Light system and method for indoor identification of shade tolerance of soybean germplasm resources

Abstract

The application discloses a light system and method for indoor identification of shade tolerance of soybean germplasm resources, and the shade tolerance of soybean to low red light and far red light ratio and low blue light two kinds of shade signals is evaluated by using low red light and far red light ratio and low blue light two kinds of indoor shade light system construction technical parameters, so that a comprehensive shade tolerance evaluation value of soybean can be obtained, excellent shade tolerance germplasm genes are excavated, and beneficial germplasm resources are provided for breeding.

Description

Technical Field

[0001] This invention relates to the field of breeding, specifically to a light system for indoor identification of shade tolerance traits in soybean germplasm resources, and also to a method for identifying shade tolerance traits in soybean germplasm resources using a light system. Background Technology

[0002] Soybean strip intercropping can cause soybeans to be shaded, triggering a shade avoidance response that alters plant morphology, including: elongated main stem, thinner stems, lodging, elongated petioles, fewer nodes, and fewer lateral branches. These changes in plant morphology lead to significant yield reductions and are detrimental to agricultural production. Therefore, it is necessary to select shade-tolerant varieties suitable for intercropping / relay cropping.

[0003] Plants sense shading signals by detecting changes in light quality through photoreceptors. Shading responses are primarily mediated by two pathways: 1) a low red to far-red light ratio (low R:FR, low R / FR), mainly mediated by phytochromes; and 2) low blue light (LBL), mainly mediated by cryptochromes. In soybeans, the shade avoidance syndromes induced by low red to far-red light ratios (Low R:FR) and low blue light (Low Blue) show significant differences. For example, low red to far-red light ratios significantly induce changes in leaf angle and petiole elongation, but hardly promote stem elongation; while low blue light significantly promotes stem elongation, but has little effect on petiole elongation. These differences indicate that soybeans respond to these different shading signals using different genes and signaling pathways. Therefore, treatments with low red and far-red light ratios and low blue light, respectively, are beneficial for identifying genes related to different shade avoidance phenotypes, elucidating their corresponding regulatory mechanisms, and providing breeders with ideas and gene resources for creating shade-tolerant germplasm. This patent innovatively establishes an indoor system for identifying soybean shade tolerance under low red light, far-red light ratio, and low blue light, addressing the needs of producers and breeders. This system offers the following advantages: 1. Controllable conditions, overcoming the uncertainties of the field environment, such as natural disasters, pests, diseases, and soil variations. 2. Overcoming the limitations of field experimental ecological zones, enabling simultaneous identification of soybean varieties in both northern and southern regions under the same conditions. 3. Decomposing the changes in different soybean traits caused by variations in light quality under shading conditions, facilitating targeted gene discovery. This indoor shade tolerance identification method allows for large-scale identification of shade tolerance traits in soybean germplasm resources, uncovering superior shade-tolerant germplasm genes, and providing valuable germplasm resources for breeding. Summary of the Invention

[0004] In view of this, one objective of the present invention is to provide an indoor illumination system for identifying shade tolerance traits in soybean germplasm resources; another objective of the present invention is to provide a method for identifying shade tolerance traits in soybean germplasm resources using the illumination system.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] 1. An indoor lighting system for identifying shade tolerance traits in soybean germplasm resources, characterized in that: the lighting system includes unshaded lighting, a low red to far-red light ratio, and low blue light;

[0007] The unshaded light quantum flux density is 334.2 μmol / m². 2 The ratio of the quantum beam flux density in the 600–700 nm wavelength range to that in the 700–780 nm wavelength range is 7.91.

[0008] The quantum flux density of the low red light to far red light ratio is 326.2 μmol / m². 2 The ratio of the quantum beam flux density in the 600–700 nm wavelength range to that in the 700–780 nm wavelength range is 0.58.

[0009] The photon flux density of low blue light is 237.5 μmol / m². 2 The ratio of the quantum beam flux density in the 600–700 nm wavelength range to that in the 700–780 nm wavelength range is 12.35.

[0010] Preferably, the unshaded light illumination has a quantum beam flux density of 134.2 μmol / m² in the wavelength range of 400–500 nm. 2 The quantum beam flux density in the wavelength range of 600–700 nm is 105.7 μmol / m². 2 The quantum beam flux density in the wavelength range of 700–780 nm is 13.37 μmol / m². 2 ·s.

