Method for breeding new variety of high-yield coptis chinensis with high content of berberine

Abstract

The application discloses a high-small-bark-cocaine-content high-yield-type new variety of Coptis chinensis breeding method, relates to the Coptis chinensis breeding technical field, and aims at solving the low yield of Coptis chinensis. The application sets a small-bark-cocaine-content hard threshold of 5.5%, combines with actual planting related indexes such as agronomy and stress resistance, and comprehensively screens germplasm, so that the screening standard is more definite, and high-quality germplasm can be accurately screened. From field cultivation to multi-dimensional identification to three rounds of reselection, the logic is closed, the operation is standard, and the high-yield, high-quality and stable Coptis chinensis excellent single strain can be effectively directionally bred.

Description

Technical Field

[0001] This invention relates to the field of Coptis chinensis breeding technology, specifically to a method for breeding a high-yielding new Coptis chinensis variety with high berberine content. Background Technology

[0002] As an important medicinal plant, the berberine content and yield of Coptis chinensis are core breeding indicators. Current Coptis chinensis breeding suffers from problems such as a narrow genetic base of germplasm resources, a single screening index lacking quantitative standards, irregular parental selection and cross-pollination management, neglect of trait stability and complementarity in progeny selection, and a lack of multi-ecological-region adaptability verification. This easily leads to uneven quality and poor adaptability of new varieties, making it difficult to balance high berberine content and high yield, thus hindering the improvement of quality and efficiency in the Coptis chinensis industry. Summary of the Invention

[0003] The purpose of this invention is to provide a method for breeding high-yielding new varieties of Coptis chinensis with high berberine content. By setting a hard threshold of 5.5% for berberine content and combining it with agronomic and stress-resistant indicators related to actual planting, the germplasm is comprehensively screened, making the screening criteria clearer and facilitating the precise selection of high-quality germplasm, which can solve the problems in the existing technology.

[0004] To achieve the above objectives, the present invention provides the following technical solution:

[0005] A method for breeding a high-yielding new Coptis chinensis variety with high berberine content, characterized by comprising the following steps:

[0006] S1: Germplasm resource collection and processing: Collect Coptis chinensis germplasm resources from different sources and with different genotypes, and perform standardized pretreatment on the collected germplasm;

[0007] S2: Initial screening of germplasm resources: The standardized pre-treated germplasm resources are evaluated by multiple indicators, and the initial screening germplasm resources are selected based on the results of the multi-indicator comprehensive evaluation.

[0008] S3: High-yield trait identification: The germplasm resources initially screened are evaluated for yield-related traits, and the parents are selected after the evaluation is completed;

[0009] S4: Parental lines and cross-pollination: Artificial cross-pollination is performed on the selected parents;

[0010] S5: Hybrid offspring cultivation: The parent plants that have completed hybridization and pollination are cultivated to obtain hybrid seedlings, which are then tested and screened.

[0011] S6: Single plant identification and re-selection: Cultivate and identify the single plants after the initial selection, and obtain superior single plant lines based on the identification results;

[0012] S7: Comparative test: Superior single plants are cultivated and tested. Based on the results of the cultivation test, strains are screened, and candidate strains are obtained after the screening is completed.

[0013] Preferably, for S1, Coptis chinensis germplasm resources from different sources and with different genotypes are collected, and the collected germplasm undergoes standardized pretreatment, including:

[0014] The germplasm resources of Coptis chinensis were collected from wild germplasm resource banks, cultivated germplasm resource banks and scientific research institutions;

[0015] The types of Coptis chinensis germplasm materials collected include seeds, seedlings, and rhizome materials;

[0016] The collected Coptis chinensis germplasm resources were subjected to standardized pretreatment;

[0017] After standardization, germplasm resources are obtained for initial screening.

[0018] Preferably, a multi-indicator comprehensive evaluation is performed on the standardized pretreated germplasm resources in S2, and preliminary germplasm resources are selected based on the results of the multi-indicator comprehensive evaluation, including:

[0019] First, confirm the comprehensive evaluation indicators for multiple indicators;

[0020] After the multi-indicator comprehensive evaluation indicators are confirmed, the germplasm resources to be screened will be numbered and randomly arranged.

[0021] The germplasm resources awaiting initial screening in the experimental area will be observed during their growth period based on a comprehensive evaluation index of multiple indicators.

[0022] After the observation of the growing season is completed, the disease situation is observed, and the degree of disease is classified according to the observation results;

[0023] Harvest period traits will be measured on germplasm resources awaiting initial screening in the experimental area. The harvest period traits will be measured in the autumn of the third year after transplanting, including the fresh weight of rhizomes per plant, the morphological index of rhizomes, and the dry weight percentage.

[0024] The germplasm resources that have been harvested and tested and are to be screened at the initial stage are dried by drying the rhizomes. After drying, the rhizomes are crushed and sieved. The berberine content of the powder of the germplasm resources to be screened at the initial stage is determined by high performance liquid chromatography.

