CN102998198A - Device and method for measuring specific growth rate of plant in-vitro root - Google Patents
Device and method for measuring specific growth rate of plant in-vitro root Download PDFInfo
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- CN102998198A CN102998198A CN2012105132313A CN201210513231A CN102998198A CN 102998198 A CN102998198 A CN 102998198A CN 2012105132313 A CN2012105132313 A CN 2012105132313A CN 201210513231 A CN201210513231 A CN 201210513231A CN 102998198 A CN102998198 A CN 102998198A
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Abstract
The invention provides a device and method for measuring a specific growth rate of a plant in-vitro root. The device comprises a biological in-vitro root reactor, a first calibrated pipe, a siphon, a waste liquid bottle and a second calibrated pipe, which are sequentially communicated with one another, wherein one end of the siphon is communicated with the lower part of the first calibrated pipe while the other end of the siphon is inserted inside the waste liquid bottle; and the second calibrated pipe is communicated with the lower part of the waste liquid bottle. The method comprises the following steps of: inoculating the plant in-vitro root in a reaction tank with the device; measuring the volume of water discharged when the size of the in-vitro root increases per day; and calculating the specific growth rate of the plant in-vitro root according to a regression equation of the weight of the in-vitro root and the volume of the corresponding discharged water. The device and the method provided by the invention can be used for measuring specific growth rates of various plant tissues such as somatic embryo, bulbs and tubers, thus having wide utilization values.
Description
Technical field
The present invention relates to bioengineering, be specifically related to a kind of apparatus and method of measuring Vitro Plant root specific growth rate.
Background technology
China is one of the abundantest country of resources of medicinal plant in the world, but caused along with the increase of market demand the excess of resources of medicinal plant is excavated, and wild plant resource reserves and output ubiquity downtrending.Resources of medicinal plant is mainly derived from field planting at present.Field production is subjected to the impact of " favourable climate and geographical position " on the one hand, and the output and quality fluctuation is large; Bring inevitably on the other hand the problems such as pesticide residual contamination.Plant tissue cell's bioreactor culture has growth rapidly, the characteristics that are easy to control, thus can replace gradually the needs of field production satisfying the market.
Because most of medicinal plant is used as medicine with root, so excised root culture is one of focus of resources of medicinal plant regeneration.During the excised root bioreactor culture, growth rate is to weigh an important indicator of condition of culture and reactor performance.In order to understand in real time the growth rhythm of Vitro Plant root, calculate specific growth rate thereby at first will draw its growth curve, i.e. the amount of growth every day.During permeation, cell can be distributed in the fluid nutrient medium equably, so as long as take out the sub-fraction cell solution, just can calculate in proportion the density of cell in the whole reactor, thereby obtain specific growth rate every day.
Can mutual winding when excised root is grown form gradually a group in reactor, can not resemble the solution that forms homogeneous the isolated cells, not only have nutrient solution so do not contain excised root in the sample that takes out, can't calculate specific growth rate.At present the index of reaction excised root growing state mainly is average growth rate, i.e. V=(final concentration-inoculum density)/growth time, but the calculating of relevant specific growth rate yet there are no report.
Summary of the invention
Purpose of the present invention is exactly in order to provide a kind of apparatus and method of measuring Vitro Plant root specific growth rate.
In order to achieve the above object, the present invention has adopted following technical scheme: a kind of device of measuring Vitro Plant root specific growth rate comprises the excised root bio-reactor, the first graduated tube, U trap, waste liquid bottle and the second graduated tube that are linked in sequence and are communicated with; The bottom of a described siphonal end and the first graduated tube connects, and the siphonal other end inserts in the waste liquid bottle; The bottom of described the second graduated tube and waste liquid bottle connects.
The device of said determination Vitro Plant root specific growth rate wherein, is connected with the snorkel with air strainer on described the first graduated tube, be connected with the snorkel with air strainer on the described waste liquid bottle.
The device of said determination Vitro Plant root specific growth rate, wherein, the precision of described the first graduated tube is 0.1 milliliter; The precision of described the second graduated tube is 1 milliliter.
