Method for screening watermelon low-temperature-resistant breeding material
By cultivating melon seedlings under different temperature and time conditions and using a variety of data analysis methods, a screening system for watermelon low-temperature breeding materials was established, which solved the problem of lack of unified methods and standards in the screening process in the existing technology, and achieved efficient and accurate screening of low-temperature resistant melon seedlings.
Patent Information
- Application Number
- CN202411987296.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-31
- Publication Date
- 2025-04-25
AI Technical Summary
In the screening process of watermelon low-temperature breeding materials in the prior art, there is a lack of unified methods and standards, which leads to the inability to effectively compare and integrate the research materials, which seriously affects the screening process and results.
A process-complete, unique insights, cost-effective and efficient watermelon low-temperature-resistant breeding material screening method is adopted. By cultivating melon seedlings under different temperature and time conditions, measuring multiple physiological and biochemical indicators, and using technical means such as Scale's mathematical method, PCA principal component analysis and OPLS-DA classification analysis, the first breeding system is established to screen low-temperature-resistant melon seedlings.
The efficient screening of watermelon low-temperature breeding materials is achieved, providing a solid foundation, which can effectively distinguish between cold-resistant groups and non-cold-resistant groups, find out key characteristics related to cold-resistant properties, and improve the screening accuracy and efficiency of breeding materials.
Smart Images
Figure CN119866927A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of plant cultivation and breeding, and particularly relates to a method for screening low-temperature-resistant breeding materials for watermelon. Background Art
[0002] Watermelon (scientific name: Citrullus lanatus (Thunb) Matsum.et Nakai) is an annual plant of the genus Citrullus in the Cucurbitaceae family, and its fruit is also a favorite fruit of people. However, watermelon production has a strong seasonality, and the listing of watermelons leads to low prices when watermelons are listed in large quantities, which seriously affects the income of melon farmers and affects their enthusiasm. If the watermelon is listed in advance, not only can people taste delicious watermelon fruits, but also the peak listing can increase the income of melon farmers, which is conducive to coordinating the supply of watermelons. However, if watermelons are to be grown outdoors in advance, the problem of watermelon resistance to low temperatures must be solved. Therefore, watermelon low temperature resistant breeding materials are the key to cultivating low temperature resistant watermelons, and are also the core content of solving the above problems. However, there is no unified production method for the selection of low temperature resistant watermelon breeding materials at present, and the setting of temperature and low temperature continuous market during the screening process is relatively chaotic, resulting in the inability to compare, summarize and integrate the research materials, which seriously affects the process and results of watermelon low temperature resistant breeding material screening. Therefore, the present invention proposes a set of complete process, unique insights, economical and efficient watermelon low temperature resistant breeding material screening system and method to solve the above problems, laying a solid foundation for watermelon low temperature resistant screening. Summary of the invention
[0003] The technical problem to be solved by the present invention is to propose a method for screening watermelon low-temperature-resistant breeding materials, which has a complete process, unique insights, and is economical and efficient.
[0004] The technical solution adopted by the present invention to solve the above technical problems is: a method for screening watermelon low temperature resistant breeding materials, the steps are as follows: S01W breeding materials, select M melon seedlings with good growth from each breeding material, and divide the same breeding material into 16 cultivation groups, where M is a multiple of 16; S02: Each group was cultivated in a complete combination of environments with temperatures of 4°C, 6°C, 8°C and 10°C and days of 1d, 4d, 7d and 10d; After S03, the melon seedlings in each culture group were measured, and the measured data included: plant height, base stem diameter, aboveground fresh weight, underground fresh weight, aboveground fresh weight / underground fresh weight, aboveground dry weight, underground dry weight, aboveground dry weight / underground dry weight, root length, root surface area, and root Na concentration; S04 presents the above data in a multi-dimensional coordinate system; S05 used the Scale mathematical method to UV standardize the data so that all variables had the same dimension; used the PCA principal component analysis method to process the data of S04, reduced the dimension of these high-dimensional data, and found the most variable principal components; used OPLS-DA to classify different cultivation groups, distinguish between cold-resistant groups and non-cold-resistant groups, and find out the key characteristics related to cold resistance; S06 established the first breeding system based on the key characteristics related to cold resistance found in S05, that is, formulated reference indicators for screening low-temperature resistant melon seedlings.
[0005] In a preferred embodiment, M is 96.
[0006] In a preferred embodiment, the data measured in step S03 also include physiological and biochemical indicators such as plant height, stem diameter, leaf area, leaf exudate conductivity, SPAD, SOD, POD, CAT, MDA and proline pro.