[0025] Germplasm resources with berberine content higher than 5.5% in the berberine content determination results were used as primary screening germplasm resources.

[0026] Preferably, yield-related traits are evaluated for the germplasm resources initially screened in S3, and parents are selected after the evaluation, including:

[0027] First, confirm the yield-related trait evaluation indicators, and at the same time, track the berberine content simultaneously;

[0028] The initial screening germplasm resources were compared with the control group based on yield-related trait evaluation indicators, and the yield increase rate was calculated. The control group was selected from the locally cultivated Coptis chinensis varieties.

[0029] Germplasm resources with a yield increase rate exceeding 15% were marked as outstanding germplasm; germplasm resources with a yield increase rate between 5% and 15% were marked as good germplasm; and germplasm resources with a yield increase rate below 5% were removed.

[0030] The outstanding and good germplasm resources from the initial screening will be used for parent selection.

[0031] Preferably, artificial hybridization pollination is performed on the selected parents in S4, including:

[0032] The selected parent plants were separated into female and male parents, and the female and male parents were transplanted to the hybridization experimental field for isolated planting.

[0033] Before isolating the plants, the female parent is emasculated and the male parent's pollen is collected.

[0034] After pollen collection, viability testing was performed, and pollen with a germination rate higher than 60% was applied to the emasculated female parent.

[0035] After pollination, the parent plants are bagged.

[0036] Confirm successful pollination 15-20 days after pollination. After successful pollination, remove the bag and label the mother plant after removing the bag.

[0037] Finally, the pollination process of the parent plants is completed.

[0038] Preferably, the parent plants that have completed hybridization and pollination in S5 are cultivated to obtain hybrid seedlings. These seedlings are then tested and screened, including:

[0039] Sow the pollinated parent plants and manage the seedlings after sowing;

[0040] After the parent lines are sown, testing and screening are carried out, including the following steps:

[0041] S501: First round of screening: The first round of screening is the initial screening of growth shape. The initial screening indicators of growth shape include: growth vigor, leaf characteristics, plant type and root system.

[0042] S502: Second round of screening: The second round of screening is berberine content detection screening. Berberine content detection screening involves testing the berberine content of the seedlings retained in the first round of screening and retaining seedlings with a berberine content ≥5.5%.

[0043] S503: Third round of screening: The third round of screening is a comprehensive trait verification screening. The comprehensive trait verification screening is to verify the comprehensive traits of the seedlings retained in the second round of screening, and retain seedlings with a stable berberine content of 5.5% or above and growth vigor higher than the average level.

[0044] Finally, hybrid seedlings were obtained.

[0045] Preferably, the selected individual plants in S6 are cultivated and identified, and superior single-plant lines are obtained based on the identification results, including:

[0046] According to the standards of the experimental area, the hybrid seedlings were planted individually in the experimental area for cultivation, and seedling management was carried out at the same time.

[0047] Then, individual plants of the hybrid seedlings were identified.

[0048] After the identification is completed, individual selection of hybrid seedlings is carried out, including the following steps:

[0049] S601: First round of re-selection: The first round of re-selection is based on basic indicators. Hybrid seedlings with berberine content ≥5.5%; single plant root and stem dry weight higher than the average level of single plants; dry weight ≥35%; and disease severity ≤1 are selected.

[0050] S602: Second round of selection: The second round of selection is a comprehensive trait ranking. The hybrid seedlings retained in the first round of selection are combined with the results of agronomic trait identification, high yield trait identification, medicinal component identification, stress resistance identification and stability identification, and the top 30% of hybrid seedlings are retained.

[0051] S603: Third round of selection: The third round of selection is a trait complementarity screening. The hybrid seedlings retained in the second round of selection are combined with the genetic background and trait performance of the male and female parents to conduct trait complementarity screening and retain the individual plants with complementary advantages.

[0052] The selected hybrid seedlings were renumbered and the parent numbers of the male and female parents were marked.

[0053] Ultimately, superior single-strain lines were obtained.

[0054] Preferably, superior individual plants from S7 are subjected to cultivation trials. Based on the results of the cultivation trials, strains are screened, and candidate strains are obtained after screening, including:

[0055] The superior individual plants were cultivated again, and the cultivation process was the same as that in step S6.

[0056] After the superior single plant is cultivated, strain selection is carried out, which includes the following steps:

[0057] S701: Superior individual plants with berberine content ≥5.5%; root and stem dry weight ≥10% higher than the control group; yield per unit area ≥8% higher than the control group; dry weight ≥35%; and disease severity ≤1 will be retained.

[0058] S702: The superior individual plants that were retained were compared in terms of yield, medicinal components, agronomic trait uniformity, and stress resistance. Based on the comparison results, the superior individual plants that performed better than the control group were retained.

[0059] S703: Confirm the growth traits and medicinal component content data of the superior individual plants retained in the previous step, and retain the superior individual plants with growth traits and medicinal component content higher than the average level.

[0060] Ultimately, a high-yielding candidate strain of Coptis chinensis was obtained.

