CN102907329A - Cultivating method for improving multiplication coefficient of shoot - Google Patents
Cultivating method for improving multiplication coefficient of shoot Download PDFInfo
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- CN102907329A CN102907329A CN 201210453551 CN201210453551A CN102907329A CN 102907329 A CN102907329 A CN 102907329A CN 201210453551 CN201210453551 CN 201210453551 CN 201210453551 A CN201210453551 A CN 201210453551A CN 102907329 A CN102907329 A CN 102907329A
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Abstract
The invention relates to a cultivating method for improving multiplication coefficient of a shoot, particularly to a cultivating method for improving multiplication coefficient of an acer mono shoot, which comprises the steps follows: 1), washing a resting shoot from an acer mono adult seed tree; 2), sterilizing a stem; 3), cultivating the sterilized stem to obtain the steam with a shoot; 4), cultivating the stem with the shoot to obtain the extended immature stem; and 5), cultivating the extended immature stem to obtain a plantlet. By adopting the resting sprouting shoot of the acer mono wild tree as the material, the research on the shoot multiplication and cultivation conditions are performed, and the cluster shoot induced cultivation system for directly multiplying the acer mono shoot is established, so as to lay a foundation of acer mono in vitro rapid propagation system. The reproductive cycle is short, the efficiency is high, the auxiliary shoot germination rate of the extended immature stem can reach 63.3%, the average height can reach 15.9 mm; and the shoot multiplication rate of the plantlet can reach 90%, the multiplication times can reach 3.19, and the shoot growth is normal.
Description
Technical field
The present invention relates to a kind of cultural method that improves the stem eye growth coefficient.
Background technology
Look wood maple (Acer mono Maxim) has another name called five jiaos of maples, look wood, for the Aceraceae maple belongs to deciduous tree.Tree vigo(u)r is graceful, is thick with leaves, and is leaf beautiful, and tender leaf is red, autumn has set in become orange or red, very attractive in appearance.In addition, look wood maple shade tolerance is better, and preferably growth also can be arranged in the factories and miness greening.Can be used as the bonsai material.At present, mainly concentrate on seeding and seedling raising, seed germination, cottage propagation, cultivating seedlings for the research of look wood maple and the aspect such as blossom and bear fruit.Organize cultivation aspect report less about look wood maple.In recent decades, the Study on tissue culture of Aceraceae has obtained larger progress.Such as the foreign scholar somatic embryo Somatic Embryogenesis of Japanese maple (A.palmatum) Immature zygotic embryo is studied, Wilhelm induces callus and indefinite bud with young shoot, plumular axis and the radicle segment of acer pseudoplatanus (A.pseudoplatanus) at the MS medium of additional BA and/or TDZ.Durkovei has set up fine stern maple (A.caudatifolium) axillalry bud and has cultivated the propagation of bud and the cultivating system of regenerating.The method of low temperature storage of utilizing Park etc. has realized the long-term germ plasm resource preservation of look wood maple suspended culture cell.Domestic scholars is organized maple and has also been done a large amount of work aspect the cultivation.As about America Acer negundo (A.saccharum), compound leaf maple (A.negundo), acer fabri (A.fabri), the tissue of white ox maple (A.mandshuricum) etc. is cultivated and rapid propagation in vitro research obtains reporting in succession.
Summary of the invention
The object of the invention provides a kind of cultural method that improves look wood maple stem eye growth coefficient.