[0007] In the preferred embodiment, in step S05, since the units of the indicators such as plant height, stem diameter, leaf area, leaf exudate conductivity, SPAD, SOD, etc. measured in step S03 are cm, mm, cm 2 , %, gm / g, etc., their dimensions and orders of magnitude are very different. Therefore, the present invention adopts a special data processing method, that is, the data of each group with different dimensions and orders of magnitude are converted with 0 as the average value and 1 as the standard deviation, so that the data of each group with different dimensions and orders of magnitude are normalized into a uniformly processable data set; secondly, PCA is used to obtain the first n principal components that can explain more than 75% of the information of the 10 measured indicators, and the load sizes of the 10 measured indicators are obtained from the load matrix under each principal component. The load under each principal component (S i , i=1, 2, n) as the candidate index; at the same time, the first n principal components are analyzed by regression analysis and saved as variables X i,u (i=1,2...n;u=1,2,3...w, where w is the number of parameter breeding materials), enter the next step of screening. Again, use the saved variable X i,U , using [X i,u -Min(X i,u )] / [Max(X i,u )-Min(X i,u )] to get the D value, and take the D value as the independent variable, S i As the dependent variable, construct Y=a1+a2*S1+a3S2...+ai*S i, if 0<Y≤0.2, it is super-sensitive to low temperature; 0.2<Y≤0.4, it is sensitive to low temperature; 0.4<Y≤0.6, it is generally resistant to low temperature; 0.6<Y≤0.8, it is resistant to low temperature; 0.8<Y≤1.0, it is extremely resistant to low temperature, which is the first breeding system for low temperature resistant breeding materials.
[0008] In a preferred embodiment, step S07 is added after step S06, including using the first breeding system to select 32 melon seedlings, dividing them into 4 culture groups on average, and placing them in culture rooms with ambient temperatures of 4°C, 6°C, 8°C and 10°C, respectively, and maintaining the humidity at 40%RH. The temperature and humidity remain constant within 7 days; the humidity is set to 40%RH, and a dry environment is maintained to enhance the stress intensity, so that the melon seedlings face greater transpiration pressure under low temperature conditions, thereby selecting plants with stronger initial cold resistance.
[0009] S08 repeats steps S03 to S05 to obtain a second breeding system.
[0010] In the preferred scheme, S09 uses the second breeding system to screen out 32 melon seedlings, divides them into 4 culture groups on average, and places them in culture rooms with ambient temperatures of 4°C, 6°C, 8°C and 10°C respectively. The temperature is periodically raised and lowered by 2°C every 12 hours, and the humidity is periodically cycled from 40%RH-60%RH-80%RH, and the time when the humidity is 40%RH is greater than the sum of the time when the humidity is 60%RH and 80%RH. The cultivation time lasts for 7 days; the humidity is increased to 60%-80%RH, and humidity targets for specific treatment groups can also be set to reduce the pressure of water transpiration on the plants in the low temperature environment, so that the melon seedlings can gradually adapt to a more humid environment.
[0011] S10 repeats steps S03 to S05 to obtain a third breeding system.
[0012] In a preferred embodiment, before obtaining the third breeding system, S11 randomly selects 6-10 melon seedlings from the cold-resistant group and the non-cold-resistant group materials in step S05 as samples for gene transcriptomics analysis, and analyzes the differentially expressed genes between the cold-resistant group and the non-cold-resistant group.
[0013] In summary, the first breeding system is suitable for the screening of most low-temperature-resistant watermelon seedling breeding materials, the second breeding system can screen out watermelon seedling breeding materials under low-temperature and dry environments, and the third breeding system can screen out low-temperature-resistant watermelon seedling breeding materials under low-temperature and fluctuating temperature and humidity. BRIEF DESCRIPTION OF THE DRAWINGS
[0014] Figure 1 is a flow chart of establishing a first breeding system of the present invention; Figure 2 is a flow chart of establishing the second breeding system of the present invention; Figure 3 It is a flow chart of establishing the third breeding system of the present invention. DETAILED DESCRIPTION
[0015] The embodiments of the present invention are further described in detail below in conjunction with the accompanying drawings. Embodiment 1
[0016] A method for screening low-temperature-resistant watermelon breeding materials, comprising the following steps: S01W breeding materials, select M melon seedlings with good growth from each breeding material, and divide the same breeding material into 16 cultivation groups, where M is a multiple of 16; S02: Each group was cultivated in a complete combination of environments with temperatures of 4°C, 6°C, 8°C and 10°C and days of 1d, 4d, 7d and 10d; After S03, the melon seedlings in each culture group were measured, and the measured data included: plant height, base stem diameter, aboveground fresh weight, underground fresh weight, aboveground fresh weight / underground fresh weight, aboveground dry weight, underground dry weight, aboveground dry weight / underground dry weight, root length, root surface area, and root Na concentration; S04 presents the above data in a multi-dimensional coordinate system; S05 used the Scale mathematical method to UV standardize the data so that all variables had the same dimension; used the PCA principal component analysis method to process the data of S04, reduced the dimension of these high-dimensional data, and found the most variable principal components; used OPLS-DA to classify different cultivation groups, distinguish between cold-resistant groups and non-cold-resistant groups, and find out the key characteristics related to cold resistance; S06 established the first breeding system based on the key characteristics related to cold tolerance found in S05.
