Malania oleifera seed germination accelerating method and application thereof

Abstract

The invention discloses a malania oleifera seed germination accelerating method and application thereof, and the method comprises the following steps: (1) applying mechanical force to malania oleifera seeds, so as to generate cracks on seed shells of malania oleifera to conduct the inner space of the seed shells and the outer part of the seed shells; the crack longitudinally extends on the seed shell and penetrates through the top of the seed shell; (2) soaking the seeds treated in the step (1) in an aqueous solution containing a dormancy breaking agent; the dormancy relieving agent comprises gibberellin and a cytokinin analogue; and (3) sowing the seeds treated in the step (2) in a breathable and water-containing culture medium to perform stratification and germination acceleration, and performing temperature control and regular irrigation in the period until the seeds germinate. According to the application, the method is applied to germination acceleration of malania oleifera seeds collected from wild malania oleifera plants in Guangxi Zhuang Autonomous Region and / or Yunnan.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of tree breeding. Specifically, it is a Malania oleifera seed germination method and its application. BACKGROUND

[0002] Malania oleifera Chun et S.Lee ex S.Lee is a evergreen tree of Olacaceae and Malania, which is a unique single-species woody oil plant in China. The seed coat of Malania oleifera is hard, the embryo physiological after-ripening period is long (75-195 days), and it is prone to rot due to long-term non-germination, which is the bottleneck problem leading to difficult germination under natural conditions. Although some studies have broken dormancy and promoted seed germination through mechanical damage and growth regulators, there are still problems of low germination rate (31.2-39.6%), long germination time (5-7 months), and uneven emergence, which cannot support the construction of Malania oleifera plantations.

[0003] In addition, there are significant differences in morphology, germination rhythm, embryo physiological after-ripening period, and sensitivity to mechanical damage and growth regulator treatment among different seed sources of Malania oleifera. There is no report on specific germination of different seed sources of Malania oleifera, which seriously hinders the process of large-scale seedling raising and artificial forest cultivation of Malania oleifera. Therefore, it is urgent to develop specific dormancy release and embryo physiological after-ripening promotion techniques for different seed sources of Malania oleifera to improve seed germination rate and shorten germination time, and provide scientific guidance for factory and standardized seedling raising. SUMMARY

[0004] Therefore, the technical problem to be solved by the present application is to provide a Malania oleifera seed germination method and its application for different seed sources, to solve the problems of low germination rate and long embryo physiological after-ripening period of existing Malania oleifera seeds. The method effectively breaks the dormancy of Malania oleifera seeds and promotes embryo physiological after-ripening to improve germination rate and germination uniformity by longitudinally breaking the seed shell, combined with disinfection, chemical treatment, and suitable germination culture conditions.

[0005] To solve the above technical problems, the present application provides the following technical solutions:

[0006] A Malania oleifera seed germination method, comprising the following steps:

[0007] (1) applying mechanical force to the Malania oleifera seed to generate a crack on the seed coat of the Malania oleifera, which leads to the space inside the seed coat and the outside of the seed coat; the crack extends longitudinally on the seed coat and penetrates the top of the seed coat; compared with the transversely extending crack, the longitudinally extending crack can extend to a closer distance to the Malania oleifera embryo, so that when the Malania oleifera seed is soaked in the aqueous solution containing the dormancy breaking agent, the dormancy breaking agent can better act on the embryo to promote the physiological after-ripening of the embryo;

[0008] (2) soaking the seed treated in step (1) in the aqueous solution containing the dormancy breaking agent; the dormancy breaking agent includes gibberellin and a cytokinin analogue;

[0009] (3) sowing the seed treated in step (2) in the breathable and water-containing culture medium for stratification germination, and controlling the temperature and air humidity of the culture environment, and irrigating regularly during the stratification process until the seed germinates.

[0010] In the above Malania oleifera seed germination method, before the mechanical force is applied to the Malania oleifera seed in step (1), the Malania oleifera seed is soaked in a disinfectant (such as a 0.5wt% potassium permanganate solution) for disinfection; the width of the crack is 0.5mm-1.0mm.

[0011] In the above Malania oleifera seed germination method, in step (2), the cytokinin analogue is 6-benzylaminopurine; in the aqueous solution, the mass fraction of the gibberellin is 0.05g / L-0.15g / L, and the mass fraction of the 6-benzylaminopurine is 0.005g / L-0.015g / L; the soaking time is 8-12h. If the soaking time is too short, the dormancy breaking agent cannot fully penetrate into the embryo tissue, which is insufficient to break dormancy, resulting in low germination rate, slow and uneven germination, and poor effect; if the soaking time is too long, the seed may be oxygen-deficient and excessively accumulate the dormancy breaking agent, which interferes with the endogenous hormone balance, resulting in inhibited seed rotting and germination, seedling malformation, and overgrowth.

