CN115281087A - Callus induction and rapid propagation method of bamboo joint tree - Google Patents
Callus induction and rapid propagation method of bamboo joint tree Download PDFInfo
- Publication number
- CN115281087A CN115281087A CN202210883164.8A CN202210883164A CN115281087A CN 115281087 A CN115281087 A CN 115281087A CN 202210883164 A CN202210883164 A CN 202210883164A CN 115281087 A CN115281087 A CN 115281087A
- Authority
- CN
- China
- Prior art keywords
- callus
- culture medium
- seeds
- bamboo joint
- rapid propagation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention belongs to the technical field of plant callus breeding, and discloses a callus induction and rapid propagation method of a bunchy tree, which comprises S1, inoculation of bunchy tree seeds: inoculating mature bamboo joint tree seeds to an MS culture medium after aseptic treatment; s2, differentiation and sprouting of the bamboo-joint tree seeds and new callus breeding: after the seeds germinate, placing the seeds in a differentiation culture medium for callus and bud induction, and subculturing until the callus differentiates to bud again and new callus; s3, rooting culture of the bamboo joint trees: cutting off callus of the differentiated sprout, inoculating the cut callus into a rooting culture medium to induce the formation of a root system, and transferring the cultured callus into a seedling hardening stage; s4, greenhouse domestication: hardening the rooted plantlets, and then cultivating the rooted plantlets to enable the rooted plantlets to be directly transplanted to a field; the method solves the problems that the breeding difficulty of the bamboo joint tree in the prior art is high, and the new root of the bamboo joint tree is easy to wither and die when the new root is induced by the existing plant root system induction scheme, and is suitable for the induction and the rapid propagation of the callus of the bamboo joint tree.
Description
Technical Field
The invention relates to the technical field of plant callus breeding, in particular to a callus induction and rapid propagation method of a bunchy tree.
Background
The bamboo joint tree (Carallia brachiata (lour.) Merr) is a mangrove family, the bamboo joint tree belongs to evergreen arbor, the height can reach 10m, the bark is smooth, and the bamboo joint tree has few cracks and is gray brown. The wood is hard and heavy, has interlaced grains, rather thick structure, large heartwood, dark reddish brown with yellow, light-colored with red and lustrous edges, and is happyGood materials for furniture, decorative wood, doors, windows, appliances, and the like; and bamboo joint tree is especially suitable for treating atmospheric pollutants, especially SO 2 The resistance grade of the gas reaches the first grade, and the bamboo joint tree is thus known as 'green diamond', so that the bamboo joint tree is an ideal new and excellent ornamental and good pollution-resistant precious tree species in gardens, the bark of the bamboo joint tree is also used for medicine, malaria can be treated, and the bamboo joint tree is a national secondary protection tree species.
But the breeding difficulty of the bamboo joint tree is high, the conventional seed seedling raising method is high in seed collection difficulty, irregular in maturity, low in breeding speed, long in seedling raising period, high in seedling sowing variation rate, difficult to maintain excellent characters of a female parent, and extremely difficult to meet market demands due to the lagged production mode; although the propagation of plants is realized by using asexual propagation modes such as tissue culture and the like in the field, the report of callus induction is not seen in the field when tissue culture is carried out by bud propagation in the tissue culture process of the Japanese tress, and secondly, the inventor discovers that the induced new roots of the Japanese tress are easy to wither according to a root system induction scheme which is commonly used for plants in the field.
Disclosure of Invention
The invention aims to provide a callus induction and rapid propagation method of a bamboo joint tree, and aims to solve the problems that breeding difficulty of the bamboo joint tree is high in the prior art, and a new root of the bamboo joint tree is easy to wither and die when a new root is induced by an existing plant root system induction scheme.