A kind of device with said determination Vitro Plant root specific growth rate is measured the method for Vitro Plant root specific growth rate, comprises the steps:
In the formula, X is final concentration, and t is incubation time, x and x
0Be inoculum density, μ is specific growth rate;
Described final concentration is to cultivate the dry weight grams of excised root when finishing and the ratio of nutrient solution volume milliliter number;
Described inoculum density is the dry weight grams of inoculation excised root and the ratio of nutrient solution volume milliliter number.
When the present invention measures the device busy of Vitro Plant root specific growth rate, be first this device to be linked in sequence on the excised root bio-reactor, again the Vitro Plant root be inoculated in the bio-reactor, cultivate.Increase along with the excised root volume, the volume that liquid in the excised root bio-reactor is increased by adventitious root arranges in the first graduated tube, when the liquid height in the first graduated tube is concordant with the U trap top, according to siphon principle, nutrient solution in the first graduated tube and the U trap can all flow in the waste liquid bottle, then, the liquid in the excised root bio-reactor is discharged in the first graduated tube with the adventitious root growth again.The amount that final excised root volume increases just equals the liquid indicated value that liquid indicated value in the second graduated tube adds the first graduated tube.Record the amount that every day, the excised root volume increased, be converted to the amount that excised root weight increases, drawn at last its growth curve, just can calculate specific growth rate according to formula.
Description of drawings
Fig. 1 is the structural representation of apparatus of the present invention;
Fig. 2 is the growth curve of radix pseudostellariae adventitious root among the embodiment;
Fig. 3 is the specific growth rate of radix pseudostellariae adventitious root among the embodiment.
Embodiment
Referring to Fig. 1, the device of the mensuration Vitro Plant root specific growth rate among the present invention comprises the excised root bio-reactor 6, the first graduated tube 2, U trap 7, waste liquid bottle 4 and the second graduated tube 3 that are linked in sequence and are communicated with; One end of U trap 7 and the bottom of the first graduated tube 2 connect, and the other end of U trap 7 inserts the bottom of waste liquid bottle 4; The second graduated tube 3 connects with the bottom of waste liquid bottle 4.Shown in the figure, 5 is the Vitro Plant root.
The present invention measures in the device of Vitro Plant root specific growth rate, all is connected with the snorkel with air strainer 1 on the first graduated tube 2 and the waste liquid bottle 4.
The present invention measures in the device of Vitro Plant root specific growth rate, and the precision of the first graduated tube is 0.1 milliliter, and what its numerical value reflected is collected liquid volume in the first graduated tube; The precision of the second graduated tube is 1 milliliter, and what its numerical value reflected is waste liquid bottle 4 interior collected liquid volumes.
The below is take the method for cultivation mensuration Vitro Plant root specific growth rate of the present invention as example further specifies of radix pseudostellariae adventitious root.
In the present embodiment, Vitro Plant root volume is 10 liters, uses this invention and has drawn the growth curve of radix pseudostellariae adventitious root in 24 days as shown in Figure 2, than growth rate as shown in Figure 3.
Concrete specific growth rate is 0.02-0.08 shown in the figure ordinate.
Claims (4)
1. a device of measuring Vitro Plant root specific growth rate is characterized in that: comprise the excised root bio-reactor, the first graduated tube, U trap, waste liquid bottle and the second graduated tube that are linked in sequence and are communicated with; The bottom of a described siphonal end and the first graduated tube connects, and the siphonal other end inserts in the waste liquid bottle; The bottom of described the second graduated tube and waste liquid bottle connects.
2. the device of mensuration Vitro Plant root specific growth rate as claimed in claim 1 is characterized in that: be connected with the snorkel with air strainer on described the first graduated tube, be connected with the snorkel with air strainer on the described waste liquid bottle.
3. the device of mensuration Vitro Plant root specific growth rate as claimed in claim 1, it is characterized in that: the precision of described the first graduated tube is 0.1 milliliter; The precision of described the second graduated tube is 1 milliliter.
