Plant factory breeding method for virus-free strawberry seedlings

Abstract

The invention belongs to the technical field of plant cultivation, and particularly relates to a plant factory breeding method of virus-free strawberry seedlings, which comprises the following steps: 1, producing breeder's seeds: performing stem tip detoxification on strawberry seedlings, and performing breeder's seed hardening and rooting; 2, breeder seed propagation: transplanting the breeder seeds into a plant factory, and carrying out breeder seed propagation; and 3, producing commercial seedlings: transferring the second-level seedlings into a seedling raising area, and carrying out large-scale breeding. In the breeding process, a phenotype recognition system based on image recognition and artificial intelligence analysis is introduced, and morphological characteristics of strawberry seedlings are dynamically monitored and digitally analyzed. The method has good replicability and generalization performance, industrialization promotion can be achieved, subjective errors caused by manual visual inspection grading are avoided through phenotype recognition, and grading consistency and efficiency are improved; data support is provided for subsequent breeding, disease and pest monitoring and accurate seedling raising; the system is linked with an environment monitoring system, and an intelligent seedling raising system integrating seedling condition, environment control and management can be constructed.

Description

Technical Field

[0001] The invention belongs to the technical field of plant cultivation, and particularly relates to a plant factory breeding method for virus-free strawberry seedlings. Background Art

[0002] With the advancement of society, the strawberry industry has also experienced rapid development, and the area of strawberry cultivation has increased year by year. However, due to the high requirements of strawberry seedlings for their growth environment, there is still no large-scale virus-free strawberry seedling production system in China.

[0003] From the perspective of strawberry seedling propagation, strawberry propagation currently generally adopts conventional cutting seedling propagation, tissue culture seedling propagation and other methods for seedling propagation. However, this traditional seedling propagation method has the following problems: (1) High virus infection rate: Conventional strawberry seedlings are mostly propagated by farmers themselves or purchased from the market, and the virus infection rate (such as strawberry mosaic virus, strawberry crinkle virus, etc.) is high, resulting in poor plant growth and reduced yield; (2) No standardized grading system: The current seedling propagation process lacks a systematic seedling grading and management system, and the phenomenon of mixing virus-free seedlings and ordinary seedlings is serious, affecting the planting efficiency; (3) Low seedling rate and long growth cycle: Although tissue culture seedlings can be virus-free, the survival rate is not high after being transferred to or raised in a nursery, and the environmental adaptability is poor, and the seedling management requirements are high; (4) Low seedling efficiency and high management cost: There is a lack of a standardized three-level propagation system, the environmental control is not precise, and the steps are not clear, resulting in a loose connection between seedlings at all levels and unstable seedling supply.

[0004] From the perspective of strawberry seedling production, the traditional tissue culture technology needs to go through the transition from "rooting in the bottle" to "seedling hardening and transplanting" during the production process of strawberry seedlings. This has three major systemic defects: (1) Low efficiency: Tissue culture seedlings need to be repeatedly transferred to the culture medium, and the production cycle of 60-75 days is too long to meet market demand; (2) Biosafety risks: The contamination rate of the open seedling hardening link exceeds 15%, resulting in 10%-20% seedling loss; (3) High cost: Manual operation accounts for 60%, and the annual energy consumption intensity reaches 1200kW·h / 10,000 plants.

[0005] The above problems have restricted the high-quality development of the strawberry industry. There is an urgent need for a scientific, efficient and systematic virus-free strawberry seedling breeding system to improve the quality of strawberry seedlings and industry benefits. Summary of the Invention

[0006] The object of the present invention is to provide a plant factory breeding method for virus-free strawberry seedlings to solve the technical problems mentioned in the above background technology.

[0007] In order to achieve the above object, the present invention discloses a plant factory breeding method for virus-free strawberry seedlings, comprising the following steps: 1. Production of original seed: Detoxify the stem tips of strawberry seedlings, and carry out hardening and rooting of original seed; 2. Expanding the stock: Transplanting the original stock into the plant factory for stock propagation; 3. Produce commercial seedlings: Move the secondary seedlings into the nursery area for large-scale breeding.

[0008] Furthermore, during the breeding process, a phenotypic recognition system based on image recognition and artificial intelligence analysis was introduced to dynamically monitor and digitally analyze the morphological characteristics of strawberry seedlings.

[0009] Furthermore, the steps of dynamically monitoring and digitally analyzing the morphological characteristics of strawberry seedlings include: (1) Model establishment: RGB cameras, hyperspectral cameras, or LiDAR are set up at key nodes in each seedling stage to acquire top view, side view, and local detail images of strawberry seedlings in real time. The large amount of collected image data is annotated, and a data model is established based on the annotated classification data. (2) Grading and screening control: Compare the actual collected images with the data model, screen out the targets and issue elimination instructions to achieve screening; Furthermore, the image data is captured under standard lighting conditions.

