Sugarcane hybrid combination selection method
The sugarcane hybrid combination selection method, which combines multi-trait evaluation and intelligent prediction models, solves the problem of blindness in the breeding process, realizes an efficient and precise breeding process, and improves the accuracy of sugarcane hybrid combinations and the genetic stability of offspring.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
- Filing Date
- 2026-02-24
- Publication Date
- 2026-04-24
AI Technical Summary
In sugarcane hybrid breeding, the selection of parental combinations based on the breeder's experience leads to significant blind spots in the screening of hybrid combinations, resulting in actual performance not meeting expectations, prolonging the breeding cycle and increasing the human and resource costs of field screening.
A method combining multi-trait evaluation and intelligent prediction was adopted. By collecting agronomic traits and genotype data of sugarcane male and female parents, a phenotypic prediction model trained by neural network was used to predict the potential phenotypic performance of hybrid combinations. Combined with value assessment and photothermal induction technology, precise hybrid combination selection was carried out, and a double control method was used to screen offspring.
It improves the accuracy of hybrid combination selection and breeding efficiency, shortens the breeding cycle, increases the yield of superior offspring, and ensures genetic stability and hybridization success rate.
Smart Images
Figure CN121909907A_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of sugarcane hybrid breeding technology, and more specifically, it relates to a method for selecting and matching sugarcane hybrid combinations. Background Technology
[0002] Sugarcane hybrid breeding refers to a crop breeding technology that uses sugarcane and its closely related species as breeding materials. It involves sexual hybridization of parents with superior traits through artificial selection, followed by segregation and screening of offspring, trait identification, and targeted breeding to cultivate high-quality new sugarcane varieties suitable for the sugar industry and bioenergy feedstock production. It is a core method for variety renewal in the sugarcane industry and is widely used in major sugarcane producing areas worldwide. Its core processes include parent selection, hybridization pollination, seedling cultivation, multi-generation selection and identification, and variety approval and promotion. Modern sugarcane hybrid breeding also integrates technologies such as molecular marker-assisted breeding, gene editing, and disease resistance identification to shorten the breeding cycle, improve selection accuracy, and focus on cultivating varieties adapted to different ecological regions and planting patterns.
[0003] Related sugarcane hybrid breeding relies on breeders' experience to select parental combinations, resulting in a certain accumulation of technology. However, this experience-driven method makes it difficult to systematically quantify the genetic effects and trait transmission patterns between parents, and it is easily affected by subjective judgment biases. This leads to a large degree of blindness in the screening of hybrid combinations, and the actual performance of a large number of hybrid offspring does not match expectations. This not only prolongs the breeding cycle but also increases the human and resource costs of field screening. Summary of the Invention
[0004] To address the problem that sugarcane hybrid breeding relies heavily on breeders' experience in selecting parental combinations, leading to significant blind spots in the selection of hybrid combinations and discrepancies between the actual performance of many hybrid offspring and expectations, which not only prolongs the breeding cycle but also increases the manpower and resource costs of field screening, this application provides a method for selecting sugarcane hybrid combinations.
[0005] This application provides a method for selecting and matching sugarcane hybrid combinations, employing the following technical solution:
[0006] A method for selecting and matching sugarcane hybrid combinations includes the following steps:
[0007] S1. Evaluation of parental traits in multiple dimensions: Genotypic data of candidate sugarcane male and female parents were analyzed based on their agronomic traits, quality traits and stress resistance traits, respectively.
[0008] S2. Intelligent selection of hybrid combinations: Based on the genotype data obtained in step S1, a phenotypic prediction model is constructed to predict the potential phenotypic performance of hybrid combinations produced by different parent pairings; the phenotypic prediction model is trained using the SNP gene data, transcriptome data, metabolome data of known parents and the phenotypic data of the combined offspring from historical breeding data as the training set, and is obtained through a neural network that includes feature enhancement and wavelet transform.
[0009] S3. Combination Value Assessment and Determination: Based on the phenotypic performance predicted in step S2, and combined with the preset breeding objectives, a value assessment is conducted, and combinations with high value scores are selected as target hybridization combinations. The value assessment prioritizes combinations whose representative phenotypes are stably inherited after combination, and focuses on assessing the predicted values of sucrose content, disease resistance, stress resistance, and yield.
[0010] S4. Pre-hybridization parent treatment and flowering period regulation: For the parent plants in the determined target hybridization combination, in the field or barrel cultivation, photoperiodic flowering induction was carried out from mid-June to early July.
[0011] S5. Artificial hybridization and pollination: In the early flowering stage of the male / female parent, place the configured male / female parent in the same hybridization cage, with the male parent's flower spikes placed above the female parent. Shake the male parent 2-3 times every morning from 8:30 to 11:30. Control the temperature of the hybridization room to be 25℃-28℃ and the humidity to be 75%-85%.
[0012] S6. Screening of hybrid offspring: Harvest hybrid seeds and cultivate seedlings. Use a double control method for screening: use the average value of all family traits as control 1, which is denoted as CK1, and use the trait value of the local main cultivated variety as control 2, which is denoted as CK2. First, eliminate families whose average trait value is lower than CK1. Then, among the selected families, select individuals with traits superior to CK2 as superior offspring.
[0013] By adopting the above technical solutions, the multi-dimensional trait evaluation steps of parents enable the systematic collection of agronomic traits and genotype data of parents, providing comprehensive input for subsequent intelligent selection and breeding. Combined with a phenotypic prediction model, which utilizes multi-omics information from historical breeding data and enhances feature extraction capabilities through neural network training, the model accurately predicts the phenotypic performance of hybrid combinations. The value assessment step focuses on key traits such as sucrose content and disease resistance, ensuring that the selected combinations meet breeding objectives. Parental treatment and flowering time regulation precisely control flowering time through photoperiod induction, creating conditions for artificial pollination. Artificial hybridization pollination employs specific time points and treatment solutions for spraying, improving pollination efficiency and success rate. Progeny selection uses a double-control method, progressively eliminating low-value families and individuals, thus improving breeding efficiency. Therefore, a highly efficient, precise, and reproducible sugarcane hybridization breeding method is obtained, shortening the breeding cycle and increasing the yield of superior progeny.
