CN114766355A

CN114766355A - Lycium ruthenicum artificial seed preparation device and regeneration factor research method thereof

Info

Publication number: CN114766355A
Application number: CN202210258586.6A
Authority: CN
Inventors: 张欣; 贝盏临
Original assignee: North Minzu University
Current assignee: North Minzu University
Priority date: 2022-03-16
Filing date: 2022-03-16
Publication date: 2022-07-22

Abstract

The invention discloses a lycium ruthenicum artificial seed preparation device and a regeneration factor research method thereof, and relates to the technical field of biological science research. According to the invention, the external driving motor is controlled to work, the first bevel gear in the transmission box body can be driven to rotate, the second bevel gear and the transmission gear drive the inner gear ring to rotate, the inner ring is driven to rotate on the inner wall of the fixing ring under the design of the rotating ring, and the experimental instrument at the top of the first mesh plate is driven to rotate, so that the contact effect of the outer wall of the experimental instrument and steam is improved, the problem of low contact rate between the sunken part of the outer wall of the experimental instrument and the steam is avoided, and the sterilization sufficiency of the device is improved.

Description

Lycium ruthenicum artificial seed preparation device and regeneration factor research method thereof

Technical Field

The invention relates to the technical field of biological science research, in particular to a lycium ruthenicum artificial seed preparation device and a regeneration factor research method thereof.

Background

Lycium ruthenicum Murr is a spiny shrub of Lycium of Solanaceae, is rich in nutrition, is rich in anthocyanin, has various benefits to human body, has the effects of protecting liver and improving eyesight, has a very good antioxidation effect, can eliminate free radicals in human body, achieves a remarkable anti-aging effect, and has a main value that the Lycium ruthenicum Murr can be planted as a shrub for water and soil conservation. The following problems exist in the prior art:

1. the existing high-pressure steam sterilization pot does not have the function of fully sterilizing experimental instruments, and the contact rate of the depressions on the outer wall of the experimental instruments and steam is low in the sterilization process, so that the sterilization sufficiency is influenced and needs to be improved;

2. the wild lycium ruthenicum murr has the advantages of severe growth environment, less distribution, very low germination rate in a natural state and limited quantity for human utilization, so that the preparation of the lycium ruthenicum murr artificial seeds is needed.

Disclosure of Invention

The invention provides a lycium ruthenicum artificial seed preparation device and a regeneration factor research method thereof, wherein one purpose is to have a function of fully sterilizing experimental instruments and solve the problem of low contact rate between depressions on the outer wall of the experimental instruments and steam; the other purpose is to solve the problem that the quantity of the wild lycium ruthenicum for human use is limited, so that the effect of improving the seed germination rate by researching the regeneration factors of the wild lycium ruthenicum is achieved.

In order to solve the technical problems, the technical scheme adopted by the invention is as follows:

in a first aspect, the invention provides a lycium ruthenicum artificial seed preparation device which comprises a high-pressure steam sterilization pot body and an external circulating water cooling device, wherein a steam nozzle is arranged at the bottom of an inner cavity of the high-pressure steam sterilization pot body, a fixed lantern ring is fixedly mounted on the outer wall of the high-pressure steam sterilization pot body, a control button is arranged on the front surface of the fixed lantern ring, a steam inlet pipe orifice is arranged on the front surface of the high-pressure steam sterilization pot body, a sealing cover is detachably connected to the top of the high-pressure steam sterilization pot body, a pressure relief valve pipe is fixedly connected to the top of the sealing cover, a steam treatment mechanism is arranged on the top of the sealing cover, and a sufficient sterilization mechanism is arranged in the inner cavity of the high-pressure steam sterilization pot body.

Steam treatment mechanism includes the sealing ring, sealing ring fixed mounting is at the top of sealed lid, the dismantlement formula is connected with the connecting pipe on the inner wall of sealing ring, the top fixedly connected with buckling pipe of connecting pipe, the bottom fixedly connected with condenser pipe of buckling pipe, the equal fixedly connected with water pipe interface in both sides of condenser pipe, external circulating water cooling device's outlet pipe and left water pipe interface threaded connection, external circulating water cooling device's inlet tube and the water pipe interface threaded connection on right side, the splitter box has all been seted up to the both sides of condenser pipe inner wall.

