阅读说明：本技术 一种提高牛大力生根率的培养方法 (A kind of cultural method improving beautiful millettia root rooting rate ) 是由 郑彬江 周国权 郑彬旭 罗志超 于 2019-07-22 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,依次包括以下步骤：步骤一,取牛大力幼苗茎尖组织进行无菌分离,并在固体培养基中培养14天；步骤二,将步骤一中得到的组织转移至含有细胞分裂素的第一液体培养基中,以150-200rpm进行振荡培养28天得到叶芽；步骤三,将步骤二中得到的叶芽转移至盛有不含细胞分裂素的第二液体培养基的锥形瓶中,在搅拌并通入空气的条件下培养21天,使叶芽组织再生并诱导生根。该培养方法能够有效提高牛大力的生根率,并避免褐化现象。(The present invention relates to a kind of cultural methods of beautiful millettia root, successively the following steps are included: step 1, takes beautiful millettia root seedling stem-tip tissue to carry out sterile separation, and cultivate 14 days in solid medium；Tissue obtained in step 1 is transferred in the first fluid nutrient medium containing the basic element of cell division by step 2, obtains leaf bud within shaken cultivation 28 days with 150-200rpm progress；Leaf bud obtained in step 2 is transferred in the conical flask for filling the culture medium of the second liquid without the basic element of cell division by step 3, is cultivated 21 days under conditions of stirring and being passed through air, and leaf bud regeneration and root induction are made.The cultural method can effectively improve the rooting rate of beautiful millettia root, and avoid browning.)

1. a kind of cultural method for improving beautiful millettia root rooting rate, which is characterized in that in turn include the following steps:

Step 1 takes beautiful millettia root seedling stem-tip tissue to carry out sterile separation, and cultivates 14 days in solid medium, the solid Culture medium contains MS salt, 0.5-1 mg/L BAP, 10-20g/L sucrose and 2-10g/L agar, and the pH of solid medium is 5.8；

Tissue obtained in step 1 is transferred in the first fluid nutrient medium containing the basic element of cell division, with 150- by step 2 200rpm carries out obtaining within shaken cultivation 28 days leaf bud, and first fluid nutrient medium is BAP containing 0.1-5mg/L and 10g/L sugarcane The MS culture medium of sugar, the pH of the first fluid nutrient medium are 5.8；

Leaf bud obtained in step 2 is transferred to the taper for filling the culture medium of the second liquid without the basic element of cell division by step 3 It in bottle, is cultivated 21 days under conditions of stirring and being passed through air, makes leaf bud regeneration and root induction, the second liquid training It supports base and contains 20-30g/L sucrose and 5-8g/L agar, the pH of second liquid culture medium is 5.8.

2. cultural method as described in claim 1, which is characterized in that the condition of root induction described in step 3 are as follows: temperature It is 25 DEG C -28 DEG C, humidity 90%-95%, photon illumination is 75 μm of ol/ m ² / s -90μmol/ m ² / s。

3. cultural method as claimed in claim 1 or 2, which is characterized in that the capacity of conical flask described in step 3 is 1500- 1800ml, the volume of second liquid culture medium are 1000-1200ml.

Technical field

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

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.

Culture of rootage is the key step of Plant Tissue Breeding, is in general after plant generation culture, using appropriate Culture medium cooperate a certain concentration and ratio hormone induction plant establishment process, using cultural method in the prior art train Beautiful millettia root is educated, culture period is long, and the size of the bud of growth is uneven, and base portion is hard and is difficult to divide, and rooting rate is low, and is transplanted to soil Survival rate is low in earth, and therefore, how to provide a kind of raising beautiful millettia root rooting rate and the cultural method of transplanting survival rate is this field Technical problem urgently to be resolved.

Summary of the invention

The purpose of the present invention is to provide a kind of cultural methods for effectively improving beautiful millettia root rooting rate and transplanting survival rate.

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

Step 1 takes beautiful millettia root seedling stem-tip tissue to carry out sterile separation, and cultivates 14 days in solid medium, the solid Culture medium contains MS salt, 0.5-1 mg/L BAP, 10-20g/L sucrose and 2-10g/L agar, and the pH of solid medium is 5.8；

Tissue obtained in step 1 is transferred in the first fluid nutrient medium containing the basic element of cell division, with 150- by step 2 200rpm carries out obtaining within shaken cultivation 28 days leaf bud, and first fluid nutrient medium is BAP containing 0.1-5mg/L and 10g/L sugarcane The MS culture medium of sugar, the pH of the first fluid nutrient medium are 5.8；

Leaf bud obtained in step 2 is transferred to the taper for filling the culture medium of the second liquid without the basic element of cell division by step 3 It in bottle, is cultivated 21 days under conditions of stirring and being passed through air, makes leaf bud regeneration and root induction, the second liquid training It supports base and contains 20-30g/L sucrose and 5-8g/L agar, the pH of second liquid culture medium is 5.8.

Preferably, the condition of root induction described in step 3 are as follows: temperature is 25 DEG C -28 DEG C, humidity 90%-95%, light Sub- illumination is 75 μm of ol/ m ² / s -90μmol/ m ² / s。

Preferably, the capacity of conical flask described in step 3 is 1500-1800ml, and the volume of second liquid culture medium is 1000-1200ml。

The invention has the benefit that

The content of BAP is 0.1-5mg/L in the first fluid nutrient medium in the present invention, if the content of BAP is less than lower limit value, leaf Former base is not grown, therefore can only obtain individual plants；If the content of BAP is more than upper limit value, the formation of phyllopodium can be inhibited, from And leaf bud cannot be obtained.

After beautiful millettia root seedling stem-tip tissue in the present invention is first grown to a certain degree in solid medium, transfer to Fluid nutrient medium, if carrying out shaken cultivation in liquid medium first, plant tissue can float the table of liquid medium within On face, and impingement vessel wall leads to death in oscillation.

Compared with solid medium, the speed of growth of plant tissue in liquid medium faster, and is studied through the present invention It was found that first being cultivated in the fluid nutrient medium containing the basic element of cell division after a certain period of time, transfer to without the basic element of cell division It is cultivated in fluid nutrient medium, rooting rate can be effectively improved.

Specific embodiment