A method for improving the germination rate of Keteleeria fortunei var. cyclolepis seeds by changing environmental factors

Abstract

The present invention discloses a method for improving the germination rate of Keteleeria fortunei var. cyclolepis seeds by changing environmental factors, which comprises the following steps: (1) seed preparation; (2) seed disinfection; (3) seed soaking treatment: preparing a seed soaking solution for variable-temperature seed soaking; (4) treatment of germination substrate; the germination substrate includes coarse-grained activated carbon, debris, fine sand, peat, vermiculite, and rock wool, and the coarse-grained activated carbon is obtained by soaking with a mixed preparation of plant growth hormones after high-temperature disinfection; (5) germination treatment: performing constant-temperature and constant-humidity germination in a germination chamber; the germination chamber includes an air conditioner, an atomization device, and a germination substrate. After seed soaking, the seeds are buried in the germination substrate at a fixed depth for germination, and the temperature and humidity of the germination chamber are controlled by the air conditioner and the atomization device during germination. The method of the present invention performs variable-temperature seed soaking on the seeds with the seed soaking solution, the coarse-grained activated carbon slowly releases the mixed preparation of plant growth hormones during germination, and the germination chamber is provided with an air conditioner and an atomization device to control the temperature and humidity of germination, so that the germination rate of Keteleeria fortunei var. cyclolepis seeds is above 74.98%.

Description

Technical Field

[0001] The present invention relates to the technical field of plant propagation, and particularly relates to a method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors. Background Art

[0002] Keteleeria cyclolepis is a perennial evergreen arbor plant of the genus Keteleeria, which is a unique tree species in China. It is mainly distributed in the southeastern part of Yunnan, Guizhou, the northwestern and eastern parts of Guangxi, the northern part of Guangdong, the southern part of Hunan, the southwestern part of Jiangxi, and the southwestern part of Zhejiang. Keteleeria cyclolepis has an elegant and beautiful tree shape and can be used as a garden greening tree species. Moreover, the bark of Keteleeria cyclolepis has the medicinal effects of promoting eruption, detumescence, and setting bones. At present, Keteleeria cyclolepis has become a vulnerable species and has been included in the "Red Data Book of Chinese Plants" and the "Atlas of Rare and Endangered Plants in Guangdong Province". The main reason is that the germination rate of Keteleeria cyclolepis is low and the germination time is long. There are the following difficulties in the germination process of Keteleeria cyclolepis seeds:

[0003] (1) The seed vigor of Keteleeria cyclolepis seeds will gradually be lost over time. Therefore, in the prior art, the sowing time of Keteleeria cyclolepis seeds is from February to March of the following year after the seeds are mature. However, it is relatively cold from February to March and the temperature difference is large, resulting in a long germination time and a low germination rate of Keteleeria cyclolepis seeds.

[0004] (2) For the soaking treatment, normal temperature soaking is generally carried out with clean water, which makes the soaking time of Keteleeria cyclolepis seeds too long. It generally takes 4 - 5 days to complete. Overlong soaking will cause the seeds to become inactivated or even rot.

[0005] (3) During the germination acceleration process, stratification germination is carried out using sand and gravel. The water storage capacity and fertilizer storage capacity of sand and gravel are poor, and they cannot provide water and nutrients for the germination of Keteleeria cyclolepis seeds during the germination acceleration process, resulting in a long germination acceleration time and a low germination rate.

[0006] The method of the present invention performs variable temperature soaking on Keteleeria cyclolepis seeds with a soaking solution, and then uses the coarse-grained activated carbon and debris soaked with a plant growth mixture preparation to make a germination acceleration substrate. The coarse-grained activated carbon can slowly release the plant growth hormone mixture preparation during the germination acceleration process. The germination acceleration chamber is equipped with an air conditioner and an atomization device to control the temperature and humidity for germination acceleration, providing a constant temperature and humidity germination acceleration environment for Keteleeria cyclolepis seeds, effectively promoting the storage, degradation, and transformation of soluble sugars and soluble proteins during the germination process of Keteleeria cyclolepis seeds, inhibiting the production of malondialdehyde during the germination process of Keteleeria cyclolepis seeds. The changes in the contents of soluble sugars, soluble proteins, and malondialdehyde in the seeds determine the germination speed and germination rate of Keteleeria cyclolepis seeds. At the same time, it overcomes the disadvantages that the germination of Keteleeria cyclolepis seeds is not timely and the seeds are inactivated due to the influence of environmental factors during germination. The method of the present invention has the advantages of saving land resources, high speed, high quality, being technology-intensive, and being convenient for intensive management and factory production.

[0007] The information disclosed in this background section is only intended to enhance the overall understanding of the background of the present invention and should not be regarded as an admission or any form of suggestion that this information constitutes prior art already known to those of ordinary skill in the art. Summary of the Invention

[0008] An object of the present invention is to provide a method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors, thereby overcoming the disadvantages of untimely germination of Keteleeria cyclolepis seeds and seed inactivation caused by environmental factors during germination.

