CN109089612B - Material culture method for preparing sugarcane wild species chromosome sample - Google Patents
Material culture method for preparing sugarcane wild species chromosome sample Download PDFInfo
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- CN109089612B CN109089612B CN201810979174.5A CN201810979174A CN109089612B CN 109089612 B CN109089612 B CN 109089612B CN 201810979174 A CN201810979174 A CN 201810979174A CN 109089612 B CN109089612 B CN 109089612B
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- 239000000463 material Substances 0.000 title claims abstract description 53
- 240000000111 Saccharum officinarum Species 0.000 title claims abstract description 35
- 235000007201 Saccharum officinarum Nutrition 0.000 title claims abstract description 35
- 210000000349 chromosome Anatomy 0.000 title claims abstract description 27
- 238000012136 culture method Methods 0.000 title claims abstract description 9
- 241000894007 species Species 0.000 title description 4
- 241000196324 Embryophyta Species 0.000 claims abstract description 29
- 238000004382 potting Methods 0.000 claims abstract description 28
- 240000005382 Saccharum spontaneum Species 0.000 claims abstract description 26
- 235000014704 Saccharum spontaneum Nutrition 0.000 claims abstract description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000012258 culturing Methods 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 8
- 239000002689 soil Substances 0.000 claims description 36
- 244000025254 Cannabis sativa Species 0.000 claims description 34
- 239000010451 perlite Substances 0.000 claims description 32
- 235000019362 perlite Nutrition 0.000 claims description 32
- 239000002985 plastic film Substances 0.000 claims description 10
- 229920003023 plastic Polymers 0.000 claims description 8
- 238000005520 cutting process Methods 0.000 claims description 4
- 229920006255 plastic film Polymers 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 3
- 238000011160 research Methods 0.000 abstract description 10
- 230000035699 permeability Effects 0.000 abstract description 8
- 230000002380 cytological effect Effects 0.000 abstract description 6
- 230000012010 growth Effects 0.000 abstract description 5
- 230000006872 improvement Effects 0.000 abstract description 5
- 210000001082 somatic cell Anatomy 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 230000002786 root growth Effects 0.000 abstract description 4
- 239000003895 organic fertilizer Substances 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 3
- 230000001737 promoting effect Effects 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 27
- 239000004744 fabric Substances 0.000 description 6
- 238000007901 in situ hybridization Methods 0.000 description 6
- 241000209051 Saccharum Species 0.000 description 5
- 230000000392 somatic effect Effects 0.000 description 5
- 230000002759 chromosomal effect Effects 0.000 description 4
- 240000003433 Miscanthus floridulus Species 0.000 description 3
- 241000209504 Poaceae Species 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 239000010455 vermiculite Substances 0.000 description 3
- 235000019354 vermiculite Nutrition 0.000 description 3
- 229910052902 vermiculite Inorganic materials 0.000 description 3
- 240000007171 Imperata cylindrica Species 0.000 description 2
- 241001230286 Narenga Species 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 239000000919 ceramic Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000031864 metaphase Effects 0.000 description 2
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- 241000234642 Festuca Species 0.000 description 1
- 206010025421 Macule Diseases 0.000 description 1
- 241000878007 Miscanthus Species 0.000 description 1
- 240000000249 Morus alba Species 0.000 description 1
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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
- 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
- A01G9/028—Multi-compartmented pots
-
- 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
- A01G22/55—Sugar cane
-
- 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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
-
- 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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
-
- 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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/22—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing plant material
Abstract
The invention discloses a material culture method for preparing a sugarcane wild-species chromosome sample, and relates to the technical field of biology. According to the culture method of the wild sugarcane root tip somatic cell chromosome material, a special potting culture basin is designed, so that the method is convenient and practical, and meanwhile, white tender root tips can be collected conveniently to prepare sugarcane root tip somatic cell chromosome samples; culturing root tip materials of wild sugarcane resource plants in a potting mode, guaranteeing air permeability and water permeability of the bottom and organic fertilizer required by growth, promoting root growth, and facilitating preparation of wild sugarcane root tip somatic cell chromosome samples for cytology study; meanwhile, through cytological research on wild resources of the sugarcane, scientific basis is provided for further utilization of the wild resources in innovation of the germplasm and improvement of varieties of the sugarcane, and development of the sugarcane planting production industry is promoted.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a material culture method for preparing a sugarcane wild-species chromosome sample.
