa by utilizing natural seawater comprises the following steps:
step 1, collecting seed material
Selecting seed materials of 'Long Cheng No. 1, Long Cheng No. 3, Long Cheng No. 4, YL (non-breed name)', judging the activity level of the seeds by a method of calculating the germination rate of the seeds by taking the radicle breaking through at least 2mm of the seed coat as a standard, and taking Chenopodium quinoa seeds with high activity level (standard germination rate 100%) as the materials of the technology;
step 2, seed selection
After the seed materials are collected, the seeds which are not damaged, have consistent size and are mature and full are manually screened in a laboratory and stored in a refrigerator at the temperature of minus 20 ℃ for a test station;
step 3, preparing natural seawater series concentration gradient solution
The natural seawater is taken from red island regions (N36 degrees 13 '7' and E120 degrees 8 '9') of Qingdao city, Shandong province, a series of concentration gradients are set, and the concentration gradients are respectively 0%, 20%, 40%, 60%, 70%, 80%, 90% and 100%, and the solutions with corresponding concentrations are prepared;
step 4, measuring physicochemical properties of the solution #
Measuring the pH value and the conductivity value of the natural seawater series concentration gradient solution;
step 5, seed Germination test #
Taking the selected quinoa seeds as materials, taking natural seawater series concentration gradient solutions as germination media, and determining the salt tolerance of different quinoa seeds in the germination process; #
Step 6, analyzing germination characteristics and salt tolerance of seeds
Judging seed germination by taking the condition that the radicle breaks through at least 2mm of seed coat as a standard, counting the number of germinated seeds, and calculating related indexes of seed germination characteristics and related indexes of salt tolerance;
step 7, determining the concentration of the natural seawater suitable for screening the salt-tolerant germplasm
Determining the proper concentration condition of the natural seawater for the germination of the quinoa seeds according to the change of germination characteristic related indexes and salt tolerance related indexes of different quinoa seeds under the stress of the natural seawater by combining statistical analysis;
step 8, screening and determining salt-resistant materials
And screening and determining the quinoa salt-tolerant material by comparing the salt tolerance differences of different quinoa seeds under the stress of natural seawater.
2. The method of claim 1, wherein, in step 1, seed material is collected
This procedure requires the collection of different quinoa seed materials. Seeds with high initial activity level are selected as test samples, and the germination characteristics and the change of salt tolerance indexes of the seeds under the stress of natural seawater are compared to determine the proper concentration of natural seawater and salt-tolerant materials screened by the quinoa salt-tolerant germplasm.
3. The method of claim 1, wherein step 3, natural seawater series concentration gradient solution preparation
The natural seawater stress concentration is an important factor influencing the germination characteristics of seeds and the change of salt tolerance indexes.
4. The method according to claim 1, wherein, in step 4, solution physicochemical property determination
The basic data of physicochemical properties of natural seawater solutions with different concentrations is very important for screening and researching the salt-tolerant germplasm of quinoa.
