CN109197474B - Method for rapidly screening peanut varieties with high oil content - Google Patents

Abstract

The invention discloses a method for rapidly screening peanut varieties with high oil content, and belongs to the technical field of flower breeding. The method for rapidly screening peanut varieties with high oil content in the present invention is as follows: sowing the peanuts to be screened, performing drought stress treatment 5 weeks after sowing, measuring the water potential of peanut leaves after drought stress, and importing the water potential value into the constructed relationship between oil content and water potential of peanuts. In the relational model, the oil content of peanuts is obtained; if the oil content is more than 55%, it is a high-oil peanut material, and if the oil content is less than 55%, it is discarded. The method of the invention for rapidly screening peanut varieties with high oil content is simple and easy to operate, and overcomes the problems of long time for oil content determination, complicated operation procedures, consumption of a lot of manpower, material resources and financial resources in conventional methods. In addition, the method for rapidly screening peanut varieties with high oil content of the present invention is not limited by seasons and spaces, and compared with the traditional method, the numerical value is not much different. The method for rapidly screening peanut varieties with high oil content of the present invention has high accuracy and reliable results.

Description

A method for rapid screening of peanut varieties with high oil content

technical field

The invention belongs to the technical field of flower breeding, and in particular relates to a method for rapidly screening peanut varieties with high oil content.

Background technique

Peanut is one of the important oil crops in my country and occupies an important position in agriculture and national economy. More than 50% of the peanuts produced in my country are used for oil extraction. Generally, the oil content of peanut varieties is about 50%, and the oil content of more than 55% is high-oil varieties or high-oil materials. Every 1% increase in oil content can increase the net profit. Therefore, cultivating new high-oil peanut varieties has become one of the main breeding goals of the majority of breeders. However, peanuts lack resources of high-oil varieties, and it is difficult to obtain high-oil progeny by cross-breeding. Therefore, the breeding of high-yield and high-oil varieties has always been a difficult problem to solve. The use of mutagenesis technology can cause gene mutation or chromosomal variation, so as to obtain high-oil mutants. However, the identification of high-oil mutants needs to be planted in the field, and can only be detected after they are mature and harvested and dried. At present, the determination of the oil content of peanut kernels is generally carried out by the residual method in qualified units such as the Oilseeds and Products Quality Supervision, Inspection and Testing Center of the Ministry of Agriculture. The determination method is cumbersome and complicated, requires a long processing time, and spends a lot of manpower, material and financial resources, and the oil content of the peanuts harvested each year can only be measured once. In order to select high oil body to breed new high oil cultivars, the oil content should be measured for each generation during the breeding process.

SUMMARY OF THE INVENTION

In view of the problems existing in the prior art, the purpose of the present invention is to provide a method for rapidly screening peanut varieties with high oil content, which can effectively shorten the breeding cycle.

In order to achieve the above object, the technical scheme of the present invention is:

The construction method of the relationship model between peanut oil content and water potential, the steps are as follows:

(1) Select multiple peanut materials of the same line, and each peanut material is divided into two parts, one part is planted in the field to determine the oil content of the kernels after mature harvest, and the other part is planted in flower pots to determine drought Leaf water potential after stress;

(2) The measuring method of field sowing and oil content is:

Choose sandy loam soil, apply enough base fertilizer, and apply 100 catties of compound fertilizer with 15% of N, P, and K content per acre, sow seeds in early May, with a sowing depth of 3-4cm, cover with mulch to keep moisture, increase temperature, and prevent weeds. Pay attention during growth. Water in time in drought, drain in time in waterlogging, harvest pods and sun-dry after maturity, and measure the oil content of kernels.

