CN107779409A - A kind of sweet potato scurf bacterium fast breeding culture medium - Google Patents
A kind of sweet potato scurf bacterium fast breeding culture medium Download PDFInfo
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- CN107779409A CN107779409A CN201711169493.1A CN201711169493A CN107779409A CN 107779409 A CN107779409 A CN 107779409A CN 201711169493 A CN201711169493 A CN 201711169493A CN 107779409 A CN107779409 A CN 107779409A
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The invention discloses a kind of fluid nutrient medium for promoting sweet potato scurf bacterium fast breeding, belong to the culture medium technical field culture medium and be named as potato yeast culture medium i.e. PDY（Potato Dextrose Yeast Medium, PDY）, by the way that a certain amount of dusty yeast is added into potato fluid nutrient medium（Potato Dextrose Broth Medium, PDB）In, it is prepared through autoclave sterilization.Its each component and dosage are：The g of peeled potatoes 200, glucose or the g of sucrose 20, the g of dusty yeast 2, the L of pure water 1.The culture medium prepares quick, economical, conveniently, the growth rate of its sweet potato scurf bacterium is 5.5 times of PDB culture mediums, the characteristics of with sweet potato scurf bacterium growth rate is significantly improved, the rapid, high volume breeding of black mole germ can be achieved, enough germ amounts can be provided for indoor and field resistance appraisal in a short time, shorten germ and expand numerous time, improve operating efficiency.
Description
Technical field
The invention belongs to bioengineering field, and in particular to a kind of culture medium for promoting sweet potato scurf bacterium propagation.
Background technology
Sweet potato scurf（Sweet potato mole disease）It is by Deuteromycotina sweet potato hair chain spore
(MonilochaetesinfuscansEll. et Halst. ex Harter) caused by, have in each sweet potato producing region in China
Occur.The disease only encroaches under earth's surface that potato is climing and potato wedge, does not go deep into potato meat, on tuber yield substantially without influence, but potato wedge is fallen ill
Moisture is easily lost afterwards, in storage period easy drying shrinkage, influences quality and edibility.Especially the commodity for eating sweet potato raw is influenceed
Greatly, have a strong impact on that sweet potato eats the development of industry raw.At present, sweet potato variety tar spot Resistance Identification work has caused sweet potato to educate
Kind unit and the attention of potato seed seedling enterprise.Field resistance appraisal needs substantial amounts of black mole germ, and black mole germ is big at present
The fluid nutrient medium that amount propagation uses is PDB, but germ growth rate on the culture medium is extremely slow, is unfavorable for field and resists
The development of property appraisal.
The content of the invention
It is an object of the invention to provide a kind of culture medium for promoting sweet potato scurf bacterium propagation, effectively solves sweet potato scurf
Bacterium growth rate is slow, cultivation cycle length, restricts the problem of anti-disease enzyme work is carried out.
To solve problem above, the invention provides a kind of sweet potato scurf bacterium fast breeding culture medium-potato yeast training
Support base（Potato Dextrose Yeast Medium, PDY）.
The present invention is achieved through the following technical solutions, a kind of sweet potato scurf bacterium fast breeding culture medium, pass through by
A certain amount of dusty yeast is added in potato fluid nutrient medium, and after packing, 121 DEG C of autoclave sterilizations are prepared；
The sweet potato scurf bacterium fast breeding culture medium, its each raw material and its dosage are：The g of peeled potatoes 200, glucose
Or the g of sucrose 20, the g of dusty yeast 2, the L of pure water 1；Specific preparation method is as follows：
Step 1）Potato is cleaned into peeling, 200 g is weighed and is cut into small pieces and be put into pot, adds pure water 1L, is heated to seething with excitement, after
It is continuous to boil 20-30min；
Step 2）Filtered while hot with 2 layers of gauze, abandon filter residue, filtrate supplements pure water to 1L；
Step 3）The g of dusty yeast 2, glucose or the g of sucrose 20 are weighed, pours into the filtrate of heat, is sufficiently stirred using glass bar mixed
It is even；
Step 4）On demand, the culture medium prepared is distributed into triangular flask or test tube, after packing, in test tube mouth or three
Tampon beyond the Great Wall on the flask mouth of angle, newspaper of being double-baged outside bottleneck tampon, is bandaged with rubber band, in 121 DEG C of autoclave sterilizations
20 min are made.