5. The method of claim 1, comprising the steps of:
step 1, collecting seed material
In order to ensure the practicability of screening research and application of the salt-tolerant seeds of the chenopodium quinoa, the activity level of the seeds is judged by a method of calculating the germination rate by taking the at least 2mm of radicle breakthrough seed coat as a standard, and the chenopodium quinoa seeds with the germination rate of 100 percent are selected as test materials;
step 2, seed selection
In order to ensure the accuracy of research results and eliminate errors caused by uneven seed quality, the invention selects seeds which are not damaged, have consistent sizes and are mature and plump for testing;
step 3, preparing natural seawater series concentration gradient solution
According to the series of concentration gradients of 0%, 20%, 40%, 60%, 70%, 80%, 90% and 100%, the volumes of the natural seawater and the distilled water corresponding to the prepared solutions with various concentrations are respectively calculated. Measuring natural seawater and distilled water with corresponding volumes by using a measuring cylinder, uniformly mixing, and preparing into solutions with different concentrations;
step 4, measuring the physical and chemical properties of the solution
The pH value and the conductivity value of the solution with each concentration are measured by a pH meter and a conductivity meter respectively. And (3) cleaning the electrode, drying by using filter paper, inserting into the solution to be measured, and reading and recording when the number displayed by the pH meter or the conductivity meter is stable. When the conductivity is measured, if the reading exceeds the maximum measuring range of the conductivity meter, the solution needs to be diluted by a certain multiple, and then the measurement is carried out again;
step 5, seed germination test
Carefully selecting 50 mature and plump chenopodium quinoa seeds without damage and with consistent size, placing the seeds into a culture dish with a cover of 110mm multiplied by 110mm and containing 3 layers of filter paper, wetting the filter paper with 10mL of natural seawater with various concentrations, and wetting the filter paper with distilled water as a control. Each treatment was repeated 3 times. Placing the culture dish in an illumination incubator (GXZ-380B-LED) to culture under the constant temperature and dark condition of 25 ℃, observing the germination condition of the seeds every 12h, judging the germination of the seeds by taking the criterion that the radicle breaks through at least 2mm of the seed coat, and counting the number of the germinated seeds until 7d of germination;
step 6, analyzing germination characteristics and salt tolerance of seeds
Respectively calculating the average germination time (MGT), germination vigor (GE), germination rate (GP), Germination Index (GI), Salt Tolerance Index (STI) and Relative Salt Damage Rate (RSDR) of the chenopodium quinoa seeds according to a formula;
MGT (days), wherein n is the number of newly germinated seeds at time t, t is the number of days from seed arrangement to counting, and Σ n is the total number of all germinated seeds;
GE (%) ═ number of seeds germinated on day 3/total number of seeds tested) × 100%;
GP (%) (N/N) x 100%, N being the total number of all germinated seeds in the first 7 days, N being the total number of test seeds;
GI ═ Σ (Gt/Dt), Gt is the number of germination in t days, Dt is the corresponding number of germination days;
STI ═ Σ (GP + GE + GI)/mn, m is the number of calculation index terms, n is the number of salt concentration gradients;
RSDR (%) - (control germination rate-treatment germination rate)/control germination rate × 100%;
the experimental data were statistically analyzed for significance using Excel 2010 and SPSS17.0 software and plotted using sigmaplot10.0 software. Results are expressed as mean ± sem;
step 7, determining the concentration of the natural seawater suitable for screening the salt-tolerant germplasm
According to the result change of the indexes related to the seed germination characteristics and the indexes related to salt tolerance, determining the stress concentration corresponding to the inflection points with obvious differences as the proper concentration condition of the natural seawater for salt tolerance germination of the quinoa seeds;
step 8, screening and determining salt-resistant materials
Screening the quinoa salt-tolerant materials according to the comparison of the salt tolerance differences of different quinoa seed materials under the stress of natural seawater.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010917060.5A CN112005657A (en) | 2020-09-03 | 2020-09-03 | Method for screening salt-tolerant germplasm of quinoa by utilizing natural seawater |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010917060.5A CN112005657A (en) | 2020-09-03 | 2020-09-03 | Method for screening salt-tolerant germplasm of quinoa by utilizing natural seawater |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112005657A true CN112005657A (en) | 2020-12-01 |