[0009] To achieve the above object, the present invention provides a method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors, which is characterized by including the following steps:

[0010] (1) Seed preparation;

[0011] (2) Seed disinfection;

[0012] (3) Seed soaking treatment: Prepare a seed soaking solution for variable-temperature seed soaking;

[0013] (4) Germination substrate treatment: The germination substrate includes coarse-grained activated carbon, debris, fine sand, peat, vermiculite, and rock wool, and the coarse-grained activated carbon is obtained by soaking it in a mixed preparation of plant growth hormones after high-temperature disinfection;

[0014] (5) Germination treatment: Carry out constant-temperature and constant-humidity germination in a germination chamber; the germination chamber includes an air conditioner, an atomization device, and a germination substrate. After soaking, the seeds are buried in the germination substrate at a certain depth for germination, and the temperature and humidity of the germination chamber are controlled by the air conditioner and the atomization device during germination.

[0015] Preferably, in the above technical solution, in step (1), the seed preparation is to remove impurities and then select seeds without damage and diseases by water selection.

[0016] Preferably, in the above technical solution, in step (2), the seed disinfection is to soak the seeds in a 0.15 - 0.3% potassium permanganate solution for 10 - 25 minutes and then rinse them with distilled water 3 - 5 times.

[0017] Preferably, in the above technical solution, the seed soaking treatment method in step (3) specifically includes the following steps:

[0018] (1) Preparation of the seed soaking solution: The seed soaking solution includes a solution concentration of 4 - 8 g / L KNO 3 , 10 - 15 g / L CaCl 2 , 10 - 15 g / L ZnSO 4 , 0.2% - 1% NH 4 NO 3, 100 - 300 mg / L gibberellin solution, 100 - 500 mg / L salicylic acid, 500 - 1000 ppm wood vinegar liquid, 3% - 10% H 2 O 2 ;

[0019] (2) Seed soaking treatment: Place the selected Keteleeria fortunei var. cyclolepis seeds in step (1) in a container filled with the seed soaking solution, cover it with a small-pore gauze and press it down. Place the container in a water bath at a constant temperature of 40 - 50 °C and heat it for 2 - 6 h, then move the container to 20 - 30 °C for cooling treatment for 2 - 6 h; then place it in a water bath at a constant temperature of 40 - 50 °C and heat it for 2 - 6 h again.

[0020] 1. Preferably, in the above technical solution, the germination substrate in step (4) includes 50% - 70% coarse-grained activated carbon, 7% - 24% debris, 8% - 23% fine sand, 1% - 5% peat, 1% - 5% vermiculite, 1% - 5% rock wool, among which,

[0021] the coarse-grained activated carbon includes one or several mixtures of granular activated carbon and coconut shell activated carbon;

[0022] the debris includes one or several mixtures of sawdust and planer powder.

[0023] Preferably, in the above technical solution, the high-temperature disinfection temperature of the coarse-grained activated carbon in step (4) is 100 °C, and the disinfection time is 2 - 4 minutes.

[0024] Preferably, in the above technical solution, the plant growth hormone mixture for soaking the coarse-grained activated carbon in step (4) includes 0.1 - 0.5 mg / L 2,4-dichlorophenoxyacetic acid (2,4-D), 0.05 - 0.2 mg / L brassinolide, 0.1 - 0.3 mg / L 6-benzylaminopurine (6-BA), 0.05 - 0.3 mg / L kinetin (KT), 0.4 - 0.7 mg / L rooting powder (GGR 6 ).

[0025] Preferably, in the above technical solution, the soaking time of the coarse-grained activated carbon with the plant growth hormone mixture in step (4) is 6 - 18 h.

[0026] Preferably, in the above technical solution, the method of the germination treatment in step (4) is to bury the completed seed-soaked Keteleeria fortunei var. cyclolepis seeds at a depth of 1 - 5 cm on the germination substrate.

[0027] Preferably, in the above technical solution, in step (5), the temperature and humidity of the germination chamber are controlled by an air conditioner and an atomizing device during germination. The temperature control by the air conditioner is to keep the temperature of the germination chamber at 15 - 30 °C using the air conditioner, and the humidity control by the atomizing device is to control the relative humidity of the germination chamber at 15 - 30% using the atomizing device.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] (1) The method of the present invention for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors controls the environmental factors during germination, and the germination time is flexible, avoiding the inactivation of Keteleeria cyclolepis seeds after missing the germination season.

[0030] (2) The seeds of Keteleeria cyclolepis are soaked at variable temperatures with a soaking solution to improve seed vigor and reduce the soaking time.

[0031] (3) Coarse-grained activated carbon can slowly release a plant growth hormone mixture during the germination process, providing plant growth hormones for the germination of Keteleeria cyclolepis seeds, promoting the germination and growth of bud points, and improving the germination uniformity and germination rate. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 is a diagram showing the observation of the germination situation by taking out seeds during the germination process of Keteleeria cyclolepis seeds in Example 1 of the present invention;

[0033] Figure 2 is a diagram showing the germination of Keteleeria cyclolepis seeds in Example 1 of the present invention;

[0034] Figure 3 is a diagram showing the Keteleeria cyclolepis seedlings obtained by germination in Example 1 of the present invention;

[0035] Figure 4 is a diagram showing the germinated seedlings of Keteleeria cyclolepis seeds with a germination substrate obtained in Example 1 of the present invention;

[0036] Figure 5 is a diagram showing the change in the soluble sugar content of Keteleeria cyclolepis seeds during the germination process in Example 1 and Comparative Examples 1-4;

[0037] Figure 6 is a diagram showing the change in the soluble protein content of Keteleeria cyclolepis seeds during the germination process in Example 1 and Comparative Examples 1-4;

[0038] Figure 7 is a diagram showing the change in the malondialdehyde content of Keteleeria cyclolepis seeds during the germination process in Example 1 and Comparative Examples 1-4. DETAILED DESCRIPTION OF THE INVENTION

[0039] The following describes in detail the specific embodiments of the present invention in conjunction with specific embodiments, but it should be understood that the protection scope of the present invention is not limited by the specific embodiments.