Should it is an advantage of the invention that：The culture medium PDY of the present invention can improve the multiplication rate of sweet potato scurf bacterium, reduce
Expand the time needed for numerous black mole germ and workload.The culture medium preparation manipulation process is easy, easy to learn, and it is easy to prepare required raw material
Obtain and cheap, there is good economy, be adapted to large-scale production and use.
Embodiment
With reference to specific embodiment, the invention will be further described.
Embodiment 1
（1）Pathogenicbacteria separation and preparation：
From field, classical symptom potato wedge occurs for collection sweet potato scurf, is separated by tissue isolation and obtains black mole germ, and according to
Koch's Postulates tieback potato wedge is identified, and germ passes through morphology and molecular biology identification；It will separate and identification is correctly black
Mole germ activates 2 weeks in 28 DEG C on PDA plate culture medium；By cultured black mole germ flat board aseptic water washing, obtain
Spore suspension, and by spore suspension concentration dilution to 1.0 × 106Individual/ml or so.
（2）The preparation of culture medium：
PDB culture mediums：1st, potato is cleaned into peeling, weighs 200 g and be cut into small pieces and be put into pot, added water 1L, be heated to seething with excitement,
Maintain 20-30min；2nd, filtered while hot with 2 layers of gauze, abandon filter residue, filtrate supplements pure water to 1L；3rd, glucose or sucrose are weighed
20 g, pour into the filtrate of heat, mixing is sufficiently stirred using glass bar；4th, the culture medium prepared is distributed into Erlenmeyer flask
It is interior, every bottle of 50 ml, after packing, the tampon beyond the Great Wall on conical flask mouth, newspaper of being double-baged outside tampon, with rubber band bag
Tie, in 121 DEG C of min of autoclave sterilization 20.
PDY culture mediums：1st, potato is cleaned into peeling, weighs 200 g and be cut into small pieces and be put into pot, added pure water 1L, add
Heat continues to boil 20-30min to seething with excitement；2nd, filtered while hot with 2 layers of gauze, abandon filter residue, filtrate supplements pure water to 1L；3rd, claim
The g of dusty yeast 2, glucose or the g of sucrose 20 are taken, pours into the filtrate of heat, mixing is sufficiently stirred using glass bar；4th, will prepare
Culture medium be distributed into Erlenmeyer flask, every bottle of 50 ml, after packing, the cotton beyond the Great Wall on test tube mouth or conical flask mouth
Fill in (or foam plug, cap test tube etc.), newspaper of being double-baged outside tampon, bandaged with rubber band, gone out in 121 DEG C of HTHPs
The min of bacterium 20.
（3）Test method：
It is 1.0 × 10 by 50 μ L concentration in the superclean bench of ultraviolet disinfection6Individual/ml sweet potato scurf bacterium spore suspensions
It is inoculated in 50 ml above two culture mediums, 28 DEG C of 150 d of rpm isothermal vibrations culture 10, with having dried the filter paper mistake weighed
Filter, thalline is put into filter paper together after drying box 80 DEG C of drying and processings 24 h, the indoor h of moisture regain 2 to weigh and calculate thalline and done
Weight.Often handle 6 repetitions.
（4）Result of the test：
As a result it is as shown in table 1.It can be seen from Table 1 that 50 μ L sweet potato scurf bacterium spore suspensions are inoculated in 50 ml PDY
After 28 DEG C of 150 d of rpm isothermal vibrations culture 10 of culture medium, the g of dry cell weight average out to 0.3812；And with PDB under the same terms
Medium culture, its dry cell weight are only 0.0691 g.The biomass obtained using PDY medium cultures is the 5.5 of PDB culture medium
Times, both differences reach the pole level of signifiance.The characteristics of with sweet potato scurf bacterium growth rate is significantly improved, tar spot can be achieved
The rapid, high volume breeding of bacterium, can provide enough germ amounts for indoor and field resistance appraisal in a short time, shorten
Germ expands numerous time, improves operating efficiency.
The sweet potato scurf bacterium of table 1 breeds efficiency comparison on PDB and PDY culture mediums
* lowercase letter significance of difference in 0.05 level, capitalization represent the significance of difference in 0.01 level.
