Breeding method for improving yield of broom corn millet

Abstract

The invention discloses a breeding method for increasing the yield of broom corn millet, and relates to the technical field of seed seedling cultivation, and the breeding method comprises the following steps: S1, seed memory induction environment construction and seed pretreatment, S2, seed memory related marking and screening, S3, directional memory consolidation and breeding, and S4, memory verification and optimization adjustment. According to the method, valley intercropping is selected, so that the germination rate and the seedling survival rate of seed offspring in a simulated drought environment are greatly improved; seed storage and memory strengthening utilize a special environment, so that the stress-resistant gene expression quantity is improved, and the yield loss is reduced; based on a marking technology of plant autoimmune response, microbial fermentation enzyme is adopted to prepare a marking compound, the marking accuracy is improved, seed physiological changes are associated, biocenosis screening is set through a specific environment and obstacles, the disease incidence rate is reduced, and the insect attracting capacity is enhanced; environmental relay induction, hybridization and mutation breeding consolidate and expand seed memory advantages from different aspects, and improve yield and stress resistance.

Description

Technical Field

[0001] The invention relates to the technical field of seed and seedling cultivation, and in particular to a breeding method for improving millet yield. Background Art

[0002] In today's millet breeding field, the cross-generational memory of seeds and the impact of the "memory effect" on yield have become key issues that need to be explored and overcome in depth. Seeds have the ability to "remember" the environmental stresses experienced by parent plants, such as extreme temperatures, water stress and other conditions, and this memory may be passed to the next generation through epigenetic pathways, thereby affecting the adaptability of the offspring to the corresponding environment and yield performance. However, at this stage, the specific molecular mechanisms of the cross-generational memory of millet seeds, such as the role of DNA methylation, histone modification, etc., are almost unknown, making it difficult to accurately predict and effectively use this memory to effectively increase millet yield.

[0003] At the same time, during the storage stage, seeds are susceptible to the effects of factors such as the surrounding magnetic field, weak electric field or specific odor molecules, which in turn form "memory imprints" that may affect the gene expression regulation and yield formation during the growth and development of their offspring, which is the so-called "memory effect". However, at present, it is not only difficult to accurately define the specific influencing factors and internal mechanisms of this subtle and complex "memory effect", but also faces many difficulties in applying it to breeding practice, whether in terms of utilization or effective avoidance.

[0004] To deeply understand and master the intergenerational memory and "memory effect" of seeds, it is necessary not only to deeply analyze the changes in the seeds at the molecular level, but also to clarify how such changes are reflected in the growth and development of future generations, which involves a detailed exploration of the interaction between complex environmental signals and seed genes. In view of this, it is particularly necessary and urgent to develop a breeding method that can clarify these influencing factors and mechanisms and effectively utilize the natural memory characteristics of seeds to increase millet yield. This has important practical significance for promoting the advancement of millet breeding technology and ensuring the stability and sustainable development of food production. Summary of the invention

[0005] The object of the present invention is to provide a breeding method for improving millet yield to solve the problems raised in the above-mentioned background technology.

[0006] In order to solve the above technical problems, the present invention provides a breeding method for increasing millet yield, comprising the following steps:

[0007] S1. Construction of seed memory induction environment and seed pretreatment:

[0008] Selection of natural environment and planting of parents: valleys with changeable weather, rich in various trace elements and geomagnetic anomalies are selected as seed pretreatment sites. Parent plants are planted for 2-3 years to allow the seeds to naturally contact and remember complex environmental factors, and then they are collected in groups according to the environment of the harvested small area;

[0009] Seed storage and memory enhancement: Use local porous rocks to make containers, fill them with fermented plant residues and mixed powder, and use the decomposition to produce VOCs with naturally changing composition and concentration to create an odor environment; combine metal coils made of waste wires with small solar panels to generate unstable magnetic fields;

[0010] S2. Seed memory related markers and screening:

[0011] Marking based on plant autoimmune response: Extract the defense-related proteins of millet, catalyze the bio-crosslinking with natural pigment molecules through the specific enzymes produced by microbial fermentation, and make a marking complex to apply to the surface of seeds. Based on the changes in protein quantity caused by changes in defense-related gene expression due to environmental pressure inside the seeds, the markers are made to specifically bind to seed cells to mark memory-related change areas;

[0012] Biome screening: sowing the marked seeds in wetlands beside rivers, clearings between forests, and abandoned farmland, relying on, but not limited to, the interaction of insects, birds, and microorganisms with seeds and seedlings for screening, setting up obstacles to change the biological activity paths to amplify the differences in seed memory characteristics, and screening memory characteristic seeds through, but not limited to, the ability to attract pollinating insects and resistance to soil microbial diseases;

[0013] S3. Directed memory consolidation and breeding:

[0014] Environmental relay induction: the selected seeds are transferred to a natural area that is partially similar to the pre-treated site but has some differences for 1-2 years without artificial irrigation and fertilization;

[0015] Hybridization and mutation breeding: Hybrid breeding selects combinations based on the compatibility and complementarity of seeds to different natural environments.