[0040] Example 1

[0041] A method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors, comprising the following steps:

[0042] (1) Seed preparation: After removing impurities, select 100 seeds without damage and disease by water selection.

[0043] (2) Seed disinfection: Soak the seeds in 0.15% potassium permanganate solution for 25 minutes, and then rinse them 3 times with distilled water.

[0044] (3) Seed soaking treatment: In a sterile room, use 4 g / L KNO 3 , 10 g / L CaCl 2 , 10 g / L ZnSO 4 , 0.2% NH 4 NO 3 , 300 mg / L gibberellin solution, 100 mg / L salicylic acid, 500 ppm wood vinegar liquid, 3% H 2 O 2 to prepare a seed soaking solution. Pour the seed soaking solution into a seed soaking container, then place the selected Keteleeria fortunei var. cyclolepis seeds in the container with the seed soaking solution, cover it with a small-pore screen and press it down. Place the container in a water bath at a constant temperature of 40 °C and heat it for 6 hours, then move the container to 20 °C for cooling treatment for 2 hours; then place it in a water bath at a constant temperature of 40 °C and heat it for 6 hours again.

[0045] (4) Germination substrate treatment: High-temperature disinfect the granular activated carbon at a high-temperature disinfection temperature of 100 °C for 2 minutes. Transfer the disinfected granular activated carbon to a sterile room, and after cooling, soak the granular activated carbon in a plant growth hormone mixture preparation composed of 0.1 mg / L 2,4-D, 0.05 mg / L brassinolide, 0.1 mg / L 6-BA, 0.05 mg / L KT, and 0.4 mg / L GGR6 for 18 hours; after the soaking treatment, take 50% of the granular activated carbon and mix it evenly with 24% sawdust, 23% fine sand, 1% peat, 1% vermiculite, and 1% rock wool.

[0046] (5) Germination treatment: Bury the soaked Keteleeria fortunei var. cyclolepis seeds at a depth of 1 cm on the germination substrate; during the entire germination treatment period, keep the temperature of the germination chamber at 15 °C with an air conditioner, and control the relative humidity of the germination chamber at 15% with a fogging device.

[0047] Germinate for 30 days, count the number of germinated Keteleeria fortunei var. cyclolepis seeds and calculate the germination rate.

[0048] Among them, the number of germinated seeds: Based on the seed plumule growing 0.1 cm.

[0049] Seed germination rate = Number of germinated seeds / Number of seeds

[0050] Average germination rate ≈ 3.14 * β1 + 4.86 * β2 - 16.1 * β3

[0051] β1 = Maximum soluble sugar content during germination - Soluble sugar content on the 30th day

[0052] β2 = Maximum soluble protein content during germination - Soluble protein content on the 30th day

[0053] β3 = Maximum malondialdehyde content during germination

[0054] Example 2

[0055] A method for improving the germination rate of Keteleeria fortunei var. cyclolepis seeds by changing environmental factors, comprising the following steps:

[0056] (1) Seed preparation: After removing debris, select 100 seeds without damage and disease by water selection method;

[0057] (2) Seed disinfection: Soak the seeds in 0.20% potassium permanganate solution for 20 minutes, and rinse with distilled water 3 times;

[0058] (3) Seed soaking treatment: In a sterile room, use 5g / L KNO 3 , 11g / L CaCl 2 , 11g / L ZnSO 4 , 0.3% NH 4 NO 3 , 150mg / L gibberellin solution, 200mg / L salicylic acid, 600ppm wood vinegar liquid, 4% H 2 O 2 to prepare a seed soaking solution. Inject the seed soaking solution into a seed soaking container, then place the selected Keteleeria fortunei var. cyclolepis seeds in the container filled with the seed soaking solution, cover them with a small-pore gauze and press them down. Place the container in a water bath at 40°C for constant temperature heating for 6 hours, then move the container to 30°C for cooling treatment for 2 hours; then place it in a water bath at 40°C for constant temperature heating for 6 hours;

[0059] (4) Germination substrate treatment: High-temperature disinfect the granular activated carbon at a high-temperature disinfection temperature of 98°C for 4 minutes. Transfer the disinfected granular activated carbon to a sterile room, and after cooling, soak the granular activated carbon with a plant growth hormone mixture preparation composed of 0.2mg / L 2,4-D, 0.1mg / L brassinolide, 0.2mg / L 6-BA, 0.1mg / L KT, and 0.45mg / L GGR6 for 15 hours; after the soaking treatment, take 55% granular activated carbon and mix it evenly with 20% sawdust, 19% fine sand, 2% peat, 2% vermiculite, and 2% rock wool;

[0060] (5) Germination treatment: Bury the soaked Keteleeria fortunei var. cyclolepis seeds into the germination substrate at a depth of 2 cm; during the entire germination treatment period, keep the temperature of the germination chamber at 20°C with an air conditioner, and control the relative humidity of the germination chamber at 15% with a atomizing device.

[0061] Germinate for 30 days, count the number of germinated seeds of Keteleeria cyclolepis Flous and calculate the germination rate.