[0012] The garlic pear seed germination method, in step (3), during the stratification germination process, the culture medium is irrigated with a growth promoter solution; in the growth promoter solution, the mass fraction of potassium dihydrogen phosphate is 1.0 g / L-2.0 g / L, the mass fraction of boric acid is 0.5 g / L-1.5 g / L; the mass fraction of zinc sulfate is calculated based on zinc sulfate heptahydrate, and the mass fraction of zinc sulfate heptahydrate is 0.25 g / L-0.75 g / L. Potassium can activate various metabolic processes in the plant cell growth system, thereby accelerating cell division and differentiation; boron promotes sugar transport to the embryo and enhances embryo vitality; zinc increases the content of indole acetic acid (IAA) in the embryo and promotes embryo differentiation. When the concentrations of the three substances are too low, they cannot fully provide nutrition and achieve the effect of activating metabolic processes. When the concentrations of potassium and zinc are too high, it will cause the dehydration, browning and necrosis of embryo and root tip cells, and when the concentration of boron is too high, it will inhibit the differentiation of the embryo and the extension of the root system. By adding specific concentrations of potassium dihydrogen phosphate, boric acid and zinc sulfate in the aqueous solution, the physiological after-ripening of garlic pear seeds can be effectively promoted and germination can be promoted.

[0013] The garlic pear seed germination method, in step (3), the culture medium is sterilized river sand; the environmental temperature during stratification germination is 25-30 DEG C, and the air relative humidity is 70%-80%; during the stratification germination process, the water content of the river sand is controlled to be 28wt%-32wt% by irrigation.

[0014] The garlic pear seed germination method, for the garlic pear seed source with a physiological after-ripening period of the seed embryo less than or equal to 75 days in a natural state: in step (1), the width of the crack is 0.5 mm-1.0 mm; in step (2), in the aqueous solution, the mass fraction of gibberellin is 0.05 g / L-0.15 g / L, and the mass fraction of 6-benzylaminoadenine is 0.005 g / L-0.015 g / L; the soaking time is 8-12 h; in step (3), the environmental temperature during stratification germination is 25-30 DEG C, and the air relative humidity is 70%-80%; during the stratification germination process, the water content of the river sand is controlled to be 28wt%-32wt% by irrigation.

[0015] In the seed germination accelerating method of Malania oleifera seeds, for the Malania oleifera seeds from the source with the physiological after-ripening period of the seed embryo being greater than 75 days in the natural state, in step (1), the width of the crack is 0.5 mm to 1.0 mm; in step (2), in the aqueous solution, the mass fraction of gibberellin is 0.05 g / L to 0.15 g / L, and the mass fraction of 6-benzylaminopurine is 0.005 g / L to 0.015 g / L; the soaking time is 8 to 12 hours; in step (3), the environmental temperature during the stratification germination is 25 to 30 DEG C, and the relative humidity of air is 70% to 80%; the culture medium is irrigated with the growth promoter solution during the stratification germination, and the water content of the culture medium is controlled to be 28 wt% to 32 wt%; in the growth promoter solution, the mass fraction of potassium dihydrogen phosphate is 1.0 g / L to 2.0 g / L, the mass fraction of boric acid is 0.5 g / L to 1.5 g / L, and the mass fraction of zinc sulfate is calculated based on zinc sulfate heptahydrate, and the mass fraction of zinc sulfate heptahydrate is 0.25 g / L to 0.75 g / L.

[0016] In the seed germination accelerating method of Malania oleifera seeds, in step (2), in the aqueous solution, the mass fraction of gibberellin is 0.10 g / L, and the mass fraction of 6-benzylaminopurine is 0.010 g / L; the soaking time is 10 hours.

[0017] In the seed germination accelerating method of Malania oleifera seeds, in step (3), the environmental temperature during the stratification germination is 25 DEG C, and the relative humidity of air is 80%; and the water content of the river sand is controlled to be 30 wt% by irrigation during the stratification germination.

[0018] Application of the seed germination accelerating method of Malania oleifera seeds, the seed germination accelerating method is applied to the Malania oleifera seeds collected from the wild Malania oleifera plants in Guangxi Zhuang Autonomous Region and Yunnan.

