CN110521594A - A kind of cultural method improving beautiful millettia root rooting rate - Google Patents
A kind of cultural method improving beautiful millettia root rooting rate Download PDFInfo
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- CN110521594A CN110521594A CN201910659510.2A CN201910659510A CN110521594A CN 110521594 A CN110521594 A CN 110521594A CN 201910659510 A CN201910659510 A CN 201910659510A CN 110521594 A CN110521594 A CN 110521594A
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- beautiful millettia
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- millettia root
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- 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
- A01H4/001—Culture apparatus for tissue culture
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- 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
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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
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- Botany (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Cultivation Of Plants (AREA)
Abstract
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.
Description
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
Embodiment 1
The present embodiment beautiful millettia root cultural method 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, 0.5mg/L BAP, 20g/L sucrose and 5g/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 by step 2, with
180rpm carries out obtaining within shaken cultivation 28 days leaf bud, and first fluid nutrient medium is BAP containing 3mg/L and 10g/L sucrose
MS culture medium, the pH of the first fluid nutrient medium are 5.8；
Leaf bud obtained in step 2 is transferred to the 1000ml second liquid culture medium filled without the basic element of cell division by step 3
1800ml conical flask in, cultivated 21 days under conditions of stirring and being passed through air, make leaf bud regeneration and in temperature
25 DEG C, humidity 95%, photon illumination is 80 μm of ol/ m 2 Root induction under conditions of/s, the second liquid culture medium contain
20g/L sucrose and 6g/L agar, the pH of second liquid culture medium are 5.8.
The rooting rate of beautiful millettia root is 97% in the present embodiment, and does not occur browning during the growth process.
Comparative example 1
This comparative example beautiful millettia root cultural method 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, 0.5mg/L BAP, 20g/L sucrose and 5g/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 by step 2, with
180rpm carries out obtaining within shaken cultivation 28 days leaf bud, and first fluid nutrient medium is BAP containing 3mg/L and 10g/L sucrose
MS culture medium, the pH of the first fluid nutrient medium are 5.8；
Leaf bud obtained in step 2 is transferred to the 1000ml second liquid culture medium filled without the basic element of cell division by step 3
1800ml conical flask in, cultivated 21 days under conditions of stirring and being passed through air, make leaf bud regeneration and in temperature
25 DEG C, humidity 95%, photon illumination is 80 μm of ol/ m 2 Root induction under conditions of/s, the second liquid culture medium contain
20g/L sucrose, 0.5mg/L BAP and 6g/L agar, the pH of second liquid culture medium are 5.8.
The rooting rate of beautiful millettia root is 75% in the present embodiment, after joined BAP in second liquid culture medium, the life of beautiful millettia root
The decline of root rate.
Comparative example 2
This comparative example beautiful millettia root cultural method 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, 0.5mg/L BAP, 20g/L sucrose and 5g/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 by step 2, with
180rpm carries out obtaining within shaken cultivation 28 days leaf bud, and first fluid nutrient medium is the MS culture medium of the sucrose containing 10g/L, the
The pH of one fluid nutrient medium is 5.8；
Leaf bud obtained in step 2 is transferred to the 1000ml second liquid culture medium filled without the basic element of cell division by step 3
1800ml conical flask in, cultivated 21 days under conditions of stirring and being passed through air, make leaf bud regeneration and in temperature
25 DEG C, humidity 95%, photon illumination is 80 μm of ol/ m 2 Root induction under conditions of/s, the second liquid culture medium contain
20g/L sucrose and 6g/L agar, the pH of second liquid culture medium are 5.8.
The rooting rate of beautiful millettia root is 60% in the present embodiment, is free of BAP, the rooting rate of beautiful millettia root in the first fluid nutrient medium
Decline.
Comparative example 3
This comparative example beautiful millettia root cultural method 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, 0.5mg/L BAP, 20g/L sucrose and 5g/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 by step 2, with
180rpm carries out obtaining within shaken cultivation 28 days leaf bud, and first fluid nutrient medium is BAP containing 6mg/L and 10g/L sucrose
MS culture medium, the pH of the first fluid nutrient medium are 5.8；
Leaf bud obtained in step 2 is transferred to the 1000ml second liquid culture medium filled without the basic element of cell division by step 3
1800ml conical flask in, cultivated 21 days under conditions of stirring and being passed through air, make leaf bud regeneration and in temperature
25 DEG C, humidity 95%, photon illumination is 80 μm of ol/ m 2 Root induction under conditions of/s, the second liquid culture medium contain
20g/L sucrose and 6g/L agar, the pH of second liquid culture medium are 5.8.
The rooting rate of beautiful millettia root is 45% in the present embodiment, BAP too high levels in the first fluid nutrient medium, the life of beautiful millettia root
The decline of root rate.
Comparative example 4
This comparative example beautiful millettia root cultural method 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 the first solid medium, described
First solid medium contains MS salt, 0.5mg/L BAP, 20g/L sucrose and 5g/L agar, and the pH of the first solid medium is
5.8；
Tissue obtained in step 1 is transferred in the second solid medium containing the basic element of cell division, described the by step 2
Two solid mediums are the MS culture medium of BAP containing 3mg/L and 10g/L sucrose, and the pH of solid medium is 5.8；It is solid second
After being cultivated 70 days in body culture medium, it is inoculated on third solid medium, third solid medium contains MS salt, 0.3
Mg/L BAP, 20g/L sucrose and 3g/L agar, the pH of the second culture medium are 5.8；
Step 3 after cultivating 28 days in third solid medium, makes leaf bud regeneration and root induction.
The rooting rate of beautiful millettia root is 70% in the present embodiment, only when being cultivated in solid medium, the rooting rate of beautiful millettia root compared with
It is low.
Claims (3)
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.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103039361A (en) * | 2013-01-11 | 2013-04-17 | 广西壮族自治区药用植物园 | Method for directly inducing cluster buds of milletia speciosa champ seeds and quickly breeding seedlings |
| CN106171985A (en) * | 2016-07-13 | 2016-12-07 | 湖南科技学院 | A kind of liquid shallow quick-breeding method of Rhizoma Zingiberis Recens |
-
2019
- 2019-07-22 CN CN201910659510.2A patent/CN110521594A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103039361A (en) * | 2013-01-11 | 2013-04-17 | 广西壮族自治区药用植物园 | Method for directly inducing cluster buds of milletia speciosa champ seeds and quickly breeding seedlings |
| CN106171985A (en) * | 2016-07-13 | 2016-12-07 | 湖南科技学院 | A kind of liquid shallow quick-breeding method of Rhizoma Zingiberis Recens |
Non-Patent Citations (3)
| Title |
|---|
| 朱建华等: "《植物组织培养使用技术》", 30 January 2002 * |
| 陈心启: "《陈心启说兰花》", 30 November 2018 * |
| 黄碧兰等: "牛大力茎段组织培养技术研究", 《安徽农业科学》 * |
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Application publication date: 20191203 |