BIOMECHANICAL MEASURING TECHNICAL METHOD FOR MAIZE SEED RADICLE AND COLEORHIZA SEPARATION

Abstract

Disclosed is a biomechanical measuring technical method for maize seed radicle and coleorhiza separation, which is characterized by comprising the following operations: (1) seed sample preparation; (2) anterior tissue cutting; (3) radicle and coleorhiza separation; (4) coleorhiza sample acquisition; (5) coleorhiza sample fixation; (6) puncture force measurement; (7) information storage and analysis. The operation (3) includes the development of a maize radicle and coleorhiza separation device, and the operation (5) includes the development of a maize coleorhiza sample carrier. The present disclosure has the beneficial effects of providing direct biomechanical evidence for the research on the coleorhiza weakening regulation and control mechanism of the maize seed germination, and simultaneously providing reference for measuring the coleorhiza weakening biological force of the gramineous plant seeds.

Claims

1. A biomechanical measuring technical method for maize seed radicle and coleorhiza separation, which is characterized by comprising the following operations: (1) seed sample preparation; (2) anterior tissue cutting; (3) radicle and coleorhiza separation; (4) coleorhiza sample acquisition; (5) coleorhiza sample fixation; (6) puncture force measurement; (7) information storage and analysis; the operation (3) includes the development of a maize radicle and coleorhiza separation device, which is characterized by comprising a separator rotor (4), a miniature electric drill (25) and a glass rotating tube (10); the front part of the separator rotor (4) comprises a separator cap (1); the middle part of the separator rotor (4) comprises a connecting cap shaft (2), a connecting cap shaft thread (3), a connecting tail shaft (5), a connecting tail shaft thread (14); the separator cap (1) comprises a glass rotating tube sleeve (11), a separator cap inner cavity (12), a separator cap inner cavity thread (13); the electric drill fixing shaft (8) comprises a separator tail (6), a separator tail antiskid stripe (7), a glass rotating tube telescopic control button (9), a separator tail inner cavity (15), a separator tail inner cavity thread (16), an electric drill fixing shaft clamping strip (17); the interior of the separator rotor (4) has a glass rotating tube sleeve (11), a rubber ring (18), a separator transfer head inner groove (19), a tube stabilizer (20), a tube stabilize sleeve (21), a compression cap (22), a spring (23), a tube stabilizer buckle joint (24); the miniature electric drill (25) comprises a rotor fixing clamp (26), a rotor fixer (27), an elastic ring (28), an elastic ring antiskid stripe (29), a speed change controller (30), a power switch (31), an electric drill fixing bayonet (32) and a battery (33); the battery (33) has a battery antiskid stripe (34), a charging interface (35) at the bottom, a charging plug (36) and a power cord (37); the operation (5) includes the development of a maize coleorhiza sample carrier comprises a transparent module (42) and a gasket (40); and the transparent module (42) is processed and manufactured on the basis of a 3D printing technology; the gasket (40) is fixed on the transparent module (42) and is provided with a gasket hole (41) which corresponds to a sample placing hole (43) on the top of the transparent module (42); in the operation (1), the seeds are germinated by adopt two germination methods of covering paper germination and rolling paper germination; in the operation (2), the seed is transected with a scalpel, the anterior tissue of seed containing radicle and coleorhiza is retained and placed on wet filter paper for later use, and the posterior tissue of seed is discarded; in the operation (3), accord to that characteristics of the maize variety and the inn diameter of the coleorhiza of a sample to be tested, and the glass rotating tube is arranged in the radicle and coleorhiza separator; the radicle is separated from the coleorhiza by rotating the glass rotating tube, and is moved out from coleorhiza; before being used, the opening of the glass rotating tube is slightly dipped with a lubricant; and particularly, when the radicle and coleorhiza are tightly connected at the initial stage of seed germination, the glass rotating tube needs to be slowly rotated forward; in the operation (4), after that radicle is removed from the coleorhiza, the coleorhiza in the seed anterior tissue is completely peel off by a scalpel, forceps and the like under a stereoscope; the tissue is soaked with sterile water by a dropper to facilitate stripping and avoid damaging the integrity of the coleorhiza; in the operation (5), the coleorhiza sample is transferred to a special coleorhiza sample carrier; in the operation (6), the tissue sample carrier is fixed on a sample carrier bed; a small amount of sterile water is dripped on a gasket before measurement to ensure that sample is wet; then the seed biomechanical measurement system is utilized to measure the puncture force of the coleorhiza sample; the measuring parameters are as follows: the needle diameter is 0.5 mm, the needle moving speed is 30 mm.Math.min.sup.−1, the test ambient temperature is 15-20° C., and the completion time is within 30 min; after biomechanical measurement, the tissue sample carrier is cleaned, the needle is unloaded, and various system components return to the original position; in the operation (7), the seed biomechanical measurement information is stored and the target data is statistically analyzed.