[0017] In the embodiment, M is 96.
[0018] In the embodiment, the data measured in step S03 also include physiological and biochemical indicators such as plant height, stem diameter, leaf area, leaf exudate conductivity, SPAD, SOD, POD, CAT, MDA and proline pro.
[0019] In the embodiment, in step S05, since the units of the indicators such as plant height, stem diameter, leaf area, leaf exudate conductivity, SPAD, SOD, etc. measured in step S03 are cm, mm, cm 2, %, gm / g, etc., their dimensions and orders of magnitude are very different. Therefore, the present invention adopts a special data processing method, that is, the data of each group with different dimensions and orders of magnitude are converted with 0 as the average value and 1 as the standard deviation, so that the data of each group with different dimensions and orders of magnitude are normalized into a uniformly processable data set; secondly, PCA is used to obtain the first n principal components that can explain more than 75% of the information of the 10 measured indicators, and the load sizes of the 10 measured indicators are obtained from the load matrix under each principal component. The load under each principal component (S i , i=1, 2, n) as the candidate index; at the same time, the first n principal components are analyzed by regression analysis and saved as variables X i,u (i=1,2...n;u=1,2,3...w, where w is the number of parameter breeding materials), enter the next step of screening. Again, use the saved variable X i,U , using [X i,u -Min(X i,u )] / [Max(X i,u )-Min(X i,u )] to get the D value, and take the D value as the independent variable, S i As the dependent variable, construct Y=a1+a2*S1+a3S2...+ai*S i , if 0<Y≤0.2, it is super-sensitive to low temperature; 0.2<Y≤0.4, it is sensitive to low temperature; 0.4<Y≤0.6, it is generally resistant to low temperature; 0.6<Y≤0.8, it is resistant to low temperature; 0.8<Y≤1.0, it is extremely resistant to low temperature, which is the first breeding system for low temperature resistant breeding materials. Embodiment 2
[0020] Adding step S07 after step S06 comprises using the first breeding system to select 32 melon seedlings, dividing them into 4 culture groups on average, and placing them in culture rooms with ambient temperatures of 4°C, 6°C, 8°C and 10°C, respectively, maintaining the humidity at 40%RH, and keeping the temperature and humidity constant within 7 days; setting the humidity to 40%RH, maintaining a dry environment to enhance the stress intensity, so that the melon seedlings face greater transpiration pressure under low temperature conditions, thereby selecting plants with stronger initial cold resistance.
[0021] S08 repeats steps S03 to S05 to obtain a second breeding system. Embodiment 3
[0022] S09 used the second breeding system to select 32 melon seedlings, divided them into 4 cultivation groups, and placed them in cultivation rooms with ambient temperatures of 4℃, 6℃, 8℃ and 10℃ respectively. The temperature was periodically raised and lowered by 2℃ every 12 hours, and the humidity was periodically cycled from 40%RH-60%RH-80%RH. The time when the humidity was 40%RH was greater than the sum of the time when the humidity was 60%RH and 80%RH. The cultivation time lasted for 7 days. The humidity was increased to 60%-80%RH to reduce the pressure of water transpiration on the plants in the low temperature environment, so that the melon seedlings could gradually adapt to a more humid environment.
[0023] S10 repeats steps S03 to S05 to obtain a third breeding system. Embodiment 4
[0024] S11 Before obtaining the third breeding system, in step S05, 6-10 melon seedlings are randomly selected from the cold-resistant group and the non-cold-resistant group materials as samples for gene transcriptomics analysis, and the differentially expressed genes between the cold-resistant group and the non-cold-resistant group are analyzed.
[0025] The best embodiment of the present invention has been described, and various changes or modifications made by those skilled in the art will not depart from the scope of the present invention.