[0011] The quantum beam flux density, with a low-red to far-red light ratio in the wavelength range of 400–500 nm, is 108.4 μmol / m³. 2 The quantum beam flux density in the 600–700 nm wavelength range is 52.91 μmol / m². 2 The quantum beam flux density in the wavelength range of 700–780 nm is 91.72 μmol / m². 2 ·s;

[0012] The low blue light has a quantum beam flux density of 10.96 μmol / m³ in the wavelength range of 400–500 nm. 2 The quantum beam flux density in the 600–700 nm wavelength range is 117.9 μmol / m². 2The quantum beam flux density in the wavelength range of 700–780 nm is 9.548 μmol / m². 2 ·s.

[0013] Preferably, the unshaded lighting of the present invention consists of 6 full-spectrum white LEDs of 16W each with a color temperature of 5000K, 2 red LEDs of 20W each with a wavelength of 660nm, and 3 blue LEDs of 22W each with a wavelength of 460nm.

[0014] The low red light to far red light ratio is composed of 6 full-spectrum white LEDs with a color temperature of 5000K, 3 red LEDs with a wavelength of 730nm and a color temperature of 20W, and 2 blue LEDs with a wavelength of 460nm.

[0015] The low blue light consists of nine 16W, 5000K full-spectrum white LEDs, two 20W, 660nm wavelength red LEDs, and a yellow filter film installed under the lamp tubes.

[0016] 2. A method for identifying shade tolerance traits in soybean germplasm resources using the aforementioned light system, comprising the following steps:

[0017] (1) Select seedlings with similar growth on the 10th day after sowing and cultivate them under white light. Start shading treatment under the light system 14 days after sowing for 35 days, and measure soybean seedling traits during the period.

[0018] (2) Calculate the average value of each trait for each variety under no shade, low red light to far red light ratio and low blue light, and calculate the shade tolerance coefficient STC by using the following formula. The closer the STC is to 1, the more shade tolerant it is.

[0019]

[0020] Among them, TTV wl TTV represents the trait value obtained under unshaded light. sh This represents the ratio of low red light to far red light or the trait value obtained under low blue light treatment.

[0021] Preferably, the traits of this invention are stem height, stem diameter, number of nodes, number of effective lateral branches, petiole length, and petiole angle.

[0022] The beneficial effects of this invention are as follows: This invention discloses a method for identifying shade tolerance traits in soybean germplasm resources. By using technical parameters for constructing two indoor shading lighting systems—low red light to far-red light ratio and low blue light ratio—and evaluating the shade tolerance of soybeans to these two shading signals, a comprehensive shade tolerance evaluation value for soybeans can be obtained, and excellent shade-tolerant germplasm genes can be discovered, providing beneficial germplasm resources for breeding. Detailed Implementation

[0023] The present invention will be further described below with reference to specific embodiments, so that those skilled in the art can better understand and implement the present invention, but the embodiments are not intended to limit the present invention.

[0024] Example 1: Construction of a shade-tolerant system

[0025] Using an LED controllable lighting system to simulate full sunlight without shading and two shading signals—low red to far-red light ratio and low blue light—we measured the phenotypic changes of soybean seedlings (germination to V2 stage, 18 days) under shading conditions, and screened and evaluated shade-tolerant varieties.

[0026] The artificial climate chamber is set at a constant temperature of 25°C and is exposed to full sunlight 24 hours a day.

[0027] The plant cultivation rack has three layers, each measuring 133cm in length, 55cm in width, and 92cm in height.

[0028] Seedling trays: 5×10 holes, 11 cm deep; fill the bottom with water and fertilizer; raise the position of the seedling trays so that they are 30 cm away from the light source.

[0029] White light unshaded illumination (WL) setup: 6 full-spectrum white LEDs (model: SSR110-016-2000, 16W, color 5000K), 2 red LEDs (model: UH-BLDG320R, 660nm, 20W), and 3 blue LEDs (model: UH-BDLDG1320B, 460nm, 22W).

[0030] Low red light to far red light ratio (LR / FR) lighting setup: 6 full-spectrum white LEDs (model: SSR110-016-2000, 16W, color 5000K), 3 far red LEDs (model: UH-BLDG1320FR, 730nm, 20W), and 2 blue LEDs (model: UH-BDLDG1320B, 460nm, 22W).