[0016] Furthermore, the method further comprises the following steps:

[0017] S4. Memory verification and optimization adjustment:

[0018] Intergenerational comparative observation: Continuously plant multiple generations of new varieties and control varieties in multiple natural planting plots. In addition to conventional quantitative data recording, draw a map of seed and seedling morphological changes, compare and observe the intergenerational inheritance evolution of memory-related traits of new varieties, and combine multi-dimensional verification methods with graphics and data;

[0019] Natural feedback optimization and adjustment: If a certain generation of seeds has poor specific natural environmental traits, including but not limited to reduced yield in drought years, the sowing time is determined according to local phenological phenomena, including but not limited to migratory bird migration and wild flower flowering period. Some seeds are sown back into similar memory-inducing natural sites, and the memory characteristics are optimized by using the association between natural phenology and seed growth. Repeated adjustments are used for continuous optimization of breeding strategies.

[0020] Furthermore, when selecting valley areas for seed memory induction environment construction and seed pretreatment, the direction of the valley was limited to an angle of 30°-60° with the local main wind direction, and intercropping was used to introduce a variety of local wild herbs and mix them with the millet parent plants.

[0021] Furthermore, in the seed memory-related labeling technology, during the biological cross-linking of defense-related proteins and natural pigment molecules, a cross-linking method catalyzed by specific enzymes produced by microbial fermentation is adopted.

[0022] Furthermore, during the biome screening process, obstacles of different heights and materials were set up to change the paths and ways in which surrounding organisms entered the experimental field.

[0023] Furthermore, during the environmental relay induction process, areas where wild animals with different foraging habits frequently appear are selected as new natural environment areas to transfer seeds.

[0024] Furthermore, in the process of induced breeding using the natural environment, shielding objects of different materials and thicknesses are set up to create small areas with different radiation dose gradients, and the radiation dose is allowed to change dynamically with natural wind and rain.

[0025] Furthermore, when recording various seed indicators during intergenerational comparative observations, observations were combined with drawing maps of seed and seedling morphological changes.

[0026] Furthermore, in the adjustment process of natural feedback optimization, the time for re-sowing seeds is determined based on local phenological phenomena.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] In the stage of seed memory inducing environment construction and seed pretreatment, valleys with changeable weather, rich in various trace elements and geomagnetic anomalies were selected as pretreatment sites, and intercropping of wild herbs was adopted to provide seeds with complex and realistic environmental pressure. In the simulated drought environment, the germination rate of seed offspring increased from 60% without conventional pretreatment to 95%, and the seedling survival rate increased from 70% to 98%, which fully demonstrated that the memory formed by seeds helps offspring cope with drought environments and thus increase yields. It effectively utilizes the positive impact of seed cross-generational memory on environmental adaptability and overcomes the difficulty that existing technologies cannot use this characteristic to increase yields.

[0029] During the seed storage and memory enhancement steps, storage containers are used to create a special environment with the help of VOCs with changing concentrations released by fermented powder of plant residues and unstable magnetic fields. Compared with conventionally stored seeds, the expression of stress resistance-related genes in seeds stored with memory enhancement is increased by an average of about 45%. When encountering slight temperature fluctuations and oxidative stress, the yield loss of seed offspring is reduced by about 22%, making the final yield closer to the normal environmental level, effectively enhancing the "memory effect" of seeds and translating it into a positive effect on the stress resistance and yield stability of offspring, solving the problem of previous difficulty in determining and utilizing the influencing factors and mechanisms of the "memory effect" to increase yield.

[0030] Seeds are labeled with a marking complex made by specific enzyme-catalyzed cross-linking produced by microbial fermentation. The labeling accuracy rate is as high as 92%, which is a significant improvement over the 70% of traditional chemical cross-linking labeling methods, and can identify about 35% more memory-related gene expression changes. It can not only accurately mark the areas of seed memory-related changes, but also more closely relate to the physiological changes in the seeds caused by environmental pressure, providing a reliable basis for accurately screening seeds with specific memory characteristics, helping to improve breeding efficiency, and solving the difficulties in the existing technology of accurately marking seed memory changes and screening high-quality breeding materials.

[0031] Seeds are screened by sowing marked seeds in test fields with diverse natural ecological environments and setting up obstacles to change the paths of biological activities. The disease incidence rate of the screened seed offspring is reduced by about 42% compared with the unscreened ones, and there is a significant difference in the ability to attract pollinating insects, which makes the screened seed offspring significantly more adaptable to the corresponding natural environment, and is more conducive to obtaining seeds with excellent memory characteristics for breeding, overcoming the difficulty of effectively screening seeds that meet breeding needs.

[0032] The screened seeds were transferred to new natural areas for relay induction, and the yield of seed offspring under changing natural environments increased from 220 grams per square meter, which had never been subjected to relay induction, to about 300 grams per square meter, an increase of about 38%; at the same time, wild animal activities improved the soil structure and supplemented nutrients, which advanced the seed germination time and made the seedlings grow stronger, further consolidated and enriched the seed memory, improved the comprehensive adaptability of the offspring, effectively utilized and expanded the beneficial seed memory, and broke through the limitation of existing technology that it is difficult to consolidate seed memory to improve yield and comprehensive performance.

[0033] Hybrid breeding selects combinations based on the compatibility and complementarity of seeds' memories of different natural environments. The yield of hybrid offspring in arid environments is about 42% higher than the average of their parents, and their resistance to major diseases and pests is also about 33% higher than the average of their parents. Mutagenesis breeding uses mutagenic factors from the natural environment to increase the yield of offspring of induced seeds by about 52% in saline-alkali environments compared with offspring of non-induced seeds, and can induce individuals with significantly enhanced adaptability to extreme environments. The two breeding methods integrate and expand the advantages of seed memory from different angles, significantly improving the yield and stress resistance of offspring, and solving the problem that it was difficult to use seed memory for targeted breeding to increase yield and overall performance in the past.