Background
Sugarcane (Saccharum spp.) is a tall plant of the Saccharum genus (Saccharum) of the Gramineae (Gramineae) sugarcane subfamily (Saccharastrae). Some species of the sugarcane subfamily are closely related to sugarcane and have important utilization value for sugarcane breeding, such as the genus festuca (Erianthus), miscanthus (Miscantus), scion (Sclerostachya), hegaku (Narenga) and Imperata (Imperata). These genus of plants are collectively referred to as "Saccharum complex" along with Saccharum. Because of the excellent properties such as strong tillering, strong stress resistance, barren tolerance and the like in the plants, the plants have potential utilization value for cultivating new varieties of sugarcane or improving original varieties, and are continuously researched and utilized as germplasm resources of the sugarcane. Among them, cut hand density (s.spontaneum l.), macula (e.arcdinaceus retz.), keca (e.furvus Ness), heba (n.porphyrocomb (Hance ex Trimen) Box), wujiqus (m.floridulus (lab.) warb.ex schum et Laut.), miscanthus (m.sinensis anderss.), and yunnan keong (e.rockikeng) have a broad utility prospect in the innovation and variety improvement of sugarcane germplasm resources, which has been the focus of acquisition and utilization of sugarcane germplasm resources, and cytological studies on these resource plants have been the important basis for their utilization in sugarcane breeding.
At present, when the cytology research of the wild resources of the sugarcane is carried out, as the wild nature of the plants of the resources is stronger and the rooting is deeper, the root tips are difficult to collect under the conventional preservation condition, the collected root tips are older and the quantity is less, and the purpose of cytology research is difficult to achieve, thereby restricting the innovation of the germplasm of the sugarcane, the improvement of the varieties and the development of the sugarcane planting production industry.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a material culture method for preparing a wild sugarcane chromosome sample, designs a special pot culture basin, cultures root tip materials of wild sugarcane resource plants in a pot culture mode, ensures air permeability and water permeability of the bottom and organic fertilizers required by growth, promotes root growth, and facilitates preparation of a wild sugarcane root tip chromosome sample for cytology study.
In order to achieve the technical purpose and the technical effect, the invention is realized by the following technical scheme:
a material culture method for preparing a sugarcane wild-species chromosome sample, comprising the following steps:
1) Digging the wild sugarcane resource clone material from a resource garden, cutting off all leaves of the material, wherein the material is provided with tillering bud points and 3-5 old roots;
2) Filling impurity-free soil into the inner container of the special pot culture basin to 1/3 of the total volume, planting the sample into the inner container, covering the soil to 1/2 of the total volume, watering thoroughly, covering a transparent plastic film, and then culturing in a greenhouse at about 30 ℃ to enable seedlings to germinate;
3) When the material grows out 8-12 tillers and the height reaches 75-85cm, lifting the liner together with soil and material plants to be completely taken out from the pot culture basin;
4) Filling perlite and sun-dried turf grass which are uniformly mixed in a pot culture basin in a ratio of 1:3, filling the perlite and the sun-dried turf grass to 1/3-1/2 of the volume of the flowerpot, re-planting the plant with soil taken out into the pot culture basin through the inner container, and then placing the water fully poured into the pot culture basin to be cultured in a greenhouse at about 30 ℃;
5) After 15-20 days of culture, a large number of white tender root tips grow at the bottom of a pot culture basin, and the inner container is lifted to collect a sample from the bottom of the inner container; filling perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3 into a pot culture basin after collection is completed, and repeatedly culturing.
Further, the seedlings in the step 2) germinate for 12-15 days, the plastic film is removed until the seedlings grow to 14-16cm, the seedlings are continuously cultured in a greenhouse at about 30 ℃ for 30-35 days, 8-12 tillers grow out of the materials, and the height reaches 75-85cm.