[0014] Preferably, the training method for the phenotypic prediction model in step S2 includes: numerically mapping the SNP gene data, performing discrete Fourier transform with a fixed window to identify protein coding regions and enhance features, denoising the high-frequency features obtained by wavelet transform, performing inverse wavelet transform on the low-frequency features, and constructing a graph structure by combining transcriptome and metabolome data.
[0015] By adopting the above technical solutions, the use of discrete Fourier transform to process SNP gene data can effectively identify protein-coding regions and enhance sequence features, thereby improving the model's ability to interpret genetic information. Wavelet transform reduces noise in high-frequency features to decrease data interference, while inverse transform of low-frequency features preserves the main signals, ensuring feature stability. By combining transcriptomic and metabolomic data to construct a graph structure and integrating multi-omics association information, the model can capture the complex network relationship between gene expression and phenotype. Therefore, a robust and highly accurate phenotype prediction model is obtained, providing a reliable basis for hybrid combination selection.
[0016] Preferably, when evaluating the combined value in step S3, the principle of equal hybridization is also applied: parents with similar proportions of wild species bloodlines are selected for pairing, and the total proportion of wild species bloodlines used to breed offspring is controlled between 12.5% and 25%.
[0017] By adopting the above technical solution, the principle of equal hybridization is used to prioritize the selection of parents with similar bloodlines in wild species for pairing, which can maintain the balance of the genetic background and avoid the disorder of phenotypic segregation in offspring due to excessive bloodline differences. The total bloodline ratio of wild species is controlled between 12.5% and 25%, which is based on the trade-off between the stress resistance of wild species and the high yield of cultivated species, ensuring that offspring retain the excellent resistance of wild species without sacrificing yield potential. Therefore, hybrid offspring with high genetic stability are obtained, reducing breeding risks and increasing the success rate of combinations.
[0018] Preferably, in step S4, the photoperiodic flowering induction process needs to be carried out in a dedicated light and temperature control greenhouse, with the day and night temperature controlled at 18-30℃ and the relative humidity maintained at 60%-80%.
[0019] By adopting the above technical solutions, the use of a light- and temperature-controlled greenhouse for photoperiod induction can accurately simulate natural light and temperature conditions, avoiding interference from external environmental fluctuations; the day and night temperature is controlled at 18-30℃, a temperature range based on research on the optimal temperature for sugarcane flower bud differentiation, ensuring balanced metabolic activity; the relative humidity is maintained at 60%-80%, preventing pollen inactivation due to excessively low humidity or disease caused by excessively high humidity; therefore, a consistent and controllable flowering induction environment is obtained, promoting the synchronization of flowering periods of the parent plants and laying the foundation for hybrid pollination.
[0020] Preferably, in step S4, the induction scheme is: per m2 Install one 5W incandescent lamp, 2.0m away from the sugarcane canopy, in a double row. Alternatively, use red and blue light, or far-infrared lamps with a wavelength of 730nm, spaced 7m apart. 2 Install 1-2 lamps, 2.0m away from the sugarcane canopy; 450nm blue light lamps every 30m. 2 Install 1-2 lamps, 2.0m away from the sugarcane canopy; 5W incandescent lamps every 30m. 2 Install 5 to 10 lamps, spaced apart, at a distance of 2.0m from the sugarcane canopy;
[0021] From early June, the plants were placed in the light induction chamber at 5:00 PM daily, with the lights turned on and the door closed. The lights were turned off between 6:15 PM and 6:30 PM. The lights were turned on again at 6:00 AM the following day and removed from the chamber at 8:00 AM, with the lights turned off again. The total daily light exposure was 735-750 minutes. The temperature was maintained at 18-30℃ and the humidity at 60-80%. After 120 days of induction, the induction duration was decreased by 30-60 seconds each day until the light exposure reached 720 minutes. Then, the induction continued with a fixed 720 minutes of light exposure daily until the plants began to develop ears and flower.
[0022] By adopting the above technical solution, this scheme can effectively regulate the photoperiodic response of sugarcane through multi-source configuration and time-series control. Incandescent lamps provide full-spectrum illumination to simulate the natural day length, while far-red lamps promote the morphological transformation of photosensitive pigments in sugarcane to enhance flowering gene expression. Blue lamps with a wavelength of 450nm enhance photosynthetic efficiency and regulate stomatal behavior. At the same time, the installation distance is set at 2.0m to ensure uniform light exposure to the canopy. Subsequently, double-row illumination and intermittent placement are used to avoid local shading affecting light signal reception. The initial illumination duration is set slightly higher than 12 hours to simulate the critical day length, and is gradually reduced to simulate seasonal changes to induce flower bud differentiation. Temperature and humidity ranges are maintained to keep cell metabolic homeostasis and prevent pollen inactivation or disease growth. Therefore, it achieves the effect of providing a controllable environment for the synchronous flowering of the parent plants, ensuring the smooth progress of the hybridization and pollination stage, and thus improving the breeding efficiency of offspring.
[0023] Preferably, in step S4, the culture soil used for parent stock cultivation is made from the following raw materials by weight percentage: 10%–15% pine cones, 0.1%–1% boric acid powder, 5%–10% well-rotted pig manure, 0.2%–1% seaweed powder, 0.05%–0.2% microsilica powder, 1%–5% glucose powder, and 0.08%–0.2% indolebutyric acid, with the remainder being red soil. The culture soil needs to be sterilized at 120°C for 2 hours before use.