[0031] Low blue light (LBL) lighting setup: 9 full-spectrum white LEDs (model: SSR110-016-2000, 16W, color 5000K), 2 red LEDs (model: UH-BLDG320R, 660nm, 20W), with a yellow filter (model: no.101, Lee Filters, CA) installed under the lamps.

[0032] Table 1 shows the light intensity of key wavelength bands in the unshaded, low red light to far red light ratio, and low blue light experimental groups.

[0033] Table 1. Simulated Illumination Values ​​under Indoor Shading

[0034]

[0035] * Elevate the plant stand by 50cm, measure the light intensity at a distance of 30cm from the light tube, using a HiPoint HR-350 spectrometer.

[0036] The light value was measured directly below the lamp tube at a distance of 35cm, and the unit was μmol·m. -2 ·s -1 .

[0037] In the table, PPFD refers to the photon flux density, which is the amount of light received per unit area per time in the range of 380–780 nm.

[0038] sub-number.

[0039] PFD-B: Quantum beam flux density in the blue field, with a quantum beam density in the range of 400–500 nm.

[0040] PFD-R: Quantum beam flux density in the red domain, quantum beam density in the range of 600-700 nm.

[0041] PFD-FR: Photonic quantum beam flux density in the far-infrared field, photonic quantum beam density in the range of 700-780nm.

[0042] Definition of soybean seedling plant architecture-related traits:

[0043] 1) Main stem length: from the point where the plant emerges from the soil to the meristem at the apex of the main stem;

[0044] 2) Stem diameter: Measure the distance from the soil emergence point to half the distance from the cotyledon;

[0045] 3) Length of petiole of compound leaf: The distance between the leaf pulvinus (the part of the petiole that attaches to the stem) and the base of the leaflet;

[0046] 4) Angle between compound leaf and petiole: The angle between the petiole of the compound leaf and the main stem;

[0047] 5) Leaf area per leaf: Measured by taking a photo using the "ShuTu" mobile app;

[0048] 6) Growth and development status;

[0049] 7) Lodging: When the height of the top of the stem of a naturally growing plant is less than half (50%) of the length of the main stem, the plant is considered to be lodging.

[0050] 8) Lodging rate (ALP%): The rapid elongation and weak stems caused by the shade avoidance response can prevent plants from growing upright and cause them to fall over.

[0051] Record the percentage of plants that have collapsed: Calculation formula: C = n / m (n is the number of collapsed plants, m is the total number of plants)

[0052] Example 2: Identification of Shade Tolerance in Soybeans

[0053] (1) Trait data detection

[0054] Soybean seeds meeting the quality requirements for original soybean seeds or first-grade improved varieties as specified in GB 4404.2 were selected. Eighteen plants of each variety were used per test, including 6 plants of LR / FR, 6 plants of LBL, and 6 plants of Baiguang WL as control groups. Each variety was tested 2-3 times. The final data volume for each variety was n = 15-18 / light treatment.

[0055] Soybeans were first germinated in seedling trays using sterilized soil. Ten days after sowing, 18 seedlings of similar growth were selected and transplanted into growing pots (20×20 cm, 15 cm high), 2 seedlings / pot (9 pots / seed), and cultivated under white light. Shading treatment began 14 days after sowing. The WL (3 pots) remained under the white light control group; the LR / FR treatment was transferred to the low red to far-red light ratio shading treatment group; and the LBL treatment was transferred to the low blue light shading treatment group. Shading treatment was carried out during the 3rd to 8th week of the plant's vegetative growth period.

[0056] T0: Before the transfer and shading treatment (D14), where D is the number of days after sowing and T is the number of days of light treatment.

[0057] T3(D17): Measure the height of the main stem, the length of the petiole of the compound leaf, and the angle of the petiole of the compound leaf.

[0058] T21(D35): Measure the main stem height, stem diameter, number of nodes, number of effective lateral branches, petiole length of four-petioles and petiole angle of five-petioles, area of ​​four-petioles, chlorophyll content of five-petioles, number of non-wilted compound leaves, lodging, and record the developmental stage.

[0059] (2) Shade tolerance data analysis:

[0060] a. Shade tolerance coefficient (STC):

[0061] Based on the data of each individual trait, the average value of each trait for each variety under white light and shade is calculated, and the original data is converted using the following formula (1).

[0062]

[0063] TTV wl Represents natural light property value, TTV sh This represents the shaded state value.