[0010] Furthermore, in step (1), the machine learning framework YOLOv8 is used for model training.

[0011] Furthermore, the classification data includes: seedling height, number of leaves, leaf area index, number of runners, branch angle, internode distance, seedling color, disease spots, curled leaves, color, and dwarfing.

[0012] Furthermore, during the dynamic monitoring and digital analysis of the morphological characteristics of strawberry seedlings, all identification data and individual seedlings are coded and bound, and uploaded to the seedling database to achieve traceability management of individual plants.

[0013] Furthermore, when detoxifying the stem tips of strawberry seedlings, the environmental control conditions were: temperature 25 / 22℃±1℃, humidity 60~70%, and carbon dioxide concentration 350±50 μL / L; During the hardening and rooting of the original seedlings, the environmental control is as follows: the temperature in the primary culture stage is controlled at 25 / 22℃±1℃, the humidity is 60~70%, the carbon dioxide concentration is 350±50μL / L, and the white light LED has a light intensity of 50~55±10μmol / m 2 / s, photoperiod 16 / 8h; from adventitious bud proliferation to rooting stage, temperature 25 / 18℃±1℃, humidity 70%, carbon dioxide concentration 350±50µL / L and later 700±50µL / L, white light LED, light intensity 150±10μmol / m 2 / s, photoperiod 16 / 8h; During the seedling hardening stage, the temperature was 21 / 19℃±1℃ and 16.4℃±1℃ in the later stage, the humidity was 80%, the carbon dioxide concentration was 350±50μL / L, and the white light LED had a light intensity of 90±10μmol / m 2 / s, photoperiod 12 / 12h.

[0014] Furthermore, when propagating the original species, the environmental control is as follows: temperature 25 / 20℃±1℃, humidity 75%±5%, carbon dioxide concentration 800±50µL / L, white light LED, light intensity 300±10μmol / m 2 / s, photoperiod 12 / 12h.

[0015] Furthermore, when producing commercial seedlings, the environmental control is as follows: the temperature is 25 / 20℃±1℃, the humidity is 75%±5%, the carbon dioxide concentration is 800±50µL / L, and the white light LED has a light intensity of 280±10μmol / m 2 / s, photoperiod 12 / 12h.

[0016] Compared with the prior art, the plant factory breeding method of the present invention for virus-free strawberry seedlings has the following advantages: (1) The strawberry seedlings obtained by the method of the present invention have higher quality and environmental adaptability: through virus-free treatment and graded breeding, the virus infection rate is effectively reduced, and the health level and survival rate of the strawberry seedlings are significantly improved; the virus-free seedlings are domesticated in stages, have strong adaptability, grow fast after transplanting, and have a high survival rate.

[0017] (2) The method of the present invention has high economic benefits: high-quality strawberry seedlings can increase per-acre yield, improve fruit quality and market competitiveness, and reduce disease prevention and control costs.

[0018] (3) The method of the present invention facilitates the construction of a standardized system: the three-level seedling cultivation system is clear, the process is standardized, and it is easy to manage, monitor and trace.

[0019] (4) The method of the present invention has good replicability and scalability, is suitable for large-scale commercial strawberry seedling production, and can be promoted industrially.

[0020] (5) In the present invention, phenotypic identification avoids subjective errors caused by manual visual grading, thereby improving grading consistency and efficiency; the visual recognition system supports rapid screening and high-throughput evaluation of massive seedlings, thereby improving the scale of seedling cultivation; it provides data support for subsequent breeding, pest and disease monitoring, and precise seedling cultivation; and it is linked to the environmental monitoring system to construct a three-in-one intelligent seedling cultivation system of "seedling condition + environmental control + management". DETAILED DESCRIPTION

[0021] The technical solution of the present invention is described in detail below through specific embodiments.

[0022] A plant factory breeding method for virus-free strawberry seedlings is a three-level seedling system for breeding virus-free strawberry seedlings in a plant factory. It aims to integrate the rooting and hardening links of strawberry tissue culture seedlings through precise environmental control and innovative facility applications, thereby improving the rooting rate, seedling index, survival rate after transplanting, and the number and quality of runners of strawberry original seedlings, shortening the seedling cycle, reducing labor costs and energy consumption, and thus improving the production efficiency of high-quality strawberry seedlings. The system consists of three stages: original seed production, original seed expansion, and commercial seedling cultivation. The core of the seedling system is "three-level stratification, virus-free passage, and traceable management" to ensure that the entire process from source to commercial seedling is virus-free and degenerate-free. Specifically, it includes the following steps: 1. Production of original seeds (first-grade seedling production) (1) Purchase strawberry seedlings from outside, sterilize the stem tips, and cultivate them into virus-free test tube seedlings under sterile conditions.