2. The biomechanical measuring technical method for maize seed radicle and coleorhiza separation according to claim 1, which is characterized in that: in the operation (5), the coleorhiza sample is fixed through the gasket hole (41) and the sample placing hole (43) on the top of the transparent module (42); in the process of coleorhiza sample puncture force measurement, the metal needle (38) sequentially passes through the gasket hole (41), the sample placing hole (43), the coleorhiza (39) and the needle outlet hole (44).

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly explain the embodiments of the present disclosure or the technical solutions in the prior art, the following will briefly introduce the drawings that need to be used in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present disclosure. For those of ordinary skill in the art, other drawings may be obtained according to these drawings without any creative efforts.

[0028] In FIGS. 2 and 3, the meaning of each number is as follows: the separator cap 1, the connecting cap shaft 2, the connecting cap shaft thread 3, the separator rotor 4, the connecting tail shaft 5, the separator tail 6, the separator tail antiskid stripe 7, the electric drill fixing shaft 8, the glass rotating tube telescopic control button 9, the glass rotating tube 10, the glass rotating tube sleeve 11, the separator cap inner cavity 12, the separator cap inner cavity thread 13, the connecting tail shaft thread 14, the separator tail inner cavity 15, the separator tail inner cavity thread 16, the electric drill fixing shaft clamping strip 17, the rubber ring 18, the separator transfer head inner groove 19, the tube stabilizer 20, the tube stabilize sleeve 21, the compression cap 22, the spring 23, the tube stabilizer buckle joint 24, the miniature electric drill 25, the rotor fixing clamp 26, the rotor fixer 27, the elastic ring 28, the elastic ring antiskid stripe 29, the speed change controller 30, the power switch 31, the electric drill fixing bayonet 32, the battery 33, the battery antiskid stripe 34, the charging interface 35, the charging plug 36, the power cord 37, the metal needle 38, the coleorhiza 39, the gasket 40, the gasket hole 41, the transparent module 42, the sample placing hole 43 and the needle outlet hole 44.

[0029] FIG. 1 is a flow technical scheme of the operation of the present disclosure.

[0030] FIG. 2 is a schematic view showing the separation of anterior and posterior of seed of the present disclosure.

[0031] FIG. 3 is a perspective view of the structure of the separator rotor of maize radicle and coleorhiza separator of the present disclosure.

[0032] FIG. 4 is a perspective view of the internal structure of the separator rotor of the maize radicle and coleorhiza separator of the present disclosure.

[0033] FIG. 5 is a perspective view of the structure of the miniature electric drill matched with the maize radicle and coleorhiza separator of the present disclosure.

[0034] FIG. 6 is a diagram of maize seed of the present disclosure after radicle and coleorhiza separation.

[0035] FIG. 7 is a perspective view of coleorhiza sample fixation and puncture force measurement of the present disclosure.

[0036] FIG. 8 shows the seed biomechanical measuring system of the present disclosure.