[0024] By adopting the above technical solutions, pine cones provide a loose structure to improve air permeability, boric acid powder supplements trace elements to promote pollen development, well-rotted pig manure serves as an organic fertilizer source to slowly release nutrients, seaweed powder is rich in algal hormones to enhance stress resistance, microsilica powder strengthens cell walls to prevent lodging, glucose powder provides a carbon source for microbial activity, and indolebutyric acid stimulates root growth; red soil serves as the main substrate to ensure stability, and sterilization at 120℃ for 2 hours eliminates pathogens; these raw materials work synergistically to create a suitable rhizosphere environment, thus obtaining healthy and strong parent plants, providing a physiological basis for flowering induction.
[0025] Preferably, in step S5, a treatment solution is sprayed onto the stigma of the female parent before placing the male parent's flower spike. The sprayed treatment solution is made from the following raw materials in weight percentages: 0.01% to 0.05% N,N-dimethylthiourea, 0.15% to 0.3% sodium nitrophenolate, and 0.03% to 0.05% boron-molybdenum mixture, with the remainder being sterile water.
[0026] By adopting the above technical solution, N,N-dimethylthiourea is used as a permeability regulator to enhance stigma adhesion, sodium nitrophenolate is used as a plant growth regulator to promote pollen tube elongation, boron-molybdenum mixture provides boron to ensure pollen viability and molybdenum to participate in nitrogen metabolism; sterile water is used as a solvent to avoid contamination; and the treatment solution is sprayed on the stigma before pollination, which can optimize the pollination interface microenvironment. Therefore, the pollination and seed setting rate and seed quality are improved.
[0027] Preferably, before performing artificial hybridization pollination in step S5, the process further includes a step of sterilizing the female parent with warm water, wherein the sterilization conditions are 50°C warm water treatment for 3 to 5 minutes.
[0028] By adopting the above technical solution, the female parent is treated with 50℃ warm water for 3-5 minutes to kill the male reproductive organs. This temperature and time range is based on pollen heat sensitivity research and can effectively destroy the activity of the female parent pollen without damaging the stigma's accepting ability. The male killing step is performed before pollination, which completely avoids interference from self-pollination and ensures the purity of hybridization. Therefore, pure hybrid offspring are obtained, ensuring the genetic authenticity of the breeding materials.
[0029] Preferably, in step S6, the seedling substrate for hybrid seeds is prepared by mixing nutrient soil: organic matter: river sand in a volume ratio of 6:3:1, and then sterilized at 120°C for 2 hours. When the seedlings have grown to 3-5 true leaves, they are transplanted.
[0030] By adopting the above technical solutions, the use of nutrient soil to provide basic nutrients, organic matter to improve soil fertility, and river sand to enhance drainage; a 6:3:1 volume ratio to balance water retention and aeration requirements; and 120℃ sterilization for 2 hours to prevent seedling diseases; seedlings are transplanted when they have 3-5 true leaves, at which time root development is optimal and transplant stress is minimal; therefore, uniform seedlings with high survival rates are obtained, accelerating the selection process for offspring.
[0031] Preferably, in step S6, the double-control screening method is as follows: three replicates are investigated for each family, with 10 to 60 plants in each replicate, and the average trait value An is calculated; if An ≥ CK1, the family is selected, otherwise the whole family is eliminated; among the selected families, the individual plant traits must be better than CK2 in order to be considered as superior individual plants and advance to the next breeding stage.
[0032] By adopting the above technical solution, the sample size of 10 to 60 plants per family is used, which is based on statistical principles to ensure the representativeness of the average value; the family average value CK1 is used as the initial screening threshold to eliminate low-value families and improve the overall level; the main cultivated variety CK2 is used as the standard for individual plants to select superior individuals; and the double control method gradually narrows the screening range. Therefore, an efficient and objective offspring selection system is obtained, which improves the accuracy and efficiency of breeding.
[0033] In summary, this application has the following beneficial effects:
[0034] 1. Because this application adopts a scheme that combines multi-trait evaluation with intelligent prediction, by collecting agronomic traits and genotype data of the parents and using a phenotypic prediction model trained with multiple sets of data to predict the performance of hybrid combinations, it overcomes the blindness of relying on experience for selection and achieves the effect of improving the accuracy of hybrid combination selection and breeding efficiency.
[0035] 2. The method of this application integrates multiple steps such as controlling the bloodline ratio of wild species, precise induction of light and temperature, application of special culture soil formula and pollination treatment solution, so as to improve the growth and development environment of the parents while ensuring the purity of hybridization, and to achieve efficient selection of offspring by using a double control screening method. Therefore, it achieves the effect of improving the success rate of hybridization and ensuring the stable inheritance of excellent traits.
[0036] 3. The method of this application, by applying the principle of equal hybridization in the value assessment stage, prioritizes the selection of parents with similar wild species bloodlines for pairing, and controls the total wild species bloodline of the offspring within the range of 12.5% to 25%, thereby avoiding excessive bloodline differences that could lead to disordered trait segregation and maintaining genetic background balance. Therefore, this method improves the genetic stability and consistency of hybrid offspring, reduces breeding risks and increases the success rate of combinations. Attached Figure Description
[0037] Figure 1 This is a flowchart of a sugarcane hybrid combination selection method proposed in this application. Detailed Implementation
[0038] The present application will be further described in detail below with reference to the accompanying drawings and embodiments.
[0039] Technical concept:
[0040] Related sugarcane hybrid breeding relies on breeders' experience to select parental combinations, resulting in a certain accumulation of technology. However, this experience-driven method makes it difficult to systematically quantify the genetic effects and trait transmission patterns between parents, and it is easily affected by subjective judgment biases. This leads to a large degree of blindness in the screening of hybrid combinations, and the actual performance of a large number of hybrid offspring does not match expectations. This not only prolongs the breeding cycle but also increases the human and resource costs of field screening.