[0023] Environmental control: When preparing the shoot tip, the temperature is 25 / 22℃±1℃ (the temperature control range is minimum 22℃ and maximum 25℃, with a deviation of ±1℃ allowed), the humidity is 60~70%, and the carbon dioxide concentration is 350±50 μL / L.

[0024] (2) Test the test tube seedlings for viruses to ensure they are completely virus-free; (3) The original seedlings are hardened and rooted in a special rooting box under an artificially controlled environment; each seedling is equipped with a unique code for easy traceability.

[0025] Environmental control: During the primary culture stage, the temperature was controlled at 25 / 22℃±1℃, the humidity was 60~70%, the carbon dioxide concentration was 350±50μL / L, the white light LED had a light intensity of 50~55±10μmol / m2 / s, and the photoperiod was 16 / 8h.

[0026] From the adventitious bud proliferation to the rooting stage, the temperature was 25 / 18℃±1℃, the humidity was 70%, the carbon dioxide concentration was 350±50μL / L and later it was 700±50μL / L, the white light LED had a light intensity of 150±10μmol / m 2 / s, photoperiod 16 / 8h.

[0027] During the seedling hardening stage, the temperature was 21 / 19℃±1℃ and in the later stage, it was 16.4℃±1, the humidity was 80%, the carbon dioxide concentration was 350±50μL / L, and the white light LED had a light intensity of 90±10μmol / m 2 / s, photoperiod 12 / 12h.

[0028] 2. Propagation of original seeds (secondary seedling production) The original seed is transplanted into the plant factory for propagation.

[0029] Environmental control: Temperature 25 / 20℃±1℃, humidity 75%±5%, carbon dioxide concentration 800±50μL / L, white light LED, light intensity 300±10μmol / m 2 / s, photoperiod 12 / 12h.

[0030] Real-time monitoring of pests and diseases to ensure the health of seedlings; morphological screening to eliminate weak and deformed seedlings; 3. Production of commercial seedlings (Level 3 seedling production) The secondary seedlings are moved into the nursery area, usually in a glass greenhouse, for large-scale breeding.

[0031] Environmental control: Seedling temperature 25 / 20℃±1℃, humidity 75%±5%, carbon dioxide concentration 800±50 μL / L, white light LED, light intensity 280±10μmol / m 2 / s, photoperiod 12 / 12h.

[0032] Strict screening at the seedling stage, unified standard classification, and graded sales; During the strawberry seedling breeding process, in order to achieve quality control, graded management, and precise selection of virus-free strawberry seedlings at different seedling stages, this method introduces a phenotypic recognition system based on image recognition and artificial intelligence analysis throughout the entire production process of original seedlings, original seedlings, and commercial seedlings. This system dynamically monitors and digitally analyzes the morphological characteristics of strawberry seedlings, thereby improving the standardization, intelligence, and automation of seedling breeding. Specifically, it includes the following technical links: (1) Model building RGB cameras, hyperspectral cameras, LiDAR, etc. are set up at key nodes in each seedling stage to obtain top views, side views and local detail images of strawberry seedlings in real time; the image acquisition frequency, angle and camera can be dynamically adjusted according to needs.

[0033] Detailed steps: The installation of the equipment for collecting image data depends on the test location of the target object. For example, during tissue culture, it needs to be fixed on a tripod to collect images in sections. In commercial seedling production in a glass greenhouse, it can be fixed on a truss at a certain height.

[0034] Taking RGB images as an example, the following are the steps to identify the health of leaves: (a) Under standard lighting conditions, the target is photographed from multiple angles, including top and 45° side views. The images of the same object are named and annotated. Data augmentation can be performed based on the dataset. To improve recognition accuracy, the input data is preprocessed and manually annotated.

[0035] (b) Model training was performed using the machine learning framework YOLOv8. The accuracy of the model test set ranged from 88% to 93%. In addition to detecting leaf health, other features can be extracted, including but not limited to: seedling height, leaf number, leaf area index (LAI); number of runners, branch angle, and internode spacing; physiological characteristics such as seedling color and lesion detection; and identification of deformed seedlings (features such as curled leaves, discoloration, and dwarfing are manually labeled and classified during image processing).

