CN102914350A - Simple plant callus subculture weight increase determination method - Google Patents
Simple plant callus subculture weight increase determination method Download PDFInfo
- Publication number
- CN102914350A CN102914350A CN2011103097927A CN201110309792A CN102914350A CN 102914350 A CN102914350 A CN 102914350A CN 2011103097927 A CN2011103097927 A CN 2011103097927A CN 201110309792 A CN201110309792 A CN 201110309792A CN 102914350 A CN102914350 A CN 102914350A
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- China
- Prior art keywords
- callus
- filter paper
- subculture
- weighing
- weight
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Links
- 206010020649 Hyperkeratosis Diseases 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title abstract description 10
- 239000001963 growth medium Substances 0.000 claims abstract description 11
- 235000015097 nutrients Nutrition 0.000 claims abstract description 9
- 238000005070 sampling Methods 0.000 claims abstract description 8
- 238000011081 inoculation Methods 0.000 claims description 4
- 239000002574 poison Substances 0.000 claims description 4
- 231100000614 poison Toxicity 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 238000004659 sterilization and disinfection Methods 0.000 claims description 4
- 238000012549 training Methods 0.000 claims description 4
- 238000005303 weighing Methods 0.000 abstract description 6
- 238000004161 plant tissue culture Methods 0.000 abstract 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract 1
- 241001075517 Abelmoschus Species 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a simple plant callus subculture weight increase determination method during plant tissue culture. The method includes: firstly, placing a piece of disinfected filter paper on a solidified culture medium, weighing, and dicing a to-be-subcultured callus and inoculating the diced callus on the filter paper prior to weighing, wherein the difference between two-time weighing results serves as initial weight; and after culture, sampling and weighing the grown callus, wherein a numerical value obtained by subtracting the initial weight from grown weight serves as increased weight during subculture. The culture medium supplies water and nutrients to the callus by the aid of a filter paper bridge, the callus is in non-sticky connection with the filter paper, is unattached to the culture medium, can be tweezed directly by tweezers and is less prone to crushing, and the method is simple, convenient, feasible, free of interference during weighing, accurate and reliable.
Description
Technical field
the invention belongs to the Plant Tissue Breeding research method, specifically a kind of determination techniques for Accurate Measurement callus subculture incubation growth increment.
Background technology
it is an important step of Plant Tissue Breeding work that the callus subculture is cultivated.In relevant research work, for the technique effect of different disposal to callus relatively, the comparison of being weighed of the weight before and after usually needing the callus subculture is cultivated.Callus growth on not only sticky but also soft nutrient culture media, base portion and nutrient culture media weave in, self organize and very tender and crisp, both easily caused the fragmentation of tissue in the process of sampling, cause the loss in weight, the residual media of easily carrying because of base portion again, cause weight to increase.The gaining effect " indeterminacy " that the existence of these situations makes the callus subculture cultivate.This difficult problem is undecided so far.Purpose of the present invention is created a kind of simple and easy to do exactly, and the callus subculture is cultivated the assay method of gaining effect accurately and reliably.
Summary of the invention
1. pad filter paper: at first at the poison that disappeared, on the nutrient culture media solidified, the pad lastblock, through the filter paper of sterilization, is weighed, and is recorded as G
a
.
2. inoculation is weighed: the callus stripping and slicing that will want the subculture cultivation is seeded in above filter paper, weighs, and is recorded as G
b
, G
b
with G
a
the difference initial weight that is callus.
3. cultivate: cover the bottle cap of tissue culture bottle, be placed in group training chamber and cultivate.
4. sampling is weighed: after having cultivated, from tissue culture bottle, the callus on filter paper is taken out and weighs with tweezers, be recorded as G
c
.
5. in Subculture, callus growth weightening finish (G) is calculated:
G=Ｇ
c
-G
b
-G
a
specific embodiment of the invention requires: the 1-3 link of operating process must strictly observe the sterile working rules of Plant Tissue Breeding.
creativeness of the present invention is:
1. utilize the bibulous characteristics of filter paper, by its pad, below the callus of wish cultivation, filter paper can successfully be drawn enough moisture and nutrition supply callus growth from nutrient culture media.
callus is grown on filter paper, with the nutrient culture media indirect association, each other, without being infected with, without adhesion; Between callus and filter paper, smoothly contact, also without adhesion, as long as directly take out with tweezers, very smooth and complete during sampling, without any carrying, also be difficult for broken.
purposes of the present invention:
the invention solves during the callus subculture cultivates, sampling is easily broken, and imperfect or portable foreign material disturb the problem of weighing results, for the research work of this respect provides a kind of simple and easy, accurate, reliable assay method.
The accompanying drawing explanation
fig. 1 is that the arrowleaf abelmoschus callus just has been seeded in the tissue culture bottle on the filter paper bridge.
fig. 2 is that the arrowleaf abelmoschus callus completes the tissue culture bottle that subculture is cultivated on the filter paper bridge.