Improving the cultural method of look wood maple stem eye growth coefficient realizes according to the following steps:
One, the resting shoot of getting on the adult elite stand of look wood maple is scrubbed with washing powder water, and then water is rinsed well, is cut into the stem section of 2～3cm, and every section has 1～2 axillalry bud or 1 terminal bud, again the stem section is soaked 10min in washing powder water, do not stop therebetween to stir, then under flowing water, wash 40min;
Two, on superclean bench be 75% alcohol solution dipping 30s with mass concentration after the stem section is cleaned, the 20min that then sterilizes in the mass concentration that is added with 2～3 Twen-20 is 2% NaClO uses aseptic water washing 3～5 times again;
Three, the stem section after the step 2 sterilization blots the moisture on surface with aseptic filter paper, then with tweezers perula is divested, until only remain 1～2mm bud point, again 0.5cm is respectively excised with scalpel in stem section two ends, the direction that makes progress according to the morphology upper end vertically is inserted into the stem section in the blake bottle that the MS minimal medium is housed, insertion depth is 0.5cm, then blake bottle is put in the culturing room with after the plastic seal membrana oralis sealing, and be that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate the stem sections that obtained in 2 weeks with bud under the condition of 16/8h;
The stem section of four, getting step 3 gained band bud is inoculated in the 50ml blake bottle that the tender stem elongation medium of 20ml is housed, and then is put in culturing room, is that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate the tender stem that 21～30d obtains elongation under the condition of 16/8h;
Five, the tender stem of getting step 4 gained elongation is inoculated in the 50ml blake bottle that the tender stem proliferated culture medium of 20ml is housed, and then is put in culturing room, is that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate 21～30d under the condition of 16/8h to obtain plantlet, namely finishes the cultivation that improves look wood maple stem eye growth coefficient;
Wherein tender stem elongation medium is take the MS minimal medium as minimal medium in the step 4, and also comprises the naa (NAA) of 0.1mg, the sucrose of 20g and the agar of 6g in the tender stem elongation medium of every 1L, and the pH value is 5.8;
Tender stem proliferated culture medium is take the MS minimal medium as minimal medium in the step 5, and also comprises the indolebutyric acid (IBA) of 0.1mg and the 6-benzyl aminoadenine (6-BA) of 1mg in the tender stem proliferated culture medium of every 1L.
The present invention utilizes look wood maple wild-type tree Dormant Buds branch to be material, carries out the research of bud propagation condition of culture, has set up the inducing clumping bud cultivating system of the direct proliferating way of look wood maple bud, for the foundation of look wood maple rapid propagation in vitro system is laid a good foundation.
Improve the cultural method of look wood maple stem eye growth coefficient among the present invention, the breeding cycle is short, and efficient is high, the tender stem of elongation, and the axillary bud sprouting rate can reach 63.3%, and average height can reach 15.9mm; Plantlet, the bud rate of increase can reach 90%, and the propagation multiple can reach 3.19, and elongation is normal.
Embodiment
Embodiment one: present embodiment improves the cultural method of look wood maple stem eye growth coefficient and realizes according to the following steps:
One, the resting shoot of getting on the adult elite stand of look wood maple is scrubbed with washing powder water, and then water is rinsed well, is cut into the stem section of 2～3cm, and every section has 1 axillalry bud or 1 terminal bud, again the stem section is soaked 10min in washing powder water, do not stop therebetween to stir, then under flowing water, wash 40min;
Two, on superclean bench be 75% alcohol solution dipping 30s with mass concentration after the stem section is cleaned, the 20min that then sterilizes in the mass concentration that is added with 2～3 Twen-20 is 2% NaClO uses aseptic water washing 3～5 times again;
Three, the stem section after the step 2 sterilization blots the moisture on surface with aseptic filter paper, then with tweezers perula is divested, until only remain 1～2mm bud point, again 0.5cm is respectively excised with scalpel in stem section two ends, the direction that makes progress according to the morphology upper end vertically is inserted into the stem section in the blake bottle that the MS minimal medium is housed, insertion depth is 0.5cm, then blake bottle is put in the culturing room with after the plastic seal membrana oralis sealing, and be that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate the stem sections that obtained in 2 weeks with bud under the condition of 16/8h;
The stem section of four, getting step 3 gained band bud is inoculated in the 50ml blake bottle that the tender stem elongation medium of 20ml is housed, and then is put in culturing room, is that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate the tender stem that 21d obtains elongation under the condition of 16/8h;
Five, the tender stem of getting step 4 gained elongation is inoculated in the 50ml blake bottle that the tender stem proliferated culture medium of 20ml is housed, and then is put in culturing room, is that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate 21d under the condition of 16/8h to obtain plantlet, namely finishes the cultivation that improves look wood maple stem eye growth coefficient;
Wherein tender stem elongation medium is take the MS minimal medium as minimal medium in the step 4, and also comprises the naa (NAA) of 0.1mg, the sucrose of 20g and the agar of 6g in the tender stem elongation medium of every 1L, and the pH value is 5.8;
Tender stem proliferated culture medium is take the MS minimal medium as minimal medium in the step 5, and also comprises the indolebutyric acid (IBA) of 0.1mg and the 6-benzyl aminoadenine (6-BA) of 1mg in the tender stem proliferated culture medium of every 1L.