[0037] FIG. 9 shows a coleorhiza sample puncture force measurement curves.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0038] The technical solutions in the embodiments of the present disclosure will be clearly and completely described below with reference to the drawings in the embodiments of the present disclosure. Obviously, the described embodiments are only part of the embodiments of the present disclosure, but not all of them. Based on the embodiment of the present disclosure, all other embodiments obtained by ordinary technicians in the field without creative labor are within the scope of the present disclosure.

[0039] In order to make the above objectives, features and advantages of the present disclosure more obvious and understandable, the present disclosure will be explained in further detail below with reference to the drawings and detailed description.

[0040] As shown in FIG. 1, biomechanics and molecular biology are effectively combined by measure the biological force of the seeds, so that the growth rule of the seeds can be better discover, and the related mechanism can be clarified. The present disclosure relates to a biomechanical measuring technical method for maize seed radicle and coleorhiza separation, which comprises the following operations: seed sample preparation; anterior tissue cutting; radicle and coleorhiza separation; coleorhiza sample acquisition; coleorhiza sample fixation; puncture force measurement; information storage and analysis. The present disclosure also comprises the completion of the research and development of a maize radicle and coleorhiza separator and the like to meet the requirements of related operations. The application of this technology will provide direct biomechanical evidence for the mechanism of maize seed germination regulated by coleorhiza weakening.

[0041] The present disclosure relates to a technical method for determining the biological force of maize seed radicle and coleorhiza separation, and the detailed description of the embodiments is as follow.

[0042] (1) Seed sample preparation: Biomechanical measurement of coleorhiza weakening mainly adopts two seed germination methods of covering paper germination and rolling paper germination, the details are as follows.

[0043] Seeds are randomly selected for germination and surface sterilized for ten minutes in 1% NaClO (w/v) and then wash thrice with distilled water (the coated seeds are cleaned with distilled water before surface sterilization). For rolled paper germination, two pieces of germination paper (Anchor Paper Co., St Paul, Minn., USA) are stacked and wet by distilled water. Excess water on the paper is removed by a towel, then the sterilized seeds are placed in a loose vertical roll of germination paper and incubated in a versatile environmental test chamber. The specific germination environment and seed treatment method are selected according to the scientific research purpose.

[0044] Germination by covering paper method: We take two pieces of germination paper (380 mm×255 mm) and stack them into a germination box/tray (450 mm×300 mm×90 mm), then add distilled water to fully wet them, and then lightly wipe the bed surface with sterile gauze to remove the residual liquid and air bubbles between the papers. The seeds are arranged in parallel and orderly on the germinating paper with the assistance of a seed placing plate which is an auxiliary tool for placing seed, with the paper edge distance of 2-2.5 cm. After the seeds are placed on the germination paper, a piece of moistened germination paper is covered on the seeds, then a germination box/tray cover is covered, a label is pasted on the germination box/tray and basic information such as variety name, sample number, repetition times, germination time and the like is marked, and finally the germination box/tray is placed in versatile environmental test chamber for germination.

[0045] Rolling paper germination: We sterilize the operating table with 75% alcohol solution, stack two pieces of germination paper (380 mm×255 mm), and mark the sample information (or prepare a waterproof strip for sample information) in the smaller area of the germination paper corner with an oily marker pen, such as sample name, repeat number, etc. The germinate paper is fully wet by distilled water, that paper surface is lightly wiped by sterile gauze, after the residual liquid and the air bubbles between the papers are removed. Then the seeds are arranged on the germination paper in a staggered way under the assistance of a seed placing plate. The micropylar end of each seed is in the same direction when the seeds are placed; and the paper edge distance is 5 cm. After the seeds are placed on the germination paper, a piece of moistened germination paper is covered on the seeds. Then roll up the germination papers (Note: here the sample information waterproofing strip could be placed between germination papers). Then the ends of the paper roll are fixed with rubber bands. The paper rolls are put into a plastic self-sealing bag to be sealed; and label paper is stuck on the plastic self-sealing bag or relevant information is marked by an oily marker pen, and then the plastic self-sealing bag containing the paper rolls is vertical placed in versatile environmental test chamber for germination (seed micropylar end down).

[0046] (2) Anterior tissue cutting: The seeds to be tested from the germinating paper are taken out (Note: the sampling time shall be determined according to the needs of scientific research). The seeds are transected with a scalpel (FIG. 2). The anterior tissue of the seed is retained and placed on wet filter paper for later use. The posterior tissue of the seed is discarded.

[0047] (3) Radicle and coleorhiza separation: Accord to that characteristics of the maize variety and the inn diameter of the coleorhiza of a sample to be tested, and the glass rotating tube 10 is arranged in a radicle and coleorhiza separator (The specific structure and usage of the radicle and coleorhiza separator are described below). The radicle is separated from the coleorhiza by rotating the glass rotating tube, and is moved out from coleorhiza. Before being used, the opening of the glass rotating tube is slightly dipped with a lubricant (such as liquid paraffin); and particularly, when the radicle and coleorhiza are tightly connected at the initial stage of seed germination, the glass rotating tube needs to be slowly rotated forward. The separated sample is shown in FIG. 3.

[0048] (4) Coleorhiza sample acquisition: After that radicle is removed from the coleorhiza, the coleorhiza in the seed anterior tissue is completely peel off by a scalpel, forceps and the like under a stereoscope. The tissue is soaked with sterile water by a dropper to facilitate stripping and avoid damaging the integrity of the coleorhiza.

[0049] (5) Coleorhiza sample fixation: The coleorhiza sample is transferred to a special coleorhiza sample carrier (FIG. 7). And the coleorhiza sample carrier comprises a transparent module 42 and a gasket 40; and the transparent module 42 is processed and manufactured on the basis of a 3D printing technology. The gasket 40 is fixed on the transparent module 42. And the coleorhiza sample is fixed through a gasket hole 41 and a sample placing hole 43 on the top of the transparent module 42.

[0050] (6) Puncture force measurement: The tissue sample carrier is fixed on a sample carrier bed (FIG. 8). A small amount of sterile water is dripped on a gasket 40 before measurement to ensure that sample is wet. Then the seed biomechanical measurement system (FIG. 8) is utilized to measure the puncture force of the coleorhiza sample. In the process of coleorhiza sample puncture force measurement, the metal needle 38 sequentially passes through the gasket hole 41, the sample placing hole 43, the coleorhiza 39 and the needle outlet hole 44. The measuring parameters are as follows: the needle diameter is 0.5 mm, the needle moving speed is 30 mm.Math.minute.sup.−1, the test ambient temperature is 15-20° C., and the completion time is within 30 minutes (min). After biomechanical measurement, the tissue sample carrier is cleaned, the needle is unloaded, and various system components return to the original position.

[0051] (7) Information storage and analysis: The seed biomechanical measurement system is used to obtain the information of maize coleorhiza puncture force. The information (including the puncture force measurement curves, see FIG. 9) is stored and the target data is statistically analyzed.

[0052] In the operation (3), the maize radicle and coleorhiza separator (FIG. 3, FIG. 4, FIG. 5) comprises a separator rotor 4, a miniature electric drill 25 and a glass rotating tube 10. Wherein, the separator rotor 4 comprises a separator cap 1, a connecting cap shaft 2, a connecting cap shaft thread 3, a connecting tail shaft 5, a separator tail 6, a separator tail antiskid stripe 7, an electric drill fixing shaft 8, a glass rotating tube telescopic control button 9, a glass rotating tube sleeve 11, a separator cap inner cavity 12, a separator cap inner cavity thread 13, a connecting tail shaft thread 14, a separator tail inner cavity 15, a separator tail inner cavity thread 16, an electric drill fixing shaft clamping strip 17, a rubber ring 18, a separator transfer head inner groove 19, a tube stabilizer 20, a tube stabilize sleeve 21, a compression cap 22, a spring 23, a tube stabilizer buckle joint 24. The miniature electric drill 25 comprises a rotor fixing clamp 26, a rotor fixer 27, an elastic ring 28, an elastic ring antiskid stripe 29, a speed change controller 30, a power switch 31, an electric drill fixing bayonet 32, a battery 33, a battery antiskid stripe 34, a charging interface 35, a charging plug 36 and a power cord 37.