The sufficient sterilization mechanism comprises an external drive motor, a transmission box body and a fixing ring, the external drive motor is fixedly installed on the left side of the high-pressure steam sterilization pot body, the transmission box body is fixedly installed on the inner wall of the high-pressure steam sterilization pot body, an output shaft of the external drive motor extends to a first bevel gear fixedly connected in an inner cavity of the transmission box body, the top of the first bevel gear is meshed with a second bevel gear, a transmission gear is fixedly connected to the top of the second bevel gear, the fixing ring is fixedly welded on the inner wall of the high-pressure steam sterilization pot body, the fixing ring is fixedly connected with a rotating ring in the internal rotation mode, an inner ring is fixedly connected to the inner wall of the rotating ring, and an inner toothed ring is fixedly installed on the inner wall of the inner ring.

The technical scheme of the invention is further improved as follows: the top and the equal fixedly connected with flow distribution plate in bottom of condenser pipe inner chamber, two one side fixedly connected with exchange tube that the flow distribution plate is adjacent, the curved copper pipe of left side fixedly connected with of exchange tube is one, fixed welding has the guide block on the inner wall of curved copper pipe one, the terminal delivery pipe of right side fixedly connected with of exchange tube, the left side fixedly connected with of terminal delivery pipe concentrates the copper pipe, concentrate on the outer wall of copper pipe fixedly connected with curved copper pipe two, the outer wall fixed connection of one end of concentrating the copper pipe and curved copper pipe one is kept away from to curved copper pipe two.

The technical scheme of the invention is further improved as follows: the bottom fixedly connected with drain pipe of condenser pipe, fixed mounting has the direction awl on the inner wall of drain pipe, fixedly connected with exhaust pipe chute on the outer wall of drain pipe, the bottom threaded connection of drain pipe has a water storage section of thick bamboo.

The technical scheme of the invention is further improved as follows: the inner wall of inner ring gear and drive gear's outer wall meshing, the top fixed mounting of inner ring has the connecting seat, the top dismantlement formula of connecting seat is connected with the connecting block, the inboard fixed mounting of connecting block has otter board one, the top dismantlement formula of otter board one is connected with the magnetic force and connects the foot, the top fixed mounting of magnetic force connection foot has otter board two, the fixed welding in top of otter board two has vertical base bar, the inboard fixed mounting of vertical base bar has around the ring, it has the rubber sleeve to fix the cover on the outer wall around the ring.

In a second aspect, the invention also provides a method for researching regeneration factors of the lycium ruthenicum artificial seeds, which comprises the following steps:

s1, preparing a laboratory instrument and a reagent;

s2, inducing somatic embryos;

s3, preparing artificial endosperm;

s4, manufacturing artificial seed coats.

Further: the step of S1 further includes: the experimental instrument comprises: high-pressure steam sterilization pot body, superclean bench, balance, blake bottle, culture dish, pipetting gun, digital display constant temperature water bath, rubber head burette, aurilave, burette, beaker and volumetric flask prepare the completion, include experimental reagent again: preparing MS culture medium, sucrose, 50% concentrated sulfuric acid, agar, NAA with different concentrations, 6BA, 2,4-D, 70% alcohol, 1% -2% sodium hypochlorite, 0.1% mercuric chloride, 2% sodium alginate, 3% sodium alginate, 4% sodium alginate, 2% calcium chloride and distilled water.

And further: the step S2 further includes the steps of:

step A1, cultivation of aseptic seedlings: using wild lycium ruthenicum seeds, firstly preparing an MS culture medium, packaging a laboratory instrument, and carrying out high-pressure steam sterilization by using a plurality of bottles of distilled water, wherein the sterilization conditions are as follows: sterilizing at 121 ℃ and 101kPa for 20min, taking out when the pressure is zero so as to avoid danger, soaking Lycium ruthenicum Murr seeds in 50% concentrated sulfuric acid for 5min, breaking the dormancy of the seeds, inoculating the seeds to an MS culture medium by using a plant tissue culture technology, placing all culture bottles on a specified culture rack in a culture room at the same interval, observing whether the culture medium is polluted or not and whether the seeds germinate or not every two or three days, observing, counting and recording for two weeks, finding that the seeds in all the culture mediums germinate, and finally counting the germination rate of the Lycium ruthenicum Murr to be 100%;

step A2, callus induction: inducing callus with early cultured Lycium ruthenicum Murr plant in different hormone formulas, and dedifferentiating to form somatic embryo, wherein the culture medium formula is as follows: MS +6-BA + NAA +6g/l agar +30g/l sucrose.