[0061] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0062] 1. The present invention provides a method for breeding high-yielding new varieties of Coptis chinensis with high berberine content. Through standardized pretreatment and multi-dimensional quantitative screening, germplasm quality is controlled at each level, which solves the problems of narrow genetic basis and single screening index in breeding.

[0063] 2. The present invention provides a method for breeding a high-yielding new Coptis chinensis variety with high berberine content, which constructs a standardized breeding system for the entire process, sets quantitative standards from hybridization and pollination to offspring screening, takes into account the stability and complementarity of traits, and effectively avoids variety degeneration.

[0064] 3. The present invention provides a method for breeding a high-yielding new Coptis chinensis variety with high berberine content. It involves conducting multi-ecological region adaptability tests, combining standardized propagation and quality control to ensure that the new variety has wide adaptability and high propagation purity. Attached Figure Description

[0065] Figure 1 This is a schematic diagram illustrating the breeding steps for a high-yielding new Coptis chinensis variety according to the present invention. Detailed Implementation

[0066] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0067] To address the problems in existing Coptis chinensis breeding, such as narrow genetic base of germplasm resources, single screening indicators, and lack of quantitative standards for parental selection, please refer to [link / reference needed]. Figure 1 This embodiment provides the following technical solution:

[0068] A method for breeding a high-yielding new Coptis chinensis variety with high berberine content, characterized by comprising the following steps:

[0069] S1: Germplasm resource collection and processing: Collect Coptis chinensis germplasm resources from different sources and with different genotypes, and perform standardized pretreatment on the collected germplasm;

[0070] S2: Initial screening of germplasm resources: The standardized pre-treated germplasm resources are evaluated by multiple indicators, and the initial screening germplasm resources are selected based on the results of the multi-indicator comprehensive evaluation.

[0071] S3: High-yield trait identification: The germplasm resources initially screened are evaluated for yield-related traits, and the parents are selected after the evaluation is completed;

[0072] S4: Parental lines and cross-pollination: Artificial cross-pollination is performed on the selected parents;

[0073] S5: Hybrid offspring cultivation: The parent plants that have completed hybridization and pollination are cultivated to obtain hybrid seedlings, which are then tested and screened.

[0074] S6: Single plant identification and re-selection: Cultivate and identify the single plants after the initial selection, and obtain superior single plant lines based on the identification results;

[0075] S7: Comparative test: Superior single plants are cultivated and tested. Based on the results of the cultivation test, strains are screened and candidate strains are obtained after the screening is completed.

[0076] S8: Regional adaptability test: The candidate lines obtained will be tested in different ecological regions, and the final new Coptis chinensis varieties will be selected based on the test results;

[0077] S9: Propagation of new varieties: The data of the final selected new Coptis chinensis varieties will be compiled and then propagated.

[0078] For S1, different sources and genotypes of Coptis chinensis germplasm resources were collected, and the collected germplasm underwent standardized pretreatment, including:

[0079] The germplasm resources of Coptis chinensis were collected from wild germplasm resource banks, cultivated germplasm resource banks and scientific research institutions;

[0080] The types of Coptis chinensis germplasm materials collected include seeds, seedlings, and rhizome materials;

[0081] The collected Coptis chinensis germplasm resources were subjected to standardized pretreatment, which included: impurity removal, appearance screening, pruning, disinfection and activation.

[0082] After standardization, germplasm resources are obtained for initial screening.

[0083] Specifically, through multi-channel and multi-type germplasm collection and standardized pretreatment, a solid foundation is laid for Coptis chinensis breeding and resource conservation. Germplasm is comprehensively collected from wild and cultivated germplasm resource banks and research institutions, covering different ecotypes and genotypes, greatly enriching genetic diversity, avoiding the narrow genetic base caused by a single source, and enhancing subsequent breeding potential. This multi-channel and multi-type germplasm collection and standardized pretreatment lays a solid foundation for Coptis chinensis breeding and resource conservation.

[0084] A multi-indicator comprehensive evaluation was conducted on the standardized pretreated germplasm resources in S2. Based on the results of the multi-indicator comprehensive evaluation, preliminary screening germplasm resources were selected, including:

[0085] First, the comprehensive evaluation indicators for multiple indicators are confirmed, including indicators of medicinal active ingredients, agronomic morphology, yield-related indicators, growth vigor, and stress resistance.

[0086] After the multi-indicator comprehensive evaluation indicators are confirmed, the germplasm resources to be screened will be numbered and randomly arranged.

[0087] Each germplasm material of the germplasm resources to be screened is given a unique number, and then each germplasm material is assigned to the corresponding experimental area using a randomized block design.

[0088] Among them, the experimental area selected flat, well-drained, fertile soil plots with previous crops that were not Coptis chinensis or plants of the same family as the evaluation experimental fields. The soil pH value was controlled between 5.5 and 6.5, and the shading degree was controlled between 60% and 70%. The experimental area was divided into several plots, and the area of ​​each plot was determined according to the number of germplasm resources.