[0019] The technical scheme of the present application has the following beneficial technical effects:

[0020] 1. The present application first aims at the germination characteristics of different seed sources of Allamanda cathartica, and provides different seed germination schemes accordingly. For Tianlin and Bama seed sources in Guangxi Zhuang Autonomous Region, which have a short embryonic physiological after-ripening period (less than 75 days), the present application only needs to be combined with longitudinal hull breaking and gibberellin (GA3) + 6-benzylaminopurine (6-BA) dormancy release agent treatment under normal temperature conditions, so that the cumulative germination rate of the seeds can reach 70.22% after 18 days of sowing, which is 37.79 percentage points higher than 32.43% of the traditional non-hull breaking water treatment; at the same time, the germination peak period is about 42 days earlier than the control, realizing high efficiency, short cycle and rapid germination. For Yunnan seed sources, which have an embryonic physiological after-ripening period of more than 75 days, the present application combines precise temperature control and regular application of growth promoters during stratification germination on the basis of longitudinal hull breaking + dormancy release agent, so that the germination peak of Allamanda cathartica seeds from Yunnan appears after 80 days of sowing, and the cumulative germination rate reaches 47.00% after 110 days, which is 23.00 percentage points higher than the room temperature + water control, and the germination peak period is about 30 days earlier, significantly improving the germination efficiency and synchronicity, and effectively reducing the stratification rot rate.

[0021] 2. The longitudinal hull breaking device and chemical treatment method used in the present application are simple to operate, the materials are easy to obtain, and the measures for different seed germination characteristics of different seed sources are flexible and adjustable, having good repeatability and large-scale production potential, providing reliable technical basis and popularization demonstration value for protective rescue breeding of Allamanda cathartica seed resources, directional cultivation of artificial forest and factory-like, standardized seedling raising. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 The crack direction of the Allamanda cathartica seeds in the present application is shown in the schematic diagram of longitudinal hull breaking and transverse hull breaking;

[0023] Figure 2 The photo of the Allamanda cathartica seeds after artificial hull breaking immersed in the solution containing dormancy release agent in the present application;

[0024] Figure 3 The river sand bed for stratification germination in the present application;

[0025] Figure 4 The river sand bed with Allamanda cathartica seeds buried under artificial temperature control in the present application;

[0026] Figure 5 The germination of Allamanda cathartica seeds under different stratification germination temperatures in the present application. DETAILED DESCRIPTION

[0027] 1. Study on the differences in morphology, germination rhythm and embryonic physiological after-ripening period of Allamanda cathartica seeds from different seed sources

[0028] From four provenances of Guangnan and Funing in Yunnan province, Tianlin and Bama in Guangxi Zhuang Autonomous Region, 20 seeds of Allosphenium podocarpum were randomly selected from each provenance. The seed coat was disinfected by soaking in 0.5% potassium permanganate solution for 2 hours and then rinsing with sterile water, and then dried in the shade. The length of the seed stalk, the longitudinal diameter of the seed core, the transverse diameter of the seed core, the longitudinal diameter of the seed kernel, and the transverse diameter of the seed kernel were measured accurately using a vernier caliper. The dry weight of the seed core and the dry weight of the seed kernel were measured using a balance. The core shape index and kernel shape index were calculated by dividing the longitudinal diameter by the transverse diameter. Among them, "seed core" refers to the Allosphenium podocarpum seed with shell, "seed stalk length" refers to the length of the sharp protrusion at the bottom of the Allosphenium podocarpum seed shell, "seed core longitudinal diameter" refers to the distance from the core stalk to the top of the core, "seed core transverse diameter" refers to the diameter of the widest part of the core perpendicular to the longitudinal diameter, "seed kernel longitudinal diameter" refers to the distance from the bottom of the seed kernel to the top of the seed kernel, and "seed kernel transverse diameter" refers to the diameter of the widest part of the seed kernel perpendicular to the longitudinal diameter.

[0029] The morphological characteristics of Allosphenium podocarpum seeds from different provenances are shown in Table 1. There were significant differences in the morphological characteristics of Allosphenium podocarpum seeds from Yunnan and Guangxi Zhuang Autonomous Region. The dry weight of the seed core, the transverse diameter of the seed kernel, and the dry weight of the seed kernel of Allosphenium podocarpum seeds from Guangnan and Funing in Yunnan were significantly greater than those from Tianlin and Bama in Guangxi Zhuang Autonomous Region (P<0.05). According to the data of kernel dry weight / core dry weight, the seed shell of Allosphenium podocarpum seeds from Guangxi Zhuang Autonomous Region was thicker.