4. the method with the device mensuration Vitro Plant root specific growth rate of mensuration Vitro Plant root specific growth rate claimed in claim 1 is characterized in that, comprises the steps:
Step 1 is got the Vitro Plant root of many parts of different volumes, is dipped into respectively in the graduated cylinder that nutrient solution is housed, and according to the volume of the water that arranges, draws the volume of every part of Vitro Plant root;
Step 2, at 60 ℃ of lower oven dry Vitro Plant roots, weighing gets its dry weight;
Step 3, take the volume of Vitro Plant root as horizontal ordinate, weight is the ordinate mapping, and calculates regression coefficient, i.e. the corresponding weight of Vitro Plant root of unit volume;
Step 4 is inoculated into Vitro Plant root sample in the excised root bio-reactor and cultivates, and records the volume that the Vitro Plant root arranges water every day;
Step 5 arranges the volume of water with the Vitro Plant root and regression coefficient that step 3 is calculated multiplies each other, and draws the dry weight of corresponding Vitro Plant root;
Step 6 is made growth curve chart;
Step 7 is in conjunction with growth curve chart, according to formula
Calculate specific growth rate,
In the formula, X is final concentration, and t is incubation time, x and x
0Be inoculum density, μ is specific growth rate;
Described final concentration is to cultivate the dry weight grams of excised root when finishing and the ratio of nutrient solution volume milliliter number;
Described inoculum density is the dry weight grams of inoculation excised root and the ratio of nutrient solution volume milliliter number.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210513231.3A CN102998198B (en) | 2012-12-04 | 2012-12-04 | Device and method for measuring specific growth rate of plant in-vitro root |
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| CN201210513231.3A CN102998198B (en) | 2012-12-04 | 2012-12-04 | Device and method for measuring specific growth rate of plant in-vitro root |
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| CN102998198A true CN102998198A (en) | 2013-03-27 |
| CN102998198B CN102998198B (en) | 2014-10-15 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107109336A (en) * | 2014-11-06 | 2017-08-29 | 奥林巴斯株式会社 | Cell culture system and cell culture container |
| CN107567934A (en) * | 2017-10-30 | 2018-01-12 | 聊城大学 | A kind of pruning administrative skill of large cherry tree |
Citations (4)
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| KR20060013907A (en) * | 2004-08-09 | 2006-02-14 | 대한민국(관리부서:농촌진흥청) | Method of mass-propagation of allium victorialis var. platyphyllum makino |
| US7005298B1 (en) * | 1999-08-27 | 2006-02-28 | University Of Guelph | Micropropagation and production of phytopharmaceutical plants |
| CN1788551A (en) * | 2005-12-08 | 2006-06-21 | 上海交通大学 | Plant regeneration method of vinca from hairly root |
| CN101457205A (en) * | 2009-01-08 | 2009-06-17 | 上海交通大学 | Siphoning type intermittent spraying bioreactor |
-
2012
- 2012-12-04 CN CN201210513231.3A patent/CN102998198B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7005298B1 (en) * | 1999-08-27 | 2006-02-28 | University Of Guelph | Micropropagation and production of phytopharmaceutical plants |
| KR20060013907A (en) * | 2004-08-09 | 2006-02-14 | 대한민국(관리부서:농촌진흥청) | Method of mass-propagation of allium victorialis var. platyphyllum makino |
| CN1788551A (en) * | 2005-12-08 | 2006-06-21 | 上海交通大学 | Plant regeneration method of vinca from hairly root |
| CN101457205A (en) * | 2009-01-08 | 2009-06-17 | 上海交通大学 | Siphoning type intermittent spraying bioreactor |
Non-Patent Citations (2)
| Title |
|---|
| GUIRONG WANG等: "Influence of Mist Intervals and Aerotion Rate on growth and Second Metabolite Production of Peseudostellaria heterophylla Adventitious Roots in a Siphon-mist Bioreactor", 《BIOTECHNOLOGY AND BIOPROCESS ENGINEERING》, vol. 15, no. 6, 31 December 2010 (2010-12-31), pages 1059 - 1064 * |
| 邱垂源等: "C2C12细胞体外微型灌注生物反应器培养初探", 《广东药学院学报》, vol. 23, no. 1, 28 February 2007 (2007-02-28), pages 59 - 62 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107109336A (en) * | 2014-11-06 | 2017-08-29 | 奥林巴斯株式会社 | Cell culture system and cell culture container |
| CN107567934A (en) * | 2017-10-30 | 2018-01-12 | 聊城大学 | A kind of pruning administrative skill of large cherry tree |
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| CN102998198B (en) | 2014-10-15 |
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