CN113016547A - Floating seedling method for tobacco gene editing material strain fine comparison - Google Patents
Floating seedling method for tobacco gene editing material strain fine comparison Download PDFInfo
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- CN113016547A CN113016547A CN202110237212.1A CN202110237212A CN113016547A CN 113016547 A CN113016547 A CN 113016547A CN 202110237212 A CN202110237212 A CN 202110237212A CN 113016547 A CN113016547 A CN 113016547A
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- 239000000463 material Substances 0.000 title claims abstract description 25
- 238000010362 genome editing Methods 0.000 title claims abstract description 23
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Images
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/45—Tobacco
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/006—Labels or label holders specially adapted for plant receptacles
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
- A01G9/029—Receptacles for seedlings
- A01G9/0291—Planting receptacles specially adapted for remaining in the soil after planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
- A01G9/029—Receptacles for seedlings
- A01G9/0293—Seed or shoot receptacles
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
- A01G9/029—Receptacles for seedlings
- A01G9/0295—Units comprising two or more connected receptacles
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Botany (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Toxicology (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Cultivation Receptacles Or Flower-Pots, Or Pots For Seedlings (AREA)
Abstract
The invention relates to a floating seedling method in the fine comparison of tobacco gene editing material strains. The non-woven fabric used in the invention is a polylactic acid material, can be degraded in soil after being transplanted, has no harmful degradation product residue after being degraded, and is safe and environment-friendly. Compared with the common field production, the fine screening of the edited materials has higher requirements on the uniformity of the growth vigor of the plants, and the growth vigor is uniform and regular, so that the errors caused by the inconsistent growth and development can be reduced to the maximum extent, and the gene efficacy is highlighted.
Description
Technical Field
The invention belongs to the technical field of tobacco floating seedling, and particularly relates to a floating seedling method in the fine comparison of tobacco gene editing material strains.
Background
The floating seedling raising is an agricultural planting technology which is mainly used for plants such as green-leaf vegetables, tobacco and the like with short growing period, can ensure the growth consistency of the plants, avoids the problem of soil diseases and insect pests caused by the traditional cultivation mode, and is widely used for greenhouse planting all over the world. The floating seedling is utilized to carry out the cultivation production of the plants in the greenhouse, which can bring greater yield and higher quality, prolong the harvest time, and limit and reduce the use amount of pesticides. Most of floating seedling raising techniques are to adopt a foaming seedling raising hole tray to produce, peat, vermiculite and other soilless culture media are added into the seedling raising tray, the seedling raising tray is placed into a water bed after seeding, and small holes are reserved at the bottom of the seedling raising tray so as to be beneficial to the absorption of plant roots to moisture and nutrition.
In the tobacco planting process, seedling culture is a key step, tobacco seedlings grow neatly and robustly, root systems are intact during transplanting, the growth uniformity of the transplanted tobacco can be effectively improved, the tobacco can be conveniently and uniformly baked subsequently, and the quality of tobacco leaves is improved. At present, the floating seedling raising technology is widely applied to the tobacco seedling raising technology and is popularized as a mature technology in China. Part 1 of the national standard GB/T25241.1-2010 intensive tobacco seedling culture technical specification: the floating seedling culture specifies each specific flow of the tobacco floating seedling culture in detail.
However, the above-mentioned standards are aimed at the rough and extensive planting in field production, and for the comparison of fine strains in the process of gene editing material cultivation, it is required to minimize the growth and development differences between different strains caused by the inconsistency of external environments. Only if the transplanted tobacco seedlings are uniform and regular, the subsequent growth and growth of tobacco plants can be ensured to be consistent, and the action rules of different editing genes on the properties, the metabolic components and the like of the tobacco plants are highlighted. Therefore, the gene editing material strain has higher requirements on planting and growth uniformity than the rough planting of field tobacco production. The conventional method is adopted for seedling raising, when seedlings are transplanted and pulled out, the situation that the seedling root systems are damaged due to adhesion (or scattering) of the substrate and the seedling tray often occurs, so that the transplanted tobacco seedlings have large difference in seedling returning period and irregular growth vigor, and interference can be caused to subsequent fine screening.