[0041] This application discloses a method for selecting and matching sugarcane hybrid combinations, comprising the following steps: S1, multi-dimensional trait evaluation of parents: for candidate sugarcane male and female parents, their agronomic traits, quality traits and stress resistance traits are investigated and their genotype data are analyzed; S2, intelligent selection and matching of hybrid combinations; S3, evaluation and determination of combination value; S4, pre-hybridization treatment of parents and regulation of flowering period; S5, artificial hybridization pollination; S6, screening of hybrid offspring.
[0042] This application adopts a scheme that combines multi-trait evaluation with intelligent prediction. By collecting agronomic traits and genotype data of the parents, and using a phenotypic prediction model trained with multiple sets of data to predict the performance of hybrid combinations, it overcomes the blindness of relying on experience for matching and achieves the effect of improving the accuracy of hybrid combination matching and breeding efficiency.
[0043] Example 1: This example provides a method for selecting and matching sugarcane hybrid combinations, comprising the following steps:
[0044] S1. Multidimensional evaluation of parental traits: Genotypic data of candidate sugarcane male and female parents were analyzed based on their agronomic traits, quality traits, and stress resistance traits.
[0045] S2. Intelligent selection of hybrid combinations: Based on the genotype data obtained in step S1, a phenotypic prediction model is constructed to predict the potential phenotypic performance of hybrid combinations produced by different parent pairings; the phenotypic prediction model is trained using the SNP gene data, transcriptome data, metabolome data of known parents and the phenotypic data of the combined offspring from historical breeding data as the training set, and is obtained through a neural network that includes feature enhancement and wavelet transform.
[0046] The training method for the phenotypic prediction model described in step S2 includes: numerically mapping the SNP gene data, performing discrete Fourier transform with a fixed window to identify protein coding regions and enhance features, denoising the high-frequency features obtained by wavelet transform, performing inverse wavelet transform on the low-frequency features, and constructing a graph structure by combining transcriptome and metabolome data.
[0047] S3. Combination Value Assessment and Determination: Based on the phenotypic performance predicted in step S2, and combined with the preset breeding objectives, a value assessment is conducted, and combinations with high value scores are selected as target hybridization combinations. The value assessment prioritizes combinations whose representative phenotypes are stably inherited after hybridization, and focuses on assessing the predicted values of sucrose content, disease resistance, stress resistance, and yield.
[0048] Among them, the principle of equal hybridization is also applied: priority is given to selecting parents with similar proportions of wild species bloodlines for pairing, and the total proportion of wild species bloodlines used to breed offspring is controlled at 12.5%.
[0049] S4. Pre-hybridization parent treatment and flowering period regulation: For the parent plants in the determined target hybridization combination, photoperiodic flowering induction was carried out in the field or in barrel cultivation from mid-June to early July.
[0050] The process must be conducted in a dedicated, temperature- and light-controlled greenhouse, maintaining a daytime and nighttime temperature of 18°C and a relative humidity of 60%. The photoperiodic flowering induction scheme is as follows: install one 5W incandescent lamp per square meter, 2.0m away from the sugarcane canopy, in double rows; or use red and blue light, with one 730nm far-red lamp installed every 7 square meters, 2.0m away from the sugarcane canopy; one 450nm blue lamp installed every 30 square meters, 2.0m away from the sugarcane canopy; and five 5W incandescent lamps installed every 30 square meters, spaced apart, 2.0m away from the sugarcane canopy. Starting from early June, enter the photoinduction room at 17:00 daily, turn on the lights and close the door, and turn off the lights from 18:15 to 18:30. The lights were turned on at 06:00 the next day and removed from the light induction chamber at 08:00. The total light duration was 735 minutes per day. The temperature was 18℃ and the humidity was 60%. After 120 days of induction, the induction duration was reduced by 30 seconds each day until the light duration reached 720 minutes. Then, the light duration was fixed at 720 minutes per day for continued induction until the ears began to develop and the flowers bloomed. The culture soil used for parent breeding was made from the following raw materials by weight percentage: 10% pine cones, 0.1% boric acid powder, 5% well-rotted pig manure, 0.2% seaweed powder, 0.05% silica powder, 1% glucose powder, and 0.08% indolebutyric acid. The remainder was red soil. The culture soil was sterilized at 120℃ for 2 hours before use.
[0051] S5. Artificial hybridization and pollination: At the early flowering stage of the male / female parent, place the configured male / female parent in the same hybridization cage, with the male parent's flower spike above the female parent. Shake the male parent 2-3 times every morning from 8:30 to 11:30. Control the temperature of the hybridization room to be 25℃-28℃ and the humidity to be 75%-85%.
[0052] The process includes spraying a treatment solution onto the stigma of the female parent before placing the male parent's flower spike. The treatment solution is made from the following raw materials by weight percentage: 0.01% N,N-dimethylthiourea, 0.15% sodium nitrophenolate, and 0.03% boron-molybdenum mixture, with the remainder being sterile water. The process also includes a step of sterilizing the female parent with warm water at 50°C for 3 minutes.
[0053] S6. Screening of hybrid offspring: Harvest hybrid seeds and cultivate seedlings. Use a double control method for screening: use the average value of all family traits as control 1, which is denoted as CK1, and use the trait value of the local main cultivated variety as control 2, which is denoted as CK2. First, eliminate families whose average trait value is lower than CK1. Then, among the selected families, select individuals with traits superior to CK2 as superior offspring.
