阅读说明：本技术 一种牛大力的培养方法 (A kind of cultural method of beautiful millettia root ) 是由 周国权 郑彬江 郑彬旭 罗志超 于 2019-06-21 设计创作，主要内容包括：本发明涉及一种牛大力的培养方法,通过对牛大力幼苗的前期浸泡处理,以及在培养基中加入邻羟基苯甲酸甲酯、牛大力幼苗根部提取液,可防止培养期间培养物褐变死亡,从而提高增殖效率,并且能够有效地诱导培养物的细胞分裂,可以促进愈伤组织的形成,提高愈伤组织生长效率。(The present invention relates to a kind of cultural methods of beautiful millettia root, pass through immersion treatment early period to beautiful millettia root seedling, and Methyl Salicylate, beautiful millettia root seedling root extracting solution are added in the medium, it can prevent culture Necrosis during culture, to improve proliferation efficiency, and it is capable of the cell division of effectively Induced cultures, calli induction can be promoted, improve callus growth efficiency.)

1. a kind of cultural method of beautiful millettia root, which comprises the steps of: take the beautiful millettia root seedling stem sprouted 14 days Portion is washed with distilled water 30 minutes after stripping epidermis, is then dipped in the Benza that concentration is 8% 2 to 6 seconds, Then concentration be 60% alcoholic solution in impregnates 1 to 3 second, then concentration be 10% liquor natrii hypochloritis in immersion 30 to 40 minutes, the seedling stem after immersion treatment is scratched into wound with cutter, then wound is put into the first culture medium downward Middle carry out callus tissue culture, first culture medium contain MS salt, 0.5-1 mg/L Methyl Salicylate, 0.3 mg/L BAP, 1 mg/L NAA, 20-30g/L sucrose and 5-10g/L agar, the pH of the first culture medium are 5.8, are trained in the first culture medium After supporting 70 days, it is inoculated on the second culture medium, the second culture medium contains MS salt, vitamin, 0.3 mg/L BAP, 1 mg/L NAA, 20-30g/L sucrose and 3-6g/L agar, the pH of the second culture medium are 5.8, after cultivating 30-60 days in the second culture medium, Callus is transferred in HSM culture medium again and is regenerated, after rudiment, no hormone MS culture medium is transferred to, then after 7-10 days It is transplanted into soil.

2. the cultural method of beautiful millettia root as described in claim 1, which is characterized in that also big containing ox in first culture medium The extracting solution of power seedling root, the extracting method of the extracting solution are as follows: the root of beautiful millettia root seedling is shredded, and is put into methanol It extracts, is then concentrated under reduced pressure to remove methanol overnight in solution, add butyl acetate and be sirred and separated into water layer and the first vinegar Acid butyl ester layer takes above-mentioned butyl acetate layer to be added in 5% sodium bicarbonate solution, and continuing to be sirred and separated is water layer and the second acetic acid Butyl ester layer takes the second butyl acetate layer to obtain extracting solution.

3. the cultural method of beautiful millettia root as claimed in claim 2, which is characterized in that the additive amount for stating extracting solution is 0.5-2ml/ L。

Technical field

The application belongs to technical field of plant culture, and in particular to a kind of cultural method of beautiful millettia root.

Background technique

Beautiful millettia root, alias pig's feet large bamboo hat with a conical crown and broad brim, Jin Zhonggen,Beautiful millettia root, hang upside down Jin Zhong, energetically potato.It is a kind of medicinal material.For pulse family pulse family Millettia plant.Beautiful millettia root is in the ascendant in China's not yet industrialization, so many farmers have often violated in understanding Mistake, take for once planting beautiful millettia root successfully good harvest.Hardly realize, beautiful millettia root there are many different germplasm product sources, no With beautiful millettia root provenance, morphological character and having a certain difference property of yield, choosing can successfully have a good harvest to the kind of high yield of fine quality； Conversely, then may failure.Beautiful millettia root seedling period management is not scientific, will seriously affect the harvest in later period, this is fatefulue science Reasonable tree crown can manufacture more photosynthates and convey to root, promote quickly expanding for rhizome.

More serious browning will appear in the process using the mistake that cultural method in the prior art cultivates beautiful millettia root, Brown stain even will occur in 4-6 weeks, seriously affected the growth efficiency of beautiful millettia root, therefore, how to provide a kind of effective Inhibiting beautiful millettia root to occur the inoculation method of browning during the growth process is a technical problem to be solved urgently.

Summary of the invention

The purpose of the present invention is to provide a kind of beautiful millettia root inoculation methods for effectively inhibiting browning.

Specifically, beautiful millettia root cultural method of the invention in turn includes the following steps:

The beautiful millettia root seedling stem sprouted 14 days is taken, is washed with distilled water 30 minutes after stripping epidermis, is then dipped in concentration It is 2 to 6 seconds in 8% Benza, is then impregnated 1 to 3 second in the alcoholic solution that concentration is 60%, then be in concentration It is impregnated 30 to 40 minutes in 10% liquor natrii hypochloritis, the seedling stem after immersion treatment is scratched into wound with cutter, Then wound is put into downward in the first culture medium and carries out callus tissue culture, first culture medium contains MS salt, 0.5-1 Mg/L Methyl Salicylate, 0.3 mg/L BAP, 1 mg/L NAA, 20-30g/L sucrose and 5-10g/L agar, the first training The pH for supporting base is 5.8, after cultivating 70 days in the first culture medium, is inoculated on the second culture medium, the second culture medium contains MS salt, vitamin, 0.3 mg/L BAP, 1 mg/L NAA, 20-30g/L sucrose and 3-6g/L agar, the pH of the second culture medium are 5.8, after being cultivated 30-60 days in the second culture medium, then callus is transferred in HSM culture medium and is regenerated, after rudiment, transfer To no hormone MS culture medium, the earth that buries then is transplanted after 7-10 days.

Preferably, the extracting solution in first culture medium also containing beautiful millettia root seedling root, the extraction of the extracting solution Method are as follows: the root of beautiful millettia root seedling is shredded, and is put into methanol solution and extracts overnight, is then concentrated under reduced pressure to remove first Alcohol adds butyl acetate and is sirred and separated into water layer and the first butyl acetate layer, above-mentioned butyl acetate layer is taken to be added to 5% carbon In sour hydrogen sodium solution, continuing to be sirred and separated is water layer and the second butyl acetate layer, and the second butyl acetate layer is taken to be extracted Liquid.

Preferably, the additive amount of the extracting solution is 0.5-2ml/L.

The beneficial effects of the present invention are: 1) improve later period callus group by immersion treatment early period to beautiful millettia root seedling The proliferation rate knitted；2) by the way that Methyl Salicylate is added in the medium, effectively inhibit the browning in incubation existing As；3) by the way that beautiful millettia root seedling root extracting solution is added in the medium, the degree of oxidation of culture medium is effectively reduced.By with Upper technological means, the present invention can prevent culture Necrosis during culture, to improve proliferation efficiency, and can be effectively The cell division of Induced cultures can promote calli induction, improve callus growth efficiency.

Specific embodiment