Further: the step S3 further includes the steps of: the preparation of the artificial endosperm is to screen out a culture medium formula suitable for the germination and growth of the embryoid, finally, the screened culture medium is added into an embedding medium, different crops and different propagules have different requirements on the culture medium, in a preparation pre-experiment of the artificial endosperm of lycium ruthenicum, compared with an MS culture medium, the 1/2MS culture medium has better effect, so a 1/2MS culture medium is selected, on the basis, NAA with the largest influence on the germination rate of the artificial seeds and 2% sodium alginate are selected for the experiment, a proper amount of sucrose is also added into the artificial endosperm, and the culture medium formula is as follows: the artificial endosperm is prepared by performing two-factor three-level test on MS + NAA + sodium alginate.

Further: the step S4 further includes the steps of: the method for forming the gel-coated material by using the hydrogel method through ion transformation or temperature mutation is an operation method of a commonly used ion exchange method embedded by sodium alginate, and comprises the following steps of: firstly, adding sodium alginate with different concentrations into an MS culture medium to prepare gel, adding a certain proportion of embryoid, then fully and uniformly mixing the culture medium and the sodium alginate, sucking up the embryoid and the gel by using a dropper, then dropping the mixture into a 2% CaCl2 solution to stay for 10-15min, completely combining the surface of the embryoid with the gel to form a round artificial seed with certain rigidity, rinsing the artificial seed with sterile water for 20min, stopping the reaction, then inoculating the prepared artificial seed onto a culture dish containing an MS solid culture medium, sealing the culture dish, placing the culture dish into a constant-temperature culture box for culture, and counting the result after one week.

Due to the adoption of the technical scheme, compared with the prior art, the invention has the technical progress that:

1. the invention provides a lycium ruthenicum artificial seed preparation device and a regeneration factor research method thereof.

2. The invention provides a lycium ruthenicum artificial seed preparation device and a regeneration factor research method thereof, wherein lycium ruthenicum artificial seeds are prepared by researching the regeneration factors of lycium ruthenicum, so that the geographical limitation is eliminated, the germination rate of the lycium ruthenicum seeds is greatly improved, the production of the artificial seeds is not limited by seasons, indoor production and expanded propagation can be realized all the year round, seed sources are provided for agricultural production in time, and the economic benefit can be increased.

3. The invention provides a lycium ruthenicum artificial seed preparation device and a regeneration factor research method thereof, wherein a connecting pipe, a bending pipe, a condensing pipe, a water pipe interface, a diversion trench and an exchange pipe are combined, the connecting pipe is connected to the inner wall of a sealing ring, steam discharged by a pressure relief valve pipe enters an inner cavity of the condensing pipe through the guide of the bending pipe, the work of an external circulating water cooling device is controlled, through the design of the water pipe interface and the diversion trench, cold water can be driven to circulate in the inner cavity of the condensing pipe, and the circulating water is subjected to cold and heat exchange with the steam through the exchange pipe, so that the function of condensing the steam is realized, the problem that the steam causes the improvement of indoor moisture is avoided, the stability of an indoor environment is ensured, the performance of other experimental work is facilitated, and the reliability of the device is improved.

Drawings

FIG. 1 is a schematic view of the structure of the autoclave body according to the present invention;

FIG. 2 is a schematic view of the internal structure of the autoclave body according to the present invention;

FIG. 3 is an enlarged schematic view of the present invention at configuration A;

FIG. 4 is a schematic view of a part of the structure of a first net plate according to the present invention;

FIG. 5 is a schematic view of the steam processing mechanism of the present invention;

FIG. 6 is a schematic cross-sectional view of an exchange tube of the present invention;

FIG. 7 is a flowchart of the study on factors for regenerating Lycium ruthenicum Murr.

In the figure: 1. a high-pressure steam sterilization pot body; 11. a steam nozzle; 2. a fixed collar; 3. a control button; 4. steam enters the pipe orifice; 5. a sealing cover; 6. a pressure relief valve tube;

7. a steam treatment mechanism; 71. a seal ring; 72. a connecting pipe; 73. bending the tube; 74. a condenser tube; 75. a water pipe connector; 76. a shunt slot; 77. a flow distribution plate; 78. exchanging pipes; 781. bending a first copper pipe; 782. a guide block; 783. concentrating the copper pipe; 784. bending a copper pipe II; 785. a terminal exhaust pipe; 79. a drain pipe; 791. an exhaust inclined pipe; 792. a water storage cylinder;

8. a sufficient sterilization mechanism; 81. an external drive motor; 82. a transmission box body; 83. a first bevel gear; 84. a second bevel gear; 85. a transmission gear; 86. a fixing ring; 87. a rotating ring; 88. an inner ring; 881. a connecting seat; 882. connecting blocks; 883. a first screen plate; 884. a magnetic connecting pin; 885. a second screen plate; 886. a vertical base rod; 887. a surrounding ring; 888. a rubber sleeve; 89. an inner gear ring.