[0054] The seedling substrate for hybrid seeds is prepared by mixing nutrient soil, organic matter, and river sand in a volume ratio of 6:3:1, and sterilized at 120℃ for 2 hours. Seedlings are transplanted when they have 3-5 true leaves. The double-control screening method is as follows: three replicates are investigated for each family, with 10 plants in each replicate, and the average trait An is calculated. If An ≥ CK1, the family is selected; otherwise, the entire family is eliminated. Among the selected families, the traits of individual plants must be better than CK2 to be considered as superior individual plants and advance to the next breeding stage.
[0055] Example 2: This example provides a method for selecting and matching sugarcane hybrid combinations, comprising the following steps:
[0056] S1. Multidimensional evaluation of parental traits: Genotypic data of candidate sugarcane male and female parents were analyzed based on their agronomic traits, quality traits, and stress resistance traits.
[0057] S2. Intelligent selection of hybrid combinations: Based on the genotype data obtained in step S1, a phenotypic prediction model is constructed to predict the potential phenotypic performance of hybrid combinations produced by different parent pairings; the phenotypic prediction model is trained using the SNP gene data, transcriptome data, metabolome data of known parents and the phenotypic data of the combined offspring from historical breeding data as the training set, and is obtained through a neural network that includes feature enhancement and wavelet transform.
[0058] The training method for the phenotypic prediction model described in step S2 includes: numerically mapping the SNP gene data, performing discrete Fourier transform with a fixed window to identify protein coding regions and enhance features, denoising the high-frequency features obtained by wavelet transform, performing inverse wavelet transform on the low-frequency features, and constructing a graph structure by combining transcriptome and metabolome data.
[0059] S3. Combination Value Assessment and Determination: Based on the phenotypic performance predicted in step S2, and combined with the preset breeding objectives, a value assessment is conducted, and combinations with high value scores are selected as target hybridization combinations. The value assessment prioritizes combinations whose representative phenotypes are stably inherited after hybridization, and focuses on assessing the predicted values of sucrose content, disease resistance, stress resistance, and yield.
[0060] Among them, the principle of equal hybridization is also applied: priority is given to selecting parents with similar proportions of wild species bloodlines for pairing, and the total proportion of wild species bloodlines used to breed offspring is controlled at 18.75%.
[0061] S4. Pre-hybridization parent treatment and flowering period regulation: For the parent plants in the determined target hybridization combination, photoperiodic flowering induction was carried out in the field or in barrel cultivation from mid-June to early July.
[0062] The process must be conducted in a dedicated, temperature- and light-controlled greenhouse, maintaining a daytime and nighttime temperature of 24°C and a relative humidity of 70%. The photoperiodic flowering induction scheme is as follows: one 5W incandescent lamp per square meter, 2.0m from the sugarcane canopy, in double rows; or red and blue light, with one 730nm far-red lamp per 7 square meters, 2.0m from the sugarcane canopy; one 450nm blue lamp per 30 square meters, 2.0m from the sugarcane canopy; and seven 5W incandescent lamps per 30 square meters, spaced apart, 2.0m from the sugarcane canopy. Starting in early June, the light induction chamber is entered daily at 17:00, lights are turned on and the door is closed, and lights are turned off between 18:15 and 18:30. The following day... Lights were turned on at 6:00 AM and removed from the light induction chamber at 8:00 AM, with a total daily light duration of 742.5 minutes. The temperature was 24℃ and the humidity was 70%. After 120 days of induction, the induction duration was decreased by 45 seconds each day until the light duration reached 720 minutes. Then, the induction was continued with a fixed daily light duration of 720 minutes until the ears began to develop and the flowers bloomed. The culture soil used for parent breeding was made from the following raw materials by weight percentage: 12.5% pine cones, 0.55% boric acid powder, 7.5% well-rotted pig manure, 0.6% seaweed powder, 0.125% silica fume, 3% glucose powder, and 0.14% indolebutyric acid, with the remainder being red soil. The culture soil was sterilized at 120℃ for 2 hours before use.
[0063] S5. Artificial hybridization and pollination: At the early flowering stage of the male / female parent, place the configured male / female parent in the same hybridization cage, with the male parent's flower spike above the female parent. Shake the male parent 2-3 times every morning from 8:30 to 11:30. Control the temperature of the hybridization room to be 25℃-28℃ and the humidity to be 75%-85%.
[0064] The process includes spraying a treatment solution onto the stigma of the female parent before placing the male parent's flower spike. The treatment solution is made from the following raw materials by weight percentage: 0.03% N,N-dimethylthiourea, 0.225% sodium nitrophenolate, and 0.04% boron-molybdenum mixture, with the remainder being sterile water. The process also includes a step of sterilizing the female parent with warm water at 50°C for 4 minutes.
[0065] S6. Screening of hybrid offspring: Harvest hybrid seeds and cultivate seedlings. Use a double control method for screening: use the average value of all family traits as control 1, which is denoted as CK1, and use the trait value of the local main cultivated variety as control 2, which is denoted as CK2. First, eliminate families whose average trait value is lower than CK1. Then, among the selected families, select individuals with traits superior to CK2 as superior offspring.
[0066] The seedling substrate for hybrid seeds is prepared by mixing nutrient soil, organic matter, and river sand in a volume ratio of 6:3:1, and sterilized at 120℃ for 2 hours. Seedlings are transplanted when they have 3-5 true leaves. The double-control screening method is as follows: three replicates are investigated for each family, with 35 plants in each replicate, and the average trait An is calculated. If An ≥ CK1, the family is selected; otherwise, the entire family is eliminated. Among the selected families, the traits of individual plants must be better than CK2 to be considered as superior individual plants and advance to the next breeding stage.
[0067] Example 3: This example provides a method for selecting and matching sugarcane hybrid combinations, comprising the following steps:
[0068] S1. Multidimensional evaluation of parental traits: Genotypic data of candidate sugarcane male and female parents were analyzed based on their agronomic traits, quality traits, and stress resistance traits.