In order to achieve the above purpose, the invention provides the following technical scheme:
the callus induction and rapid propagation method of the Japanese cypress comprises the following steps:
s1, inoculation of the bamboo joint tree seeds: removing pulp from mature Japanese shrub fruits, leaving seeds, performing aseptic treatment on the seeds, and inoculating the seeds to an MS culture medium;
s2, differentiation and budding of the seeds of the bamboo-joint tree and new callus breeding: the method comprises the following steps of planting seeds in an MS culture medium for germination, taking out aseptic seedlings when the hypocotyls are 3-5 cm long, shearing the hypocotyls to be 0.5-2 cm long in an aseptic environment, then placing the hypocotyls in a differentiation culture medium for callus and bud induction, germinating and callus simultaneously on the hypocotyls of the bamboo joint tree, forming more callus on one section contacting the culture medium, timely separating after the callus is formed, transferring the callus into the differentiation culture medium for subculture, and enabling the callus to differentiate again to sprout and form new callus;
s3, rooting culture of the bunchy trees: when the stem of the sprout differentiated in the S2 grows to 0.5-1 cm, cutting off callus at the base part of the stem, inoculating the stem with the sprout into a rooting culture medium to induce the formation of a root system, carrying out dark culture for one week, then carrying out illumination culture, when 2-4 roots grow on each stem and the root system grows to be 0.25-0.5 cm long, transferring the stem into an MS culture medium to continue growing until the root system grows to be 1-2 cm, and transferring into a seedling hardening stage;
s4, greenhouse domestication: closing a bottle to harden the rooted plantlets transferred to the hardening stage in the S3, opening the bottle to harden the plantlets, soaking the rooted plantlets in 1000 times of carbendazim solution for 3-5 minutes, fishing out, airing and transplanting the rooted plantlets, spraying 1000 times of carbendazim solution on the transplanted plantlets, culturing the rooted plantlets, and transplanting the rooted plantlets to a field for normal field water and fertilizer management when the height of the plantlets is more than 10cm or the number of leaves is more than 8; thereby completing the callus induction and rapid propagation of the bamboo joint tree.
Furthermore, in S1, S2 and S3, agar powder in the MS culture medium is changed into 6g/L plant gel, the rest components and the concentration are unchanged, and the pH value of the culture medium is 5.8-6.0.
Further, in S1, the step of aseptically processing the seed is: washing with clean tap water, soaking and cleaning with 75% alcohol, and cleaning twice with double distilled water; and then soaking for 30min by using a 1% sodium hypochlorite solution, shaking clockwise once every 10min, shaking for 1min every time, and cleaning twice by using double distilled water after the treatment is finished, wherein the cleaning time is not less than 3min every time.
Further, in S2, the culture conditions for germination of seeds in MS medium are: the ambient temperature is 28 +/-2 ℃, the illumination time is 12h/d, and the light intensity is 3000 lx-5000 lx.
Further, in S2, the differentiation medium is MS medium supplemented with 1.5mg/L ZT zeatin and 0.3mg/L IAA.
Further, in S3, the rooting medium is MS medium added with 7mg/L of indolebutyric acid.
Further, in S4, the closed-bottle hardening time of the seedlings of the opposite roots is 7 days, and the open-bottle hardening time of the seedlings of the opposite roots is 3 days; the matrix for transplanting the rooted plantlets is river sand and peat soil with the volume ratio of 1.
Further, in S4, the culture conditions for the plantlets are: the air humidity is more than 90%, the ambient temperature is 28 +/-2 ℃, the illumination time is 12h/d, and the light intensity is 3000lx.
The technical scheme has the beneficial effects that:
the invention can not only induce the sprout, but also induce the bamboo tree to form callus, thus realizing the purpose of propagation; the method solves the problems that the breeding difficulty of the bamboo-joint tree in the prior art is high, and the new root of the bamboo-joint tree is easy to wither when induced by the existing plant root system induction scheme.
Detailed Description
The present invention will be described in further detail with reference to the following embodiments:
the callus induction and rapid propagation method of the Japanese cypress comprises the following steps:
s1, inoculation of the bamboo joint tree seeds: removing pulp from mature bamboo joint fruit, leaving seeds, washing the seeds with clean tap water, soaking and cleaning with 75% alcohol, and cleaning twice with double distilled water; soaking for 30min by using a 1% sodium hypochlorite solution, shaking clockwise once every 10min, shaking for 1min every time, cleaning twice by using double distilled water after treatment, cleaning for not less than 3min every time, and inoculating to an MS culture medium after aseptic treatment;
s2, differentiation and budding of the seeds of the bamboo-joint tree and new callus breeding: the method comprises the steps that seeds are planted and germinated in an MS culture medium under the conditions that the environmental temperature is 28 +/-2 ℃, the illumination time is 12h/d, and the light intensity is 3000 lx-5000 lx, when the embryonic axis is 3-5 cm long, aseptic seedlings are taken out, the embryonic axis is cut to be 0.5-2 cm long in the aseptic environment, then the embryonic axis is placed in a differentiation culture medium to be induced into callus and buds, the differentiation culture medium is formed by adding 1.5mg/L ZT zeatin and 0.3mg/L IAA to the MS culture medium, the embryonic axis of a bunchy tree can simultaneously bud and callus, the callus is formed in a section which is in contact with the culture medium, and the callus is timely separated and transferred to the differentiation culture medium to be subjected to subculture after the callus is formed, so that the callus differentiates to bud and form new callus again;
s3, rooting culture of the bunchy trees: when the stem of the sprout differentiated in the S2 grows to 0.5-1 cm, cutting off callus at the base part of the stem, inoculating the stem with the sprout into a rooting culture medium to induce the formation of a root system, wherein the rooting culture medium is an MS culture medium added with 7mg/L indolebutyric acid, carrying out dark culture for one week, then carrying out illumination culture, when 2-4 roots grow on each stem and the root system grows to 0.25-0.5 cm, transferring the stem into the MS culture medium to continue growing until the root system grows to 1-2 cm, and transferring into a seedling hardening stage;
s4, greenhouse domestication: the rooting plantlets in the seedling hardening stage in the S3 are firstly hardened in a closed bottle for 7 days, then hardened in an open bottle for 3 days, then soaked in 1000 times of carbendazim solution for 3-5 minutes, fished out, aired and dried, and then transplanted into river sand and peat soil matrixes with the volume ratio of 1; thereby completing the callus induction and rapid propagation of the bamboo joint tree.