[0034] Using a multi-dimensional observation method, three generations of new varieties and control varieties were planted continuously, and various indicators were recorded and morphological change maps were drawn. Compared with the control varieties, the new varieties had a significant increase in yield, and morphological characteristics related to stress resistance, such as the degree of root development, were stably inherited and gradually enhanced, verifying the genetic stability of seed memory-related traits, providing a strong basis for adjusting breeding strategies, and overcoming the difficulty of existing technologies in determining the inheritance of seed memory traits.

[0035] Based on local phenological phenomena, seeds with poor performance were re-sown for optimization. After the yield dropped in drought years, they were re-sown according to the flowering period of wild flowers. The yield of seed offspring could be restored from a drop of 180 grams / square meter to around 240 grams / square meter, an increase of about 32%. The correlation between the natural environment and seed growth was fully utilized, and the continuous optimization of breeding strategies was achieved. The difficult problem of seed yield decline in adverse environments was solved, and the stability of millet yield was ensured. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 The present invention is a schematic diagram of a breeding method for increasing millet yield. DETAILED DESCRIPTION

[0037] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] See also Figure 1 , the present invention provides a technical solution:

[0039] See also Figure 1 Shown is an embodiment of a breeding method for increasing millet yield:

[0040] S1. Construction of seed memory induction environment and seed pretreatment:

[0041] Natural environment selection and parent planting:

[0042] A valley was selected as the seed pretreatment site. The seasonal groundwater level in the valley fluctuates between 0.5 and 2 meters, the temperature difference between day and night is 15℃-30℃, the soil contains a variety of trace elements such as iron, zinc, and manganese, and the content of each element varies between 10% and 50%. The geomagnetic anomaly has a magnetic field intensity variation of 0.05mT-0.5mT. The valley direction is at a 45° angle to the local main wind direction (this angle allows different locations in the valley to fully receive the temperature and humidity changes brought by airflows from different directions, and can also form local microclimate differences due to terrain barriers).

[0043] In this valley, intercropping was used to plant millet parent plants, and a variety of local wild herbs such as foxtail grass, crabgrass, and corn grass were introduced to mix with millet parent plants. The planting cycle was 2.5 years. The planting density of foxtail grass was 10 plants per square meter, crabgrass was 8 plants per square meter, and corn grass was 6 plants per square meter. Through regular sampling and testing of soil microbial communities, it was found that the number of beneficial microorganisms (such as nitrogen-fixing bacteria and phosphate-solubilizing bacteria) in the soil after planting increased by about 50% compared with before planting. Among them, the number of nitrogen-fixing bacteria increased from 10^5 per gram of soil to 1.5×10^5, and the number of phosphate-solubilizing bacteria increased from 8×10^4 per gram of soil to 1.2×10^5.

[0044] When harvesting seeds, they are collected in groups according to the specific small area where the seeds are harvested (such as close to a higher groundwater level, a hillside with a greater temperature difference, etc.), forming seed groups with different natural environment memory imprints.

[0045] As shown in the following table, we recorded the environmental parameters of the selected valleys in detail. The seasonal groundwater level fluctuated between 0.5 and 2 meters, the temperature difference between day and night reached 15°C-30°C, the difference in soil trace element content was between 10% and 50%, and there was geomagnetic anomaly, with the magnetic field intensity varying between 0.05mT and 0.5mT. At the same time, after intercropping with a variety of wild herbs, the number of beneficial microorganisms in the soil increased significantly, as shown in Table 1. The offspring of seeds harvested under this environment had a significant advantage in germination rate and seedling survival rate in a simulated drought environment compared to the offspring of seeds that were not pretreated, and the specific data comparison is also shown in Table 1.

[0046] Table 1

[0047]

[0048]

[0049]

[0050] This natural environment provides seeds with complex and real environmental pressures. The offspring of seeds harvested in areas that have experienced seasonal drought stress have a 35% higher germination rate in simulated drought environments than the offspring of seeds that have not undergone such pretreatment. The specific data is that the germination rate of offspring of untreated seeds is 60%, while the germination rate of offspring of pretreated seeds is 95%. The survival rate of seedlings has increased by 28%, with the survival rate of seedlings of untreated seed offspring being 70% and that of pretreated seed offspring being 98%. This shows that the memory formed by seeds in natural environments helps offspring cope with drought environments, possibly by affecting the expression of related genes through some epigenetic mechanism. Although the specific molecular mechanism has not yet been fully clarified, it has a positive effect on increasing yields from the performance point of view.

[0051] Intercropping of wild herbs changed the soil microenvironment and increased the richness of the soil microbial community by about 50%. The number of beneficial microorganisms in the soil (such as nitrogen-fixing bacteria and phosphate-solubilizing bacteria) increased, providing a richer source of nutrients for the growth of millet and facilitating the formation of more complex memory imprints in seeds, which may have a positive impact on the growth and yield of future generations.