(3) The method of planting in flowerpot and measuring water potential is:

①Preparation of sowing substrate: The nutrient bowl substrate is selected from sandy loam and vermiculite in a ratio of 4:1, and an appropriate amount of compound fertilizer (N, P, K=1:1:1) is added according to the fertility of the soil and stirred evenly;

②Sowing: choose a flowerpot with an inner diameter of 25cm, and the substrate loaded is equal to the upper edge of the flowerpot. The seeds are sown in the nutrition bowl in early May, and the sowing depth is 3-4cm; water 600mL;

③Management

The above-mentioned nutritional bowl is placed in a natural environment, and watered according to the moisture content, 600mL each time;

④Treatment of drought stress

Carry out drought stress treatment after sowing 5 weeks, namely do not water when this watering according to moisture condition, and drought stress treatment is 2 days;

⑤ Take the discs of peanut leaves after drought stress, and measure the water potential of the leaves.

The relationship formula y=ax+b was constructed with the measured water potential of peanut leaves after drought stress as the x-axis and the oil content of peanut kernels as the y-axis.

A method for rapidly screening peanut varieties with high oil content, the steps are:

①Preparation of sowing substrate: The nutrient bowl substrate is selected from sandy loam and vermiculite in a ratio of 4:1, and an appropriate amount of compound fertilizer (N, P, K=1:1:1) is added according to the fertility of the soil and stirred evenly;

②Sowing: Choose a flowerpot with an inner diameter of 25cm, and the substrate to be loaded is the same as the upper edge of the flowerpot. In early May, the seeds of a certain peanut variety obtained by mutagenesis to obtain multiple mutants were sown in the nutrient pot, and the sowing depth was 3 -4cm; water 600mL;

③Management

The above-mentioned nutritional bowl is placed in a natural environment, and watered according to the moisture content, 600mL each time;

④Treatment of drought stress

Carry out drought stress treatment after sowing 5 weeks, namely do not water when this watering according to moisture condition, and drought stress treatment is 2 days;

⑤ Take the discs of peanut leaves after drought stress, and measure the water potential of the leaves;

⑥ Import the measured water potential value of peanut leaves into the constructed relationship model of peanut oil content and water potential to obtain the peanut oil content;

⑦Select mutants with oil content above 55% as high-oil materials, and discard those with oil content less than 55%.

On the basis of the above scheme, the method for measuring the water potential of the peanut leaf is:

a. Take the discs of peanut leaves after drought stress, take 10 discs of peanut leaves and place them in a 1mL gradient water potential sucrose solution, shake well and let stand for 20min;

b. Determine the refractive index of the sucrose solution after soaking the leaves, and compare it with the refractive index of the original sucrose solution:

If the refractive index of a certain sucrose solution is the same as that of the original sucrose solution, the measured value is the refractive index of the peanut mutant;

If the refractive index of a sucrose solution is different from that of the original sucrose solution, the measured refractive index of the two adjacent sucrose solutions after soaking the leaves is one larger than that of the original sucrose solution and one smaller than that of the original sucrose solution. The average of the coefficients is the refractive index of the peanut mutant;

c. Bring the measured refractive index of a peanut mutant after drought stress treatment into the relationship between the refractive index of sucrose solution and the water potential, and the obtained water potential is the water potential of the peanut mutant leaves.

On the basis of the above scheme, the water potentials of the gradient water potential sucrose solution are: -1.0MPa, -1.2MPa, -1.4MPa, -1.6MPa, respectively.

On the basis of the above scheme, the relationship between the refractive index of sucrose solution and the water potential is to measure the refractive index of sucrose solutions with different water potentials, and the refractive index is used as the x-axis and the water potential as the y-axis to construct a relational formula of y=ax+b.

The advantages of the technical solution of the present invention:

1. The present invention utilizes the determination of the water potential of the leaves of the peanuts after drought stress treatment, and the identification of the peanut kernel high-oil material. It overcomes the problems of conventional methods, such as long time for oil content determination, complicated operation procedures, and a lot of human, material, and financial resources. Not limited by season and space.

2. The present invention utilizes the determination of leaf water potential of peanuts after drought stress treatment to identify peanut kernel high-oil materials, which can be used as high-oil identification materials no matter which season is planted or where the peanut materials are planted. It overcomes the problems of inaccurate oil content of peanut kernels from Hainan Jiadai in breeding and poor seed fullness of greenhouse Jiadai, which cannot be used as conventional oil content determination materials.