In S1, S2 and S3, agar powder in all MS culture media is changed into 6g/L plant gel, the rest components and the concentration are unchanged, and the pH value of the culture media is 5.8-6.0.
The above description is only an example of the present invention, and the common general knowledge of the technical solutions or characteristics known in the solutions is not described herein too much. It should be noted that, for those skilled in the art, without departing from the technical solution of the present invention, several variations and modifications can be made, and these should also be considered as the protection scope of the present invention, which will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
Claims (8)
1. The callus induction and rapid propagation method of the Japanese cypress is characterized by comprising the following steps:
s1, inoculation of the bamboo joint tree seeds: removing pulp from mature Japanese shrub fruits, leaving seeds, performing aseptic treatment on the seeds, and inoculating the seeds to an MS culture medium;
s2, differentiation and sprouting of the bamboo-joint tree seeds and new callus breeding: the method comprises the following steps of planting seeds in an MS culture medium for germination, taking out aseptic seedlings when the hypocotyls grow to 3-5 cm, shearing the hypocotyls to be 0.5-2 cm in an aseptic environment, placing the hypocotyls in a differentiation culture medium for callus and bud induction, germinating the hypocotyls of the bunchy tree and callus at the same time, forming the callus at one section which is in contact with the culture medium, timely separating the formed callus and transferring the callus into the differentiation culture medium for subculture, and enabling the callus to differentiate again to sprout and form new callus;
s3, rooting culture of the bamboo joint trees: when the stem of the sprout differentiated in the S2 grows to 0.5-1 cm, cutting off callus at the base part of the stem, inoculating the stem with the sprout into a rooting culture medium to induce the formation of a root system, carrying out dark culture for one week, then carrying out illumination culture, when 2-4 roots grow on each stem and the root system grows to be 0.25-0.5 cm long, transferring the stem into an MS culture medium to continue growing until the root system grows to be 1-2 cm, and transferring into a seedling hardening stage;
s4, greenhouse domestication: closing a bottle to harden the rooted plantlets transferred to the hardening stage in the S3, opening the bottle to harden the plantlets, soaking the rooted plantlets in 1000 times of carbendazim solution for 3-5 minutes, fishing out, airing and transplanting the rooted plantlets, spraying 1000 times of carbendazim solution on the transplanted plantlets, culturing the rooted plantlets, and transplanting the rooted plantlets to a field for normal field water and fertilizer management when the height of the plantlets is more than 10cm or the number of leaves is more than 8; thereby completing the callus induction and rapid propagation of the bamboo joint tree.
2. The callus induction and rapid propagation method of a Japanese ardisia according to claim 1, characterized in that: in S1, S2 and S3, agar powder in the MS culture medium is changed into 6g/L plant gel, other components and concentration are unchanged, and the pH value of the culture medium is 5.8-6.0.
3. The callus induction and rapid propagation method of a bamboo joint tree according to claim 1, characterized in that: in S1, the steps of aseptic processing of the seeds are: washing with clean tap water, soaking and cleaning with 75% alcohol, and cleaning twice with double distilled water; and then soaking for 30min by using a 1% sodium hypochlorite solution, shaking clockwise once every 10min, shaking for 1min every time, and cleaning twice by using double distilled water after the treatment is finished, wherein the cleaning time is not less than 3min every time.
4. The callus induction and rapid propagation method of a Japanese ardisia according to claim 1, characterized in that: in S2, the culture conditions for seed germination in MS medium are: the ambient temperature is 28 +/-2 ℃, the illumination time is 12h/d, and the light intensity is 3000 lx-5000 lx.