Further, perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3 are filled in the pot culture pot, wherein the perlite is clean perlite or vermiculite which is not used, and the turf grass is grass turf grass which is trimmed and sun-dried repeatedly.
Furthermore, in the step 4), the plants are planted in the pot culture basin again through the inner container, and the surface soil is slightly dried and needs to be immediately watered to the bottom of the basin, so that a small amount of water flows out.
The wild sugarcane root tip somatic cell chromosome material prepared by the method is used for chromosome counting, karyotype analysis, genome In Situ Hybridization (GISH) and Fluorescence In Situ Hybridization (FISH) analysis after sampling treatment.
The invention further aims to provide a special pot culture basin for preparing sugarcane wild-species chromosome sample materials, which comprises a pot culture basin and an inner container, wherein the pot culture basin is a cylindrical basin body, a base is arranged at the lower end and provided with a through hole and is arranged on a basin pad, the inner container is arranged in the pot culture basin, the inner container is a hollow cylinder with the diameter being 1-2cm smaller than that of the pot culture basin, a cross-shaped bracket and grid cloth are arranged at the bottom, and a handle with central symmetry is arranged at the upper end;
scale marks are designed on the inner walls of the pot culture basin and the inner container;
the pot culture basin is a ceramic basin, and the inner container is made of plastic with the thickness of 0.3-0.5 cm.
Use of a special potting culture pot for preparing a sugarcane wild-species chromosomal sample material for culturing a wild sugarcane root tip somatic chromosomal material, comprising:
placing the inner container in a pot culture basin, filling 1/3 of impurity-free soil in the inner container, planting the prepared sample on the soil, covering the soil to 1/2 of the total volume, watering thoroughly, covering a transparent plastic film, and placing in a greenhouse at about 30 ℃ for culture to enable seedlings to germinate;
when the material grows out 8-12 tillers and the height reaches 75-85cm, the inner container is taken out from the pot culture basin through the handle on the inner container, and meanwhile, soil and material plants in the inner container are completely preserved;
filling perlite and sun-dried turf grass which are uniformly mixed in a pot culture basin in a ratio of 1:3, filling the perlite and the sun-dried turf grass to 1/3-1/2 of the volume of a flowerpot, re-planting the inner container of the plant with soil taken out into the pot culture basin, and then placing the pot culture basin with sufficient water in a greenhouse at about 30 ℃ for culture;
after 15-20 days of culture, a large number of white tender root tips grow at the bottom of the pot culture basin, and a sample can be collected from the bottom of the liner layer by lifting the liner; filling perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3 into a pot culture basin after collection is completed, and repeatedly culturing.
The beneficial effects of the invention are as follows: according to the invention, a potting mode is adopted to culture root tip materials of wild sugarcane resource plants, perlite and dried turf grass which are uniformly mixed in a potting culture basin are filled in a 1:3 mode to ensure air permeability and water permeability of the bottom and organic fertilizer required by growth, root growth is promoted, a wild sugarcane root tip somatic chromosome sample is conveniently prepared for cytology research, a special potting culture basin is designed, the special potting culture basin comprises the potting culture basin and a liner, the liner is filled with impurity-free soil and then is planted with the sample for culture, after the liner is placed on the potting culture basin after the perlite and the dried turf grass which are uniformly mixed in a 1:3 mode are filled in the potting culture basin, a large amount of white young root tips of wild sugarcane resources can grow on the bottom of the potting culture basin through grid cloth on the lower side of the liner, and when the pot is used, the sample is collected only by lifting the liner on the bottom of the pot, so that the sample is conveniently obtained; simultaneously, repeated culture is carried out for a plurality of times, so that enough and tender root tips are provided for cytological researches (chromosome counting, karyotype analysis, GISH and FISH analysis) of sugarcane resource plants, and meanwhile, innovation and variety improvement of sugarcane germplasm are continuously carried out through cytological researches on wild sugarcane resources, so that the development of sugarcane planting production industry is promoted.