Embodiment
below by specific embodiment, the present invention will be further described.
embodiment 1: arrowleaf abelmoschus callus subculture is cultivated weightening finish and is measured:
1. pad filter paper: at first at the poison that disappeared, on the nutrient culture media solidified, the pad lastblock, through the filter paper of sterilization, is weighed, and is recorded as G
a
.
2. inoculation is weighed: the arrowleaf abelmoschus callus stripping and slicing that will want the subculture cultivation is seeded in above filter paper, weighs, and is recorded as G
b
, G
b
with G
a
the difference initial weight that is callus.
3. cultivate: cover the bottle cap of tissue culture bottle, be placed in group training chamber and cultivate 15 days.
4. sampling is weighed: after having cultivated, from tissue culture bottle, the callus on filter paper is taken out and weighs with tweezers, be recorded as G
c
.
5. in Subculture, callus growth weightening finish (G) is calculated:
G=Ｇ
c
-G
b
-G
a
embodiment 2: the Rice Callus subculture is cultivated weightening finish and is measured:
1. pad filter paper: at first at the poison that disappeared, on the nutrient culture media solidified, the pad lastblock, through the filter paper of sterilization, is weighed, and is recorded as G
a
.
2. inoculation is weighed: the Rice Callus stripping and slicing that will want the subculture cultivation is seeded in above filter paper, weighs, and is recorded as G
b
, G
b
with G
a
the difference initial weight that is callus.
3. cultivate: cover the bottle cap of tissue culture bottle, be placed in group training chamber and cultivate 18 days.
4. sampling is weighed: after having cultivated, from tissue culture bottle, the callus on filter paper is taken out and weighs with tweezers, be recorded as G
c
.
5. in Subculture, callus growth weightening finish (G) is calculated:
G=Ｇ
c
-G
b
-G
a
the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention.For a person skilled in the art, the present invention can have change and change.Within the spirit and principles in the present invention all, any modification of doing, improvement etc., within all should being included in protection scope of the present invention.
Claims (5)
1. pad filter paper: at first at the poison that disappeared, on the nutrient culture media solidified, the pad lastblock, through the filter paper of sterilization, is weighed, and is recorded as G
a.
2. inoculation is weighed: the callus stripping and slicing that will want the subculture cultivation is seeded in above filter paper, weighs, and is recorded as G
b, G
bwith G
athe difference initial weight that is callus.
3. cultivate: cover the bottle cap of tissue culture bottle, be placed in group training chamber and cultivate.
4. sampling is weighed: after having cultivated, from tissue culture bottle, the callus on filter paper is taken out and weighs with tweezers, be recorded as G
c.
5. in Subculture, callus growth weightening finish (G) is calculated:
G=G
c-G
b-G
a 。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103097927A CN102914350A (en) | 2011-10-13 | 2011-10-13 | Simple plant callus subculture weight increase determination method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103097927A CN102914350A (en) | 2011-10-13 | 2011-10-13 | Simple plant callus subculture weight increase determination method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102914350A true CN102914350A (en) | 2013-02-06 |
Family
ID=47612821
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011103097927A Pending CN102914350A (en) | 2011-10-13 | 2011-10-13 | Simple plant callus subculture weight increase determination method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102914350A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1631101A (en) * | 2003-12-22 | 2005-06-29 | 刘文革 | Excised mutagenesis tetraploid method of water melon and ploidy early stage certification technique |
| KR100795708B1 (en) * | 2006-12-26 | 2008-01-17 | 주식회사 알앤엘바이오 | Isolation and Culture Method of Adult Stem Cells Derived from Human Amniotic Epithelial Cells |
| CN101126085A (en) * | 2007-05-22 | 2008-02-20 | 上海光兆植物速生技术有限公司 | Plant cell space embarkation method |
| CN102301957A (en) * | 2011-07-26 | 2012-01-04 | 四川农业大学 | Callus culture method by taking Rhizoma Paridis roots and stems as explants |
-
2011
- 2011-10-13 CN CN2011103097927A patent/CN102914350A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1631101A (en) * | 2003-12-22 | 2005-06-29 | 刘文革 | Excised mutagenesis tetraploid method of water melon and ploidy early stage certification technique |
| KR100795708B1 (en) * | 2006-12-26 | 2008-01-17 | 주식회사 알앤엘바이오 | Isolation and Culture Method of Adult Stem Cells Derived from Human Amniotic Epithelial Cells |
| CN101126085A (en) * | 2007-05-22 | 2008-02-20 | 上海光兆植物速生技术有限公司 | Plant cell space embarkation method |
| CN102301957A (en) * | 2011-07-26 | 2012-01-04 | 四川农业大学 | Callus culture method by taking Rhizoma Paridis roots and stems as explants |
Non-Patent Citations (1)
| Title |
|---|
| 曹景林等: "棉花高效体细胞胚发生及同步控制培养体系研究", 《作物学报》 * |
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Application publication date: 20130206 |