[0062] Among them, the number of germinated seeds: Take the radicle of the seed growing 0.1 cm as the standard.

[0063] Seed germination rate = Number of germinated seeds / Number of seeds

[0064] Average germination rate ≈ 3.14 * β1 + 4.86 * β2 - 16.1 * β3

[0065] β1 = Maximum content of soluble sugar during germination - Content of soluble sugar on the 30th day

[0066] β2 = Maximum content of soluble protein during germination - Content of soluble protein on the 30th day

[0067] β3 = Maximum content of malondialdehyde during germination

[0068] Example 3

[0069] A method for improving the germination rate of Keteleeria cyclolepis Flous seeds by changing environmental factors, comprising the following steps:

[0070] (1) Seed preparation: After removing impurities, select 100 seeds without damage and disease by water selection.

[0071] (2) Seed disinfection: Soak the seeds in 0.25% potassium permanganate solution for 15 minutes for disinfection, and rinse 4 times with distilled water.

[0072] (3) Seed soaking treatment: In a sterile room, use 6 g / L KNO 3 , 12 g / L CaCl 2 , 12 g / L ZnSO 4 , 0.4% NH 4 NO 3 , 200 mg / L gibberellin solution, 300 mg / L salicylic acid, 700 ppm wood vinegar liquid, 6% H 2 O 2 to prepare a seed soaking solution, inject the seed soaking solution into a seed soaking container, then place the selected Keteleeria cyclolepis Flous seeds in the container filled with the seed soaking solution, cover them with a small-pore wire mesh and press them down, place the container in a water bath at 45 °C for constant temperature heating for 4 hours, then move the container to 20 °C for cooling treatment for 2 hours; then place it in a water bath at 45 °C for constant temperature heating for 4 hours.

[0073] (4) Pretreatment of germination substrate: The granular activated carbon is subjected to high-temperature disinfection at a temperature of 96°C for 6 minutes. After the disinfection is completed, the granular activated carbon is transferred to a sterile room. After cooling, it is soaked in a mixed preparation of plant growth hormones prepared from 0.2 mg / L 2,4-D, 0.1 mg / L brassinolide, 0.2 mg / L 6-BA, 0.1 mg / L KT, and 0.45 mg / L GGR for 12 hours; after the soaking treatment, 60% of the granular activated carbon is taken and mixed evenly with 15% sawdust powder, 15% fine sand, 3% peat, 3% vermiculite, and 4% rock wool; 6 After the soaking treatment, 60% of the granular activated carbon is taken and mixed evenly with 15% sawdust powder, 15% fine sand, 3% peat, 3% vermiculite, and 4% rock wool;

[0074] (5) Germination treatment: The seeds of Keteleeria fortunei var. cyclolepis that have completed seed soaking are buried in the germination substrate at a depth of 3 cm; during the entire germination treatment period, the temperature of the germination chamber is maintained at 25°C with an air conditioner, and the relative humidity of the germination chamber is controlled at 20% with a fogging device.

[0075] Germinate for 30 days, count the number of germinated seeds of Keteleeria fortunei var. cyclolepis and calculate the germination rate.

[0076] Among them, the number of germinated seeds: based on the radicle of the seed growing 0.1 cm.

[0077] Seed germination rate = Number of germinated seeds / Number of seeds

[0078] Average germination rate ≈ 3.14 * β1 + 4.86 * β2 - 16.1 * β3

[0079] β1 = Maximum content of soluble sugar during germination - Content of soluble sugar on the 30th day

[0080] β2 = Maximum content of soluble protein during germination - Content of soluble protein on the 30th day

[0081] β3 = Maximum content of malondialdehyde during germination

[0082] Example 4

[0083] A method for improving the germination rate of Keteleeria fortunei var. cyclolepis seeds by changing environmental factors, including the following steps:

[0084] (1) Seed preparation: After removing impurities, the seeds are selected by water selection to obtain 100 seeds without damage and disease;

[0085] (2) Seed disinfection: The seeds are soaked in a 0.25% potassium permanganate solution for 15 minutes for disinfection and rinsed 4 times with distilled water;

[0086] (3) Seed soaking treatment: In a sterile room, use 7 g / L KNO 3 , 13 g / L CaCl 2 , 13 g / L ZnSO 4 , 0.5% NH4 NO 3 、 250 mg / L gibberellin solution, 400 mg / L salicylic acid, 800 ppm wood vinegar liquid, 8% H 2 O 2 Prepare a seed soaking solution, inject the seed soaking solution into a seed soaking container, then place the selected Keteleeria fortunei var. cyclolepis seeds in the container filled with the seed soaking solution, cover them with a small pore gauze net and press them down, place the container in a water bath at a constant temperature of 45 °C and heat for 6 h, then move the container to 25 °C for cooling treatment for 4 h; then place it in a water bath at a constant temperature of 45 °C and heat for 4 h;

[0087] (4) Germination medium treatment: High-temperature sterilize coconut shell activated carbon, the high-temperature sterilization temperature is 94 °C, and the sterilization time is 6 minutes. Transfer the sterilized coconut shell activated carbon to a sterile room, and after cooling, soak the coconut shell activated carbon with a plant growth hormone mixture prepared from 0.3 mg / L 2,4-D, 0.15 mg / L brassinolide, 0.25 mg / L 6-BA, 0.15 mg / L KT, 0.5 mg / L GGR 6 for 9 h; after the soaking treatment, take 65% coconut shell activated carbon and mix it evenly with 11% planer pollen material, 11% fine sand, 4% peat, 4% vermiculite, and 5% rock wool;

[0088] (5) Germination promotion treatment: Bury the soaked Keteleeria fortunei var. cyclolepis seeds into the germination medium at a depth of 4 cm; during the entire germination promotion treatment period, keep the temperature of the germination chamber at 25 °C with an air conditioner, and control the relative humidity of the germination chamber at 25% with a atomizing device.