[0030] Table 1 Variations in morphological characteristics of Allosphenium podocarpum seeds from different provenances

[0031]

[0032] About 150 Allosphenium podocarpum seeds were taken from each of the four provenances, and they were divided into two categories according to Yunnan and Guangxi Zhuang Autonomous Region. A total of 296 seeds were from Guangxi Zhuang Autonomous Region, and 330 seeds were from Yunnan. On October 12, 2023, untreated Allosphenium podocarpum seeds were directly sown in sterilized river sand, and the germination of the seeds was observed. The emergence of the root tip was counted as germination, and the number of germinated seeds was counted every 30 days until the number of newly germinated seeds was less than 1% of the total number of seeds. The physiological after-ripening period of the seed embryo was determined. The statistical results are shown in Table 2.

[0033] Table 2 Differences in germination rhythm and physiological after-ripening period of Allosphenium podocarpum seeds from different provenances

[0034]

[0035] As shown in Table 2, garlic seeds from Guangxi Zhuang Autonomous Region began germinating within 30 days of sowing, with a cumulative germination rate of 32.43% after 60 days. In contrast, garlic seeds from Yunnan Province only germinated in small numbers between 30 and 60 days after sowing, with a cumulative germination rate of only 22.73% after 120 days. This indicates that under natural conditions, there are significant differences in the physiological after-ripening period of garlic seeds from different sources. Furthermore, garlic seeds from Yunnan Province exhibit a prominent problem of an excessively long physiological after-ripening period and a low germination rate, necessitating different treatments for different sources to improve germination rates.

[0036] 2. Screening of mechanical shell-breaking methods and dormancy-breaking agents for seeds from Tianlin and Bama seed sources in Guangxi Zhuang Autonomous Region.

[0037] In this embodiment, to address the problem of low germination rate of garlic seeds from Tianlin and Bama in Guangxi Zhuang Autonomous Region, seed dormancy is broken through a dual approach of physical shell breaking and dormancy-breaking agent treatment.

[0038] First, the effects of different seed-breaking treatments on promoting the breaking of dormancy in garlic clove seeds were tested, such as... Figure 1 As shown, without damaging the endosperm, use a wooden stick to crack the seed coat longitudinally or transversely, creating conductive cracks on the seed coat (i.e., cracks that connect the space inside the seed coat to the outside of the seed coat). Figure 1 The seeds in the upper left and lower left corners are garlic cloves that have cracked horizontally, while the seeds in the upper right and lower right corners are garlic cloves that have cracked vertically. The direction in which the crack on the seed coat of the seed in the upper right corner extends is "vertical".

[0039] Then, two plant growth regulators, gibberellin (GA3) and 6-benzylaminopurine (6-BA), were prepared into an aqueous solution and used to soak the cracked garlic seeds to test the dormancy-breaking effect of these two plant growth regulators on garlic seed dormancy. In this embodiment, gibberellin and 6-BA are the dormancy-breaking agents.

[0040] Gibberellin was treated with a concentration gradient of 0 g / L, 0.05 g / L, 0.10 g / L, and 0.15 g / L; 6-benzylaminopurine was treated with a concentration gradient of 0 g / L, 0.005 g / L, 0.010 g / L, and 0.015 g / L; garlic seeds were treated with the above dormancy-breaking agents for 10 hours (e.g., ...). Figure 2 As shown), seeds were sown in river sand (river sand bed) sterilized with a 0.5% potassium permanganate solution for stratification and germination. The river sand was kept moist and well-aerated throughout the stratification and germination process, with the moisture content (mass fraction of water in the river sand) controlled at approximately 30 wt%. Figure 3 This is a river sand bed containing garlic clove seeds.

[0041] Table 3 summarizes the germination of seeds of Melia toosendan from Tianlin and Bama in Guangxi Zhuang Autonomous Region. The control group (without cracking the seed coat, only soaked with water) started to germinate 30 days after sowing, and the cumulative germination rate was 32.43% at 60 days after sowing. The treatment of cracking the seed coat (whether horizontally or vertically) and soaking with water could slightly advance the germination time, but the cumulative germination rate did not significantly increase. For the seeds with horizontal cracking of the seed coat and treatment with dormancy-breaking agents (GA30.10 g / L + 6-BA 0.010 g / L), more seeds started to germinate at 18 days after sowing, and the cumulative germination rate was 46.08%-52.04% at 30-60 days after sowing. The cumulative germination rate at 60 days after sowing was 19.61 percentage points higher than that of the control (not cracked + water), and the germination peak period was about 30 days earlier. In the treatment group with vertical cracking of the seed coat and the same dormancy-breaking agents, the cumulative germination rate was 70.22% at 18 days after sowing, which was 37.79 percentage points higher than that of the control (not cracked + water) at 60 days after sowing, and the germination peak period was about 42 days earlier.