[0089] The germplasm resources to be screened in the experimental area were observed during their growth period according to a multi-index comprehensive evaluation system. The growth period included the seedling stage, vigorous growth stage, budding stage, flowering stage, and rhizome enlargement stage.

[0090] Growth period observation data include emergence period, emergence rate, plant height, number of leaves, and leaf area;

[0091] After the observation of the growing season is completed, the disease situation is observed, and the degree of disease is classified according to the observation results;

[0092] Among them, level 0 means no disease spots; level 1 means the disease spot area accounts for less than 5% of the leaf area; level 2 means the disease spot area accounts for 5% to 25%; level 3 means the disease spot area accounts for 25% to 50%; and level 4 means the disease spot area accounts for more than 50%.

[0093] Harvest period traits will be measured on germplasm resources awaiting initial screening in the experimental area. The harvest period traits will be measured in the autumn of the third year after transplanting, including the fresh weight of rhizomes per plant, the morphological index of rhizomes, and the dry weight percentage.

[0094] The germplasm resources that have been harvested and tested and are to be screened at the initial stage are dried by drying the rhizomes. After drying, the rhizomes are crushed and sieved. The berberine content of the powder of the germplasm resources to be screened at the initial stage is determined by high performance liquid chromatography.

[0095] Germplasm resources with berberine content higher than 5.5% in the berberine content determination results were used as primary screening germplasm resources.

[0096] Specifically, before the initial screening, five major evaluation indicators, including medicinal active ingredients and agronomic morphology, are clearly defined. Simultaneously, standardized pre-treated germplasm resources are uniquely numbered and randomly arranged in blocks. This ensures comprehensive and complete evaluation dimensions while guaranteeing fair experimental conditions for each germplasm type. Standardized conditions such as soil and shading in the experimental fields are uniformly regulated, and standard plots are divided, continuing the standardized treatment logic described earlier to eliminate interference from environmental differences. This ensures consistent evaluation environments for germplasm from different sources, improving the comparability of screening results. Based on clearly defined multi-stage observation requirements throughout the growth period and a clear disease grading standard, the growth status and disease severity of the germplasm are systematically recorded. This achieves precise quantification of each indicator and a clear understanding of the germplasm's overall growth performance. By setting a hard threshold of 5.5% berberine content, combined with agronomic and stress-resistance indicators relevant to actual planting, a comprehensive screening of germplasm is conducted, making the screening criteria clearer and facilitating the precise selection of high-quality germplasm.

[0097] For the germplasm resources initially screened in S3, yield-related traits were evaluated, and after the evaluation, parents were selected, including:

[0098] First, the yield-related trait evaluation indicators were confirmed. These indicators included fresh weight of single rhizome, dry weight of single rhizome, rhizome fullness, number of rhizome branches, rhizome length, rhizome thickness, dry weight, yield per unit area, and aboveground biomass. At the same time, the berberine content was tracked synchronously.

[0099] The initial screening germplasm resources were compared with the control group based on yield-related trait evaluation indicators, and the yield increase rate was calculated. The control group was selected from the locally cultivated Coptis chinensis varieties.

[0100] Germplasm resources with a yield increase rate exceeding 15% were marked as outstanding germplasm; germplasm resources with a yield increase rate between 5% and 15% were marked as good germplasm; and germplasm resources with a yield increase rate below 5% were removed.

[0101] Parental selection will be conducted on outstanding and good germplasm resources from the initial screening.

[0102] The parent selection criteria are as follows: among the outstanding and good germplasm, those with berberine content ≥5.5%, rhizome dry weight ≥35%, and single plant rhizome dry weight in the top 20% are selected as parents.

[0103] Specifically, before screening parent lines, nine yield-related assessment indicators, such as fresh weight of single plant rhizomes and yield per unit area, are clearly defined, while berberine content is tracked simultaneously. This ensures that the yield assessment dimensions are comprehensive and without omissions, while also taking into account the core medicinal value. Using the locally cultivated Coptis chinensis variety as a control group, the pre-screened germplasm is compared with the control group, and the yield increase rate is calculated. Continuing the quantitative evaluation logic mentioned above, the germplasm grading standards for different yield increase ranges are clarified to eliminate interference caused by varietal differences. By setting clear yield increase rate thresholds, the pre-screened germplasm is graded, labeled, and screened, eliminating germplasm with poor yield increase effects, focusing on high-quality germplasm resources, and improving the efficiency of parent line screening. Based on the rigid standards of berberine content ≥5.5%, dry weight ≥35%, and top 20% of single plant rhizome dry weight, outstanding and good germplasm is selected as parent lines, ensuring both the medicinal value and yield potential of the parent lines, as well as their excellent quality.

[0104] To address the problems in existing Coptis chinensis hybridization breeding, such as low success rate of artificial pollination, high risk of self-pollination contamination, single and lack of quantitative standards for screening hybrid offspring, and neglect of trait stability and complementarity, please refer to [the relevant documentation / reference]. Figure 1 This embodiment provides the following technical solution:

[0105] Artificial hybridization and pollination will be carried out on the selected parents in S4, including:

[0106] The selected parent plants were separated into female and male parents, and the female and male parents were transplanted to the hybridization experimental field for isolated planting.