[0089] Promote germination for 30 d, count the number of germinated Keteleeria fortunei var. cyclolepis seeds and calculate the germination rate.

[0090] Among them, the number of germinated seeds: Take the emergence of the seed embryo by 0.1 cm as the standard.

[0091] Seed germination rate = number of germinated seeds / number of seeds

[0092] Average germination rate ≈ 3.14 * β1 + 4.86 * β2 - 16.1 * β3

[0093] β1 = maximum content of soluble sugar during germination promotion - content of soluble sugar on the 30th day

[0094] β2 = maximum content of soluble protein during germination promotion - content of soluble protein on the 30th day

[0095] β3 = maximum content of malondialdehyde during germination promotion

[0096] Example 5

[0097] A method for improving the germination rate of Keteleeria fortunei var. cyclolepis seeds by changing environmental factors, comprising the following steps:

[0098] (1) Seed preparation: After removing impurities, select 100 seeds without damage or disease by water selection.

[0099] (2) Seed disinfection: Soak the seeds in 0.30% potassium permanganate solution for 10 minutes, and then rinse them 5 times with distilled water.

[0100] (3) Seed soaking treatment: In a sterile room, use 8 g / L KNO 3 , 14 g / L CaCl 2 , 14 g / L ZnSO 4 , 0.8% NH 4 NO 3 , 300 mg / L gibberellin solution, 500 mg / L salicylic acid, 900 ppm wood vinegar liquid, 9% H 2 O 2 to prepare a seed soaking solution. Pour the seed soaking solution into a seed soaking container, and then place the selected Keteleeria fortunei var. cyclolepis seeds in the container with the seed soaking solution, cover them with a small-pore gauze net and press them down. Place the container in a water bath at a constant temperature of 50 °C and heat for 2 hours, then move the container to 25 °C for cooling treatment for 4 hours; then place it in a water bath at a constant temperature of 50 °C and heat for 2 hours again.

[0101] (4) Germination substrate treatment: Sterilize the coconut shell activated carbon at a high temperature. The high-temperature sterilization temperature is 92 °C and the sterilization time is 8 minutes. Transfer the sterilized coconut shell activated carbon to a sterile room, and after cooling, soak the coconut shell activated carbon with a plant growth hormone mixture prepared from 0.4 mg / L 2,4-D, 0.2 mg / L brassinolide, 0.3 mg / L 6-BA, 0.2 mg / L KT, and 0.6 mg / L GGR 6 for 6 hours; after the soaking treatment, take 65% coconut shell activated carbon and mix it evenly with 10% sawdust powder, 10% fine sand, 5% peat, 5% vermiculite, and 5% rock wool.

[0102] (5) Germination promotion treatment: Bury the soaked Keteleeria fortunei var. cyclolepis seeds at a depth of 5 cm on the germination substrate; during the entire germination promotion treatment, use an air conditioner to keep the temperature in the germination chamber at 30 °C, and use atomization equipment to control the relative humidity in the germination chamber at 30%.

[0103] Promote germination for 30 days, count the number of germinated Keteleeria fortunei var. cyclolepis seeds and calculate the germination rate.

[0104] Among them, the number of germinated seeds: Take the emergence of the seed embryo by 0.1 cm as the standard.

[0105] Seed germination rate = Number of germinated seeds / Number of seeds

[0106] Average germination rate ≈ 3.14 * β1 + 4.86 * β2 - 16.1 * β3

[0107] β1 = Maximum soluble sugar content during germination - Soluble sugar content on the 30th day

[0108] β2 = Maximum soluble protein content during germination - Soluble protein content on the 30th day

[0109] β3 = Maximum malondialdehyde content during germination

[0110] Example 6

[0111] A method for improving the germination rate of Keteleeria fortunei var. cyclolepis seeds by changing environmental factors, comprising the following steps:

[0112] (1) Seed preparation: After removing impurities, select 100 seeds without damage and disease by water selection;

[0113] (2) Seed disinfection: Soak the seeds in 0.30% potassium permanganate solution for 10 minutes, and rinse with distilled water 5 times;

[0114] (3) Seed soaking treatment: In a sterile room, use 8 g / L KNO 3 , 15 g / L CaCl 2 , 15 g / L ZnSO 4 , 1% NH 4 NO 3 , 300 mg / L gibberellin solution, 500 mg / L salicylic acid, 1000 ppm wood vinegar liquid, 10% H 2 O 2 Prepare a seed soaking solution, inject the seed soaking solution into a seed soaking container, then place the selected Keteleeria fortunei var. cyclolepis seeds in the container filled with the seed soaking solution, cover with a small pore gauze net and press down, place the container in a water bath at 50 °C for constant temperature heating for 2 h, then move the container to 30 °C for cooling treatment for 6 h; then place it in a water bath at 50 °C for constant temperature heating for 2 h;