Disclosure of Invention
The invention aims to provide a floating seedling method for tobacco gene editing material strain fine comparison, which aims to solve the problems that during seedling raising by adopting a conventional method, a matrix is adhered to a seedling tray (or the matrix is scattered) to damage a seedling root system when seedlings are transplanted and pulled out, so that the transplanted tobacco seedlings have large seedling return period difference and irregular growth condition, and the interference on subsequent fine screening comparison is caused.
The invention is realized by the following technical scheme:
a floating seedling method in the fine comparison of tobacco gene editing material strains comprises the following steps:
s1, determining the number and position of a floating tray where the gene editing seeds are located, the number of dibbling, seedling raising responsible persons, planning time and seedling raising places;
s2, preparing a nursery pond according to the GB/T25241.1-2010 standard;
s3, filling a matrix;
s4, printing the floating plate number and a two-dimensional code label containing the seed number and the floating plate information by using a bar code printer, attaching the printed floating plate number to a ground inserting label, and inserting the ground inserting label in the central position of the seedling raising plate; the seed label is stuck on the tag label, and the tag label is hung at a set position of the seedling tray in a penetrating way;
s5, checking that the seed number on the freezing storage pipe is consistent with the seed number of the label on the floating plate before sowing;
s6, sowing a set number of seeds in each seedling hole of the seedling tray, wherein the number of the sowed seeds is larger than that of the coated seeds;
s7, multiplying the seeds by the multiplication ratio, and multiplying the seeds by the multiplication ratio of 27-45;
s8, referring to GB/T25241.1-2010, making seedling management measures;
s9, when 1 true leaf grows, thinning and final singling are completed at one time.
Preferably, the matrix loading in step S3 includes the steps of:
s31, executing according to the specification in YC/T310;
s32, selecting a flat and sanitary site as a tray loading site, and wetting the substrate before tray loading to ensure that the water content of the substrate reaches 30-40%;
s33, selecting a seedling tray with set seedling holes, paving a layer of non-woven fabric on the seedling tray, pressing the non-woven fabric into each seedling hole, filling a matrix into each seedling hole, and cutting the non-woven fabric according to the seedling holes of the seedling tray by using a cutter.
Preferably, the non-woven fabric is a spiral fiber polylactic acid non-woven fabric, holes with the diameter of 0.2-1.2 mm are formed in the non-woven fabric, the non-woven fabric has good elasticity and water permeability, and water and fertilizer absorption and root penetration in the growth and development process are not hindered.
Preferably, the cutter comprises an iron sheet welded with sharp slices and a left handle and a right handle on the iron sheet, and the cutter is provided with blades which are matched with the seedling culture holes of the seedling tray and arranged in a grid shape.
Preferably, the iron sheet floats a set specification design according to the standard of 162 nursery holes, and long being 660mm, wide being 340mm, sharp sliced iron sheet is according to horizontal 8 rows, and perpendicular 17 rows cross welding arrange, and horizontal sliced length is 660mm, and two adjacent horizontal sliced intervals are 8mm, and vertical sliced length is 340mm, and two adjacent vertical sliced intervals are 8 mm.
Preferably, in the propagation dibbling, the seeds on the left are in five rows from the second row, and the seeds on the right are in five rows from the second row on the right.
Preferably, the seedling management comprises seedling water, nutrient solution, temperature and humidity management measures.
Preferably, the tobacco seedlings grow to be suitable for transplanting seedlings and then are transplanted, the tobacco seedlings are pulled out of the seedling tray by holding the edges of the non-woven fabrics with hands, and the root systems of the tobacco seedlings can be kept complete due to the fact that the non-woven fabrics are arranged between the substrate and the seedling tray and wrapped by the non-woven fabrics.
The invention has the beneficial effects that:
in the invention, the floating tray number and the label containing the seed number are printed by the bar code printer for identification, thereby ensuring the ordering of seedling culture and being not easy to make mistakes, being convenient for code scanning information tracing in the subsequent material planting process and being beneficial to realizing intelligent and information management.