Of course, it is not necessary for any one product to practice the invention to achieve all of the advantages set forth above at the same time.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram of a special pot culture basin according to an embodiment of the invention;
FIG. 2 is a schematic view of a pot culture basin and a liner according to an embodiment of the present invention;
FIG. 3 is a sample diagram of a material prepared by culturing a wild sugarcane chromosome sample according to an embodiment of the present invention;
in the drawings, the list of components represented by the various numbers is as follows:
1-pot culture basin, 2-inner container, 3-basin pad, 4-bracket, 5-grid cloth, 6-base and 7-scale mark.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
As shown in fig. 1-2:
a material culture method for preparing a sugarcane wild-species chromosome sample, comprising the following steps:
1) Digging the wild sugarcane resource clone material from a resource garden, cutting off all leaves of the material, wherein the material is provided with tillering bud points and 3-5 old roots;
2) Filling impurity-free soil into the inner container 2 of the special pot culture basin 1 to 1/3 of the total volume, planting a sample into the inner container 2, covering the soil to 1/2 of the total volume, watering thoroughly, covering a transparent plastic film, and then placing the pot culture basin in a greenhouse at about 30 ℃ for culture to enable seedlings to germinate;
3) When the material grows out 8-12 tillers and the height reaches 75-85cm, lifting the liner 2 together with soil and material plants to be completely taken out from the potting culture basin 1;
4) Filling perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3 into a potting culture basin 1, filling the perlite and the sun-dried turf grass to 1/3-1/2 of the volume of a flowerpot, re-planting the plant taken out of the flowerpot and with soil into the potting culture basin 1 through an inner container 2, and placing sufficient water into a greenhouse at about 30 ℃ for culture;
5) After 15-20 days of culture, a large number of white tender root tips grow at the bottom of the pot culture basin 1 (shown in figure 3), the liner 2 is lifted up, and a sample is collected from the bottom of the liner 2; filling perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3 into a pot culture basin 1 after collection is completed, and repeatedly culturing.
And 2) 12-15 days after the seedlings germinate, removing the plastic film until the seedlings grow to 14-16cm, and continuously culturing the seedlings in a greenhouse at about 30 ℃ for 30-35 days, wherein 8-12 tillers grow out of the material, and the height reaches 75-85cm.
The pot culture basin 1 is filled with perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3, wherein the perlite is clean perlite or vermiculite which is not used, and the turf grass is grass which is trimmed and sun-dried repeatedly.
In the step 4), the plants are planted into the pot culture basin 1 again through the inner container, and in the culture process, the surface soil is slightly dried and needs to be immediately watered to the bottom of the basin, and a small amount of water flows out.
The wild sugarcane root tip somatic cell chromosome material prepared by the method is used for chromosome counting, karyotype analysis, genome In Situ Hybridization (GISH) and Fluorescence In Situ Hybridization (FISH) analysis after sampling treatment.
Example 2
As shown in fig. 1-2: the pot culture basin comprises a pot culture basin 1 and an inner container 2, wherein the pot culture basin 1 is a cylindrical basin body, a base 6 is arranged at the lower end and provided with a through hole and is arranged on a basin pad 3, the inner container 2 is arranged in the pot culture basin 1, the inner container 2 is a hollow cylinder with the diameter being 1-2cm smaller than that of the pot culture basin 1, a cross-shaped bracket 4 and a grid cloth 5 are arranged at the bottom, and a handle with central symmetry is arranged at the upper end and is opened;
scale marks 7 are arranged on the inner walls of the pot culture basin 1 and the inner container 2;
the pot culture basin 1 is a ceramic basin, and the inner container 2 is made of plastic with the thickness of 0.3-0.5 cm.