[0115] (4) Germination substrate treatment: High-temperature disinfect the coconut shell activated carbon, the high-temperature disinfection temperature is 90 °C, and the disinfection time is 10 minutes. Transfer the disinfected coconut shell activated carbon to a sterile room, and after cooling, soak the coconut shell activated carbon with a plant growth hormone mixture prepared from 0.4 mg / L 2,4-D, 0.2 mg / L brassinolide, 0.3 mg / L 6-BA, 0.2 mg / L KT, 0.6 mg / L GGR 6 for 6 h; After the soaking treatment, take 70% coconut shell activated carbon and mix it evenly with 7% sawdust, 8% fine sand, 5% peat, 5% vermiculite, and 5% rock wool;

[0116] (5) Germination treatment: Bury the soaked Keteleeria fortunei var. cyclolepis seeds into the germination substrate at a depth of 5 cm; During the entire germination treatment period, keep the temperature of the germination chamber at 30 °C with an air conditioner, and control the relative humidity of the germination chamber at 30% with a atomizing device.

[0117] Germinate for 30 days, count the number of germinated seeds of Keteleeria cyclolepis and calculate the germination rate.

[0118] Among them, the number of germinated seeds: based on the radicle of the seed growing 0.1 cm.

[0119] Seed germination rate = number of germinated seeds / number of seeds

[0120] Average germination rate ≈ 3.14 * β1 + 4.86 * β2 - 16.1 * β3

[0121] β1 = maximum content of soluble sugar during germination - content of soluble sugar on the 30th day

[0122] β2 = maximum content of soluble protein during germination - content of soluble protein on the 30th day

[0123] β3 = maximum content of malondialdehyde during germination

[0124] Comparative example 1

[0125] Steps (1) and (2) are the same as those in Example 1; the difference is that,

[0126] (3) Seed soaking treatment: Inject clean water into the seed soaking container in the sterile room, then place the selected Keteleeria cyclolepis seeds in the container filled with the seed soaking solution, cover them with a small-pore gauze and press them down, and place the container at 25 °C for seed soaking for 18 h;

[0127] (4) Germination substrate treatment: Disinfect the river sand at high temperature, the high temperature disinfection temperature is 90 °C, and the disinfection time is 10 minutes. Transfer the disinfected river sand to the germination chamber as the germination substrate;

[0128] (5) Germination treatment: Bury the soaked Keteleeria cyclolepis seeds at a depth of 1 cm on the germination substrate; during the whole germination treatment period, use the air conditioner and atomization equipment to simulate the temperature of 10 °C and humidity of 10% in February and March.

[0129] Comparative example 2

[0130] Steps (1), (2), (4), and (5) are the same as those in Example 1; the difference is that,

[0131] (3) Seed soaking treatment: Use 4 g / L KNO 3 , 10 g / L CaCl 2 , 300 mg / L gibberellin solution, 100 mg / L salicylic acid, 500 ppm wood vinegar solution, 3% H 2 O 2Prepare it into a seed soaking solution, inject the seed soaking solution into a seed soaking container, then place the selected Keteleeria fortunei var. cyclolepis seeds in the container filled with the seed soaking solution, cover it with a small-pore gauze and press it down, and place the container in a water bath at a constant temperature of 40 °C and heat it for 14 h.

[0132] Comparative Example 3

[0133] Steps (1), (2), (3), and (5) are the same as those in Example 1; the difference is that,

[0134] (4) Treatment of germination substrate: High-temperature sterilize the nano-activated carbon at a high-temperature sterilization temperature of 100 °C for 2 minutes, transfer the sterilized nano-activated carbon to a sterile room, and after cooling, soak the nano-activated carbon in a plant growth hormone mixture prepared from 0.1 mg / L 6-BA, 0.05 mg / L KT, and 0.4 mg / L GGR6 for 18 h; use the soaked nano-activated carbon as the germination substrate.

[0135] Comparative Example 4

[0136] Steps (1), (2), (3), and (5) are the same as those in Example 1; the difference is that,

[0137] (4) Treatment of germination substrate: High-temperature sterilize the granular activated carbon at a high-temperature sterilization temperature of 100 °C for 2 minutes, transfer the sterilized granular activated carbon to a sterile room, and after cooling, soak the granular activated carbon in a plant growth hormone mixture prepared from 1 mg / L 2,4-D, 1 mg / L brassinolide, 1 mg / L 6-BA, 1 mg / L KT, and 1 mg / L GGR6 for 18 h; after the soaking treatment, take 50% of the granular activated carbon and mix it evenly with 24% sawdust, 23% fine sand, 1% peat, 1% vermiculite, and 1% rock wool.

[0138] The above Examples 1-6 and Comparative Examples 1-4 were germinated and sprouted for 30 days, and three replicates were set for each experiment. The contents of soluble sugar, soluble protein, and malondialdehyde in the seeds were measured and recorded at 0 d, 5 d, 10 d, 15 d, 20 d, 25 d, and 30 d of sprouting, as shown in Tables 1, 2, and 3; the germination numbers of Keteleeria fortunei var. cyclolepis seeds were counted and the germination rates were calculated at 30 d of sprouting, as shown in Tables 4 and 5.