Detailed Description

The present invention is further illustrated in detail below with reference to examples:

example 1

In a first aspect, as shown in fig. 1-7, the invention provides an artificial lycium ruthenicum seed preparation device, which comprises a high-pressure steam sterilization pot body 1 and an external circulating water cooling device, wherein a steam nozzle 11 is arranged at the bottom of an inner cavity of the high-pressure steam sterilization pot body 1, a fixed lantern ring 2 is fixedly installed on the outer wall of the high-pressure steam sterilization pot body 1, a control button 3 is arranged on the front surface of the fixed lantern ring 2, a steam inlet pipe orifice 4 is arranged on the front surface of the high-pressure steam sterilization pot body 1, a sealing cover 5 is detachably connected to the top of the high-pressure steam sterilization pot body 1, a pressure relief valve pipe 6 is fixedly connected to the top of the sealing cover 5, a steam treatment mechanism 7 is arranged at the top of the sealing cover 5, a sufficient sterilization mechanism 8 is arranged in the inner cavity of the high-pressure steam sterilization pot body 1, the steam treatment mechanism 7 comprises a sealing ring 71, and the sealing ring 71 is fixedly installed at the top of the sealing cover 5, the inner wall of the sealing ring 71 is detachably connected with a connecting pipe 72, the top of the connecting pipe 72 is fixedly connected with a bent pipe 73, the bottom of the bent pipe 73 is fixedly connected with a condenser pipe 74, both sides of the condenser pipe 74 are fixedly connected with water pipe interfaces 75, a water outlet pipe of an external circulating water cooling device is in threaded connection with the water pipe interface 75 on the left side, a water inlet pipe of the external circulating water cooling device is in threaded connection with the water pipe interface 75 on the right side, both sides of the inner wall of the condenser pipe 74 are provided with splitter boxes 76, the sufficient sterilization mechanism 8 comprises an external driving motor 81, a transmission box 82 and a fixing ring 86, the external driving motor 81 is fixedly arranged on the left side of the high-pressure steam sterilization pot body 1, the transmission box 82 is fixedly arranged on the inner wall of the high-pressure steam sterilization pot body 1, and a bevel gear 83 is fixedly connected in an inner cavity of the transmission box 82 extending from an output shaft of the external driving motor 81, the top of the first bevel gear 83 is engaged with a second bevel gear 84, the top of the second bevel gear 84 is fixedly connected with a transmission gear 85, a fixed ring 86 is fixedly welded on the inner wall of the high-pressure steam sterilization pot body 1, the inner part of the fixed ring 86 is rotatably connected with a rotating ring 87, the inner wall of the rotating ring 87 is fixedly connected with an inner ring 88, the inner wall of the inner ring 88 is fixedly provided with an inner toothed ring 89, the outer driving motor 81 is controlled to work, the bevel gear I83 in the transmission box body 82 can be driven to rotate, the inner toothed ring 89 is driven to rotate through the transmission of the bevel gear II 84 and the transmission gear 85, the inner ring 88 is driven to rotate on the inner wall of the fixed ring 86 under the design of the rotating ring 87, and then the experimental apparatus that drives a 883 top of otter board rotates, promotes the contact effect of experimental apparatus outer wall and steam, avoids the lower problem of the contact rate of the depressed part of experimental apparatus outer wall and steam, promotes the sufficiency of this device sterilization.