[0042] Table 3 Differences in seed germination of Melia toosendan from Guangxi Zhuang Autonomous Region under different mechanical cracking and dormancy-breaking agent treatments

[0043]

[0044]

[0045] 3. Temperature regulation and growth promoter screening for Yunnan Guangnan and Funing provenances

[0046] In this example, to address the problems of long physiological after-ripening period and low seed germination rate of Melia toosendan seeds from Guangnan and Funing provenances in Yunnan, further tests were conducted on the promotion of temperature and growth promoters during the stratification and germination process on the basis of the vertical physical cracking of the seed coat + 0.10 g / L gibberellin solution + 0.010 g / L 6-benzylaminopurine solution to break seed dormancy in "2. Mechanical cracking of seeds and dormancy-breaking agent screening for Tianlin and Bama provenances in Guangxi Zhuang Autonomous Region".

[0047] In this embodiment, the seeds of Elaeagnus oldhami from Yunnan provenance were subjected to longitudinal hull breaking and dormancy breaking agent (0.10 g / L gibberellin + 0.010 g / L 6-benzylaminopurine) soaking treatment. Then, when stratification and germination were carried out, the optimal concentrations of three growth promoters were first screened at room temperature, and the mass fraction of potassium dihydrogen phosphate solution was set to 0.5 g / L, 1.0 g / L, 1.5 g / L and 2.0 g / L, the mass fraction of boric acid solution was set to 0.5 g / L, 1.0 g / L, 1.5 g / L and 2.0 g / L, and the mass fraction of zinc sulfate heptahydrate solution was set to 0.25 g / L, 0.5 g / L, 0.75 g / L and 1.00 g / L. Each treatment was irrigated with growth promoter solution once a week, and the irrigation amount was 20-25 L per square meter. The room temperature culture and water irrigation were used as controls, and 100 seeds were treated in each treatment. The sand was kept moist and ventilated during the whole stratification and germination process, and the water content (mass fraction of water in sand) was controlled at about 30 wt%. In this embodiment, the room temperature was the temperature from October to the next January in Guangzhou, i.e. 9-30°C on average.

[0048] Table 4 Difference in germination of Elaeagnus oldhami seeds from Yunnan provenance treated with different concentrations of growth promoters at room temperature

[0049]

[0050] As shown in Table 4, on the basis of the longitudinal hull breaking + dormancy breaking agent treatment carried out in the foregoing, the physiological after-ripening period of Elaeagnus oldhami seeds from Yunnan provenance was significantly shortened and the germination rate of Elaeagnus oldhami from Yunnan provenance was improved by perfusion with growth promoter solution during stratification and germination. The control group (room temperature + water) did not show germination 30 days after sowing, and only 8.00% showed germination 60 days after sowing. However, all the growth promoter treatment groups showed germination 30 days after sowing, except that the 1.00 g / L mass fraction zinc sulfate heptahydrate solution treatment showed germination inhibition. The other growth promoter treatments effectively shortened the germination period and significantly improved the subsequent seed germination rate.

[0051] Therefore, the optimal concentrations of the three growth promoters were screened in this embodiment, i.e. potassium dihydrogen phosphate (KH2PO4, mass fraction 1.5 g / L in solution), boric acid (H3BO3, mass fraction 1.0 g / L in solution) and zinc sulfate heptahydrate (ZnSO4·7H2O, mass fraction 0.50 g / L in solution).

[0052] The three substances were sequentially dissolved in water, mixed after dissolution to obtain growth promoters, and a series of temperature gradients 15°C, 20°C, 25°C and 30°C were set for temperature treatment, and the stratification and germination were carried out in an air humidity of 80% (see Table 5). Figure 4The seeds of some of the treatment groups were treated at different temperatures. The seeds were irrigated with a growth promoting agent solution once a week, with the amount of irrigation being 20-25 L per square meter. The seeds were incubated at room temperature and irrigated with water as a control. The sand was kept moist and well aerated during the entire stratification process, with the water content being controlled at about 30 wt%. The seeds of each treatment group were 100 seeds.

[0053] Table 5: Differences in seed germination of Yunnan provenance Elaeagnus oldhamii seeds treated at different temperatures and with a growth promoting agent

[0054] Treatment Germination statistics 45 days after sowing 80 days after sowing 110 days after sowing 15°C + growth stimulant Cumulative germination rate (%) 8.00 17.00 27.00 20°C + growth stimulant Cumulative germination rate (%) 9.00 21.00 29.00 25°C + growth stimulant Cumulative germination rate (%) 12.00 37.00 47.00 30°C + growth stimulant Cumulative germination rate (%) 9.00 31.00 39.00 Room temperature + clean water Cumulative germination rate (%) 1.00 11.00 24.00