[0107] Before isolating the plants, the female parent is emasculated and the male parent's pollen is collected.

[0108] After pollen collection, viability testing was performed, and pollen with a germination rate higher than 60% was applied to the emasculated female parent.

[0109] After pollination, the parent plants are bagged.

[0110] Confirm successful pollination 15-20 days after pollination. After successful pollination, remove the bag and label the mother plant after removing the bag.

[0111] Finally, the pollination process of the parent plants is completed.

[0112] Specifically, before hybridization, the parent plants are separated into female and male lines and transplanted to a hybridization experimental field for isolated planting. At the same time, the female line is emasculated and the pollen of the male line is collected to avoid interference from natural hybridization and ensure the purity of the pollination material. After pollen collection, viability testing is performed, and only pollen with a germination rate higher than 60% is selected for pollination. Following the quantitative standard logic mentioned earlier, pollen with insufficient viability is eliminated to ensure the pollination success rate. After pollination, the female line is bagged to prevent contamination by exogenous pollen. After 15-20 days, the bag is removed and labeled after confirming successful pollination. The steps are standardized and traceable to ensure the clear origin of the hybrid offspring.

[0113] The parent plants that have completed hybridization and pollination in S5 are then cultivated to obtain hybrid seedlings. These seedlings are then tested and screened, including:

[0114] After pollination, the parent plants are sown. Seedling management is carried out after sowing, which includes water management, temperature management, thinning, seedling establishment, topdressing, weeding, loosening the soil and pest and disease control.

[0115] After the parent lines are sown, testing and screening are carried out, including the following steps:

[0116] S501: First round of screening: The first round of screening is the initial screening of growth shape. The initial screening indicators of growth shape include: growth vigor, leaf characteristics, plant type and root system.

[0117] S502: Second round of screening: The second round of screening is berberine content detection screening. Berberine content detection screening involves testing the berberine content of the seedlings retained in the first round of screening and retaining seedlings with a berberine content ≥5.5%.

[0118] S503: Third round of screening: The third round of screening is a comprehensive trait verification screening. The comprehensive trait verification screening is to verify the comprehensive traits of the seedlings retained in the second round of screening, and retain seedlings with a stable berberine content of 5.5% or above and growth vigor higher than the average level.

[0119] Finally, hybrid seedlings were obtained.

[0120] Specifically, after hybridization and pollination, the parent plants are sown and comprehensive seedling management, including water and temperature control, is carried out to provide a suitable environment for seedling growth and reduce the impact of seedling diseases and pests. A three-round gradient screening is adopted. The first round focuses on growth traits and eliminates seedlings with abnormal growth, continuing the quantitative screening logic mentioned above to ensure that the remaining seedlings have excellent growth. The second round specifically tests the berberine content, with ≥5.4% as the threshold to ensure the medicinal value of the seedlings. The third round reviews the comprehensive traits and selects individuals with stable medicinal content and outstanding growth.

[0121] For S6, the selected individual plants were cultivated and identified, and superior single-plant lines were obtained based on the identification results, including:

[0122] According to the standards of the experimental area, the hybrid seedlings were planted individually in the experimental area for cultivation, and seedling management was carried out at the same time.

[0123] Then, individual plant identification was performed on the hybrid seedlings, including agronomic traits identification, high-yield traits identification, medicinal components identification, stress resistance identification, and stability identification.

[0124] Among them, the agronomic trait identification involves measuring the agronomic traits of each individual plant, including plant height, number of leaves, single leaf area, root and stem length, root and stem thickness, number of root and stem branches, root and stem fullness, fibrous root density, and aboveground biomass; the yield trait identification is based on yield-related trait evaluation indicators, focusing on identifying the yield potential of individual plants, including fresh weight of root and stem of individual plants, dry weight of root and stem of individual plants, and dry weight percentage. The determination methods are consistent with those for the harvest period traits; the medicinal component identification involves determining the berberine content in the rhizomes of each individual plant; the stress resistance identification involves comprehensively assessing the stress resistance of individual plants by combining the observation records of pests and diseases during the cultivation period, with identification indicators including disease resistance, shade tolerance, and moisture tolerance; the stability identification involves comparing the growth traits and effective component content data of individual plants to analyze their trait stability, focusing on observing the interannual changes in rhizome growth rate and berberine content, and selecting individual plants with stable growth traits, small fluctuations in berberine content, and no obvious trait degeneration;

[0125] After the identification is completed, individual selection of hybrid seedlings is carried out, including the following steps:

[0126] S601: First round of re-selection: The first round of re-selection is based on basic indicators. Hybrid seedlings with berberine content ≥5.5%; single plant root and stem dry weight higher than the average level of single plants; dry weight ≥35%; and disease severity ≤1 are selected.

[0127] S602: Second round of selection: The second round of selection is a comprehensive trait ranking. The hybrid seedlings retained in the first round of selection are combined with the results of agronomic trait identification, high yield trait identification, medicinal component identification, stress resistance identification and stability identification, and the top 30% of hybrid seedlings are retained.