Example 2

As shown in fig. 1 to 7, on the basis of embodiment 1, the present invention provides a technical solution: preferably, the top and the bottom of the inner cavity of the condenser tube 74 are both fixedly connected with the splitter plates 77, one adjacent side of the two splitter plates 77 is fixedly connected with the exchange tube 78, the left side of the exchange tube 78 is fixedly connected with a first bent copper tube 781, a guide block 782 is fixedly welded on the inner wall of the first bent copper tube 781, the right side of the exchange tube 78 is fixedly connected with a tail end discharge tube 785, the left side of the tail end discharge tube 785 is fixedly connected with a concentrated copper tube 783, the outer wall of the concentrated copper tube 783 is fixedly connected with a second bent copper tube 784, one end of the second bent copper tube 784, which is far away from the concentrated copper tube 783, is fixedly connected with the outer wall of the first bent copper tube 78, the bottom of the condenser tube 74 is fixedly connected with a drain tube 79, a guide cone is fixedly installed on the inner wall of the drain tube 7811, the outer wall of the drain tube 79 is fixedly connected with an inclined exhaust tube 791, the bottom of the drain tube 792 is in threaded connection with the inner wall of the water storage tube 792, which is meshed with the outer wall of the transmission gear 85, the top of inner ring 88 is fixedly provided with a connecting seat 881, the top of connecting seat 881 is detachably connected with a connecting block 882, the inner side of connecting block 882 is fixedly provided with a first screen plate 883, the top of first screen plate 883 is detachably connected with a magnetic connecting pin 884, the top of magnetic connecting pin 884 is fixedly provided with a second screen plate 885, the top of second screen plate 885 is fixedly welded with a vertical base rod 886, the inner side of vertical base rod 886 is fixedly provided with a surrounding ring 887, the outer wall of surrounding ring 887 is fixedly sleeved with a rubber sleeve 888, firstly, a connecting pipe 72 is connected on the inner wall of a sealing ring 71, steam discharged by a pressure relief valve pipe 6 enters the inner cavity of a condenser pipe 74 through the guide of a bending pipe 73, the work of an external circulating water cooling device is controlled, through the design of a water pipe connector 75 and a diversion trench 76, cold water can be driven to circulate through the inner cavity of the condenser pipe 74, the circulating water exchanges heat with the steam through a exchanging pipe 78, and then realize carrying out the function of condensation to steam, avoid steam to lead to the problem that indoor moisture promoted, guarantee indoor environment's stationarity further promotes the heat exchange efficiency of steam and water through the design of bent copper pipe 781, guide block 782, concentrated copper pipe 783, terminal delivery pipe 785 and bent copper pipe two 784.

Example 3

In a second aspect, as shown in fig. 1-7, the present invention further provides a method for researching regeneration factors of artificial lycium ruthenicum seeds, wherein the method for researching regeneration factors of artificial lycium ruthenicum seeds comprises the following steps:

s1, preparing experimental instruments and reagents, specifically:

the experimental instrument comprises: high-pressure steam sterilization pot body 1, superclean bench, balance, blake bottle, culture dish, pipetting gun, digital display constant temperature water bath, rubber head burette, aurilave, burette, beaker and volumetric flask prepare the completion, include experimental reagent again: preparing an MS culture medium, cane sugar, 50% concentrated sulfuric acid, agar, NAA with different concentrations, 6BA, 2,4-D, 70% alcohol, 1% -2% sodium hypochlorite, 0.1% mercuric chloride, 2% sodium alginate, 3% sodium alginate, 4% sodium alginate, 2% calcium chloride and distilled water;

s2, inducing somatic embryos, specifically:

step A1, cultivation of aseptic seedlings: using wild lycium ruthenicum seeds, firstly preparing an MS culture medium, packaging a laboratory instrument, and carrying out high-pressure steam sterilization by using a plurality of bottles of distilled water, wherein the sterilization conditions are as follows: sterilizing at the temperature of 121 ℃ for 20min, taking out the lycium ruthenicum seeds when the pressure is zero so as to avoid danger, soaking the lycium ruthenicum seeds in 50% concentrated sulfuric acid for 5min, breaking the dormancy of the seeds, inoculating the lycium ruthenicum seeds to an MS culture medium by using a plant tissue culture technology, placing all culture bottles on a specified culture rack in a culture room at the same interval, observing whether the culture medium is polluted or not and whether the seeds germinate or not every two or three days, observing, counting and recording for two weeks to find that the seeds in all the culture medium germinate, and finally counting the germination rate of the lycium ruthenicum to be 100%;

step A2, callus induction: inducing callus with early cultured Lycium ruthenicum Murr plant in different hormone formulas, and dedifferentiating to form somatic embryo, wherein the culture medium formula is as follows: MS +6-BA + NAA +6g/l agar and 30g/l sucrose, and the specific formula is shown in the following table:

formulation of	6-BA mg/L	NAA mg/L
			1	0.5	0.5
2	0.5	1
			3	0.5	1.5
4	1	0.5
			5	1	1
6	1	1.5
			7	1.5	0.5
8	1.5	1
			9	1.5	1.5