Example 3
As shown in fig. 1-2: use of a special potting culture pot for preparing a sugarcane wild-species chromosomal sample material for culturing a wild sugarcane root tip somatic chromosomal material, comprising:
placing the inner container 2 in a pot culture basin 1, filling 1/3 of impurity-free soil in the inner container 2, planting the prepared sample on the soil, covering the soil to the position of 1/2 of the total volume, watering thoroughly, covering a transparent plastic film, and placing in a greenhouse at about 30 ℃ for culture to enable seedlings to germinate;
when the material grows out 8-12 tillers and the height reaches 75-85cm, the inner container is taken out of the pot culture basin 1 through a handle on the inner container 2, and meanwhile, soil and material plants in the inner container 2 are completely stored;
filling perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3 into a potting culture basin 1, filling the perlite and the sun-dried turf grass to 1/3-1/2 of the volume of a flowerpot, re-planting the inner container 2 of the plant with soil taken out into the potting culture basin 1, and placing sufficient water into a greenhouse at about 30 ℃ for culture;
after 15-20 days of culture, a large number of white tender root tips grow at the bottom of the pot culture basin 1, and the inner container 2 is lifted up to collect samples from the bottom of the inner container 2 layer; filling perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3 into a potting culture basin 1 after collection is completed, and repeatedly culturing;
the specific implementation steps are as follows:
material preparation:
preparing a pot culture pot, unused clean perlite (or vermiculite) and grass of Gramineae which is trimmed and sufficiently dried as described in the above example 2;
digging the wild sugarcane resource clone material from a resource garden, carrying out tillering bud points and partial old roots, and cutting out all leaves of the material;
the operation steps are as follows:
placing the inner container 2 in a pot culture basin 1, filling 1/3 of impurity-free soil in the inner container 2, planting the prepared sample on the soil, covering the soil to the position of 1/2 of the total volume, watering thoroughly, covering a transparent plastic film, and placing in a greenhouse at about 30 ℃ for culture to enable seedlings to germinate;
15 days after the seedlings germinate, the seedlings grow to 15cm, the plastic film is removed, the seedlings are continuously cultured in a greenhouse at about 30 ℃ for 30 days, 10 tillers grow out of the material, and the height reaches 80cm;
the inner container is taken out of the pot culture basin 1 through the handle on the inner container 2, meanwhile, soil and material plants in the inner container 2 are completely preserved, in the taking-out process, the lower half part of the basin body is lightly taken by hands until the soil in the basin is loosened, when the inner container 2 is taken out, the inner container is lightly taken and put, damage to plant leaves, root systems and the like is prevented, meanwhile, the soil is not loosened, and the soil and material plants in the inner container 2 are completely preserved, and meanwhile, the use is convenient;
filling perlite and sun-dried turf grass which are uniformly mixed in a pot culture basin 1 in a ratio of 1:3, filling the perlite and sun-dried turf grass to 1/3 of the volume of a flowerpot, re-planting an inner container 2 for taking out plants with soil into the pot culture basin 1, and then placing enough water for watering into a greenhouse at about 30 ℃ for culture, wherein proper watering is paid attention during the period, the root tips are long and old due to lack of water, but rotten roots are generated due to excessive watering, namely, the surface soil is slightly dried and immediately watered, and the water is a small amount of water for each watering for the bottom flow of the pot;
after 20 days of culture, a large number of white tender root tips grow at the bottom of the pot culture basin 1, the inner container 2 is lifted up to collect samples from the bottom of the inner container 2 layer, a large number of white tender root tips of wild sugarcane resources can pass through the mesh cloth 5 at the lower side of the inner container 2 to grow at the bottom 1 of the pot culture basin, namely after the inner container 2 is lifted up, a large number of white tender root tips are directly exposed, the chromosome materials of the wild sugarcane root tips are collected for chromosome counting, nuclear analysis, genome In Situ Hybridization (GISH) and Fluorescence In Situ Hybridization (FISH) analysis, a large number of white tender root tips are conveniently collected for research,
and filling perlite and dried turf grass which are uniformly mixed in a ratio of 1:3 into a pot culture basin 1 after collection is completed, repeating the operation and repeatedly culturing to obtain more experimental samples, and sampling for multiple times.
Example 4
In 2012-2017, the inventor cultures root tips of more than 200 parts of sugarcane wild germplasm resource materials by using the method provided by the invention to obtain a large number of tender root tips (50-100 root tip growth condition pictures of the materials after actual culture), and the materials used for experiments after treatment have a large number of metaphase division phases (sugarcane wild species metaphase chromosome pictures) with evenly dispersed chromosomes and a large number, so that the requirements of somatic chromosome counting and karyotype analysis can be completely met.