The advantages of using non-woven fabrics to wrap the substrate: as the gene editing tobacco seeds belong to precious breeding materials, the germination rate of atypical coated seeds is slightly lower than that of the coated seeds, and the growth of the tobacco seeds in the seedling stage is weaker. The non-woven fabric seedling raising water permeability and air permeability are good, the seedlings are not easy to rot roots, the growth vigor is better, and the survival rate is higher.
The hole of the seedling tray is covered with the polylactic acid spiral fiber non-woven fabric, and the non-woven fabric can effectively avoid the condition that the root system is damaged due to the adhesion (or the scattering) of the matrix and the seedling tray when the seedling is transplanted and pulled out. The non-woven fabric is used for wrapping, so that the root system of each transplanted tobacco seedling can be ensured to be complete and undamaged. The transplanted tobacco seedlings have short seedling returning period and good consistency of the seedling returning period, and the subsequent growth vigor of the tobacco plants is more uniform and regular, thereby being very beneficial to the refined observation of characters in the subsequent material screening comparison and the refined comparison of the metabolic component difference of tobacco leaves. The non-woven fabric used in the invention is a polylactic acid material, is a spiral fiber polylactic acid non-woven fabric, is provided with holes with the diameter of 0.2-1.2 mm, has good elasticity and water permeability, and does not hinder root penetration in the growth process. Moreover, the transplanted non-woven fabric can be degraded in soil, no harmful degradation product is left after degradation, and the method is safe and environment-friendly.
The gene editing tobacco is a precious breeding material, the quantity of the tobacco gene editing material in propagation and planting is large, the links are multiple, the process is long, and no corresponding operation standard exists.
Drawings
FIG. 1 is a top view of a tool for filling a floating seedling substrate;
FIG. 2 is a front view of a tool for filling the floating seedling substrate;
FIG. 3 is a left side view of a tool for filling the floating seedling substrate;
FIG. 4 is a diagram of an example of a label of a dibbling float plate;
FIG. 5 is a schematic view of floating seedling dibbling;
FIG. 6 is a schematic diagram of the seedling-remaining position of the floating nursery tray.
1. Seedling tray, 2, seedling growing hole, 3, float a serial number, 4, two-dimensional code, 5, seed serial number.
Detailed Description
The technical solutions of the present invention are described in detail below by examples, and the following examples are only exemplary and can be used only for explaining and explaining the technical solutions of the present invention, but not construed as limiting the technical solutions of the present invention.
As shown in FIGS. 1 to 3, the present embodiment provides a filling scheme and a tool for floating seedling substrate, which mainly comprises
Matrix: performing as specified in YC/T310; substrate dishing principle: uniform and consistent, and moderate tightness.
A flat and sanitary field is selected as a tray loading field, and the substrate is moistened before the tray loading so that the water content of the substrate reaches 30-40%.
For a large number of standard 162-hole seedling trays used for field planting, a layer of non-woven fabric is laid on the empty floating tray. The nonwoven is then pressed against each aperture such that the nonwoven is held against each aperture wall. Each well is filled with substrate, and the nursery wells not filled with substrate are inspected and filled while preventing too tight a fill. Then a special cutter is used for cutting, and the special cutter mainly comprises an iron sheet welded with sharp slices and a left handle and a right handle on the iron sheet. The special cutter is characterized in that the cutter is provided with blades which can be matched with the seedling tray cavities and are arranged in a grid shape. The seedling tray is made of foam material, and the non-woven fabric can be cut according to the cavity of the seedling tray by using the cutter to put the cutter on the seedling tray, press the cutter downwards and slightly slide forwards and backwards and leftwards and rightwards.
The specification of the iron sheet is designed according to the specification of a standard floating plate with 162 holes, the length is 660mm, and the width is 340 mm.
The arrangement mode of the sharp slices is that the sharp slices are arranged in a cross welding mode according to 8 rows in the transverse direction and 17 rows in the vertical direction, the length of the transverse slices is 660mm, and the distance between the slices is 8 mm; the length of the vertical slices is 340mm, and the spacing between the slices is also 8 mm.