S3, preparing artificial endosperm, which specifically comprises the following steps: the preparation of the artificial endosperm is to screen out a culture medium formula suitable for the germination and growth of the embryoid, finally, the screened culture medium is added into an embedding medium, different crops and different propagules have different requirements on the culture medium, in a preparation pre-experiment of the artificial endosperm of lycium ruthenicum, compared with an MS culture medium, the 1/2MS culture medium has better effect, so a 1/2MS culture medium is selected, on the basis, NAA with the largest influence on the germination rate of the artificial seeds and 2% sodium alginate are selected for the experiment, a proper amount of sucrose is also added into the artificial endosperm, and the culture medium formula is as follows: the MS + NAA + sodium alginate is used for carrying out two-factor three-level test to prepare the artificial endosperm, and the formula table of the artificial endosperm is as follows:

level of	NAA(mg/L)	Sodium alginate (%)
			1	0.5	2
2	1	3
			3	1.5	4

S4, manufacturing artificial seed coats, which specifically comprises the following steps: the method for forming the gel-coated material by using the hydrogel method through ion transformation or temperature mutation is an operation method of a commonly used ion exchange method embedded by sodium alginate, and comprises the following steps of: firstly, adding sodium alginate with different concentrations into an MS culture medium to prepare jelly, adding a certain proportion of embryoid, then fully and uniformly mixing the culture medium and the sodium alginate, sucking up the embryoid and the gel by using a dropper, then dropping the mixture into a 2% CaCl2 solution to stay for 10-15min, completely combining the surface of the embryoid with the jelly to form a round artificial seed with certain rigidity, rinsing the artificial seed for 20min by using sterile water, terminating the reaction, then inoculating the prepared artificial seed onto a culture dish containing an MS solid culture medium, sealing the culture dish and placing the culture dish into a constant-temperature culture box for culture, counting the results after one week, and inducing callus with different concentrations as follows:

formulation(s)	Number of blades	Wound healing	Browning	Wound healing rate	Browning rate
							1	18	14	2	78％	14％
2	12	9	2	75％	22％
						3	14	12	6	86％	50％
4	17	15	5	88％	30％
						5	15	9	4	60％	44％
6	13	9	4	70％	44％
						7	14	8	4	57％	50％
8	15	4	2	27％	50％
						9	18	8	4	44％	50％
Is totaled	136	88	33	65％	38％

And (4) analyzing results: from the above graph, callus was induced with MS +1mg/L6-BA +0.5mg/LNAA +6g/L agar +30g/L sucrose. Callus produces very fast and shows emerald green, do not take place vitrification phenomenon, it reaches 88% to count the callus rate at last, the brown ization rate is low to 30%, from this data can find out, induced callus effect is better, make the development of later stage experiment go on smoothly, in different concentration sodium alginate solution, the germination rate of artifical seed is different, the gradient that this experiment set up respectively is 2%, 3%, 4%, the result shows that 2% sodium alginate germination rate is higher, so it is comparatively suitable to bury black fruit matrimony vine with 2% sodium alginate mechanism, the influence of sodium alginate to artifical seed germination is as follows:

sodium alginate concentration	Number of artificial seeds	Number of sprouts	Germination rate
					2％	12	10	83.3％
3％	9	7	77.7％
				4％	10	7	72.7％

The effect of NAA on germination of artificial seeds is given in the following table:

NAA concentration (mg/L)	Number of artificial seeds	Number of sprouts	Percentage of germination
				0.5	9	7	77.7％
1	11	9	81.8％
				1.5	11	8	72.7％

The working principle of the device for preparing the lycium ruthenicum artificial seeds and the research method of the regeneration factors thereof is described in detail below.

As shown in fig. 1-7, the sealing cover 5 is opened, then the testing apparatus is placed in the inner cavity of the rubber sleeve 888, the output pipe of the steam generator is connected to the steam inlet pipe orifice 4 in advance, then the sealing cover 5 is closed, and the high-temperature steam sterilization treatment can be performed on the testing apparatus through the high-pressure steam sterilization pot body 1, during the treatment process, the external driving motor 81 is controlled to work to drive the testing apparatus at the top of the magnetic connecting pin 884, so as to improve the sterilization sufficiency, the connecting pipe 72 is connected to the inner wall of the sealing ring 71 in advance, and then the external circulating water cooling device is controlled to work to drive cold water to circulate through the inner cavity of the condensation pipe 74, and the circulating water is subjected to heat and cold exchange with the steam through the exchange pipe 78, so as to realize the function of condensing the steam.