[0069] S2. Intelligent selection of hybrid combinations: Based on the genotype data obtained in step S1, a phenotypic prediction model is constructed to predict the potential phenotypic performance of hybrid combinations produced by different parent pairings; the phenotypic prediction model is trained using the SNP gene data, transcriptome data, metabolome data of known parents and the phenotypic data of the combined offspring from historical breeding data as the training set, and is obtained through a neural network that includes feature enhancement and wavelet transform.
[0070] The training method for the phenotypic prediction model described in step S2 includes: numerically mapping the SNP gene data, performing discrete Fourier transform with a fixed window to identify protein coding regions and enhance features, denoising the high-frequency features obtained by wavelet transform, performing inverse wavelet transform on the low-frequency features, and constructing a graph structure by combining transcriptome and metabolome data.
[0071] S3. Combination Value Assessment and Determination: Based on the phenotypic performance predicted in step S2, and combined with the preset breeding objectives, a value assessment is conducted, and combinations with high value scores are selected as target hybridization combinations. The value assessment prioritizes combinations whose representative phenotypes are stably inherited after hybridization, and focuses on assessing the predicted values of sucrose content, disease resistance, stress resistance, and yield.
[0072] Among them, the principle of equal hybridization is also applied: priority is given to selecting parents with similar proportions of wild species bloodlines for pairing, and the total proportion of wild species bloodlines used to breed offspring is controlled at 25%.
[0073] S4. Pre-hybridization parent treatment and flowering period regulation: For the parent plants in the determined target hybridization combination, photoperiodic flowering induction was carried out in the field or in barrel cultivation from mid-June to early July.
[0074] The process must be conducted in a dedicated, temperature- and light-controlled greenhouse, maintaining a daytime and nighttime temperature of 30°C and a relative humidity of 80%. The photoperiodic flowering induction scheme is as follows: one 5W incandescent lamp per square meter, positioned 2.0m from the sugarcane canopy, in double rows; or red and blue light, with two 730nm far-red lamps per 7 square meters, positioned 2.0m from the sugarcane canopy; two 450nm blue lamps per 30 square meters, positioned 2.0m from the sugarcane canopy; and ten 5W incandescent lamps per 30 square meters, spaced apart, positioned 2.0m from the sugarcane canopy. Starting in early June, the light induction chamber is entered daily at 17:00, the lights are turned on and the door is closed, and the process continues from 18:15 to 18:30. Lights were turned off at 0:00 the next day; the lights were turned on again at 06:00, and the plants were removed from the light induction chamber at 08:00, with a total light duration of 750 minutes per day; the temperature was 30℃ and the humidity was 80%. After 120 days of induction, the induction duration was reduced by 60 seconds each day until the light duration reached 720 minutes. Then, the induction was continued with a fixed 720 minutes of light per day until the plants began to develop ears and flower. The culture soil used for parent breeding was made from the following raw materials by weight percentage: 15% pine cones, 1% boric acid powder, 10% well-rotted pig manure, 1% seaweed powder, 0.2% silica powder, 5% glucose powder, and 0.2% indolebutyric acid, with the remainder being red soil. The culture soil was sterilized at 120℃ for 2 hours before use.
[0075] S5. Artificial hybridization and pollination: At the early flowering stage of the male / female parent, place the configured male / female parent in the same hybridization cage, with the male parent's flower spike above the female parent. Shake the male parent 2-3 times every morning from 8:30 to 11:30. Control the temperature of the hybridization room to be 25℃-28℃ and the humidity to be 75%-85%.
[0076] The process includes spraying a treatment solution onto the stigma of the female parent before placing the male parent's flower spike. The treatment solution is made from the following raw materials by weight percentage: 0.05% N,N-dimethylthiourea, 0.3% sodium nitrophenolate, and 0.05% boromolybdenum mixture, with the remainder being sterile water. The process also includes a step of sterilizing the female parent with warm water at 50°C for 5 minutes.
[0077] S6. Screening of hybrid offspring: Harvest hybrid seeds and cultivate seedlings. Use a double control method for screening: use the average value of all family traits as control 1, which is denoted as CK1, and use the trait value of the local main cultivated variety as control 2, which is denoted as CK2. First, eliminate families whose average trait value is lower than CK1. Then, among the selected families, select individuals with traits superior to CK2 as superior offspring.
[0078] The seedling substrate for hybrid seeds is prepared by mixing nutrient soil, organic matter, and river sand in a volume ratio of 6:3:1, and sterilized at 120℃ for 2 hours. Seedlings are transplanted when they have 3-5 true leaves. The double-control screening method is as follows: three replicates are investigated for each family, with 60 plants in each replicate, and the average trait An is calculated. If An ≥ CK1, the family is selected; otherwise, the entire family is eliminated. Among the selected families, the traits of individual plants must be better than CK2 to be considered as superior individual plants and advance to the next breeding stage.
[0079] Comparative Example 1
[0080] The comparative example refers to the content of Example 1, except that when performing the combined value assessment in step S3, the total bloodline ratio of the wild species used is controlled at 32.5%, and the rest is the same as in Example 1.
[0081] Comparative Example 2
[0082] The comparative example refers to the content of Example 1, except that in step S4, the day and night temperature is controlled at 34℃ / 20℃ during the photoperiodic flowering induction process, and the rest is the same as in Example 1.
[0083] Comparative Example 3
[0084] The comparative example refers to the content of Example 1, except that in step S4, the relative humidity is maintained at 50% during the photoperiodic flowering induction process, and the rest is the same as in Example 1.
[0085] Comparative Example 4
[0086] The comparative example refers to the content of Example 1, except that in step S4, the weight percentage of boric acid powder in the culture soil used for parent breeding is 1.3%, and the rest is the same as in Example 1.