[0055] As shown in Table 5, in this embodiment, on the basis of the aforementioned longitudinal hull breaking + dormancy breaking agent treatment, by performing temperature regulation during stratification and incubation, and irrigating with a growth promoting agent solution, the physiological after-ripening period of the seeds of Yunnan provenance Elaeagnus oldhamii was significantly shortened, and the germination rate of the seeds of Yunnan provenance Elaeagnus oldhamii was increased. All of the temperature regulation + growth promoting agent groups had observable germination 45 days after sowing, while the control group (room temperature + water) had only 1.00% germination, indicating that temperature regulation and irrigation with a growth promoting agent significantly broke seed dormancy, promoted physiological after-ripening and germination. Photographs of some of the germinated Elaeagnus oldhamii seeds in each temperature treatment of this embodiment are shown in Figure 5

[0056] The cumulative germination rate of the 25°C + growth promoting agent group was 12.00% at 45 days, which was higher than that of the 15°C (8.00%), 20°C (9.00%) and 30°C (9.00%) groups. By 80 days after sowing, the cumulative germination rate of this group was 37.00%, which was much higher than that of the 15°C (17.00%), 20°C (21.00%) and 30°C (31.00%) groups, and was the highest among all of the treatment groups. By 110 days after sowing, the cumulative germination rate of this group reached 47.00%, which was 23.00 percentage points higher than that of the control group (24.00%), and was also 8-20 percentage points higher than that of the other temperature treatment groups, indicating that 25°C was the most suitable temperature. As can be seen from Figure 5

[0057] Under the condition of 15°C, although some of the Elaeagnus oldhamii seeds broke dormancy (8.00% of the seeds germinated at 45 days), the highest cumulative rate was only 27.00% at a later stage, indicating that low temperature limited subsequent physiological maturation. At 30°C, the initial performance was similar to that at 20°C (9.00% of the seeds germinated at 45 days), but by 80 days, the cumulative germination rate reached 31.00%, which was lower than that of the 25°C group, indicating that excessively high temperature might inhibit seed germination of Elaeagnus oldhamii.

[0058] ​​As can be seen from the comparison with the control group, the treatment of simple room temperature + water did not reach 24.00% until 110 days, while the 25°C + accelerant group reached 37.00% at 80 days and 47.00% at 110 days, indicating that temperature regulation and growth accelerant can advance the germination peak by about 30 days and greatly improve the germination rate of the Allium cepa L. seeds from Yunnan.

[0059] 4. Accelerated germination of Allium cepa L. seeds from Tianlin County, Baise City, and Bama County, Hechi City, Guangxi Zhuang Autonomous Region (1) Seed disinfection and soaking

[0060] The seeds were soaked in 0.5wt% potassium permanganate solution (as a disinfectant) for 2.0h to remove surface microorganisms; then washed with sterile water for three times and drained for standby. The purpose of this step is mainly to disinfect, and also to make the seed coat absorb water and soften to some extent, which facilitates the subsequent accelerated germination operation. This is because: the Allium cepa L. seed coat contains a large number of microorganisms inside and outside, and the seed kernel is rich in oil, which is easy to rot during the accelerated germination process if not disinfected. In addition, if the soaking time is too short, the disinfection will not be complete, and if the soaking time is too long, the embryo may be damaged.

[0061] (2) Longitudinal hull breaking treatment

[0062] A sterile wooden stick was used to lightly tap the seeds, and a longitudinal crack was knocked on the seed coat without damaging the endosperm, and the crack must penetrate the top of the seed coat. For example, Figure 1 The right side is the seed that has completed the longitudinal hull breaking treatment, Figure 1 The lower right shows the crack penetrating the top of the seed coat. For example, Figure 1 The seed in the upper right corner is an example. The funiculus of Allium cepa L. is located at the bottom of the seed coat ( Figure 1 upper part of the seed in the upper right corner), and the embryo is located at the top ( Figure 1 lower part of the seed in the upper right corner) to 1-3mm inside the shell. The width of the crack is about 0.5-1.0mm, and when the crack is too wide, it means that the tapping force is too large, and the seed kernel may be damaged. When the crack is too narrow, it means that the tapping force is small, and the crack may not penetrate the top of the seed coat.

[0063] (3) Soaking in solution containing dormancy breaking agent

[0064] The broken hull seeds were soaked in an aqueous solution containing gibberellin (GA3, mass fraction 0.10g / L) and 6-benzylaminopurine (6-BA, mass fraction 0.010g / L) (this solution is the solution of the dormancy breaking agent) for 10.0h at room temperature (25±2℃). Gibberellin and 6-BA are dormancy breaking agents that can break the dormancy of Allium cepa L. seeds. For Guangxi Zhuang Autonomous Region, only longitudinal hull breaking + dormancy breaking agent is needed to achieve the desired effect of germination time and germination rate (the treated Allium cepa L. seeds should be fully germinated).