The present invention has been described in general terms, but it will be apparent to those skilled in the art that modifications and improvements can be made based on the present invention. Therefore, modifications or improvements are within the scope of the invention without departing from the spirit of the inventive concept.

Claims

1. The utility model provides an artifical seed preparation facilities of lycium ruthenicum, includes high pressure steam sterilization pot body (1) and external circulating water cooling device, its characterized in that: the steam sterilizing boiler comprises a high-pressure steam sterilizing boiler body (1), and is characterized in that a steam nozzle (11) is arranged at the bottom of an inner cavity of the high-pressure steam sterilizing boiler body (1), a fixed sleeve ring (2) is fixedly installed on the outer wall of the high-pressure steam sterilizing boiler body (1), a control button (3) is arranged on the front surface of the fixed sleeve ring (2), a steam inlet pipe orifice (4) is arranged on the front surface of the high-pressure steam sterilizing boiler body (1), a sealing cover (5) is detachably connected to the top of the high-pressure steam sterilizing boiler body (1), a pressure relief valve pipe (6) is fixedly connected to the top of the sealing cover (5), a steam treatment mechanism (7) is arranged on the top of the sealing cover (5), and a sufficient sterilizing mechanism (8) is arranged in the inner cavity of the high-pressure steam sterilizing boiler body (1);

the steam treatment mechanism (7) comprises a sealing ring (71), the sealing ring (71) is fixedly installed at the top of the sealing cover (5), a connecting pipe (72) is detachably connected to the inner wall of the sealing ring (71), a bent pipe (73) is fixedly connected to the top of the connecting pipe (72), a condenser pipe (74) is fixedly connected to the bottom of the bent pipe (73), water pipe interfaces (75) are fixedly connected to two sides of the condenser pipe (74), a water outlet pipe of an external circulating water cooling device is in threaded connection with the water pipe interface (75) on the left side, a water inlet pipe of the external circulating water cooling device is in threaded connection with the water pipe interface (75) on the right side, and diversion grooves (76) are formed in two sides of the inner wall of the condenser pipe (74);

the full sterilization mechanism (8) comprises an external driving motor (81), a transmission box body (82) and a fixing ring (86), the external driving motor (81) is fixedly installed on the left side of the high-pressure steam sterilization pot body (1), the transmission box body (82) is fixedly installed on the inner wall of the high-pressure steam sterilization pot body (1), an output shaft of the external driving motor (81) extends into an inner cavity of the transmission box body (82) and is fixedly connected with a first bevel gear (83), the top of the first bevel gear (83) is meshed with a second bevel gear (84), the top of the second bevel gear (84) is fixedly connected with a transmission gear (85), the fixing ring (86) is fixedly welded on the inner wall of the high-pressure steam sterilization pot body (1), the internal rotation of the fixing ring (86) is connected with a rotating ring (87), and the inner wall of the rotating ring (87) is fixedly connected with an inner ring (88), an inner gear ring (89) is fixedly arranged on the inner wall of the inner ring (88).

2. The device of claim 1, wherein the device comprises: the top and the equal fixedly connected with flow distribution plate (77) in bottom of condenser pipe (74) inner chamber, two flow distribution plate (77) adjacent one side fixedly connected with exchange tube (78), the curved copper pipe of the left side fixedly connected with of exchange tube (78) (781), fixed welding has guide block (782) on the inner wall of curved copper pipe (781), terminal delivery pipe (785) of the right side fixedly connected with of exchange tube (78), copper pipe (783) is concentrated to the left side fixedly connected with of terminal delivery pipe (785), the curved copper pipe of fixedly connected with is two (784) on the outer wall of concentrated copper pipe (783), the outer wall fixed connection of the one end of concentrated copper pipe (783) and curved copper pipe (781) is kept away from in curved copper pipe two (784).

3. The device of claim 1, wherein the device comprises: the bottom fixedly connected with drain pipe (79) of condenser pipe (74), fixed mounting has the direction awl on the inner wall of drain pipe (79), fixedly connected with exhaust pipe chute (791) on the outer wall of drain pipe (79), the bottom threaded connection of drain pipe (79) has water storage cylinder (792).

4. The device of claim 1, wherein the device comprises: the inner wall of interior ring gear (89) and the outer wall meshing of drive gear (85), the top fixed mounting of inner ring (88) has connecting seat (881), the top dismantlement formula of connecting seat (881) is connected with connecting block (882), the inboard fixed mounting of connecting block (882) has otter board (883), the top dismantlement formula of otter board (883) is connected with magnetic force and connects foot (884), the top fixed mounting of magnetic force connection foot (884) has otter board two (885), the fixed welding in top of otter board two (885) has vertical base pole (886), the inboard fixed mounting of vertical base pole (886) has around ring (887), fixed cover has cup jointed rubber sleeve (888) on the outer wall of around ring (887).