[0087] Comparative Example 5
[0088] The comparative example refers to the content of Example 1, except that in step S5, before placing the male parent's flower spike, a treatment solution is sprayed onto the stigma of the female parent. The weight percentage of N,N-dimethylthiourea in the sprayed treatment solution is 0.065%, and the rest is the same as in Example 1.
[0089] Comparative Example 6
[0090] The comparative example is based on the content of Example 1, except that before artificial hybridization pollination in step S5, the female parent is subjected to a hot water treatment to kill the males. The male-killing conditions are 50°C hot water treatment for 6.5 minutes. The rest of the content is the same as in Example 1.
[0091] Performance testing
[0092] Sample preparation: In this experiment, three groups of hybrid offspring samples were prepared using the sugarcane hybridization combination selection method of Examples 1-3. At the same time, six groups of control samples were prepared using the comparative examples 1-6. Each group of samples went through the entire process of parent selection, parent evaluation, intelligent selection, flowering period regulation, artificial pollination and offspring screening, and finally obtained a hybrid offspring population that could be tested for subsequent performance comparison and detection.
[0093] Phenotypic prediction accuracy testing: First, a validation set was constructed based on historical breeding data, containing real phenotypic data of known parents and their offspring. Then, the phenotypic prediction models of Examples 1-3 and Comparative Examples 1-6 were applied to predict the phenotypes of the hybrid combinations in the validation set, obtaining predicted values. Finally, linear regression analysis was performed between the predicted values and the actual observed values, and the coefficient of determination R² was calculated to evaluate the prediction accuracy. The testing standard referenced the phenotypic prediction model validation guidelines published by the International Plant Breeding Association, requiring an R² of not less than 0.7 to be considered valid.
[0094] Hybridization seed set rate test: After artificial pollination, the total number of pollinated flowers and the number of successfully set seeds for each hybrid combination were counted, and the seed set rate percentage was calculated. The experiment was repeated three times and the average value was taken to reduce environmental errors. The test standard is based on the agricultural industry standard NY / T1305-2020 "Technical Specifications for Hybrid Crop Breeding".
[0095] Determination of sucrose content in offspring: After harvesting mature stems of hybrid offspring, juice was extracted using a pressing method. The sucrose content was then calculated using a digital refractometer and calibrated by iodometric titration. Ten plants were randomly sampled from each family, and measurements were taken three times, with the average value taken. The testing standard followed the International Committee for Unified Methods for Sugar Analysis (ICUMSAGS1-2018).
[0096] Progeny smut resistance testing: During the peak season for the disease, the hybrid progeny population was artificially inoculated with a suspension of smut pathogen spores. After inoculation, the plants were incubated at a low temperature for 21 days. The proportion of diseased plants was counted and the resistance index was calculated. The testing standard was based on the national plant quarantine standard GB / T28068-2011 "Technical Specification for Identification of Sugarcane Disease Resistance". A resistance index higher than 80% was considered highly resistant.
[0097] Genetic trait uniformity testing: Genotypic analysis of the hybrid progeny population was performed using SSR molecular markers to obtain the coefficient of variation (COP) of major agronomic traits. Subsequently, the within-population standard deviations of plant height, stem diameter, and sucrose content were examined; lower COPs indicate higher genetic uniformity. The testing standard adopted the ISTA (International Convention on Plant Breeding) guidelines, requiring that the COP of major traits not exceed 15% to confirm stability.
[0098] Table 1: Comparison of Performance Test Data for Examples and Comparative Examples
[0099]
[0100] Example Conclusion:
[0101] As can be seen from Examples 1-3 and Comparative Example 1, and Table 1, controlling the proportion of wild species lineage helps maintain the balance of the genetic background and avoids disordered phenotypic segregation caused by excessive differences in lineage, thereby improving the genetic stability and consistency of hybrid offspring. However, an excessively high proportion of wild species lineage disrupts this balance, which corroborates the role of the principle of equal hybridization in improving genetic combinations in this application.
[0102] Based on Examples 1-3 and Comparative Example 2, and in conjunction with Table 1, it can be seen that the regulation of day and night temperatures during photoperiod induction helps to promote flower bud differentiation and balanced metabolic activity, ensuring the synchronization of flowering periods of the parent plants. However, in Comparative Example 2, excessively high day and night temperatures caused environmental stress and interfered with the flowering physiological mechanism, confirming the influence of light and temperature conditions on the hybridization success rate in this application.
[0103] Combining Examples 1-3 and Comparative Example 3 with Table 1, it can be seen that maintaining a stable relative humidity helps ensure pollen viability and disease control, providing a suitable pollination microenvironment; while in Comparative Example 3, excessively low relative humidity leads to pollen inactivation and decreased stress resistance, emphasizing the synergistic relationship between humidity control and hybridization seed set rate.
[0104] Based on Examples 1-3 and Comparative Example 4, and in conjunction with Table 1, it can be seen that the appropriate addition of boric acid powder to the potting soil can supplement trace elements and promote pollen development and root health; while the excessive boric acid powder content in Comparative Example 4 disrupts the nutrient balance and inhibits plant growth, which indirectly confirms the synergistic effect among the components of the potting soil formula in this application.
[0105] Combined with Examples 1-3 and Comparative Example 5 and Table 1, it can be seen that the use of N,N-dimethylthiourea in the treatment solution can improve stigma adhesion and pollen tube elongation, thereby enhancing the pollination effect; however, excessively high concentrations can interfere with normal physiological processes, demonstrating the regulatory relationship between the components of the treatment solution and pollination efficiency.
[0106] Combining Examples 1-3 and Comparative Example 6 with Table 1, it can be seen that controlling the time of male sterilization in warm water can eliminate the risk of self-pollination without damaging stigma activity and ensure hybridization purity; while the excessively long male sterilization time in Comparative Example 6 caused tissue heat damage, confirming the role of the male sterilization step of the present invention in protecting the offspring's genetics.