3. The present invention utilizes the determination of leaf water potential after peanut drought stress treatment, and identification of the oil content of peanut kernels, which provides the possibility for peanut high-oil breeding, speeding up the breeding process, and shortening the breeding period. Overcome the problems of high-oil peanut breeding, the use of conventional methods to determine the oil content, can only be measured once a year, can not be added, and the breeding years are long.

Description of drawings

Fig. 1 Standard curve of refractive index and water potential of sucrose solutions with different water potentials;

Fig. 2 The standard curve of peanut water potential-oil content of Huayu 20 strain;

Fig. 3 Standard curve of peanut water potential-oil content of Huayu 22 strain;

Fig. 4 Standard curve of peanut water potential-oil content of Luhua No. 11 line.

Detailed ways

Terms used in the present invention generally have the meanings commonly understood by those of ordinary skill in the art unless otherwise specified.

The present invention will be described in further detail below with reference to specific embodiments and data. The following examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention in any way.

Example 1

The construction method of the relationship model between peanut oil content and water potential, the steps are as follows:

(1) Select multiple peanut materials of Huayu No. 20, Huayu No. 22 and Luhua No. 11 respectively, each peanut material is divided into two parts, one part is planted in the field, and used to determine the oil content of the kernels after ripening and harvesting , and the other part was planted in flowerpots to measure leaf water potential after drought stress;

(2) The measuring method of field sowing and oil content is:

Choose sandy loam soil, apply enough base fertilizer, and apply 100 catties of compound fertilizer with 15% of N, P, and K content per acre, sow seeds in early May, with a sowing depth of 3-4cm, cover with mulch to keep moisture, increase temperature, and prevent weeds. Pay attention during growth. Water in time when it is dry, drain water when it is flooded, and harvest the pods and dry them when they are ripe.

(3) The method of planting in flowerpot and measuring water potential is:

①Preparation of sowing substrate: The nutrient bowl substrate is selected from sandy loam and vermiculite in a ratio of 4:1, and an appropriate amount of compound fertilizer (N, P, K=1:1:1) is added according to the fertility of the soil and stirred evenly;

②Sowing: choose a flowerpot with an inner diameter of 25cm, and the substrate loaded is equal to the upper edge of the flowerpot. The seeds are sown in the nutrition bowl in early May, and the sowing depth is 3-4cm; water 600mL;

③Management

The above-mentioned nutritional bowl is placed in a natural environment, and watered according to the moisture content, 600mL each time;

④Treatment of drought stress

Carry out drought stress treatment after sowing 5 weeks, namely do not water when this watering according to moisture condition, and drought stress treatment is 2 days;

⑤ Take the discs of peanut leaves after drought stress, and measure the water potential of the leaves.

Determination of water potential in peanut leaves:

Preparation of sucrose solutions with different water potentials

According to the sucrose concentration corresponding to the known water potential of the sucrose solution, sucrose solutions of different mass concentrations were prepared (Table 1).

Table 1 Water potential corresponding to different quality sucrose solutions

Sucrose (g/100mL) 13.34 15.91 18.44 20.99 Water potential (MPa) -1.0 -1.2 -1.4 -1.6

Preparation of standard curves of refractive index and water potential of sucrose solutions with different water potentials

Measure the refractive index of each water potential sucrose solution on an Abbe refractometer (Table 2), make a standard curve between the refractive index and water potential, the standard curve formula: y=ax+b, y: water potential, x: refractive index, a and b is an automatically generated constant, which is related to temperature, etc. In this experiment, the room temperature is about 25°C. According to the refractive index and water potential, the standard curve is calculated as: y=-70.367x+93.962, correlation coefficient R ² =0.9992;

Table 2 Determination of the refractive index of sucrose solutions of various water potentials on Abbe refractometer

Water potential (MPa) -1.0 -1.2 -1.4 -1.6 refractive index 1.3496 1.3526 1.3552 1.3582

The standard curves of the refractive index and water potential of the sucrose solutions with different water potentials are shown in FIG. 1 .