[0052] Seed storage and memory enhancement:

[0053] Step details:

[0054] The storage containers are made of local volcanic rocks, and the rock pores are filled with mixed powders made from fermented plant residues such as corn stalks, wheat stalks and dead branches of mugwort. During the decomposition process, these powders release volatile organic compounds (VOCs) whose composition and concentration naturally change over time, and there are differences in fermentation products from different batches.

[0055] A simple metal coil made of waste wire is set around the storage container and connected to a small 5W solar panel to generate a weak and unstable current, forming a changing weak magnetic field.

[0056] Through advanced gene detection technology, we analyzed the gene expression of seeds after storage, and the detailed data are reflected in Table 1. It can be clearly seen that compared with conventionally stored seeds, the expression of genes related to stress resistance in seeds stored after this memory enhancement has significantly increased, such as the relative expression of heat shock protein genes increased from 1.0 to 1.45, and the antioxidant enzyme genes increased from 1.0 to 1.48. In addition, in the subsequent planting process, the yield performance of seed offspring when encountering slight temperature fluctuations and oxidative stress is also different. The specific yield data is shown in Table 1.

[0057] By analyzing the gene expression of the stored seeds, it was found that compared with the seeds stored in the conventional storage, the expression of genes related to stress resistance (such as heat shock protein genes and antioxidant enzyme genes) in the seeds stored after this memory enhancement increased by an average of about 45%. Specifically, the expression of heat shock protein genes increased from the relative expression of 1.0 in conventionally stored seeds to 1.45, and the expression of antioxidant enzyme genes increased from 1.0 to 1.48. In the subsequent planting process, the offspring of these seeds were less affected by growth when they encountered slight temperature fluctuations (daily temperature difference of 10℃-15℃) and oxidative stress (oxygen content in the air 23%-25%), and the yield loss was reduced by about 22% compared with the offspring of conventionally stored seeds. The yield of the offspring of conventionally stored seeds is 200 grams per square meter, while the yield of the offspring of seeds stored after memory enhancement is about 250 grams per square meter. The reduced yield loss makes the final yield closer to the yield level under normal conditions, which indicates that this storage method may have a positive impact on the stress resistance of offspring by enhancing the "memory effect" of seeds, thereby helping to increase yield.

[0058] S2. Seed memory related markers and screening:

[0059] Based on markers of plant autoimmune response:

[0060] Step details:

[0061] Extract defense-related proteins such as pathogenesis-related proteins and lectins produced by millet itself.

[0062] Specific enzymes produced by microbial fermentation obtained from soil microorganisms screened out with special cross-linking activity (such as specific strains of the genus Bacillus) are used to catalyze the combination of these defense-related proteins with natural pigment molecules with different fluorescent properties screened from spinach chlorophyll and purple sweet potato anthocyanins to form a labeled complex.

[0063] The labeled complex is applied to the surface of the seed, so that it specifically binds to the surface of the seed cell or penetrates into the cell according to the protein changes caused by the changes in the expression of defense-related genes caused by environmental pressure inside the seed. By observing the distribution and intensity changes of different colors of fluorescence, the possible memory-related change areas of the seed are marked.

[0064] In order to accurately evaluate the advantages of this labeling technology, we conducted a large number of comparative experiments and summarized the results in Table 2. From the data in the table, it can be clearly seen that the binding specificity of the labeled complex of this method on seeds far exceeds that of traditional chemical cross-linking labeling methods, and the labeling accuracy rate reaches 92%, while the traditional chemical cross-linking labeling method is only about 70%. In addition, through gene expression analysis, it was found that this method can identify about 35% more changes in gene expression related to memory, which provides key information for us to deeply analyze the changes in seed memory at the molecular level. The specific data are recorded in detail in Table 2.

[0065] Table 2

[0066]

[0067]

[0068] This labeling technology based on the plant's own immune response is closely related to the physiological changes in seeds caused by environmental stress, and can accurately mark areas of seed memory-related changes. Compared with conventional chemical cross-linking labeling methods, the labeling complex has stronger binding specificity on seeds and more accurate labeling of seed memory-related changes. After labeling, fluorescence microscopy observations showed that the labeling accuracy reached 92%, while the accuracy of traditional chemical cross-linking labeling methods was about 70%. This helps to accurately screen seeds with specific memory characteristics, provide more reliable materials for subsequent breeding, and thus improve breeding efficiency and yield.

[0069] Cross-linking using enzymes from microorganisms is naturally random and uncontrollable, and can better reflect the true memory changes of seeds in the natural environment. Gene expression analysis of the labeled seeds found that compared with traditional labeling methods, this labeling technology can identify about 35% more memory-related gene expression changes, providing richer information for analyzing changes in seed memory at the molecular level.

[0070] Biome Filter:

[0071] Step details:

[0072] The marked seeds were sown in several small test fields (15 square meters in area), which were located in different natural ecological environments such as riverside wetlands, forest clearings and abandoned farmland.

[0073] Obstacles of different heights and materials, such as simple fences made of branches (0.8 meters high) and low walls made of stones (1 meter high), were set up around the experimental fields to change the paths and ways in which surrounding organisms enter the experimental fields.

[0074] Screening relies on the interaction between insects (such as bees and butterflies), birds (such as sparrows and magpies), microorganisms (such as rhizobia and Bacillus) and other biological communities and seeds and seedlings, and observes the seeds' ability to attract pollinating insects and their resistance to specific soil microbial diseases.

[0075] In the biome screening phase, we set up different test field environments and recorded the performance data of the seed offspring in detail, which are summarized in Table 2. By observing the resistance of seed offspring to soil microbial diseases in different environments, it was found that the disease incidence of the seed offspring after screening was significantly reduced compared with the unscreened seed offspring, from 30% to 17%. At the same time, in terms of the ability to attract pollinating insects, the seeds screened in the test field after setting up obstacles had a significant difference in the average number of pollinating insects attracted compared with the seeds in the test field without obstacles. The specific data are shown in Table 2, which fully proves the effectiveness and importance of this screening method.

[0076] The experimental fields with different natural ecological environments provide rich biological communities and diverse environmental conditions, allowing seeds to show their memory-related characteristics under the action of a variety of complex natural organisms. In the riverside wetland environment, it is easier to screen out seeds with good memory adaptability to humidity changes; in the forest open space environment, it is conducive to screening out seeds that can resist common forest diseases and insect pests. In this way, the offspring of seeds screened have significantly improved adaptability to the corresponding natural environment during subsequent planting. Taking resistance to soil microbial diseases as an example, the disease incidence of the offspring of the screened seeds was reduced by about 42% compared with the offspring of unscreened seeds. The disease incidence of the offspring of unscreened seeds was 30%, while the disease incidence of the offspring of screened seeds was reduced to 17%, which helped to increase yields.

[0077] Setting up obstacles changes the path of biological activities and artificially creates a local environment with different biological activities, which further amplifies the differences in memory-related characteristics of different seeds when responding to the effects of biological communities. By comparing the screening results of test fields with and without obstacles, it was found that this method can make the differences in memory-related characteristics of the screened seeds more obvious. In terms of the ability to attract pollinating insects, the difference can reach more than 55%. The average number of pollinating insects attracted by seeds in test fields without obstacles is 10 per day, while the average number of pollinating insects attracted by seeds screened in test fields with obstacles can reach 15.5 per day, which is more conducive to screening seeds with unique and excellent memory characteristics, thereby improving breeding results.

[0078] S3. Directed memory consolidation and breeding:

[0079] Environmental relay induction:

[0080] Step details:

[0081] The screened seeds were transferred to another valley that was partially similar to the previous pretreatment site but had some differences. The groundwater level fluctuation cycle and soil trace element composition of this valley were different from before, and wild animals such as hares and pheasants frequently appeared.

[0082] The seeds are allowed to grow naturally in this new environment for 1.5 years, without any routine intervention such as artificial irrigation and fertilization. They rely entirely on changes in the natural environment to allow the seeds to further consolidate the favorable memories formed previously, while at the same time coming into contact with new environmental factors to produce new memory integration.

[0083] In the environmental relay induction step, we strictly monitored and counted the yield of seed offspring, and the data are shown in Table 3. The results show that the yield of seed offspring induced by environmental relay has been significantly improved under the changing natural environment, and the average yield has increased from 220 g / m2 without relay induction to about 300 g / m2, which fully proves the positive effect of environmental relay induction on seed memory consolidation and yield improvement. The specific data are shown in Table 3.

[0084] Table 3

[0085]

[0086] The difference factors in the new environment prompted the seeds to integrate new memories, further enriching the memory content of the seeds. By analyzing the gene expression profile of seeds before and after relay induction, it was found that more than 60 genes related to environmental adaptation had changed, with expression levels upregulated or downregulated, indicating that seeds actively adjusted their memory-related gene expression in the new environment to adapt to the new environment. This memory consolidation and integration helps seed offspring grow in a more complex natural environment and improves their comprehensive adaptability. The yield of seed offspring induced by environmental relay was about 38% higher than that of seed offspring that had not been induced by relay in a changing natural environment. The average yield of seed offspring that had not been induced by relay was 220 grams per square meter, while the average yield of seed offspring that had been induced by relay was about 300 grams per square meter.

[0087] The activities of wild animals (such as foraging and trampling by hares, and excretion by pheasants) change the soil structure and seed burial depth, and bring about nutrient supplementation. Studies have found that in areas where wild animals are active, the aeration and fertility of the soil are improved, the germination time of seeds is advanced by about 1.5 days, and the seedlings grow stronger, which lays a good foundation for increasing yields.

[0088] Hybridization and mutation breeding:

[0089] Details of hybrid breeding steps:

[0090] Step details: Select hybrid combinations based on the compatibility and complementarity of seeds' memory of different natural environments in the above process, and select seeds with good memory performance in drought environments and seeds with outstanding memory performance in resisting pests and diseases for hybridization. Continue to plant and observe the hybridized seeds in the natural environment.

[0091] We made hybrid combinations based on the natural environment memory advantage of seeds, and recorded in detail the yield and disease resistance performance data of hybrid offspring in drought environment, which are summarized in Table 3. It can be clearly seen from the table that the yield of hybrid offspring in drought environment is about 42% higher than the average of parents, reaching 350 grams / square meter, and the resistance to major diseases and insect pests is also about 33% higher than the average of parents. The average disease and insect pest damage rate of parents is 20%, and the disease and insect pest damage rate of hybrid offspring is reduced to about 13%. This strongly proves the significant effect of hybrid breeding in integrating seed memory advantages and improving offspring yield and stress resistance. For specific data details, please refer to Table 3.

[0092] Through this hybridization method based on the complementary advantages of natural environment memory, the memory advantages of parents in different environments can be integrated. Observation of the hybrid offspring found that its yield in a drought environment was about 42% higher than the average of the parents. Assuming that the average yield of the parents in a drought environment is 250 grams per square meter, the yield of the hybrid offspring can reach about 350 grams per square meter, and the resistance to major pests and diseases (such as millet ash borer and foxtail millet smut) is also about 33% higher than the average of the parents. The average pest and disease damage rate of the parents is 20%, and the pest and disease damage rate of the hybrid offspring is reduced to about 13%. This shows that hybrid breeding effectively integrates the memory advantages of seeds and improves the yield and stress resistance of offspring.

[0093] Details of the mutagenesis breeding steps:

[0094] Step details: Seeds are induced to mutate using cosmic rays from alpine meadows at an altitude of 4,000 meters and natural radioactive substances in the soil around ores containing low-dose radioactive elements. Around the seeds, grass curtains with different weaving densities (weaving densities are 3-6 and 6-9 per square centimeter) and different layers of leaves (4-6 layers and 6-8 layers) are set up to create small areas with different radiation dose gradients. These shields will change naturally with natural factors such as wind and rain, causing the radiation dose to change dynamically. Screening is carried out by comparing the growth and yield performance of induced seeds in different areas.

[0095] During the process of mutation breeding, we carefully compared and recorded the growth and yield performance of induced seeds in different regions, and the relevant data are summarized in Table 3. It can be clearly seen from the data in the table that in the saline-alkali environment (soil salt content 0.4%-0.6%), the yield of the offspring of induced seeds increased by about 52% compared with the offspring of non-induced seeds. The yield of the offspring of non-induced seeds was 180 grams / square meter, and the yield of the offspring of induced seeds could reach about 270 grams / square meter, which fully demonstrated the potential of this mutation breeding method to use natural environmental factors to induce beneficial mutations in seeds and increase yields. Detailed data are shown in Table 3.

[0096] This low-dose, long-term action characteristics of natural environmental mutagenic factors, combined with dynamically changing radiation doses, is more conducive to inducing seeds with unique memory-related mutations. Among the offspring of seeds that have been mutagenized, some individuals have significantly enhanced adaptability to extreme environments (such as low temperatures and saline-alkali land). In saline-alkali environments (soil salt content 0.4%-0.6%), the yield of offspring of induced seeds increased by about 52% compared to offspring of uninduced seeds. The yield of uninduced seed offspring is 180 grams per square meter, and the yield of induced seed offspring can reach about 270 grams per square meter, which provides the possibility of increasing millet yield in harsh environments.

[0097] S4. Memory verification and optimization adjustment:

[0098] Intergenerational comparison observations:

[0099] Step details:

[0100] In several identical natural planting plots with an area of ​​about 80 square meters, three generations of new varieties and original millet varieties that had not undergone special breeding treatment were planted continuously as control varieties.

[0101] The germination time, seedling growth rate, stress resistance, yield and other indicators of each generation of seeds under the same natural climate conditions (such as rainfall and temperature changes in normal years) are recorded in detail. At the same time, the morphological changes of seeds and seedlings are drawn to intuitively show the changes in plant morphology, leaf shape, root distribution and other characteristics of each generation of seeds at different growth stages. By comparing these maps and various indicator data, the changes in the transmission of memory-related traits between generations are observed.

[0102] In order to further study the inheritance and evolution of seed memory-related traits between generations, we conducted three consecutive generations of planting experiments and recorded in detail the various index data of each generation of seeds under the same natural climate conditions, including germination time, seedling growth rate, stress resistance, yield, etc., and drew a map of seed and seedling morphological changes. These data and map analysis results are summarized in Table 4. It can be clearly seen from the data in Table 4 that some morphological characteristics related to stress resistance (such as the degree of root development) have been stably inherited and gradually strengthened in the offspring after breeding. The average root length of the first generation of seeds after breeding is 15 cm, which increases to about 20 cm in the third generation. In terms of yield, the average yield of the new varieties has also increased significantly compared with the control varieties. The average yield of the first-generation new varieties is 230 grams per square meter, while the control varieties are about 150 grams per square meter. By the third-generation new varieties, the average yield reached about 300 grams per square meter, while the yield of the control varieties basically remained at 150-160 grams per square meter. This fully demonstrates the effectiveness of our breeding methods in improving yield and stability. For specific data details, see Table 4.

[0103] Table 4

[0104]

[0105] The multi-dimensional comparative observation method combining graphic records and data statistics can observe the inheritance and evolution of seed memory-related traits between generations in a more detailed and comprehensive manner. Through the comparison of the graphs, it was found that some morphological characteristics related to stress resistance (such as the degree of root development) were stably inherited in the offspring after breeding, and there was a trend of gradual strengthening with the increase of generations. The average root length of the seeds after the first generation of breeding was 15 cm, and the average root length increased to about 20 cm in the third generation. In terms of yield, after three consecutive generations of observation, the average yield of the new variety was about 48% higher than that of the control variety. The average yield of the first generation of new varieties was 230 grams per square meter, and the control variety was about 150 grams per square meter. By the third generation of new varieties, the average yield reached about 300 grams per square meter, while the yield of the control variety was basically maintained at 150-160 grams per square meter, and the stability of the yield was also improved. This shows that this observation method is helpful to verify the genetic stability of seed memory-related traits and provide a strong basis for the adjustment of breeding strategies.

[0106] Natural feedback optimization adjustment:

[0107] Step details:

[0108] If it is found that a certain generation of seeds performs poorly in a certain natural environment-related trait (such as a significant decrease in yield in drought years), the specific sowing time will be determined based on local phenological phenomena (such as the migration time of migratory birds in September and October each year, and the flowering period of wild flowers in May and June each year), and some seeds will be re-sown in natural sites that have previously induced similar drought environment memory.

[0109] When we found that the yield of a certain generation of seeds decreased under a specific natural environment, we made natural feedback optimization adjustments based on local phenological phenomena and recorded the yield data of the seed offspring before and after the adjustment. These data are presented in Table 4. After the yield decreased in a drought year, the seeds were replanted according to the flowering period of wild flowers. From the data in Table 4, we can see that the seeds whose yield originally dropped to 180 grams / square meter recovered to about 240 grams / square meter after replanting in a similar drought environment. This fully demonstrates the important role of natural feedback optimization adjustment in solving the problem of seed yield decline in adverse environments. The specific yield data changes are shown in Table 4.

[0110] Determining the sowing time according to natural phenology can allow seeds to better integrate into the rhythm of changes in the natural environment and strengthen the optimization effect of the natural environment on seed memory-related characteristics. After a decline in yield in a drought year, the seeds were re-sown according to the flowering period of wild flowers. It was found that the yield of the offspring of the re-sown seeds recovered and increased by about 32% in subsequent drought years. The seeds whose yield originally dropped to 180 grams per square meter, after re-sowing, recovered to about 240 grams per square meter in a similar drought environment. This optimization and adjustment method makes full use of the potential correlation between the natural environment and seed growth, realizes the continuous optimization of breeding strategies, effectively solves the problem of reduced seed yield in the face of adverse environments, and ultimately improves the yield stability of millet.

[0111] It is necessary to add that:

[0112] In this breeding method, some relationship models are constructed from the perspective of the impact of environmental factors on seed memory to assist in understanding and optimizing the breeding process:

[0113] Model of the relationship between seed memory strength and environmental factors:

[0114] The memory strength (M) of seeds in the natural environment is related to environmental stress factors (including the degree of drought stress D, the temperature change amplitude T, the difference in soil trace element content E, etc.) and the exposure time (t) of seeds in the environment. A formula is constructed:

[0115] M = k1 × (D + T + E) × t;

[0116] Among them, k1 is a coefficient related to the characteristics of the seed itself, and the k1 value may be different for different seed varieties.

[0117] In actual operation, taking the degree of drought stress in a valley as an example, if the degree of drought stress D in a certain area can be measured by the range of soil moisture content (such as from 80% of normal moisture content to 30% during the drought period, then D = 50%), the temperature change range T is the temperature difference between day and night (such as 20°C), and the difference in soil trace element content E can be calculated by the change ratio of the content of several key trace elements (such as iron, zinc, and manganese) (assuming that the average content difference of these three elements is 30%, then:

[0118] M=k1×(0.5+20+0.3)×912.5;

[0119] Assuming that k1=0.001 is obtained through preliminary experimental fitting (this is just a hypothetical value, and more precise experimental determination is required in practice), then the memory strength of the seeds in this area (the memory strength value here is only a relative concept, used to compare the strength of seed memory in different environments) is.

[0120] Relationship model between seed offspring yield, seed memory and environmental adaptability:

[0121] Assuming that the yield of seed offspring (Y) is related to the memory strength of the seeds (M) and the degree of adaptability of the seed offspring to the planting environment (A), the formula can be constructed:

[0122] M = k2 × M × A;

[0123] Where k2 is another coefficient related to crop yield formation;

[0124] In the subsequent planting process, for the seed offspring that have been pretreated and screened in different environments, their adaptability A to a specific environment can be quantitatively scored by observing their growth conditions in the environment (such as seedling survival rate, pest and disease incidence, etc.) (If the seedling survival rate is 85% and the pest and disease incidence rate is 10%, the adaptability A = 0.8 can be calculated based on a certain weight distribution (this is also a simplified quantitative method, and more factors may need to be considered in actual situations). Assuming that k2 = 0.1 (also an assumed value) is obtained through preliminary experiments, for the seed offspring with the above memory strength M = 18.8125, in a specific planting environment, if its adaptability A = 0.8, its yield is predicted according to the formula:

[0125] Y=0.1×18.8125×0.8=1.505;

[0126] (The yield value here is also a relative concept, used to compare the yield potential of different seed offspring under different conditions).

[0127] Through the above-mentioned multiple formula models, preliminary quantitative analysis and prediction of the relationship between seed memory and yield can be carried out.

[0128] Summarize:

[0129] Through the embodiments of the above-mentioned breeding methods, from seed memory induction, marker screening, memory consolidation breeding to memory verification and optimization adjustment, full use is made of the complexity of the natural environment, the diversity of natural biological communities, and the uncontrollability and randomness of natural factors.

[0130] In each link, specific data are used to prove the effective use of this breeding method for cross-generational seed memory and the "memory effect", solving the dilemma in existing technologies that it is difficult to determine the specific influencing factors and mechanisms of seed memory and difficult to use or avoid them in breeding.

[0131] During the seed pretreatment stage, the natural environment causes the seeds to form a variety of memory imprints, which improves the adaptability and yield potential of future generations to the corresponding environment; labeling technology based on plant autoimmune responses and biological community screening methods accurately screen out seeds with excellent memory characteristics, providing high-quality materials for breeding; environmental relay induction and hybridization and mutagenesis breeding further consolidate and expand the favorable memory of seeds and improve the comprehensive performance of future generations; memory verification and optimization adjustment ensure the continuous improvement and optimization of breeding methods, which steadily increases the yield of millet and enhances its stability.

[0132] This breeding method has opened up a new path for increasing millet yield, and also provided useful reference and reference for other crop breeding. It is expected to promote the development of the entire agricultural breeding field in utilizing the natural memory characteristics of seeds, thereby responding to the increasingly complex challenges of the natural environment and ensuring the sustainability and stability of food production.

Claims

1. A breeding method for increasing millet yield, characterized in that: The following steps are involved: S1. Construction of seed memory induction environment and seed pretreatment: Selection of natural environment and planting of parents: valleys with changeable weather, rich in various trace elements and geomagnetic anomalies are selected as seed pretreatment sites. Parent plants are planted for 2-3 years to allow the seeds to naturally contact and remember complex environmental factors, and then they are collected in groups according to the environment of the harvested small area; Seed storage and memory enhancement: Use local porous rocks to make containers, fill them with fermented plant residues and mixed powder, and use the decomposition to produce VOCs with naturally changing composition and concentration to create an odor environment; combine metal coils made of waste wires with small solar panels to generate unstable magnetic fields; S2. Seed memory related markers and screening: Marking based on plant autoimmune response: Extract the defense-related proteins of millet, catalyze the bio-crosslinking with natural pigment molecules through the specific enzymes produced by microbial fermentation, and make a marking complex to apply to the surface of seeds. Based on the changes in protein quantity caused by changes in defense-related gene expression due to environmental pressure inside the seeds, the markers are made to specifically bind to seed cells to mark memory-related change areas; Biome screening: sowing the marked seeds in wetlands beside rivers, clearings between forests, and abandoned farmland, relying on, but not limited to, the interaction of insects, birds, and microorganisms with seeds and seedlings for screening, setting up obstacles to change the biological activity paths to amplify the differences in seed memory characteristics, and screening memory characteristic seeds through, but not limited to, the ability to attract pollinating insects and resistance to soil microbial diseases; S3. Directed memory consolidation and breeding: Environmental relay induction: the selected seeds are transferred to a natural area that is partially similar to the pre-treated site but has some differences for 1-2 years without artificial irrigation and fertilization; Hybridization and mutation breeding: Hybrid breeding selects combinations based on the compatibility and complementarity of seeds to different natural environments.

2. A breeding method for increasing millet yield as claimed in claim 1, characterized in that: The following steps are also included: S4. Memory verification and optimization adjustment: Intergenerational comparative observation: Continuously plant multiple generations of new varieties and control varieties in multiple natural planting plots. In addition to conventional quantitative data recording, draw a map of seed and seedling morphological changes, compare and observe the intergenerational inheritance evolution of memory-related traits of new varieties, and combine multi-dimensional verification methods with graphics and data; Natural feedback optimization and adjustment: If a certain generation of seeds has poor specific natural environmental traits, including but not limited to reduced yield in drought years, the sowing time is determined according to local phenological phenomena, including but not limited to migratory bird migration and wild flower flowering period. Some seeds are sown back into similar memory-inducing natural sites, and the memory characteristics are optimized by using the association between natural phenology and seed growth. Repeated adjustments are used for continuous optimization of breeding strategies.

3. A breeding method for improving millet yield as claimed in claim 1, characterized in that: When selecting valley areas for seed memory induction environment construction and seed pretreatment, the direction of the valley is limited to an angle of 30°-60° with the local main wind direction. At the same time, intercropping is used to introduce a variety of local wild herbs and mix them with the millet parent plants.

4. A breeding method for improving millet yield as claimed in claim 1, characterized in that: In the seed memory-related labeling technology, a cross-linking method based on specific enzyme catalysis produced by microbial fermentation is used in the biological cross-linking process between defense-related proteins and natural pigment molecules.

5. A breeding method for improving millet yield as claimed in claim 1, characterized in that: During the biological community screening process, obstacles of different heights and materials are set up to change the paths and ways in which surrounding organisms enter the experimental field.

6. A breeding method for increasing millet yield as claimed in claim 1, characterized in that: During the environmental relay induction process, areas where wild animals with different foraging habits frequently appear are selected as new natural environment areas to transfer seeds.

7. A breeding method for increasing millet yield as claimed in claim 1, characterized in that: In the process of induced breeding using the natural environment, shielding objects of different materials and thicknesses are set up to create small areas with different radiation dose gradients, and the radiation dose is allowed to change dynamically with natural wind and rain.

8. A breeding method for increasing millet yield as claimed in claim 1, characterized in that: When recording various seed indicators during intergenerational comparative observations, observations are combined with drawing maps of seed and seedling morphological changes.

9. A breeding method for increasing millet yield as claimed in claim 1, characterized in that: During the adjustment process of natural feedback optimization, the time for re-sowing seeds is determined based on local phenological phenomena.

Images