Family
ID=73515705
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010917060.5A Pending CN112005657A (en) | 2020-09-03 | 2020-09-03 | Method for screening salt-tolerant germplasm of quinoa by utilizing natural seawater |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112005657A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101142917A (en) * | 2007-08-30 | 2008-03-19 | 中国农业大学 | Method for improving halophyte seed germinating potential and its special-purpose soaking liquid |
| WO2013063555A1 (en) * | 2011-10-28 | 2013-05-02 | E. I. Du Pont De Nemours And Company | Methods for the identification of genes involved in abiotic stress tolerance in plants |
| CN110226382A (en) * | 2019-05-20 | 2019-09-13 | 湖北省农业科学院经济作物研究所 | Nature seawater coerces cotton seed germination phase Tolerant salt method |
-
2020
- 2020-09-03 CN CN202010917060.5A patent/CN112005657A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101142917A (en) * | 2007-08-30 | 2008-03-19 | 中国农业大学 | Method for improving halophyte seed germinating potential and its special-purpose soaking liquid |
| WO2013063555A1 (en) * | 2011-10-28 | 2013-05-02 | E. I. Du Pont De Nemours And Company | Methods for the identification of genes involved in abiotic stress tolerance in plants |
| CN110226382A (en) * | 2019-05-20 | 2019-09-13 | 湖北省农业科学院经济作物研究所 | Nature seawater coerces cotton seed germination phase Tolerant salt method |
Non-Patent Citations (2)
| Title |
|---|
| 修妤等: ""混合盐碱胁迫对藜麦萌发期的影响"", 《山东农业科学》 * |
| 袁飞敏等: ""不同钠盐胁迫对藜麦种子萌发的影响"", 《干旱区资源与环境》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103238449B (en) | Method for rapidly identifying and selecting salt-tolerant peanut varieties in room | |
| Serraj et al. | Screening chickpea mini-core germplasm for tolerence to soil salinity | |
| CN102754592B (en) | Method for screening variety with relatively-high phosphor utilization efficiency from multiple cut-flower chrysanthemum varieties | |
| CN102187774A (en) | System for identifying and evaluating seedling resistance of malus | |
| CN112634072A (en) | Evaluation method for cold resistance of rice in seedling stage based on principal component analysis | |
| Amanov et al. | Dependence of Growth Period, Yield Elements and Grain Quality of Winter Bread Wheat Varieties and Lines on Different Soil and Climate Conditions. | |
| CN103238493A (en) | Responses of different peach stock types for persistent drought and evaluation method of drought resistance of different peach stock types | |
| Zhang et al. | The effects of water and nitrogen on the roots and yield of upland and paddy rice | |
| CN109819714A (en) | A kind of method that can largely screen rice seedling Salinity tolerance germplasm | |
| CN110036858B (en) | Method for establishing soybean seedling emergence period salt tolerance identification method and screening salt tolerance germplasm | |
| CN100495390C (en) | Computer estimation method for salt tolerance of non-halophyte herbaceous plant | |
| CN110367067A (en) | A kind of paddy drought resistance evaluation method based on more gradient multiple characters Comprehensive drought resistant indexes | |
| Staniak et al. | Prolonged drought stress induced changes in yield and physiological processes of Trifolium repens and Festulolium braunii | |
| CN112005657A (en) | Method for screening salt-tolerant germplasm of quinoa by utilizing natural seawater | |
| CN110609124A (en) | Method for measuring salt tolerance level of mulberry variety | |
| HU et al. | Genetic analysis on characteristics to measure drought resistance using Dongxiang wild rice (Oryza rufupogon Griff.) and its derived backcross inbred lines population at seedling stage | |
| Li et al. | Soil boron content and the effects of boron application on yields of maize, soybean, rice and sugarbeet in Heilongjiang Province, PR China | |
| CN110476744A (en) | Floral leaf character is increasing the application on rape group child's hair twisted in a knot-childhood pericarp area | |
| CN107581061A (en) | A kind of rape seed High Temperature during Grain Filling tolerance high frequency zone authentication method | |
| CN112273222A (en) | Cotton variety breeding method | |
| CN112005658A (en) | Method for determining salt tolerance of quinoa seeds in germination period | |
| CN110367066A (en) | A kind of paddy drought resistance evaluation method based on the comprehensive drought resistance coefficient of more gradient multiple characters | |
| Li et al. | Inter-simple sequence repeat (ISSR) markers revealed genetic diversity and population structure among 9 wild species of Clematis L. | |
| Yang et al. | Association of carbon isotope discrimination with leaf gas exchange and water use efficiency in maize following soil amendment with superabsorbent hydrogel. | |
| CN107094518A (en) | A kind of method for quantitatively calculating the intrinsic anti-low-phosphorus stress ability of plant |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201201 |
|
| RJ01 | Rejection of invention patent application after publication |