5. A method for researching regeneration factors of Lycium ruthenicum Murr artificial seeds, which is realized based on the device for preparing Lycium ruthenicum Murr artificial seeds of claims 1 to 4, and is characterized in that: the method for researching the regeneration factors of the lycium ruthenicum artificial seeds comprises the following steps:

s1, preparing a laboratory instrument and a reagent;

s2, inducing somatic embryos;

s3, preparing artificial endosperm;

s4, manufacturing artificial seed coats.

6. The method for researching regeneration factors of the Lycium ruthenicum Murr artificial seeds according to claim 5, wherein the method comprises the following steps: the step of S1 further includes: the experimental instrument comprises: high-pressure steam sterilization pot body (1), superclean bench, balance, blake bottle, culture dish, pipette gun, digital display constant temperature water bath, rubber head drip tube, aurilave, burette, beaker and volumetric flask prepare the completion, include experimental reagent again: preparing MS culture medium, sucrose, 50% concentrated sulfuric acid, agar, NAA with different concentrations, 6BA, 2,4-D, 70% alcohol, 1% -2% sodium hypochlorite, 0.1% mercuric chloride, 2% sodium alginate, 3% sodium alginate, 4% sodium alginate, 2% calcium chloride and distilled water.

7. The method for researching regeneration factors of the Lycium ruthenicum Murr artificial seeds according to claim 5, wherein the method comprises the following steps: the step S2 further includes the steps of:

step A1, culturing aseptic seedlings: using wild lycium ruthenicum seeds, firstly preparing an MS culture medium, packaging a laboratory instrument, and carrying out high-pressure steam sterilization by using a plurality of bottles of distilled water under the sterilization conditions: sterilizing at the temperature of 121 ℃ for 20min, taking out the lycium ruthenicum seeds when the pressure is zero so as to avoid danger, soaking the lycium ruthenicum seeds in 50% concentrated sulfuric acid for 5min, breaking the dormancy of the seeds, inoculating the lycium ruthenicum seeds to an MS culture medium by using a plant tissue culture technology, placing all culture bottles on a specified culture rack in a culture room at the same interval, observing whether the culture medium is polluted or not and whether the seeds germinate or not every two or three days, observing, counting and recording for two weeks to find that the seeds in all the culture medium germinate, and finally counting the germination rate of the lycium ruthenicum to be 100%;

8. The method for researching regeneration factors of lycium ruthenicum artificial seeds according to claim 5, wherein the method comprises the following steps: the step S3 further includes the steps of: the preparation of the artificial endosperm is to screen out a culture medium formula suitable for the germination and growth of the embryoid, finally, the screened culture medium is added into an embedding medium, different crops and different propagules have different requirements on the culture medium, in a preparation pre-experiment of the artificial endosperm of lycium ruthenicum, compared with an MS culture medium, the 1/2MS culture medium has better effect, so a 1/2MS culture medium is selected, on the basis, NAA with the largest influence on the germination rate of the artificial seeds and 2% sodium alginate are selected for the experiment, a proper amount of sucrose is also added into the artificial endosperm, and the culture medium formula is as follows: the artificial endosperm is prepared by performing two-factor three-level test on MS + NAA + sodium alginate.

9. The method for researching regeneration factors of the Lycium ruthenicum Murr artificial seeds according to claim 5, wherein the method comprises the following steps: the step of S4 further includes: the method for forming the gel wrapping material by using the hydrogel method through ion transformation or temperature mutation is a commonly used method for operating the ion exchange method embedded by sodium alginate, and comprises the following steps: firstly, adding sodium alginate with different concentrations into an MS culture medium to prepare gel, adding a certain proportion of embryoid, then fully and uniformly mixing the culture medium and the sodium alginate, sucking up the embryoid and the gel by using a dropper, then dropping the mixture into a 2% CaCl2 solution to stay for 10-15min, completely combining the surface of the embryoid with the gel to form a round artificial seed with certain rigidity, rinsing the artificial seed with sterile water for 20min, stopping the reaction, then inoculating the prepared artificial seed onto a culture dish containing an MS solid culture medium, sealing the culture dish, placing the culture dish into a constant-temperature culture box for culture, and counting the result after one week.