5. The callus induction and rapid propagation method of a bamboo joint tree according to claim 1, characterized in that: in S2, the differentiation medium is MS medium added with 1.5mg/L ZT zeatin and 0.3mg/L IAA.
6. The callus induction and rapid propagation method of a bamboo joint tree according to claim 1, characterized in that: in S3, the rooting culture medium is MS culture medium added with 7mg/L indolebutyric acid.
7. The callus induction and rapid propagation method of a bamboo joint tree according to claim 1, characterized in that: in S4, the closed-bottle hardening time of the seedlings with the roots is 7 days, and the open-bottle hardening time of the seedlings with the roots is 3 days; the matrix for transplanting the rooted plantlets is river sand and peat soil with the volume ratio of 1.
8. The callus induction and rapid propagation method of a Japanese ardisia according to claim 1, characterized in that: in S4, the culture conditions for the plantlets were: the air humidity is more than 90%, the ambient temperature is 28 +/-2 ℃, the illumination time is 12h/d, and the light intensity is 3000lx.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210883164.8A CN115281087A (en) | 2022-07-26 | 2022-07-26 | Callus induction and rapid propagation method of bamboo joint tree |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210883164.8A CN115281087A (en) | 2022-07-26 | 2022-07-26 | Callus induction and rapid propagation method of bamboo joint tree |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115281087A true CN115281087A (en) | 2022-11-04 |
Family
ID=83823400
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210883164.8A Pending CN115281087A (en) | 2022-07-26 | 2022-07-26 | Callus induction and rapid propagation method of bamboo joint tree |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115281087A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102227987A (en) * | 2011-05-20 | 2011-11-02 | 深圳市华美绿环境建设工程有限公司 | Method for planting mangrove plants in fresh water, |
| CN105454043A (en) * | 2015-03-09 | 2016-04-06 | 佛山市农业科学研究所 | Tissue culture virus elimination and rapid propagation method of Carallia brachiata |
-
2022
- 2022-07-26 CN CN202210883164.8A patent/CN115281087A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102227987A (en) * | 2011-05-20 | 2011-11-02 | 深圳市华美绿环境建设工程有限公司 | Method for planting mangrove plants in fresh water, |
| CN105454043A (en) * | 2015-03-09 | 2016-04-06 | 佛山市农业科学研究所 | Tissue culture virus elimination and rapid propagation method of Carallia brachiata |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111616052A (en) | Rapid propagation and sugar-free rooting culture method and application of apple rootstock catalpa bungei | |
| CN111280056A (en) | Subculture breeding method of stingless pepper tissue culture seedlings | |
| CN103430845A (en) | Strawberry tissue culturing method | |
| CN113080063B (en) | Rapid rooting method for tissue culture of coarse chaff tree | |
| CN110771365B (en) | Cutting propagation method of lingbao rhododendron | |
| CN116584380A (en) | Tissue culture method suitable for Liupu tea mountain-sill purple stock seedlings | |
| CN108811835B (en) | Rapid propagation method for citrus detoxification and micro-bud grafting | |
| CN114946655B (en) | Liupao tea seedling tissue culture method | |
| CN110999782A (en) | Tissue culture and rapid propagation method for gardenia jasminoides | |
| CN112243860B (en) | Tissue culture and rapid propagation method for Chinese parasol trees | |
| CN108391591A (en) | A kind of Golden Bell Tree tissue cultivation rapid breeding method | |
| CN1541518A (en) | Dendrobium unicum aseptic seeding and test tube seedling tecnnology | |
| CN115281087A (en) | Callus induction and rapid propagation method of bamboo joint tree | |
| CN112136526A (en) | Greenhouse castanea henryi twig cutting seedling method | |
| CN111374057A (en) | Environment-friendly and efficient micro-propagation method for cold-resistant China roses | |
| CN110537489A (en) | Test tube micro-cuttage aseptic propagation method for eggplant rootstocks | |
| CN112237141B (en) | Tissue culture propagation method of sclerotium rolfsii | |
| LU502555B1 (en) | In vitro culture method of thornless green prickly ash (zanthoxylum armatum dc) | |
| CN112655564B (en) | Method for establishing high-efficiency regeneration system of Chinese tallow tree stem segments | |
| CN112106658B (en) | Tissue culture and rapid propagation method for gelsemium elegans | |
| CN114208680B (en) | Tissue culture and rapid propagation method for sedum formosanum | |
| CN110506632B (en) | Tissue culture method of euscaphis konishii hayata | |
| CN113068614B (en) | Method for rapidly propagating high-quality seedlings of golden strelitzia | |
| CN112690215B (en) | Tissue culture method for Pixumu | |
| CN109964814B (en) | In-vitro rapid propagation method of viburnum sargentii |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |