CN104388379A - Method for obtaining sugarcane calluses from spermine - Google Patents
Method for obtaining sugarcane calluses from spermine Download PDFInfo
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- CN104388379A CN104388379A CN201410679433.4A CN201410679433A CN104388379A CN 104388379 A CN104388379 A CN 104388379A CN 201410679433 A CN201410679433 A CN 201410679433A CN 104388379 A CN104388379 A CN 104388379A
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- callus
- spermine
- sugarcane
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- calluses
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Abstract
The invention relates to a method for obtaining sugarcane calluses from spermine. According to the method, sugarcane young leaves obtained by a conventional method as explants are sliced and routinely disinfected and spermine is used as a callus inducer; according to the conventional preparation method of MS solid culture medium, 1.5mg.L<-1>-3.0mg.L<-1> spermine is added and the pH value is adjusted to 5.8-6.0; routine sterilization and inoculation are performed; the sugarcane callus is induced under normal culture conditions, the 10-day callus induction rate is up to 29%-43%, the 25-day callus induction rate is up to 77%-85% and under the same conditions, the fresh weight of the harvested callus is 1-4 times of that of 2,4-D group (CK). The callus obtained by the method is very slight in browning and hardening, and has excellent quality; the method has the advantages of high callus induction rate and high efficiency; since the spermine is cheap and the replacement of novel culture medium is not needed, a lot of labor and culture media are saved and thus the production cost can be decreased and the method is suitable for being popularized and applied in large-scale production and research.
Description
Technical field
The invention belongs to the technical field of tissue culture of Plant Biotechnology, especially a kind of spermine obtains the method for sugarcane callus.
Technical background
Sugarcane is the most important sweeting agent crops of the mankind.The fast breeding of Sugarcane Superior Variety and the research of the various modern biotechnology systems relevant to sugarcane and foundation, nearly all need first to be obtained based on callus by the dedifferentiation of tissue culture technique induction explant.Since sugarcane tissue culture technology is set up, generally all with 2 in the industry, 4-D (2,4-dichlorphenoxyacetic acid) as sugarcane callus inductor [sugarcane tissue culture Review Study [J] Guangxi hotwork science and technology yellow favour virtue 1998, (2): 33-37], [sugarcane tissue culture progress [J] Agriculture of Anhui science willow etc. 2007.35 (12): 3490-3492].But 2,4-D belongs to the benzene derivative of synthetic, have obvious toxicity and remain, very easily make the rapid brown stain of callus of just acquisition (polluting also known as phenol evil or phenol) and harden, even death, success ratio is very low.Technician has to expend a lot of time for this reason, upgrades substratum continually to desalinate the toxicity hazard [phenol evil [J] sugarcane Qin Ting person of outstanding talent in simple analysis sugarcane tissue-culture waits 1997 (02): 12-14] of 2,4-D; [2003 (2) l8-21 such as progress [J] Fujian sugarcane Lee hamming of brown stain in sugarcane tissue culture].In recent years, Lan Taoju and Chen Nian equality LFS (spirit hair element, code name PGR-08) successfully establishes technical system [a kind of method China Patent No. ZL 201010216180.9 of spirit hair element induction sugarcane callus] that is brand-new, energy high-efficiency high-quality acquisition sugarcane callus; [spirit hair element (LFS) is on impact [J] the seed Chen Nian equality 2013,32 (11): 1-2 of induction sugarcane callus].But LFS there is no manufacturer at home, import preparation price is very expensive, is difficult to apply.So the new technology exploring high-quality and efficient cheapness acquisition sugarcane callus is the focus that insider is concerned about always.Spermine (Spermine, Spm.) be the one of polyamines, there is the physiologically active of plant growth regulating substance, and be extensively present in higher plant circle, be pure natural substance [polyamines is instrumentality [J] the Plant Physiology Communications Pan Rui vehement 1985 (6): 63-68 of growth and development of plants].People, to the research of its Promoting plant growth, the aspects such as anti-adversity ability that delay senility, strengthen, have many achievements.But, not yet retrieve spermine for inducing the research data of this important biomolecule technical field of sugarcane callus.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of method that spermine obtains sugarcane callus.
The present invention solves the problems of the technologies described above with following technical scheme:
Spermine obtains a method for sugarcane callus, comprises the steps:
Obtain In The Young Leaf of Sugarcane section in conventional manner and make explant, and routine disinfection, do callus induction agent with spermine; The MS solid medium prepared routinely, adds Spm.1.5mgL
-1~ 3.0mgL
-1, adjust pH5.8 ~ 6.0; Conventional sterilant, inoculation; Under conventional culture conditions, induce sugarcane callus, 10d healing rate reaches 29%-43%, and 25d healing rate reaches 77%-85%.
The optimum concentration of spermine induction sugarcane callus is 2.0mgL
-1-2.5mgL
-1.
The present invention is used for the induction of sugarcane callus, and institute obtains the brown stain of callus and hardens all very slightly, and quality is superior; Go out more speed fast, efficiency is high, and namely 10d healing rate reaches more than 29%, 25d healing rate can reach more than 80%, gathers in the crops 1-4 times that callus fresh weight is CK group under identical conditions; Spermine low price (domestic every gram of Renminbi about 500 yuan; 1/400 of not enough spirit hair element), and inducing culture midway is without the need to more renewing substratum, saves a large amount of labour and substratum; low production cost, is suitable for applying in scientific research and large-scale production.