Take 10 peanut leaf discs after drought stress and immerse them in PCR reaction tubes containing 1 mL of sucrose solution with different water potentials of -1.0, -1.2, -1.4, and -1.6MPa respectively; put 10 circles in each tube. slice, cover, shake and let stand for 20 minutes.

The sucrose solution of each soaked leaf was taken separately, and the refractive index was measured on an Abbe refractometer. Compare the measured refractive index of the sucrose solution of each soaked leaf with the refractive index of the original sucrose solution. If the measured refractive index of the sucrose solution of a certain soaked leaf is the same as that of the original sucrose solution, the measured value is the variety (strain). ) of the refractive index;

If the refractive index of the original sucrose solution is different from that of the original sucrose solution, the measured average of the refractive indices of two adjacent water potentials, one larger than that of the original sucrose solution and the other smaller than that of the original sucrose solution, is the refractive index of the variety (strain). coefficient.

Through the above method, the oil content and refractive index of each strain of peanut material were measured as shown in Table 3. According to the standard curve y=70.367x+93.962, the refractive index was substituted into the formula, and the water potential was shown in Table 3:

Table 3 Oil content, refractive index and water potential of peanut materials of each strain

The relationship formula y=ax+b was constructed with the measured water potential of peanut leaves after drought stress as the x-axis and the oil content of peanut kernels as the y-axis. The results are shown in Figures 2 to 4:

The standard curve of oil content and water potential of Huayu 20 is: y=25.264x+90.262, and the correlation coefficient is 0.882, which is extremely significant. The standard curve of oil content and water potential of Huayu 22 is: y=14.561x+73.347, and the correlation coefficient is 0.884, which is extremely significant. The standard curve of oil content and water potential of Luhua No. 11 is: y=22.888x+84.167, and the correlation coefficient is 0.888, which is extremely significant.

Example 2

Screening of high-oil materials in Luhua 11 mutants:

The Luhua No. 11 mutant T77 (mutant obtained from ⁶⁰ Co-irradiated progeny) was selected as the test material.

Select mature and plump seeds of T77 to plant in flowerpots. The specific steps are:

①Preparation of sowing substrate: The nutrient bowl substrate is selected from sandy loam and vermiculite in a ratio of 4:1, and an appropriate amount of compound fertilizer (N, P, K=1:1:1) is added according to the fertility of the soil and stirred evenly;

②Sowing: choose a flowerpot with an inner diameter of 25cm, and the substrate loaded is equal to the upper edge of the flowerpot. The seeds are sown in the nutrition bowl in early May, and the sowing depth is 3-4cm; water 600mL;

③Management

The above-mentioned nutritional bowl is placed in a natural environment, and watered according to the moisture content, 600mL each time;

④Treatment of drought stress

Carry out drought stress treatment after sowing 5 weeks, namely do not water when this watering according to moisture condition, and drought stress treatment is 2 days;

⑤ Take the discs of peanut leaves after drought stress, and use the water potential measurement method in Example 1 to measure the water potential of the leaves.

It was determined that the water potential of T77 peanut leaves after drought stress treatment was between -1.2 and -1.4MPa, and the refractive index measured in the sucrose solution with adjacent water potentials of -1.2MPa and -1.4MPa were 1.3527 and 1.3531, respectively. The average value of the refractive index of the two is 1.3529. Substitute this refractive index value into the standard curve of the refractive index and water potential of sucrose solutions with different water potentials: y=-70.367x+93.962, the calculated water potential is -1.2375143.

Substitute the water potential value -1.2305143 of the above-mentioned T77 peanut leaves into the standard curve y=22.888x+84.167 of the peanut water potential-oil content of Luhua No. 11, and the oil content of peanuts is 55.84%. The Oilseeds and Products Quality Supervision, Inspection and Testing Center of the Ministry of Agriculture, the oil content measured by the residual method is 55.78%, and the oil content of the mutant T77 is greater than 55%, which is a high-oil peanut material; in addition, the values detected by the two methods are not much different. The method for rapidly screening high-oil materials in peanut mutants of the present invention has high accuracy and reliable results, can greatly save manpower, material and financial resources, and overcome the problem of long time for measuring oil content in conventional methods.

Example 3

Screening of high-oil material in Huayu 20 mutant:

Huayu 20 mutant 13-5 (mutant obtained from fast neutron-irradiated progeny) was selected as the test material.

Select 13-5 mature and plump seeds to plant in flowerpots. The specific steps are:

①Preparation of sowing substrate: The nutrient bowl substrate is selected from sandy loam and vermiculite in a ratio of 4:1, and an appropriate amount of compound fertilizer (N, P, K=1:1:1) is added according to the fertility of the soil and stirred evenly;

②Sowing: choose a flowerpot with an inner diameter of 25cm, and the substrate loaded is equal to the upper edge of the flowerpot. The seeds are sown in the nutrition bowl in early May, and the sowing depth is 3-4cm; water 600mL;

③Management

The above-mentioned nutritional bowl is placed in a natural environment, and watered according to the moisture content, 600mL each time;

④Treatment of drought stress

Carry out drought stress treatment after sowing 5 weeks, namely do not water when this watering according to moisture condition, and drought stress treatment is 2 days;

⑤ Take the discs of peanut leaves after drought stress, and use the water potential measurement method in Example 1 to measure the water potential of the leaves.

It was determined that the refractive index of 13-5 peanut leaves in -1.2MPa sucrose solution after drought stress treatment was 1.3526, which was the same as that of the original sucrose solution. Therefore, the refractive index of 13-5 was 1.3526. Substitute this refractive index value into the standard curve of the refractive index and water potential of sucrose solutions with different water potentials: y=-70.367x+93.962, the calculated water potential is -1.2164042MPa.

Substitute the water potential value -1.2164042MPa of the above-mentioned 13-5 peanut leaves into the standard curve y=25.26x+90.262 of peanut water potential-oil content of Huayu 20 line, and the oil content of peanut is 59.54%, while the peanut after harvest is planted in the field. The seed kernel was commissioned by the Oilseeds and Products Quality Supervision, Inspection and Testing Center of the Ministry of Agriculture, and the oil content tested by the residual method was 59.38%. It can be seen that the oil content of mutant 13-5 is greater than 55%, which is a high-oil peanut material; in addition, the two The values detected by these methods are not much different. The method of the present invention for rapidly screening high-oil materials in peanut mutants has high accuracy and reliable results, can greatly save manpower, material resources and financial resources, and overcome the problem of long time for measuring oil content in conventional methods.

The above are only preferred embodiments of the present invention, and are not intended to limit the present invention in other forms. Any person skilled in the art may use the technical content disclosed above to make changes or modifications to equivalent changes. Example. However, any simple modifications, equivalent changes and modifications made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solutions of the present invention still belong to the protection scope of the technical solutions of the present invention.

Claims (8)