The look wood maple resting shoot on the elite stand of growing up is taken from Maoershan Experimental Forest Farm of Northeast Forestry University in the present embodiment step 1 April, after branch is fetched in indoor water planting (16～22 ℃ of temperature) 3～4d, during every day change water one time, for subsequent use for testing.
Tender stem elongation medium and tender stem proliferated culture medium in the present embodiment are all through 121 ℃ of sterilization 20min, naturally coolings.
Inoculate the stem section with bud in a tender stem proliferated culture medium in the present embodiment step 4.
The tender stem of an elongation of inoculation in tender stem proliferated culture medium in the present embodiment step 5.
Embodiment two: the resting shoot of getting in the step 1 on the adult elite stand of look wood maple that is not both of present embodiment and embodiment one is scrubbed with washing powder water, and then water is rinsed well, is cut into the stem section of 3cm.Other step and parameter are identical with embodiment one.
Embodiment:
Improving the cultural method of look wood maple stem eye growth coefficient realizes according to the following steps:
One, the resting shoot of getting on the adult elite stand of look wood maple is scrubbed with washing powder water, and then water is rinsed well, is cut into the stem section of 3cm, and every section has 1 axillalry bud or 1 terminal bud, again the stem section is soaked 10min in washing powder water, do not stop therebetween to stir, then under flowing water, wash 40min;
Two, on superclean bench be 75% alcohol solution dipping 30s with mass concentration after the stem section is cleaned, the 20min that then sterilizes in the mass concentration that is added with 2 Twen-20 is 2% NaClO uses aseptic water washing 4 times again;
Three, the stem section after the step 2 sterilization blots the moisture on surface with aseptic filter paper, then with tweezers perula is divested, until only remain 1mm bud point, again 0.5cm is respectively excised with scalpel in stem section two ends, the direction that makes progress according to the morphology upper end vertically is inserted into the stem section in the blake bottle that the MS minimal medium is housed, insertion depth is 0.5cm, then blake bottle is put in the culturing room with after the plastic seal membrana oralis sealing, and be that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate the stem sections that obtained in 2 weeks with bud under the condition of 16/8h;
The stem section of four, getting step 3 gained band bud is inoculated in the 50ml blake bottle that the tender stem elongation medium of 20ml is housed, and then is put in culturing room, is that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate the tender stem that 30d obtains elongation under the condition of 16/8h;
Five, the tender stem of getting step 4 gained elongation is inoculated in the 50ml blake bottle that the tender stem proliferated culture medium of 20ml is housed, and then is put in culturing room, is that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate 30d under the condition of 16/8h to obtain plantlet, namely finishes the cultivation that improves look wood maple stem eye growth coefficient;
Wherein tender stem elongation medium is take the MS minimal medium as minimal medium in the step 4, and also comprises the naa of 0.1mg, the sucrose of 20g and the agar of 6g in the tender stem elongation medium of every 1L, and the pH value is 5.8;
Tender stem proliferated culture medium is take the MS minimal medium as minimal medium in the step 5, and also comprises the indolebutyric acid of 0.1mg and the 6-benzyl aminoadenine of 1mg in the tender stem proliferated culture medium of every 1L.
The tender stem of gained elongation in the present embodiment step 4, the axillary bud sprouting rate can reach 63.3%, and average height can reach 15.9mm.
Gained plantlet in the present embodiment step 5, the bud rate of increase can reach 90%, and the propagation multiple can reach 3.19, and elongation is normal.