[0036] (2) Grading and screening control In this method, since data annotation is manual, the initial classification of diseased or deformed seedlings is performed by human annotators. Models are then built based on the labeled classification data. A grading and screening system can identify targets based on classification (for example, by leaf area index or by leaf disease or health) and automatically remove them. This system can be integrated with automated sorting, robotic arms, or grading conveyor systems to achieve automated screening.

[0037] During the dynamic monitoring and digital analysis of strawberry seedling morphological characteristics, all identification data is linked to individual seedling codes and uploaded to the seedling database, enabling single-plant traceability management. This data can be used for statistical analysis, seedling strategy optimization, and production decision-making support.

[0038] The above description is only a preferred embodiment of the present invention and is not intended to limit the invention. Any modifications, equivalent replacements, improvements, etc. made within the design concept of the present invention should be included in the scope of protection of the present invention.

Claims

1. A plant factory breeding method for virus-free strawberry seedlings, characterized by: The steps include:

1. Production of original seed: Detoxify the stem tips of strawberry seedlings, and carry out hardening and rooting of original seed; 2. Expanding the stock: Transplanting the original stock into the plant factory for stock propagation; 3. Produce commercial seedlings: Move the secondary seedlings into the nursery area for large-scale breeding.

2. The plant factory breeding method for virus-free strawberry seedlings according to claim 1, wherein: During the breeding process, a phenotypic recognition system based on image recognition and artificial intelligence analysis was introduced to dynamically monitor and digitally analyze the morphological characteristics of strawberry seedlings.

3. The plant factory breeding method for virus-free strawberry seedlings according to claim 2, wherein: The steps for dynamic monitoring and digital analysis of the morphological characteristics of strawberry seedlings include: (1) Model establishment: RGB cameras, hyperspectral cameras, or LiDAR are set up at key nodes in each seedling stage to acquire top view, side view, and local detail images of strawberry seedlings in real time. The large amount of collected image data is annotated, and a data model is established based on the annotated classification data. (2) Grading and screening control: Compare the actual collected images with the data model, filter out the targets and issue elimination instructions to achieve screening.

4. The plant factory breeding method for virus-free strawberry seedlings according to claim 3, wherein: Image data was captured under standard lighting conditions.

5. The plant factory breeding method of virus-free strawberry seedlings according to claim 3, characterized in that: In step (1), the machine learning framework YOLOv8 is used for model training.

6. The plant factory breeding method for virus-free strawberry seedlings according to claim 3, characterized in that: The classification data include: seedling height, number of leaves, leaf area index, number of runners, branch angle, internode distance, seedling color, disease spots, leaf curling, color, and dwarfing.

7. The plant factory breeding method for virus-free strawberry seedlings according to claim 3, characterized in that: During the dynamic monitoring and digital analysis of the morphological characteristics of strawberry seedlings, all identification data and individual seedlings are coded and bound, and uploaded to the seedling database to realize traceability management of individual plants.

8. The plant factory breeding method for virus-free strawberry seedlings according to claim 1, wherein: When detoxifying the stem tips of strawberry seedlings, the environmental control conditions are: temperature 25 / 22℃±1℃, humidity 60~70%, and carbon dioxide concentration 350±50 μL / L; During the hardening and rooting of the original seedlings, the environmental control is as follows: the temperature in the primary culture stage is controlled at 25 / 22℃±1℃, the humidity is 60~70%, the carbon dioxide concentration is 350±50μL / L, and the white light LED has a light intensity of 50~55±10μmol / m 2 / s, photoperiod 16 / 8h; from adventitious bud proliferation to rooting stage, temperature 25 / 18℃±1℃, humidity 70%, carbon dioxide concentration 350±50µL / L and later 700±50µL / L, white light LED, light intensity 150±10μmol / m 2 / s, photoperiod 16 / 8h; During the seedling hardening stage, the temperature was 21 / 19℃±1℃ and 16.4℃±1℃ in the later stage, the humidity was 80%, the carbon dioxide concentration was 350±50μL / L, and the white light LED had a light intensity of 90±10μmol / m 2 / s, photoperiod 12 / 12h.

9. The plant factory breeding method for virus-free strawberry seedlings according to claim 1, characterized in that: When propagating the original species, environmental control: temperature 25 / 20℃±1℃, humidity 75%±5%, carbon dioxide concentration 800±50µL / L, white light LED, light intensity 300±10μmol / m 2 / s, photoperiod 12 / 12h.

10. The plant factory breeding method of virus-free strawberry seedlings according to claim 1, characterized in that: When producing commercial seedlings, environmental control: seedling temperature 25 / 20℃±1℃, humidity 75%±5%, carbon dioxide concentration 800±50µL / L, white light LED, light intensity 280±10μmol / m 2 / s, photoperiod 12 / 12h.