[0128] S603: Third round of selection: The third round of selection is a trait complementarity screening. The hybrid seedlings retained in the second round of selection are combined with the genetic background and trait performance of the male and female parents to conduct trait complementarity screening and retain the individual plants with complementary advantages.

[0129] The selected hybrid seedlings were renumbered and the parent numbers of the male and female parents were marked.

[0130] Ultimately, superior single-strain lines were obtained.

[0131] Specifically, the initially selected individual plants were first planted and meticulously managed according to the standards of the experimental area to provide a stable environment for the expression of individual plant traits and ensure the authenticity and reliability of the identification results. A comprehensive identification system was established, covering five dimensions: agronomy, high yield, medicinal properties, stress resistance, and stability. Indicators covered plant type, yield, core components, and stress resistance, comprehensively reflecting the overall quality of individual plants. Among these, stress resistance and stability identification were particularly crucial. Through pest and disease records and interannual variation analysis, materials with genetically stable traits were accurately screened, effectively avoiding the risk of later trait degradation. The reselection process adopted a progressive screening mechanism: the first round used a hard threshold to eliminate inferior individuals; the second round used multi-dimensional comprehensive ranking to select the best among the best; and the third round combined complementary traits based on genetic background to achieve a precise leap from individual to superior strain.

[0132] To address the issues in existing Coptis chinensis breeding techniques, such as inconsistent strain selection criteria, lack of regional adaptability evaluation, insufficient stability analysis, incomplete variety records, and difficulty in controlling propagation purity, please refer to [the relevant documentation / reference]. Figure 1 This embodiment provides the following technical solution:

[0133] For S7, superior individual plants were bred and tested. Based on the results of the bred plants, strains were screened, and candidate strains were obtained after screening, including:

[0134] The superior individual plants were cultivated again, and the cultivation process was the same as that in step S6.

[0135] After the superior single plant is cultivated, strain selection is carried out, which includes the following steps:

[0136] S701: Superior individual plants with berberine content ≥5.5%; root and stem dry weight ≥10% higher than the control group; yield per unit area ≥8% higher than the control group; dry weight ≥35%; and disease severity ≤1 will be retained.

[0137] S702: The superior individual plants that were retained were compared in terms of yield, medicinal components, agronomic trait uniformity, and stress resistance. Based on the comparison results, the superior individual plants that performed better than the control group were retained.

[0138] S703: Confirm the growth traits and medicinal component content data of the superior individual plants retained in the previous step, and retain the superior individual plants with growth traits and medicinal component content higher than the average level.

[0139] Ultimately, a high-yielding candidate strain of Coptis chinensis was obtained.

[0140] Specifically, superior individual plants are cultivated using the S6 step, continuing the standardized cultivation logic described above to ensure consistency between the cultivation environment and identification standards, reducing screening bias caused by environmental differences and ensuring data comparability. The strain selection employs a three-round progressive screening process. The first round uses clearly quantified thresholds to identify individual plants that meet core indicators, focusing on high yield and medicinal needs, eliminating inferior individuals from the source. The second round uses multi-dimensional comparisons to ensure that the remaining individual plants outperform the control group, highlighting their high-yield advantage. The third round re-examines growth traits and medicinal components, further selecting the best among the best to ensure the stability of candidate strain quality.

[0141] The candidate lines obtained from S8 will be tested in different ecological regions. Based on the test results, the final new varieties of Coptis chinensis will be selected, including:

[0142] Select 3-4 experimental areas based on climate differences, soil differences, and cultivation conditions;

[0143] The candidate lines were planted in the selected experimental areas;

[0144] The yield stability analysis and berberine content stability analysis were performed on the candidate lines in each experimental area.

[0145] Among them, the yield stability analysis is to compare the yield performance of the same candidate line at different test sites. The line with small yield differences between different sites indicates that it has good stability and low sensitivity to the environment. The berberine content stability analysis is to analyze the fluctuation of berberine content in different ecological regions. The line with small fluctuations has more stable quality.

[0146] Based on the analysis results, preliminary screening of candidate strains was conducted;

[0147] The candidate lines retained from the initial screening underwent a second screening. The screening criteria included: the average dry weight of the rootstock per plant of the candidate lines was higher than that of the control variety, and the average yield increase rate was not less than 8%; the average dry weight reduction rate of the candidate lines was not less than 35%; the average disease index of the candidate lines was not higher than that of the control variety; and the yield variation coefficient of the candidate lines did not exceed 20%.

[0148] Meanwhile, the berberine content was above 6.0%, and the fluctuation between test areas was less than 0.3%;

[0149] The result was a new variety of Coptis chinensis.