Claims (2)
1. cultural method that improves look wood maple stem eye growth coefficient is characterized in that the cultural method that improves look wood maple stem eye growth coefficient realizes according to the following steps:
One, the resting shoot of getting on the adult elite stand of look wood maple is scrubbed with washing powder water, and then water is rinsed well, is cut into the stem section of 2～3cm, and every section has 1～2 axillalry bud or 1 terminal bud, again the stem section is soaked 10min in washing powder water, do not stop therebetween to stir, then under flowing water, wash 40min;
Two, on superclean bench be 75% alcohol solution dipping 30s with mass concentration after the stem section is cleaned, the 20min that then sterilizes in the mass concentration that is added with 2～3 Twen-20 is 2% NaClO uses aseptic water washing 3～5 times again;
Three, the stem section after the step 2 sterilization blots the moisture on surface with aseptic filter paper, then with tweezers perula is divested, until only remain 1～2mm bud point, again 0.5cm is respectively excised with scalpel in stem section two ends, the direction that makes progress according to the morphology upper end vertically is inserted into the stem section in the blake bottle that the MS minimal medium is housed, insertion depth is 0.5cm, then blake bottle is put in the culturing room with after the plastic seal membrana oralis sealing, and be that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate the stem sections that obtained in 2 weeks with bud under the condition of 16/8h;
The stem section of four, getting step 3 gained band bud is inoculated in the 50ml blake bottle that the tender stem elongation medium of 20ml is housed, and then is put in culturing room, is that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate the tender stem that 21d obtains elongation under the condition of 16/8h;
Five, the tender stem of getting step 4 gained elongation is inoculated in the 50ml blake bottle that the tender stem proliferated culture medium of 20ml is housed, and then is put in culturing room, is that 25 ℃, intensity of illumination are 40 μ mol.m in temperature
-2s
-1, light/dark cycle is to cultivate 21d under the condition of 16/8h to obtain plantlet, namely finishes the cultivation that improves look wood maple stem eye growth coefficient;
Wherein tender stem elongation medium is take the MS minimal medium as minimal medium in the step 4, and also comprises the naa of 0.1mg, the sucrose of 20g and the agar of 6g in the tender stem elongation medium of every 1L, and the pH value is 5.8;
Tender stem proliferated culture medium is take the MS minimal medium as minimal medium in the step 5, and also comprises the indolebutyric acid of 0.1mg and the 6-benzyl aminoadenine of 1mg in the tender stem proliferated culture medium of every 1L.
2. a kind of cultural method that improves look wood maple stem eye growth coefficient according to claim 1 is characterized in that getting in the step 1 resting shoot that look wood maple grows up on the elite stand and scrubs with washing powder water, and then water is rinsed well, is cut into the stem section of 3cm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201210453551 CN102907329A (en) | 2012-11-13 | 2012-11-13 | Cultivating method for improving multiplication coefficient of shoot |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103404438A (en) * | 2013-08-06 | 2013-11-27 | 巴中市光雾山植物研究所 | Acer paimatum seed tissue culture method |
| CN114711143A (en) * | 2022-04-21 | 2022-07-08 | 东北林业大学 | Method for asexually and rapidly propagating acer truncatum seedlings through tissue culture |
| CN116724889A (en) * | 2023-05-31 | 2023-09-12 | 东北林业大学 | Method for in-vitro rapid propagation by using axillary buds of Acer mono |
-
2012
- 2012-11-13 CN CN 201210453551 patent/CN102907329A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103404438A (en) * | 2013-08-06 | 2013-11-27 | 巴中市光雾山植物研究所 | Acer paimatum seed tissue culture method |
| CN114711143A (en) * | 2022-04-21 | 2022-07-08 | 东北林业大学 | Method for asexually and rapidly propagating acer truncatum seedlings through tissue culture |
| CN114711143B (en) * | 2022-04-21 | 2023-08-08 | 东北林业大学 | Method for asexually and rapidly propagating Acer ginnala Maxim seedlings through tissue culture |
| CN116724889A (en) * | 2023-05-31 | 2023-09-12 | 东北林业大学 | Method for in-vitro rapid propagation by using axillary buds of Acer mono |
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Application publication date: 20130206 |