[0139] Table 1 Changes in the content of soluble sugar in seeds during the germination process of Examples 1-6 and Comparative Example 1

[0140] Time 0d 5d 10d 15d 20d 25d 30d Example 1 4.7 8 14 18.5 14 8 6.4 Example 2 4.8 9 14.7 19 14.4 8 6 Example 3 5 8.7 15 21 16 9 5.8 Example 4 5.5 9.1 16.6 21.3 15.7 8 5.3 Example 5 6 11 18.9 21.6 17 10.3 6.1 Example 6 6.3 13 19.5 22.5 16.8 11 7 Comparative Example 1 4 7.6 10 12.4 14.3 12.5 7.9 Comparative Example 2 4.3 6.7 12.8 15.6 17.5 14.3 7.9 Comparative Example 3 4.7 8.4 13 16 13.4 8.8 6.3 Comparative Example 4 4.7 6 9 13 10 8.2 6.4

[0141] Soluble sugar is an important osmotic regulatory substance in plant cells. The change in the content of soluble sugar during germination directly reflects the conversion of carbohydrates and the metabolism of organic matter in seeds, and is the key to the metabolism of organic matter and the successful germination and growth of seeds. As shown in Table 1, with the increase in the concentration of the seed soaking solution, the content of soluble sugar in the seeds before germination increases; with the increase in the concentration of the plant growth hormone mixture preparation soaked in coarse-grained activated carbon in the germination substrate, more soluble sugar is produced in the seeds during germination; temperature and humidity affect the content of soluble sugar at 30 d, and when the temperature is 25 °C and the humidity is 25%, the content of soluble sugar is 5.3, indicating that the degradation and conversion of soluble sugar are the best under this temperature and humidity. The results prove that the method of the present invention changes the environmental factors during the germination process of Keteleeria fortunei var. cyclolepis seeds by using a variable-temperature seed soaking with the seed soaking solution, slowly releasing the plant growth hormone mixture preparation with coarse-grained activated carbon during the germination process, and controlling the temperature and humidity. Compared with Comparative Examples 1-2, the composition of the seed soaking solution and the variable-temperature seed soaking used in the method of the present invention promote the synthesis of soluble sugar in the seeds before germination and advance seed germination; compared with Comparative Examples 3-4, the germination substrate of the method of the present invention uses coarse-grained activated carbon, the composition and concentration range of the plant growth hormone mixture preparation, and is more conducive to the synthesis, degradation and conversion of soluble sugar in Keteleeria fortunei var. cyclolepis seeds during the germination process, thereby promoting the faster germination of Keteleeria fortunei var. cyclolepis seeds.

[0142] Table 2 Changes in the content of soluble protein in seeds during the germination process of Examples 1-6 and Comparative Example 1

[0143]

[0144]

[0145] In the seeds in the early stage of germination, the soluble proteins increase due to the synthesis of enzymes required for the life activities of the embryo. In the later stage of seed germination, the soluble proteins in the seeds can be degraded into small molecule amino acids and other forms that are easily absorbed by the embryo under the action of related enzymes, providing the energy required for seed germination and the materials for forming new tissues. As shown in Table 2, the content of soluble proteins in the seeds before germination increased with the increase of the concentration of the soaking solution. The concentration, temperature and humidity of the slow-release plant growth hormone mixture of coarse-grained activated carbon affected the synthesis rate and synthesis amount of soluble proteins. The method of the present invention changes the environmental factors in the seed germination process of Keteleeria fortunei var. cyclolepis by using variable-temperature soaking with the soaking solution, using the slow-release plant growth hormone mixture of coarse-grained activated carbon during the germination process, and controlling the temperature and humidity. Compared with Comparative Examples 1-2, the soaking solution components and variable-temperature soaking adopted in the method of the present invention synthesized more soluble protein content in the seeds before germination; compared with Comparative Example 3, adding 2,4-D and brassinolide to the mixture of coarse-grained activated carbon and plant growth hormone in the method of the present invention can significantly promote the increase of the soluble protein content in the germination of Keteleeria fortunei var. cyclolepis seeds; compared with Comparative Example 4, the concentration range of the plant growth hormone mixture in the method of the present invention is more conducive to the faster synthesis, degradation and transformation of soluble proteins in the seeds of Keteleeria fortunei var. cyclolepis during the germination process, thus promoting the faster germination of Keteleeria fortunei var. cyclolepis seeds.

[0146] Table 3 Changes in the content of malondialdehyde in seeds during the germination process of Examples 1-6 and Comparative Example 1

[0147]

[0148]