1. a method for quick screening of high oil content peanut varieties, is characterized in that: the peanut to be screened is sown, carry out drought stress treatment after 5 weeks of sowing, measure the water potential of the peanut leaf after the drought stress, and the water potential value is imported into the constructed In the relationship model between peanut oil content and water potential, the peanut oil content is obtained; if the oil content is more than 55%, it is a high-oil peanut material, and if the oil content is less than 55%, it is discarded; The method for establishing the relationship model between the peanut oil content and water potential is: Multiple peanut materials of the same line were selected, and each peanut material was divided into two parts, one part was planted in the field to determine the oil content of the kernel after mature harvest, and the other part was planted in flower pots to determine the leaves after drought stress. water potential; The relationship formula y=ax+b was constructed with the measured water potential of peanut leaves after drought stress as the x-axis and the oil content of peanut kernels as the y-axis. 2. the method for fast screening high oil content peanut varieties according to claim 1, is characterized in that: the assay method of described field sowing and oil content is: Choose sandy loam soil, apply enough base fertilizer, and apply 100 catties of compound fertilizer with 15% of N, P, and K content per acre, sow seeds in early May, with a sowing depth of 3-4cm, cover with mulch to keep moisture, increase temperature, and prevent weeds. Pay attention during growth. Water in time in drought, drain in time in waterlogging, harvest pods and sun-dry after maturity, and measure the oil content of kernels. 3. the method for fast screening high oil content peanut varieties according to claim 1, is characterized in that: flowerpot sowing and water potential determination method are: ①Preparation of sowing substrate: The nutrient bowl substrate is selected from sandy loam and vermiculite in a ratio of 4:1, and an appropriate amount of compound fertilizer is added according to the fertility of the soil to stir evenly. In the compound fertilizer, N, P, K=1:1:1 ; ②Sowing: choose a flowerpot with an inner diameter of 25cm, and the substrate loaded is equal to the upper edge of the flowerpot. The seeds are sown in the nutrition bowl in early May, and the sowing depth is 3-4cm; water 600mL; ③Management The above-mentioned nutritional bowl is placed in a natural environment, and watered according to the moisture content, 600mL each time; ④Treatment of drought stress Drought stress treatment was carried out 5 weeks after sowing; ⑤ Take the discs of peanut leaves after drought stress, and measure the water potential of the leaves. 4. according to the method for quick screening high oil content peanut varieties described in any one of claim 1～3, it is characterized in that: step is as follows: ①Preparation of sowing substrate: The nutrient bowl substrate is selected from sandy loam and vermiculite in a ratio of 4:1, and an appropriate amount of compound fertilizer is added according to the fertility of the soil to stir evenly. In the compound fertilizer, N, P, K=1:1:1 ; ②Sowing: Choose a flowerpot with an inner diameter of 25cm, and the substrate to be loaded is the same as the upper edge of the flowerpot. In early May, the seeds of a certain peanut variety obtained by mutagenesis to obtain multiple mutants were sown in the nutrient pot, and the sowing depth was 3 -4cm; water 600mL; ③Management The above-mentioned nutritional bowl is placed in a natural environment, and watered according to the moisture content, 600mL each time; ④Treatment of drought stress Drought stress treatment was carried out 5 weeks after sowing; ⑤ Take the discs of peanut leaves after drought stress, and measure the water potential of the leaves; ⑥ Import the measured water potential value of peanut leaves into the constructed relationship model of peanut oil content and water potential to obtain the peanut oil content; ⑦Select mutants with oil content above 55% as high-oil materials, and discard those with oil content less than 55%. 5. the method for fast screening high oil content peanut varieties according to claim 4, is characterized in that: the drought stress of described peanut is: after peanut sowing 5 weeks, do not water when this watering according to soil moisture, drought stress Process 2 days. 6. The method for rapidly screening high oil content peanut varieties according to any one of claims 1 to 3, characterized in that: The method for measuring the water potential of peanut leaves after the drought stress is: a. Take the discs of peanut leaves after drought stress, take 10 discs of peanut leaves and place them in a 1mL gradient water potential sucrose solution, shake well and let stand for 20min; b. Determine the refractive index of the sucrose solution after soaking the leaves, and compare it with the refractive index of the original sucrose solution: If the refractive index of a certain sucrose solution is the same as that of the original sucrose solution, the measured value is the refractive index of the peanut material; If the refractive index of a sucrose solution is different from that of the original sucrose solution, the measured refractive index of the two adjacent sucrose solutions after soaking the leaves is one larger than that of the original sucrose solution and one smaller than that of the original sucrose solution. The average of the coefficients is the refractive index of the peanut material; c. Bring the measured refractive index of a certain peanut material after drought stress treatment into the relationship between the refractive index of sucrose solution and the water potential, and the obtained water potential is the water potential of the peanut material leaves. 7. The method for rapidly screening peanut varieties with high oil content according to claim 6, wherein the water potentials of the sucrose solution of the gradient water potential are respectively: -1.0MPa, -1.2MPa, -1.4MPa, -1.6MPa. 8. the method for fast screening high oil content peanut varieties according to claim 6, is characterized in that: the relational formula of sucrose solution refractive index and water potential is to measure the refractive index of the sucrose solution of different water potentials, and be x-axis with refractive index, Construct the relationship y=ax+b with the water potential as the y-axis.

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