[0107] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.
Claims
1. A method for selecting and matching sugarcane hybrid combinations, characterized in that, Includes the following steps: S1. Evaluation of parental traits in multiple dimensions: Genotypic data of candidate sugarcane male and female parents were analyzed based on their agronomic traits, quality traits and stress resistance traits, respectively. S2. Intelligent selection of hybrid combinations: Based on the genotype data obtained in step S1, a phenotypic prediction model is constructed to predict the potential phenotypic performance of hybrid combinations produced by different parent pairings; the phenotypic prediction model is trained using the SNP gene data, transcriptome data, metabolome data of known parents and the phenotypic data of the combined offspring from historical breeding data as the training set, and is obtained through a neural network that includes feature enhancement and wavelet transform. S3. Combination Value Assessment and Determination: Based on the phenotypic performance predicted in step S2, and combined with the preset breeding objectives, a value assessment is conducted, and combinations with high value scores are selected as target hybridization combinations. The value assessment prioritizes combinations whose representative phenotypes are stably inherited after combination, and focuses on assessing the predicted values of sucrose content, disease resistance, stress resistance, and yield. S4. Pre-hybridization parent treatment and flowering period regulation: For the parent plants in the determined target hybridization combination, in the field or barrel cultivation, photoperiodic flowering induction was carried out from mid-June to early July. S5. Artificial hybridization and pollination: At the early flowering stage of the male / female parent, place the configured male / female parent in the same hybridization cage, with the male parent's flower spikes placed above the female parent. Shake the male parent 2-3 times every morning from 8:30 to 11:
30. Control the temperature of the hybridization room to be 25℃-28℃ and the humidity to be 75%-85%. S6. Screening of hybrid offspring: Harvest hybrid seeds and cultivate seedlings. Use a double control method for screening: use the average value of all family traits as control 1, which is denoted as CK1, and use the trait value of the local main cultivated variety as control 2, which is denoted as CK2. First, eliminate families whose average trait value is lower than CK1. Then, among the selected families, select individuals with traits superior to CK2 as superior offspring.
2. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, The training method for the phenotypic prediction model described in step S2 includes: numerically mapping the SNP gene data, performing discrete Fourier transform with a fixed window to identify protein coding regions and enhance features, denoising the high-frequency features obtained by wavelet transform, performing inverse wavelet transform on the low-frequency features, and constructing a graph structure by combining transcriptome and metabolome data.
3. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, When evaluating the combined value in step S3, the principle of equal hybridization is also applied: parents with similar proportions of wild species bloodlines are preferred for pairing, and the total proportion of wild species bloodlines used to breed offspring is controlled between 12.5% and 25%.
4. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, In step S4, the photoperiodic flowering induction process needs to be carried out in a dedicated light and temperature control greenhouse, with the day and night temperature controlled at 18-30℃ and the relative humidity maintained at 60%-80%.
5. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, In step S4, the induction scheme is as follows: per m 2 Install one 5W incandescent lamp, 2.0m away from the sugarcane canopy, in a double row. Alternatively, use red and blue light, or far-infrared lamps with a wavelength of 730nm, spaced 7m apart. 2 Install 1-2 lamps, 2.0m away from the sugarcane canopy; 450nm blue light lamps every 30m. 2 Install 1-2 lamps, 2.0m away from the sugarcane canopy; 5W incandescent lamps every 30m. 2 Install 5 to 10 lamps, spaced apart, at a distance of 2.0m from the sugarcane canopy; From early June, the plants were placed in the light induction chamber at 5:00 PM daily, with the lights turned on and the door closed. The lights were turned off between 6:15 PM and 6:30 PM. The lights were turned on again at 6:00 AM the following day and removed from the chamber at 8:00 AM, with the lights turned off again. The total daily light exposure was 735-750 minutes. The temperature was maintained at 18-30℃ and the humidity at 60-80%. After 120 days of induction, the induction duration was decreased by 30-60 seconds each day until the light exposure reached 720 minutes. Then, the induction continued with a fixed 720 minutes of light exposure daily until the plants began to develop ears and flower.
6. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, In step S4, the culture soil used for parent stock cultivation is made from the following raw materials by weight percentage: 10%–15% pine cones, 0.1%–1% boric acid powder, 5%–10% well-rotted pig manure, 0.2%–1% seaweed powder, 0.05%–0.2% microsilica powder, 1%–5% glucose powder, and 0.08%–0.2% indolebutyric acid, with the remainder being red soil. The culture soil needs to be sterilized at 120°C for 2 hours before use.
7. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, In step S5, before placing the male parent's flower spike, a treatment solution is sprayed onto the stigma of the female parent. The sprayed treatment solution is made from the following raw materials in weight percentages: 0.01% to 0.05% N,N-dimethylthiourea, 0.15% to 0.3% sodium nitrophenolate, and 0.03% to 0.05% boron-molybdenum mixture, with the remainder being sterile water.
8. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, Before artificial hybridization and pollination in step S5, the process also includes a step of killing males in warm water at 50°C for 3 to 5 minutes.
9. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, In step S6, the seedling substrate for hybrid seeds is prepared by mixing nutrient soil, organic matter and river sand in a volume ratio of 6:3:1, and sterilized at 120℃ for 2 hours. When the seedlings have grown to 3-5 true leaves, they are transplanted.
10. The method for selecting and matching sugarcane hybrid combinations according to claim 1, characterized in that, In step S6, the double-control screening method is as follows: each family is investigated with 3 replicates, each replicate with 10 to 60 plants, and the average trait value An is calculated; if An ≥ CK1, the family is selected, otherwise the whole family is eliminated; among the selected families, the individual plant traits must be better than CK2 in order to be considered as excellent individual plants and advance to the next breeding stage.