[0064] Using the above method, various indicators of different soybean varieties were tested, and the values ​​of each indicator were obtained. The results are shown in Table 1.

[0065] Table 1. Detection results of different soybean varieties

[0066]

[0067] Table 2, LBL-STC

[0068]

[0069]

[0070] Table 3, LR / FR-STC

[0071]

[0072] T21_D35 represents 21 days of light processing, with a total of 35 days.

[0073] It can perform identification and analysis of individual indicators, as well as comprehensive analysis.

[0074] 1) Analysis of a single indicator

[0075] Under shaded conditions, soybeans exhibit a shade-avoidance response, resulting in elongated main stems. Therefore, when determining soybean shade tolerance, the stem height (STC) is used as a reference indicator. A higher STC (strain tolerance) than 1 indicates greater shade tolerance. Since stem height is primarily regulated by shade-tolerant varieties (LBL), shade-tolerant materials were selected by comparing stem height STC under LBL conditions. Under LBL, the white-hidden stem height STC was 1.01, indicating the most shade tolerance, with Jidou 17 showing the most sensitivity. William 82 was more shade-tolerant than Tianlong 1.

[0076] Soybean petiole length is mainly regulated by the LR / FR ratio. William82 exhibits the highest shade tolerance at a petiole length of 1.46 cm, while Heinong 11 is the most sensitive. Similarly, William82 is more shade-tolerant than Tianlong 1. Considering both main stem height and petiole length, William82 performs better under shaded conditions.

[0077] Performance of various traits under LBL

[0078] Main stem height T3_D17, white hilum is the most shade tolerant, Jidou 17 is the most sensitive;

[0079] Main stem height T21_D35, Tiefeng 15 showed the most shade tolerance, while Tianlong 1 was the most sensitive;

[0080] Stem diameter T21_D35, Dan90-702 showed the most shade tolerance, and Tiefeng15 was the most sensitive;

[0081] T21_D35 is the most shade-tolerant of the T21 and D35 varieties, while Dan 90-702 is the most sensitive.

[0082] The effective number of lateral branches is T21_D35. Jidou 12 is the most shade-tolerant, while Tiefeng 15 is the most sensitive.

[0083] The petiole length of compound leaves is T3_D17, white-navel leaves are the most shade-tolerant, and 'Jidou 17' leaves are the most sensitive.

[0084] The petiole length of the compound leaf is T21_D35. Dan 90-702 is the most shade-tolerant, while Tiefeng 9 is the most sensitive.

[0085] The angle between the petiole of the compound leaf is T3_D17. Jidou 17 is the most shade-tolerant, while William 82 is the most sensitive.

[0086] The compound leaves with five petioles have an angle of T21_D35. William82 is the most shade-tolerant, while Dan90-702 is the most sensitive.

[0087] The area of ​​compound leaves with four leaves is T21_D35. William82 is the most shade-tolerant, while Tiefeng15 is the most sensitive.

[0088] The chlorophyll content of compound leaves is T21_D35, with william82 being the most shade-tolerant and dan90-702 being the most sensitive.

[0089] The number of days required for half of the lodging to occur is T21_D35. William82, Tiefeng15, and Jidou12 are the most shade-tolerant, while Dan90-702 is the most sensitive.

[0090] Expression of various traits under LR / FR

[0091] Main stem height T3_D17, Tiefeng 15 is the most shade-tolerant, Heinong 11 is the most sensitive;

[0092] Main stem height T21_D35, Tiefeng 15 showed the most shade tolerance, while Tiefeng 9 was the most sensitive;

[0093] Stem diameter T21_D35, Tiefeng 9 showed the most shade tolerance, while Heinong 11 was the most sensitive;

[0094] T21_D35 is the most shade-tolerant of the T21 and D35 varieties, while Dan90-702 is the most sensitive.

[0095] Effective lateral branch count: T21_D35; Tianlong No. 1 is the most shade-tolerant, while White Navel is the most sensitive.

[0096] The petiole length of compound leaves is T3_D17. Tiefeng No. 9 is the most shade-tolerant, while Jidou No. 17 is the most sensitive.

[0097] The petiole length of compound leaves is T21_D35, with william82 showing the most shade tolerance and black farmer 11 being the most sensitive;

[0098] The angle of the petiole of the compound leaf is T3_D17. Heinong 11 is the most shade-tolerant, while Tianlong 1 is the most sensitive.