[0150] Specifically, before screening, 3-4 experimental areas were selected based on differences in climate, soil, etc., and candidate lines were planted simultaneously to simulate different actual cultivation scenarios and ensure experimental coverage. The focus was on conducting stability analysis of yield and berberine content, clarifying stability evaluation standards, continuing the quantitative screening logic mentioned above, and accurately judging the environmental adaptability and quality stability of the lines. The second screening set clear hard thresholds, focusing on core indicators such as yield, dry weight, and disease index, and raising the berberine content standard to 6.0%.

[0151] Data will be compiled for the final selected new Coptis chinensis varieties from S9. After compilation, propagation will be carried out, including:

[0152] The basic information of the new Coptis chinensis varieties was compiled, including source information, variety characteristics and biological characteristics;

[0153] Next, the experimental data of the new Coptis chinensis variety were compiled, including a summary of yield data, quality data, stress resistance data, and agronomic trait data.

[0154] Cultivation data were compiled based on the cultivation characteristics of the new Coptis chinensis varieties.

[0155] After all the data has been compiled, the advantages of the new Coptis chinensis varieties will be summarized, including their yield advantages, quality advantages, and resistance advantages.

[0156] Based on the advantages of the new Coptis chinensis variety, the new variety is propagated, including division propagation and seed propagation;

[0157] Purity control, health control, and trait sampling are carried out during the breeding process.

[0158] New varieties of Coptis chinensis are graded for quality before being released from the nursery.

[0159] After grading, the breeding process for high-yielding new Coptis chinensis varieties is completed.

[0160] Specifically, we organized three types of data on new varieties: basic information, experimental data, and cultivation data. We comprehensively reviewed variety information and experimental data, and summarized the advantages in yield, quality, and resistance. This ensured that the variety information was complete and traceable, while clearly highlighting the core value of the variety. Propagation was carried out using both division and seed methods, taking into account both propagation efficiency and the genetic stability of the variety, continuing the quality control logic mentioned earlier. During propagation, we strictly carried out purity, health control, and trait sampling inspections. Before leaving the nursery, we conducted quality grading to ensure that the propagated seedlings were of high quality and uniform.

[0161] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0162] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention.

Claims

1. A method for breeding a high-yielding new Coptis chinensis variety with high berberine content, characterized in that, Includes the following steps: S1: Germplasm resource collection and processing: Collect Coptis chinensis germplasm resources from different sources and with different genotypes, and perform standardized pretreatment on the collected germplasm; S2: Initial screening of germplasm resources: The standardized pre-treated germplasm resources are evaluated by multiple indicators, and the initial screening germplasm resources are selected based on the results of the multi-indicator comprehensive evaluation. S3: High-yield trait identification: The germplasm resources initially screened are evaluated for yield-related traits, and the parents are selected after the evaluation is completed; S4: Parental lines and cross-pollination: Artificial cross-pollination is performed on the selected parents; S5: Hybrid offspring cultivation: The parent plants that have completed hybridization and pollination are cultivated to obtain hybrid seedlings, which are then tested and screened. S6: Single plant identification and re-selection: Cultivate and identify the single plants after the initial selection, and obtain superior single plant lines based on the identification results; S7: Comparative test: Superior single plants are cultivated and tested. Based on the results of the cultivation test, strains are screened, and candidate strains are obtained after the screening is completed.

2. The method for breeding a high-yielding new Coptis chinensis variety with high berberine content according to claim 1, characterized in that, For S1, different sources and genotypes of Coptis chinensis germplasm resources were collected, and the collected germplasm underwent standardized pretreatment, including: The germplasm resources of Coptis chinensis were collected from wild germplasm resource banks, cultivated germplasm resource banks and scientific research institutions; The types of Coptis chinensis germplasm materials collected include seeds, seedlings, and rhizome materials; The collected Coptis chinensis germplasm resources were subjected to standardized pretreatment; After standardization, germplasm resources are obtained for initial screening.

3. The method for breeding a high-yielding new Coptis chinensis variety with high berberine content according to claim 2, characterized in that, A multi-indicator comprehensive evaluation was conducted on the standardized pretreated germplasm resources in S2. Based on the results of the multi-indicator comprehensive evaluation, preliminary screening germplasm resources were selected, including: First, confirm the comprehensive evaluation indicators for multiple indicators; After the multi-indicator comprehensive evaluation indicators are confirmed, the germplasm resources to be screened will be numbered and randomly arranged. The germplasm resources awaiting initial screening in the experimental area will be observed during their growth period based on a comprehensive evaluation index of multiple indicators. After the observation of the growing season is completed, the disease situation is observed, and the degree of disease is classified according to the observation results; Harvest period traits will be determined for germplasm resources awaiting initial screening in the experimental area. The germplasm resources that have been harvested and tested and are to be screened at the initial stage are dried by drying the rhizomes. After drying, the rhizomes are crushed and sieved. The berberine content of the powder of the germplasm resources to be screened at the initial stage is determined by high performance liquid chromatography. Germplasm resources with berberine content higher than 5.5% in the berberine content determination results were used as primary screening germplasm resources.