According to the invention, a potting mode is adopted to culture root tip materials of wild sugarcane resource plants, perlite and dried turf grass which are uniformly mixed in a potting culture basin are filled in a 1:3 mode to ensure air permeability and water permeability of the bottom and organic fertilizer required by growth, root growth is promoted, a wild sugarcane root tip somatic chromosome sample is conveniently prepared for cytology research, a special potting culture basin is designed, the special potting culture basin comprises the potting culture basin and a liner, the liner is filled with impurity-free soil and then is planted with the sample for culture, after the liner is placed on the potting culture basin after the perlite and the dried turf grass which are uniformly mixed in a 1:3 mode are filled in the potting culture basin, a large amount of white young root tips of wild sugarcane resources can grow on the bottom of the potting culture basin through grid cloth on the lower side of the liner, and when the pot is used, the sample is collected only by lifting the liner on the bottom of the pot, so that the sample is conveniently obtained; simultaneously, repeated culture is carried out for a plurality of times, so that enough and tender root tips are provided for cytological researches (chromosome counting, karyotype analysis, GISH and FISH analysis) of sugarcane resource plants, and meanwhile, innovation and variety improvement of sugarcane germplasm are continuously carried out through cytological researches on wild sugarcane resources, so that the development of sugarcane planting production industry is promoted.
In the description of the present specification, the descriptions of the terms "one embodiment," "example," "specific example," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The preferred embodiments of the invention disclosed above are intended only to assist in the explanation of the invention. The preferred embodiments are not exhaustive or to limit the invention to the precise form disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best understand and utilize the invention. The invention is limited only by the claims and the full scope and equivalents thereof.
Claims (1)
1. A material culture method for preparing a sugarcane wild-species chromosome sample is characterized by comprising the following steps of: the method comprises the following steps:
(1) Digging the wild sugarcane resource clone material from a resource garden, cutting off all leaves of the material, wherein the material is provided with tillering bud points and partial old roots;
(2) Filling impurity-free soil into the inner container of the pot culture basin to 1/3 of the total volume, planting the sample into the inner container and covering the soil to 1/2 of the total volume, watering thoroughly and covering a transparent plastic film, and then placing the pot culture basin in a greenhouse at about 30 ℃ for culture to enable seedlings to germinate;
after the seedlings germinate for 12-15 days, the seedlings grow to 14-16cm, the plastic film is removed, the seedlings are continuously cultured in a greenhouse at about 30 ℃ for 30-35 days, 8-12 tillers grow out of the materials, and the height reaches 75-85cm;
(3) When the material grows out 8-12 tillers and the height reaches 75-85cm, lifting the liner together with soil and material plants to be completely taken out from the pot culture basin;
(4) Filling perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:3 into a potting culture basin, filling the perlite and the sun-dried turf grass to 1/3-1/2 of the volume of a flowerpot, re-planting the plant with soil taken out into the potting culture basin through a liner, and then pouring enough water into a greenhouse at 30 ℃ for culture;
in the culture process of the plants after being planted into the pot culture pot again through the inner container, the surface soil is slightly dried and needs to be immediately watered to the bottom of the pot, and a small amount of water flows out;
(5) After 15-20 days of culture, a large number of white tender root tips grow at the bottom of a pot culture basin, and the inner container is lifted to collect a sample from the bottom of the inner container; filling perlite and sun-dried turf grass which are uniformly mixed in a ratio of 1:1 into a pot culture basin after collection is completed, and repeatedly culturing;
wherein, pack 1 in the culture pot cultivated in a pot: and 3, uniformly mixing perlite and sun-dried turf grass, wherein the perlite is clean perlite which is not used, and the turf grass is grass turf grass which is trimmed and sun-dried repeatedly.
Priority Applications (1)
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| 李茂枝.浅谈四川甘蔗有性杂交育种四十年的几点体会.《甘蔗糖业》.1997,第1997年卷(第第1期期),第2-8页. * |
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