[0053] The front part of the separator rotor 4 is the separator cap 1. The rear part of the separator rotor 4 is the electric drill fixing shaft 8. The separator tail inner cavity 15 of the separator cap 1 with the separator cap inner cavity thread 13 and the connecting cap shaft 2 with the connecting cap shaft thread 3 are fixed by rotating connection.

[0054] The front part of the separator cap 1 is provided with a glass rotating tube sleeve 11. Inside the separator cap 1 there is a rubber ring 18, which is behind the glass rotating tube sleeve 11 and serves to fix the glass rotating tube 10. The tube stabilizer buckle joint 24 is clamped in the separator transfer head inner groove 19 and used for fixing the tube stabilizer 20. The tube stabilize sleeve 21 is sleeved on the compression cap 22. When the spring 23 extends the tube stabilize sleeve 21 will compress the compression cap 22. The glass rotating tube telescopic control button 9 controls the compression and extension of the spring 23 so as to control the length of the glass rotating tube 10 at the front end of the glass rotating tube sleeve 11. And when that glass rotating tube 10 is arranged in the tube stabilizer 20, the glass rotary tube telescopic control button 9 can be taken down. The rear part of the separator rotor 4 is provided with the electric drill fixing shaft 8 which is provided with the electric drill fixing shaft clamping strip 17 which can be tightly fixed with the rotor fixing clamp 26 of the miniature electric drill 25.

[0055] The front part of the miniature electric drill 25 is provided with a rotor fixer 27. The rotor fixer 27 top is provided with a rotor fixing clamp 26. The rotor fixer 27 rear is provided with an elastic ring 28. The rotor fixing clamp 26 is opened or closed by rotating the elastic ring 28 through the elastic ring antiskid stripe 29. And this is very convenient for installing and removing the separator rotor 4. The middle part of the miniature electric drill 25 is provide with a power switch 31 and a speed change controller 30. The power switch 31 controls whether the miniature electric drill 25 work. The rotational direction and speed of the rotor fixing clamp 26 are controlled by changing the direction of the speed change controller 30 and changing the pressure applied to the speed change controller 30, respectively. In addition, the middle part of the miniature electric drill 25 is provided with two electric drill fixing bayonet 32. And the miniature electric drill 25 can be fixed on a specific electric drill frame through the electric drill fixing bayonet 32 as required. The rear part of the miniature electric drill 25 is provided with a battery 33. The surface of the battery 33 is provided with a battery antiskid stripe 34. And the battery antiskid stripe 34 can facilitate the unloading of the battery 33. The bottom of the battery 33 is provided with a charging interface 35. When charging, the charging plug 36 is inserted into the charging interface 35; and the power cord 37 is connected with the power supply.

[0056] There are two ways to install the glass rotating tube 10. One is to remove the glass rotating tube telescopic control button 9, and put the glass rotating tube 10 from the rear of the tube stabilizer 20. And the other is to press the glass rotating tube telescopic control button 9 to directly put the glass rotating tube 10 into the tube stabilizer 20 from the front end of the glass rotating tube sleeve 11.

[0057] In summary, the present disclosure can provide direct biomechanical (puncture force) evidence for the study of coleorhiza weakening mechanism during maize seed germination.

[0058] The above-mentioned embodiments only describe the preferred mode of the present disclosure, but do not limit the scope of the present disclosure. On the premise of not departing from the design spirit of the present disclosure, all kinds of modifications and improvements made by ordinary technicians in the field to the technical scheme of the present disclosure shall fall within the scope of protection determined by the claims of the present disclosure.