Claims (1)
- A kind of 1. sweet potato scurf bacterium fast breeding culture medium, it is characterised in that：By the way that a certain amount of dusty yeast is added into horse In bell potato fluid nutrient medium, after packing, 121 DEG C of autoclave sterilizations are prepared；The sweet potato scurf bacterium fast breeding culture medium, its each raw material and its dosage are：The g of peeled potatoes 200, glucose Or the g of sucrose 20, the g of dusty yeast 2, the L of pure water 1；Specific preparation method is as follows：Step 1）Potato is cleaned into peeling, 200 g is weighed and is cut into small pieces and be put into pot, adds pure water 1L, is heated to seething with excitement, after It is continuous to boil 20-30min；Step 2）Filtered while hot with 2 layers of gauze, abandon filter residue, filtrate supplements pure water to 1L；Step 3）The g of dusty yeast 2, glucose or the g of sucrose 20 are weighed, pours into the filtrate of heat, is sufficiently stirred using glass bar mixed It is even；Step 4）On demand, the culture medium prepared is distributed into triangular flask or test tube, after packing, in test tube mouth or three Tampon beyond the Great Wall on the flask mouth of angle, newspaper of being double-baged outside bottleneck tampon, is bandaged with rubber band, in 121 DEG C of autoclave sterilizations 20 min are made.
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| CN201711169493.1A CN107779409A (en) | 2017-11-22 | 2017-11-22 | A kind of sweet potato scurf bacterium fast breeding culture medium |
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Citations (6)
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| CN1600147A (en) * | 2003-09-22 | 2005-03-30 | 中国农业大学 | Immunity strengthened feedstuff additive of glycopeptide composite, preparation method and usage |
| CN101041859A (en) * | 2007-04-29 | 2007-09-26 | 上海市农业科学院食用菌研究所 | Mushroom 507 bacterial AFLP quick-detecting method |
| CN101186882A (en) * | 2007-12-20 | 2008-05-28 | 上海交通大学 | Method for separating metabolite capable of inducing cucumber resisting powdery mildew from molecule modifying trichoderma harzianum |
| CN101836597A (en) * | 2009-03-18 | 2010-09-22 | 西南科技大学 | New grey white variant strain with red pleurotus |
| CN103667378A (en) * | 2012-08-31 | 2014-03-26 | 武汉蜀泰科技有限公司 | Method for producing ARA grease rich in chrome |
| CN105557993A (en) * | 2015-12-29 | 2016-05-11 | 河北省农林科学院植物保护研究所 | Application of fludioxonil and compound thereof in preventing and treating diseases of sweet potato during storage |
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2017
- 2017-11-22 CN CN201711169493.1A patent/CN107779409A/en active Pending
Patent Citations (6)
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| CN1600147A (en) * | 2003-09-22 | 2005-03-30 | 中国农业大学 | Immunity strengthened feedstuff additive of glycopeptide composite, preparation method and usage |
| CN101041859A (en) * | 2007-04-29 | 2007-09-26 | 上海市农业科学院食用菌研究所 | Mushroom 507 bacterial AFLP quick-detecting method |
| CN101186882A (en) * | 2007-12-20 | 2008-05-28 | 上海交通大学 | Method for separating metabolite capable of inducing cucumber resisting powdery mildew from molecule modifying trichoderma harzianum |
| CN101836597A (en) * | 2009-03-18 | 2010-09-22 | 西南科技大学 | New grey white variant strain with red pleurotus |
| CN103667378A (en) * | 2012-08-31 | 2014-03-26 | 武汉蜀泰科技有限公司 | Method for producing ARA grease rich in chrome |
| CN105557993A (en) * | 2015-12-29 | 2016-05-11 | 河北省农林科学院植物保护研究所 | Application of fludioxonil and compound thereof in preventing and treating diseases of sweet potato during storage |
Non-Patent Citations (7)
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| C. O. OLADOYE等: "Biomolecular characterization, identification, enzyme activities of molds and physiological changes in sweet potatoes (Ipomea batatas) stored under controlled atmospheric conditions", 《JOURNAL OF ZHEJIANG UNIVERSITY-SCIENCE B》 * |
| 刘晓芸等: "甘薯黑痣病防治药剂的筛选", 《河南农业科学》 * |
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Application publication date: 20180309 |