Embodiment
Below by way of case study on implementation, technical scheme of the present invention is described further.If do not specialize, technique means used in embodiment is all the conventional means of those skilled in the art.
The method of spermine induction sugarcane callus of the present invention comprises the following steps:
Conventionally method strips sugarcane tip yolk yellow spire close to the inner portion, cuts into the section of about 0.5cm × 0.5cm, is inoculated into containing Spm.1.5mgL respectively
-1~ 3.0mgL
-1each group of modified form MS solid medium in, under conventional culture conditions, carry out sugarcane callus induction and cultivation.
In Table 1, example 1 is control group (CK); Example 2-5 is the treatment group of adding various dose Spm..Each group of explant material used, operation steps and culture condition are all identical.
Table 1.Spm. and 2,4-d induces the contrast of In The Young Leaf of Sugarcane callus effect
Note: 1. 2,4-D be the agent of conventional sugarcane callus induction in subordinate list; Spm. for the present invention innovates sugarcane callus inductor.
2. healing rate (%)=go out more spire number of slices/inoculation spire number of slices × 100%
3. browning rate (%)=brown stain spire number of slices (no matter going out whether to heal)/inoculation spire number of slices × 100%
4. how many browning degree, represent with "+".
5. the rate that hardens (%)=callus consolidation is difficult to the spire number of slices of proliferation and differentiation/go out more spire number of slices × 100%
6. the metering method of callus fresh weight is, after cultivating 25d, often group randomly draws 3 bottles, takes out cultivated material (containing former section, callus and derivative appurtenant thereof) and remove adhesion substratum in units of bottle, claims its fresh weight fast, statistical average value.
The contrast of each routine experimental data from table 1: the technology of the present invention spermine induces sugarcane callus, under identical experiment condition, go out more speed, go out more total amount, particularly callus quality, be all significantly better than traditional 2,4-D.The suitable concentration of spermine induction sugarcane callus is 2.0mgL
-1-2.5mgL
-1.
In view of spermine has many places manufacturer at home, ample supply and prompt delivery, low price, the present invention is easy to apply.
Each case study on implementation listed by table 1 has done comparatively detailed description to technical system of the present invention.But, on basis of the present invention, those skilled in the relevant art are easy to replace minimum medium MS of the present invention with other minimum medium, or the consumption of spermine is changed, even spermine and other active substance carry out composite after be used further to induce the callus of sugarcane callus and other plant.So, allly in the cultivation program of induction sugarcane and other plant callus, with the addition of spermine, all belong to the scope of protection of present invention.
Claims (2)
1. obtain a method for sugarcane callus with spermine, it is characterized in that explant is made in the In The Young Leaf of Sugarcane section obtained in conventional manner, after routine disinfection, do callus induction agent with spermine; The method of the MS solid medium prepared routinely, adds Spm.1.5mgL
-1~ 3.0mgL
-1, adjust pH 5.8 ~ 6.0; Conventional sterilant, inoculation; Under the culture condition of routine, induce sugarcane callus, 10d healing rate reaches 29%-77%, and 25d healing rate reaches 77%-85%.
2. obtain the method for sugarcane callus as claimed in claim 1 with spermine, it is characterized in that the optimum concentration of spermine induction sugarcane callus is 2.0mgL
-1-2.5mgL
-1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410679433.4A CN104388379A (en) | 2014-11-24 | 2014-11-24 | Method for obtaining sugarcane calluses from spermine |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410679433.4A CN104388379A (en) | 2014-11-24 | 2014-11-24 | Method for obtaining sugarcane calluses from spermine |
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| Publication Number | Publication Date |
|---|---|
| CN104388379A true CN104388379A (en) | 2015-03-04 |
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|---|---|---|---|
| CN201410679433.4A Pending CN104388379A (en) | 2014-11-24 | 2014-11-24 | Method for obtaining sugarcane calluses from spermine |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106258988A (en) * | 2016-10-14 | 2017-01-04 | 广西壮族自治区烟草公司百色市公司 | Method with spermine evoking tobacco callus |
| CN106359102A (en) * | 2016-10-14 | 2017-02-01 | 广西壮族自治区烟草公司百色市公司 | Method for directly inducing tobacco leaf regenerated plant by using spermine |
-
2014
- 2014-11-24 CN CN201410679433.4A patent/CN104388379A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106258988A (en) * | 2016-10-14 | 2017-01-04 | 广西壮族自治区烟草公司百色市公司 | Method with spermine evoking tobacco callus |
| CN106359102A (en) * | 2016-10-14 | 2017-02-01 | 广西壮族自治区烟草公司百色市公司 | Method for directly inducing tobacco leaf regenerated plant by using spermine |
| CN106359102B (en) * | 2016-10-14 | 2018-08-28 | 广西壮族自治区烟草公司百色市公司 | With the method for the direct evoking tobacco leaf regeneration plant of spermine |
| CN106258988B (en) * | 2016-10-14 | 2019-02-26 | 广西壮族自治区烟草公司百色市公司 | With the method for spermine evoking tobacco callus |
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Application publication date: 20150304 |