4. The method for breeding a high-yielding new Coptis chinensis variety with high berberine content according to claim 3, characterized in that, For the germplasm resources initially screened in S3, yield-related traits were evaluated, and after the evaluation, parents were selected, including: First, the yield-related trait evaluation indicators were confirmed. These indicators included fresh weight of single rhizome, dry weight of single rhizome, rhizome fullness, number of rhizome branches, rhizome length, rhizome thickness, dry weight, yield per unit area, and aboveground biomass. At the same time, the berberine content was tracked synchronously. The initial screening germplasm resources were compared with the control group based on yield-related trait evaluation indicators, and the yield increase rate was calculated. The control group was selected from the locally cultivated Coptis chinensis varieties. Germplasm resources with a yield increase rate exceeding 15% were marked as outstanding germplasm; germplasm resources with a yield increase rate between 5% and 15% were marked as good germplasm; and germplasm resources with a yield increase rate below 5% were removed. The outstanding and good germplasm resources from the initial screening will be used for parent selection.

5. The method for breeding a high-yielding new Coptis chinensis variety with high berberine content according to claim 4, characterized in that, Artificial hybridization and pollination will be carried out on the selected parents in S4, including: The selected parent plants were separated into female and male parents, and the female and male parents were transplanted to the hybridization experimental field for isolated planting. Before isolating the plants, the female parent is emasculated and the male parent's pollen is collected. After pollen collection, viability testing was performed, and pollen with a germination rate higher than 60% was applied to the emasculated female parent. After pollination, the parent plants are bagged. Confirm successful pollination 15-20 days after pollination. After successful pollination, remove the bag and label the mother plant after removing the bag. Finally, the pollination process of the parent plants is completed.

6. The method for breeding a high-yielding new Coptis chinensis variety with high berberine content according to claim 5, characterized in that, The parent plants that have completed hybridization and pollination in S5 are then cultivated to obtain hybrid seedlings. These seedlings are then tested and screened, including: Sow the pollinated parent plants and manage the seedlings after sowing; After the parent lines are sown, testing and screening are carried out, including the following steps: S501: First round of screening: The first round of screening is the initial screening of growth shape. The initial screening indicators of growth shape include: growth vigor, leaf characteristics, plant type and root system. S502: Second round of screening: The second round of screening is berberine content detection screening. Berberine content detection screening involves testing the berberine content of the seedlings retained in the first round of screening and retaining seedlings with a berberine content ≥5.5%. S503: Third round of screening: The third round of screening is a comprehensive trait verification screening. The comprehensive trait verification screening is to verify the comprehensive traits of the seedlings retained in the second round of screening, and retain seedlings with a stable berberine content of 5.5% or above and growth vigor higher than the average level. Finally, hybrid seedlings were obtained.

7. The method for breeding a high-yielding new Coptis chinensis variety with high berberine content according to claim 6, characterized in that, For S6, the selected individual plants were cultivated and identified, and superior single-plant lines were obtained based on the identification results, including: According to the standards of the experimental area, the hybrid seedlings were planted individually in the experimental area for cultivation, and seedling management was carried out at the same time. Then, individual plants of the hybrid seedlings were identified. After the identification is completed, individual selection of hybrid seedlings is carried out, including the following steps: S601: First round of re-selection: The first round of re-selection is based on basic indicators. Hybrid seedlings with berberine content ≥5.5%; single plant root and stem dry weight higher than the average level of single plants; dry weight ≥35%; and disease severity ≤1 are selected. S602: Second round of selection: The second round of selection is a comprehensive trait ranking. The hybrid seedlings retained in the first round of selection are combined with the results of agronomic trait identification, high yield trait identification, medicinal component identification, stress resistance identification and stability identification, and the top 30% of hybrid seedlings are retained. S603: Third round of selection: The third round of selection is a trait complementarity screening. The hybrid seedlings retained in the second round of selection are combined with the genetic background and trait performance of the male and female parents to conduct trait complementarity screening and retain the individual plants with complementary advantages. The selected hybrid seedlings were renumbered and the parent numbers of the male and female parents were marked. Ultimately, superior single-strain lines were obtained.

8. The method for breeding a high-yielding new Coptis chinensis variety with high berberine content according to claim 7, characterized in that, For S7, superior individual plants were bred and tested. Based on the results of the bred plants, strains were screened, and candidate strains were obtained after screening, including: The superior individual plants were cultivated again, and the cultivation process was the same as that in step S6. After the superior single plant is cultivated, strain selection is carried out, which includes the following steps: S701: Superior individual plants with berberine content ≥5.5%; root and stem dry weight ≥10% higher than the control group; yield per unit area ≥8% higher than the control group; dry weight ≥35%; and disease severity ≤1 will be retained. S702: The superior individual plants that were retained were compared in terms of yield, medicinal components, agronomic trait uniformity, and stress resistance. Based on the comparison results, the superior individual plants that performed better than the control group were retained. S703: Confirm the growth traits and medicinal component content data of the superior individual plants retained in the previous step, and retain the superior individual plants with growth traits and medicinal component content higher than the average level. Ultimately, a high-yielding candidate strain of Coptis chinensis was obtained.

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