[0065] (4) Sowing and Cultivation

[0066] The soaked seeds are sown in the river sand bed. The river sand in the river sand bed is disinfected by 0.5% potassium permanganate solution, and the water content is about 30 wt%. The thickness of the river sand is 6-10 cm, and the depth is 2-3 cm. If the thickness of the river sand is too thin, the water holding capacity of the sand bed will decrease, and if the thickness of the river sand is too thick, the permeability of the sand bed will decrease and the seeds will easily rot. If the depth is too shallow, the water in the seeds will easily evaporate, and if the depth is too deep, the permeability will be poor, resulting in slow germination or rotting.

[0067] The sown river sand bed is placed in an environment with a temperature of 25°C and a relative humidity of air for cultivation to stratify and germinate. The river sand is watered daily to keep the river sand moist (the water content is controlled at about 30 wt%), and the river sand has good permeability. No growth regulator needs to be added to the river sand bed.

[0068] 5. Accelerated Germination of Garlic Pear Seeds from Funing County and Guangnan County, Wenshan Zhuang and Miao Autonomous Prefecture, Yunnan Province

[0069] The physiological after-ripening process of garlic pear seeds from Guangnan and Funing sources in a natural state requires more than 75 days. In this embodiment, after breaking the dormancy of garlic pear seeds by hull breaking and dormancy breaking agents, precise temperature control measures are used to accelerate the process of converting starch, soluble protein, and soluble sugar in the endosperm into small molecule nutrients and promote the directional transport of these small molecule nutrients to the embryo. At the same time, exogenous growth promoters are supplemented to provide nutrients for seed embryo development, thereby systematically accelerating the physiological after-ripening process of the seed embryo and ultimately achieving the dual technical goals of shortening the seed germination period and improving the germination rate.

[0070] The specific steps of the accelerated germination method in this embodiment are as follows:

[0071] (1) Disinfection and seed soaking

[0072] The same method as that for garlic pear seeds from Tianlin County, Baise City, and Bama County, Hechi City, Guangxi Zhuang Autonomous Region is used for disinfection and seed soaking.

[0073] (2) Longitudinal hull breaking

[0074] The same method as that for garlic pear seeds from Tianlin County, Baise City, and Bama County, Hechi City, Guangxi Zhuang Autonomous Region is used for longitudinal hull breaking.

[0075] (3) Soaking with dormancy breaking agents

[0076] The broken shell seeds are placed in a water solution containing a dormancy breaking agent and soaked at room temperature (25±2℃) for 10.0 hours. The dormancy breaking agent is gibberellin (GA3, mass fraction in solution 0.10 g / L) and 6-benzylaminopurine (6-BA, mass fraction in solution 0.010 g / L).

[0077] (4) Sowing culture and temperature control and solution irrigation containing growth promoters

[0078] The sowing method and the preparation method of the river sand bed are the same as those of the seeds of the Garcinia mangostana in Tianlin County, Baise City and Bama County, Guangxi Zhuang Autonomous Region. After sowing, the river sand bed is placed in an environment with a temperature of 25℃ and an air relative humidity of 80% for stratification and germination. The river sand is watered daily to keep it moist, and the river sand is well ventilated. The solution containing growth promoters is poured into the sand bed every 7 days. The solution containing growth promoters used in this step contains potassium dihydrogen phosphate (KH2PO4, mass fraction in solution 1.5 g / L), boric acid (H3BO3, mass fraction in solution 1.0 g / L) and zinc sulfate (ZnSO4·7H2O, mass fraction in solution 0.50 g / L), and the river sand bed is controlled to maintain a water content (calculated by mass fraction) of about 30wt%.

[0079] In other embodiments, the dormancy breaking agent can be an aqueous solution composed of 0.05 g / L to 0.15 g / L gibberellin (GA3) and 0.005 g / L to 0.015 g / L 6-benzylaminopurine (6-BA). The growth promoter can be an aqueous solution composed of 1.0 g / L to 2.0 g / L potassium dihydrogen phosphate, 0.5 g / L to 1.5 g / L boric acid and 0.25 g / L to 0.75 g / L zinc sulfate (ZnSO4·7H2O). The concentrations of the components are all mass fractions, and the solvent is sterile water.

[0080] Obviously, the above embodiments are only examples for clarity and are not limiting of the embodiments. Based on the above description, those skilled in the art can make other different forms of changes or variations. Here, it is not necessary and impossible to exhaust all the embodiments. The obvious changes or variations derived therefrom are still within the protection scope of the claims of the present patent application.