[0149] Malondialdehyde is the most important product of membrane lipid peroxidation. The content of malondialdehyde in seeds reflects the damage degree of various membranes in the seeds and the activity of the free radical scavenging enzyme system in the body. As shown in Table 3, the content of malondialdehyde in the seeds before germination decreased with the increase of the concentration of the soaking solution, but when the solution in the soaking solution reached 7 g / L KNO 3 , 13 g / L CaCl 2 , 13 g / L ZnSO 4 , 0.5% NH 4 NO 3 , 250 mg / L gibberellin solution, 400 mg / L salicylic acid, 800 ppm wood vinegar liquid, 8% H 2 O 2After that, even if the concentration of the seed soaking solution increases, the malondialdehyde content in Keteleeria fortunei var. cyclolepis seeds remains in dynamic balance between 0.29 and 0.3; during germination, the malondialdehyde content in Keteleeria fortunei var. cyclolepis seeds reaches the maximum at 15 days. The generation rate and the maximum content of malondialdehyde in the seeds decrease with the increase in the concentration of the plant growth hormone mixture preparation soaked in coarse-grained activated carbon in the germination substrate. Moreover, the decline range of the maximum malondialdehyde content in Examples 1-3 is greater than that in Examples 4-6, indicating a positive correlation between the increase in the concentration of the plant growth hormone mixture preparation and the decrease in the malondialdehyde content in the seeds. The results prove that the method of the present invention changes the environmental factors during the germination process of Keteleeria fortunei var. cyclolepis seeds by using variable-temperature seed soaking with the seed soaking solution, slowly releasing the plant growth hormone mixture preparation by using coarse-grained activated carbon during the germination process, and controlling temperature and humidity. Compared with Comparative Examples 1-4, the variable-temperature seed soaking with the seed soaking solution in the method of the present invention reduces the malondialdehyde content in the seeds before germination, promotes the seeds to germinate in advance, the concentration of the plant growth hormone mixture preparation in the coarse-grained activated carbon promotes the conversion of malondialdehyde, and reduces the damage of malondialdehyde to the membrane structure of Keteleeria fortunei var. cyclolepis seeds during the germination process.

[0150] Table 4 Germination conditions and average germination rates of Keteleeria fortunei var. cyclolepis seeds in Examples 1-6

[0151]

[0152] Table 5 Germination conditions and average germination rates of Keteleeria fortunei var. cyclolepis seeds in Example 1 and Comparative Examples 1-4

[0153]

[0154] As shown in Table 4 and Table 5, the method of the present invention changes the environmental factors during the germination process of Keteleeria fortunei var. cyclolepis seeds by using variable-temperature seed soaking with the seed soaking solution, slowly releasing the plant growth hormone mixture preparation by using coarse-grained activated carbon during the germination process, and controlling temperature and humidity. Compared with Comparative Examples 1-4, the germination rate of Keteleeria fortunei var. cyclolepis seeds can be significantly increased during the germination process.

Claims

1. A method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors, characterized in that, the method comprises the following steps: S1, seed preparation; S2, seed disinfection; S3, Seed soaking treatment: Prepare a seed soaking solution for variable-temperature seed soaking. The seed soaking solution is prepared with 4 - 8 g / L KNO 3 , 10 - 15 g / L CaCl 2 , 10 - 15 g / L ZnSO4, 0.2% - 1% NH 4 NO 3 , 100 - 300 mg / L gibberellin solution, 100 - 500 mg / L salicylic acid, 500 - 1000 ppm wood vinegar solution, 3% - 10% H 2 O 2 and formulated; S4, germination substrate treatment; the germination substrate comprises 50%-70% coarse-grained activated carbon, 7%-24% debris, 8%-23% fine sand, 1%-5% peat, 1%-5% vermiculite, 1%-5% rock wool, and the coarse-grained activated carbon is obtained by soaking in a mixed preparation of plant growth hormones for 6-18 h after high-temperature disinfection, wherein the mixed preparation of plant growth hormones comprises 0.1-0.5 mg / L 2,4-dichlorophenoxyacetic acid, 0.05-0.2 mg / L brassinolide, 0.1-0.3 mg / L 6-benzylaminopurine, 0.05-0.3 mg / L kinetin, 0.4-0.7 mg / L rooting powder; the coarse-grained activated carbon comprises one or several mixtures of granular activated carbon and coconut shell activated carbon; the debris comprises one or several mixtures of sawdust and planing powder; S5, germination treatment: constant temperature and humidity germination is carried out in a germination chamber; the germination chamber comprises an air conditioner, an atomization device, and a germination substrate, and the soaked seeds are buried in the germination substrate at a fixed depth for germination, and the temperature and humidity of the germination chamber are controlled by the air conditioner and the atomization device during germination.

2. The method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors according to claim 1, characterized in that, the seed preparation in S1 is to select the seeds without damage and diseases by water selection after removing impurities.

3. The method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors according to claim 1, characterized in that, the seed disinfection in S2 is to soak and disinfect the seeds with 0.15-0.30% potassium permanganate solution for 10-25 min and rinse with distilled water 3-5 times.

4. The method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors according to claim 1, characterized in that, the soaking treatment in S3 further comprises: placing the selected Keteleeria cyclolepis seeds in S1 in a container filled with soaking solution, covering with a small-pore gauze and pressing to sink, placing the container in a water bath at 40-50 °C for constant temperature heating for 2-6 h, then moving the container to 20-30 °C for cooling treatment for 2-6 h; and then placing it in a water bath at 40-50 °C for constant temperature heating for 2-6 h.

5. The method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors according to claim 1, characterized in that, the high-temperature disinfection temperature of the coarse-grained activated carbon is 90-100 °C, and the disinfection time is 2-10 minutes.

6. The method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors according to claim 1, characterized in that, the method of the germination treatment in S5 is to bury the soaked Keteleeria cyclolepis seeds on the germination substrate at a depth of 1-5 cm.

7. The method for improving the germination rate of Keteleeria cyclolepis seeds by changing environmental factors according to claim 1, characterized in that, When germinating as described in S5, an air conditioner and an atomizing device are used to control the temperature and humidity in the germination chamber. The temperature control by the air conditioner is to keep the temperature in the germination chamber at 15 - 30 °C using the air conditioner, and the humidity control by the atomizing device is to control the relative humidity in the germination chamber at 15 - 30% using the atomizing device.

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