And aligning a cutting tool to each hole of the floating disc, and then cutting the non-woven fabric by using the cutting tool to ensure that the non-woven fabric in each hole is not adhered any more, and each hole is independently corresponding to a small piece of non-woven fabric.
The invention also relates to a floating seedling method of gene editing tobacco based on the substrate filling method and the cutting tool, which comprises the following steps:
s1, determining the floating tray number and position of the gene editing seeds, the seed counting number, the seedling responsible person, the planning time and the seedling location.
S2, preparing a seedling raising pond: performing according to GB/T25241.1-2010 standard
S3, filling the substrate, wherein the filling scheme of the floating seedling substrate and the used tools are adopted.
S4, printing the floating plate number and a two-dimensional code label (shown in figure 4) containing the seed number and the floating plate information by using a bar code printer, attaching the printed floating plate number to a ground inserting label, and inserting the ground inserting label in the central position of the seedling raising plate; the seed label is pasted on the tag label, and the tag label is hung at the positions of the first left row of the 9 th hole (L9) and the first right row of the 9 th hole (R9) of the seedling raising tray in a penetrating way.
S5, each bare tobacco seed stored in the freezing storage tube has a unique serial number, and the serial numbers of the seeds on the freezing storage tube and the serial numbers of the seeds marked on the floating plate are carefully checked before sowing to ensure consistency;
s6, the tobacco seeds belong to breeding materials, are atypical coated seeds, have a germination rate slightly lower than that of the coated seeds, and 4 seeds are sown in each hole during dibbling.
S7, multiplying propagation and dibbling: the seeds are sown by 45 times, the seeds on the left are sown in five rows from the second row, and the seeds on the right are sown in five rows from the second row on the right, as shown in FIG. 5.
S8, seedling management: with reference to GB/T25241.1-2010, measures for managing moisture, nutrient solution, temperature and humidity in the seedling stage are established.
S9, thinning period: the period is 1 true leaf period, and thinning and final singling are completed in one time.
Thinning number: taking 31 strains for molecular detection, and reserving seedlings in 5 rows (reserving seedlings from the left to L6 and reserving seedlings from the right to R6); other special cases are increased or decreased according to actual situations. When the tobacco seedlings are not enough, the tobacco seedlings are needed to be replenished on the cavities, and the tobacco seedlings with the same number are needed to be replenished, so that the numbers of the tobacco seedlings cannot be changed, which is shown in figure 6. The thinning specific method comprises the following steps: ensuring that one tobacco seedling is planted in each hole, and pulling out redundant tobacco seedlings by using tweezers to ensure that the tobacco seedlings are uniform; the thinning tool is sterilized by 75% ethanol, and the disc is sterilized once after the operation.
And S10, transplanting the tobacco seedlings after the tobacco seedlings grow to the age suitable for transplanting the seedlings, wherein the transplanting is to grasp the edges of the non-woven fabrics by hands and pull the tobacco seedlings out of the seedling trays. Because the non-woven fabrics are arranged between the substrate and the seedling tray, the tobacco seedlings are wrapped by the non-woven fabrics, the root system of each tobacco seedling can be kept complete, the root injury phenomenon frequently occurring in the traditional seedling pulling and transplanting process is avoided, the seedling returning period of the transplanted tobacco seedlings is shortened, the seedling returning period is unified, and the growth vigor of the tobacco seedlings is more uniform and neat.