[0099] The compound leaves with five petioles have an angle of T21-D35; Jidou 12 is the most shade-tolerant, while Anda Baimei is the most sensitive.

[0100] The area of ​​compound leaves with four leaves is T21_D35. Jidou 12 is the most shade-tolerant, while Heinong 11 is the most sensitive.

[0101] The chlorophyll content of compound leaves is T21_D35. Tianlong No. 1 is the most shade-tolerant, while Heinong 11 is the most sensitive.

[0102] The number of days required for half of the lodging to occur is T21_D35. Anda Bai Mei and Tie Feng No. 15 are the most shade-tolerant, while Tianlong No. 1 is the most sensitive.

[0103] 2) Comprehensive indicator analysis

[0104] Based on the STC of a single trait, principal component analysis (operated in SPSS software "Analyze-Dimensionality Reduction-Factor" to obtain KMO, Bartlett test, principal components, factor score coefficients also known as "comprehensive index", variance explained rate) can be used to obtain principal components. Then, the weights of the principal components and the normalized factor scores are multiplied and summed to obtain the comprehensive shade tolerance value.

[0105] The above-described embodiments are merely preferred embodiments provided to fully illustrate the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the claims.

Claims

1. A method for identifying shade tolerance traits in soybean germplasm resources using a light system, characterized in that, The lighting system includes unshaded lighting, a low red to far-red light ratio, and low blue light; The unshaded light quantum flux density is 334.2 μmol / m². 2 The ratio of the quantum beam flux density in the 600–700 nm wavelength range to that in the 700–780 nm wavelength range is 7.91; the unshaded illumination consists of six 16W LED full-spectrum white lamps with a color temperature of 5000K, two 20W LED red lamps with a wavelength of 660nm, and three 22W LED blue lamps with a wavelength of 460nm. The quantum flux density of the low red light to far red light ratio is 326.2 μmol / m². 2 The ratio of the quantum beam flux density in the 600–700 nm wavelength range to that in the 700–780 nm wavelength range is 0.58; the low red light to far red light ratio is composed of 6 16W LED full-spectrum white lamps with a color temperature of 5000K, 3 20W LED red lamps with a wavelength of 730nm, and 2 22W LED blue lamps with a wavelength of 460nm. The photon flux density of low blue light is 237.5 μmol / m². 2 The ratio of the quantum beam flux density in the 600–700 nm wavelength range to that in the 700–780 nm wavelength range is 12.35; the low blue light consists of nine 16W, 5000K full-spectrum white LEDs and two 20W, 660nm red LEDs, with a yellow filter film installed under the lamp tubes; The specific steps are as follows: (1) On the 10th day after sowing, select seedlings with similar growth and cultivate them under white light. On the 14th day after sowing, begin shading treatment under the light system for 35 days and measure soybean seedling traits during this period. (2) Calculate the average value of each trait for each variety under no shading light, low red light to far red light ratio and low blue light. Calculate the obtained data using the following formula to obtain the shade tolerance coefficient STC. The closer the STC is to 1, the more shade tolerant it is. in, This represents the phenotypic value obtained under unshaded light. The values ​​represent the ratio of low red light to far red light or the trait values ​​obtained under low blue light treatment; the traits are the main stem height, stem diameter, number of nodes, number of effective lateral branches, petiole length, and petiole angle.

2. The method according to claim 1, characterized in that: The unshaded light source, with a quantum beam flux density in the wavelength range of 400–500 nm, has a flux density of 134.2 μmol / m³. 2 The quantum beam flux density in the wavelength range of 600–700 nm is 105.7 μmol / m². 2 The quantum beam flux density in the wavelength range of 700–780 nm is 13.37 μmol / m². 2 .s; The quantum beam flux density, with a low-red to far-red light ratio in the wavelength range of 400–500 nm, is 108.4 μmol / m³. 2 The quantum beam flux density in the 600–700 nm wavelength range is 52.91 μmol / m². 2 The quantum beam flux density in the wavelength range of 700–780 nm is 91.72 μmol / m². 2 .s; The low blue light has a quantum beam flux density of 10.96 μmol / m³ in the wavelength range of 400–500 nm. 2 The quantum beam flux density in the 600–700 nm wavelength range is 117.9 μmol / m². 2 The quantum beam flux density in the wavelength range of 700–780 nm is 9.548 μmol / m². 2 ·s.

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