Claims

1. A method for germinating garlic clove seeds, characterized in that, Includes the following steps: (1) Apply mechanical force to the garlic seed to create cracks on the seed coat that connect the internal space of the seed coat to the external space of the seed coat; the cracks extend longitudinally on the seed coat and penetrate the top of the seed coat. (2) Soak the seeds treated in step (1) in an aqueous solution containing a dormancy-breaking agent; the dormancy-breaking agent includes gibberellin and cytokinin analogs; (3) Sow the seeds treated in step (2) in a breathable and water-containing culture medium for stratification and germination, and control the temperature and water regularly until the seeds germinate.

2. The method for germinating garlic clove seeds according to claim 1, characterized in that, In step (1), the width of the crack is 0.5 mm to 1.0 mm.

3. The method for germinating garlic clove seeds according to claim 1, characterized in that, In step (2), the cytokinin analog is 6-benzylaminopurine; in the aqueous solution, the mass-volume fraction of the gibberellin is 0.05 g / L to 0.15 g / L, and the mass-volume fraction of the 6-benzylaminopurine is 0.005 g / L to 0.015 g / L; the soaking time is 8 to 12 hours.

4. The method for germinating garlic clove seeds according to claim 1, characterized in that, In step (3), during the stratification and germination process, the culture medium is irrigated with a growth promoter solution; in the growth promoter solution, the mass volume fraction of potassium dihydrogen phosphate is 1.0 g / L to 2.0 g / L, the mass volume fraction of boric acid is 0.5 g / L to 1.5 g / L, and the mass volume fraction of zinc sulfate is calculated as zinc sulfate heptahydrate, which is 0.25 g / L to 0.75 g / L.

5. The method for germinating garlic clove seeds according to claim 1, characterized in that, In step (3), the culture medium is sterilized river sand; the ambient temperature during stratification germination is 25-30℃ and the relative humidity is 70%-80%; during stratification germination, the moisture content of the river sand is controlled to be 28wt%-32wt% by irrigation.

6. The method for germinating garlic clove seeds according to claim 1, characterized in that, For garlic seed sources with a physiological after-ripening period of less than or equal to 75 days under natural conditions: In step (1), the width of the crack is 0.5 mm to 1.0 mm; In step (2), the mass volume fraction of gibberellin in the aqueous solution is 0.05 g / L to 0.15 g / L, and the mass volume fraction of 6-benzylaminopurine is 0.005 g / L to 0.015 g / L; The soaking time is 8 to 12 hours; In step (3), the ambient temperature during stratification and germination is 25 to 30°C, and the relative humidity is 70% to 80%; During the stratification and germination process, the moisture content of the river sand is controlled to be 28 wt% to 32 wt% by irrigation water.

7. The method for germinating garlic clove seeds according to claim 1, characterized in that, For garlic seed sources with a physiological after-ripening period of more than 75 days under natural conditions: In step (1), the width of the crack is 0.5 mm to 1.0 mm; in step (2), the gibberellin in the aqueous solution has a mass-volume fraction of 0.05 g / L to 0.15 g / L, and the 6-benzylaminopurine has a mass-volume fraction of 0.005 g / L to 0.015 g / L; the soaking time is 8 to 12 hours; in step (3), the ambient temperature during stratification and germination is 25 to 30°C, and the relative humidity is... The humidity is 70%–80%; during the tiered germination process, the culture medium is irrigated with a growth promoter solution, and the water content of the culture medium is controlled to be 28wt%–32wt%; in the growth promoter solution, the mass volume fraction of potassium dihydrogen phosphate is 1.0g / L–2.0g / L, and the mass volume fraction of boric acid is 0.5g / L–1.5g / L; the mass volume fraction of zinc sulfate is calculated as zinc sulfate heptahydrate, and the mass volume fraction of zinc sulfate heptahydrate is 0.25g / L–0.75g / L.

8. The method for germinating garlic clove seeds according to claim 6 or 7, characterized in that, In step (2), the gibberellin in the aqueous solution has a mass-volume fraction of 0.10 g / L and the 6-benzylaminopurine has a mass-volume fraction of 0.010 g / L; the soaking time is 10 h.

9. The method for germinating garlic clove seeds according to claim 6 or 7, characterized in that, In step (3), the ambient temperature during stratification germination is 25°C and the relative humidity is 80%. During the stratification germination process, the moisture content of the river sand is controlled to be 30 wt% by irrigation.

10. The application of a method for germinating garlic clove seeds, characterized in that, The method described in claim 1 is applied to the germination of garlic seeds collected from wild garlic plants.

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