The foregoing shows and describes the general principles and broad features of the present invention and advantages thereof. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (8)
1. A floating seedling method in the fine comparison of tobacco gene editing material strains is characterized by comprising the following steps:
s1, determining the number and position of a floating tray where the gene editing seeds are located, the number of dibbling, seedling raising responsible persons, planning time and seedling raising places;
s2, preparing a nursery pond according to the GB/T25241.1-2010 standard;
s3, filling a matrix;
s4, printing the floating plate number and a two-dimensional code label containing the seed number and the floating plate information by using a bar code printer, attaching the printed floating plate number to a ground inserting label, and inserting the ground inserting label in the central position of the seedling raising plate; the seed label is stuck on the tag label, and the tag label is hung at a set position of the seedling tray in a penetrating way;
s5, checking that the seed number on the freezing storage pipe is consistent with the seed number of the label on the floating plate before sowing;
s6, sowing a set number of seeds in each seedling hole of the seedling tray, wherein the number of the sowed seeds is larger than that of the coated seeds;
s7, multiplying the seeds by the multiplication ratio, and multiplying the seeds by the multiplication ratio of 27-45;
s8, referring to GB/T25241.1-2010, making seedling management measures;
s9, when 1 true leaf grows, thinning and final singling are completed at one time.
2. The floating seedling method in the fine comparison of tobacco gene editing material lines as claimed in claim 1, wherein the filling of the substrate in step S3 comprises the steps of:
s31, executing according to the specification in YC/T310;
s32, selecting a flat and sanitary site as a tray loading site, and wetting the substrate before tray loading to ensure that the water content of the substrate reaches 30-40%;
s33, selecting a seedling tray with set seedling holes, paving a layer of non-woven fabric on the seedling tray, pressing the non-woven fabric into each seedling hole, filling a matrix into each seedling hole, and cutting the non-woven fabric according to the seedling holes of the seedling tray by using a cutter.
3. The floating seedling method for use in the fine comparison of tobacco gene editing material lines as claimed in claim 2, wherein the non-woven fabric is a helical fiber polylactic acid non-woven fabric having pores with a diameter of 0.2 to 1.2mm and having good elasticity and water permeability.
4. The floating seedling method in the fine comparison of tobacco gene editing material lines as claimed in claim 2, wherein the cutter is composed of an iron sheet welded with sharp slices and two left and right handles on the iron sheet, and the cutter has blades arranged in a grid shape matching with the seedling tray seedling holes.
5. The floating seedling method in the fine comparison of tobacco gene editing material lines as claimed in claim 4, wherein the iron sheets are designed according to the standard floating plate specification of 162 seedling holes, the length is 660mm, the width is 340mm, the iron sheets of the sharp slices are arranged in the cross welding mode according to 8 rows and 17 rows, the length of the transverse slices is 660mm, the distance between two adjacent transverse slices is 8mm, the length of the vertical slices is 340mm, and the distance between two adjacent vertical slices is 8 mm.
6. The floating seedling method in the fine comparison of tobacco gene editing material lines as claimed in claim 1, wherein in the expanding propagation and dibbling, the left seeds are in five rows from the second row, and the right seeds are in five rows from the second row on the right.
7. The floating seedling method in the fine comparison of tobacco gene editing material lines as claimed in claim 1, wherein the seedling management includes measures of seedling moisture, nutrient solution, temperature and humidity management.
8. The floating seedling method in the fine comparison of tobacco gene editing material lines as claimed in claim 1, wherein the tobacco seedlings are transplanted after growing to a suitable seedling age for transplantation, the transplantation is carried out by grasping the edges of the non-woven fabrics with hands and pulling the tobacco seedlings out of the seedling tray, and the tobacco seedlings are wrapped by the non-woven fabrics due to the non-woven fabrics between the substrate and the seedling tray, so that the root system integrity of each tobacco seedling can be maintained.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113287486A (en) * | 2021-07-06 | 2021-08-24 | 苏州市农业科学院 | Self-floating crop blanket and preparation method thereof |
| CN115104529A (en) * | 2022-07-27 | 2022-09-27 | 云南中烟工业有限责任公司 | Gene editing material selfing homozygous management method and information platform |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20090298888A1 (en) * | 2005-05-19 | 2009-12-03 | Wolfgang Thielert | Method of improving plant development and increasing the resistance of plants to soil-borne harmful fungi |
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| CN115104529A (en) * | 2022-07-27 | 2022-09-27 | 云南中烟工业有限责任公司 | Gene editing material selfing homozygous management method and information platform |
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