Claims
1. A method for screening low temperature resistant watermelon breeding materials, characterized in that: S01W breeding materials, select M melon seedlings with good growth from each breeding material, and divide the same breeding material into 16 cultivation groups, where M is a multiple of 16; S02: Each group was cultivated in a complete combination of environments with temperatures of 4°C, 6°C, 8°C and 10°C and days of 1d, 4d, 7d and 10d; After S03, the melon seedlings in each culture group were measured, and the measured data included: plant height, base stem diameter, aboveground fresh weight, underground fresh weight, aboveground fresh weight / underground fresh weight, aboveground dry weight, underground dry weight, aboveground dry weight / underground dry weight, root length, root surface area, and root Na concentration; S04 presents the above data in a multi-dimensional coordinate system; S05 used the Scale mathematical method to UV standardize the data so that all variables had the same dimension; used the PCA principal component analysis method to process the data of S04, reduced the dimension of these high-dimensional data, and found the most variable principal components; used OPLS-DA to classify different cultivation groups, distinguish between cold-resistant groups and non-cold-resistant groups, and find out the key characteristics related to cold resistance; S06 established the first breeding system based on the key characteristics related to cold tolerance found in S05.
2. The method for screening low temperature resistant watermelon breeding materials according to claim 1, characterized in that: M is 96.
3. A method for screening low temperature resistant watermelon breeding materials according to claim 2, characterized in that: The data measured in step S03 also include physiological and biochemical indicators such as plant height, stem diameter, leaf area, leaf exudate conductivity, SPAD, SOD, POD, CAT, MDA and proline pro.
4. The method for screening low temperature resistant watermelon breeding materials according to claim 3, characterized in that: In step S05, since the units of the indicators such as plant height, stem diameter, leaf area, leaf exudate conductivity, SPAD, and SOD measured in step S03 are cm, mm, and cm respectively, 2 , %, gm / g, etc., their dimensions and orders of magnitude are very different. Therefore, the present invention adopts a special data processing method, that is, the data of each group with different dimensions and orders of magnitude are converted with 0 as the average value and 1 as the standard deviation, so that the data of each group with different dimensions and orders of magnitude are normalized into a uniformly processable data set; secondly, PCA is used to obtain the first n principal components that can explain more than 75% of the information of the 10 measured indicators, and the load sizes of the 10 measured indicators are obtained from the load matrix under each principal component. The load under each principal component (S i , i=1, 2, n) as the candidate index; at the same time, the first n principal components are analyzed by regression analysis and saved as variables X i,u (i=1,2...n;u=1,2,3...w, where w is the number of parameter breeding materials), enter the next step of screening. Again, use the saved variable X i,U , using [X i,u -Min(X i,u )] / [Max(X i,u )-Min(X i,u )] to get the D value, and take the D value as the independent variable, S i As the dependent variable, construct Y=a1+a2*S1+a3S2...+ai*S i , if 0<Y≤0.2 low temperature super sensitive type; 0.2<Y≤0.4 low temperature sensitive type; 0.4<Y≤0.6General low temperature resistant type; 0.6<Y≤0.8 low temperature resistant type; 0.8<Y≤1.0 Extremely low temperature resistant type, used as the first breeding system for low temperature resistant breeding materials.
5. The method for screening low temperature resistant watermelon breeding materials according to claim 3, characterized in that: Step S07 is added after step S06, comprising selecting 32 melon seedlings using the first breeding system, dividing them into 4 culture groups on average, and placing them in culture rooms with ambient temperatures of 4°C, 6°C, 8°C and 10°C, respectively, maintaining the humidity at 40%RH, and keeping the temperature and humidity constant within 7 days.
6. S08 Repeat steps S03 to S05 to obtain a second breeding system.
7. The method for screening low temperature resistant watermelon breeding materials according to claim 5, characterized in that: S09 used the second breeding system to select 32 melon seedlings, divided them into 4 culture groups, and placed them in culture rooms with ambient temperatures of 4°C, 6°C, 8°C and 10°C respectively. The temperature was periodically raised and lowered by 2°C every 12 hours, and the humidity was periodically cycled from 40%RH-60%RH-80%RH. The time when the humidity was 40%RH was greater than the sum of the time when the humidity was 60%RH and 80%RH. The cultivation time lasted for 7 days.
8. S10 Repeat steps S03 to S05 to obtain a third breeding system.
9. The method for screening low temperature resistant watermelon breeding materials according to claim 6, characterized in that: S11 Before obtaining the third breeding system, in step S05, 6-10 melon seedlings are randomly selected from the cold-resistant group and the non-cold-resistant group materials as samples for gene transcriptomics analysis, and the differentially expressed genes between the cold-resistant group and the non-cold-resistant group are analyzed.