CN100445372C - Differentiation culture medium adapted for indica rice isolated culture - Google Patents
Differentiation culture medium adapted for indica rice isolated culture Download PDFInfo
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- CN100445372C CN100445372C CNB2006101417027A CN200610141702A CN100445372C CN 100445372 C CN100445372 C CN 100445372C CN B2006101417027 A CNB2006101417027 A CN B2006101417027A CN 200610141702 A CN200610141702 A CN 200610141702A CN 100445372 C CN100445372 C CN 100445372C
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Abstract
The invention discloses a differentiation culture medium preparing method and application of rice, which comprises the following parts: 2800-3000mg/L KNO3,350-450mg/L (NH4)2SO4,300-400mg/L KH2PO4, 180-200mg/L MgSO4 .7H2, 170-200mg/L CaCl2 .2H2, 45-55mg/L FeSO4 .7H2O, Na2EDTA 60-74mg/, 80-100mg/L MnSO4 .4H2, 15-25mg/L ZnSO4 .7H2O, 25-30mg/L H3BO,6.0-8.0mg/L KI, 0.2-0.3mg/L CuSO4 .5H2O,2.0-3.0mg/L Na2MoO4 .2H2O, 0.2-0.3mg/L CoCl2 .6H2O,2.0-3.0mg/L aminoacetic acid, 0.5-1.0mg/L VB1, 1.0-1.5mg/L VB6,1.0-1.5mg/L niacin,100-150mg/L inositol,300-500mg/L glutamine,500-800mg/L proline,30-50g/L maltose,2mg/L 6-aminopurine benzyl,2mg/L activator,0.2mg/L heteroauxin,0.2mg/L 1-naphthalene acetic acid,5g/L vegetable glue with pH value at 6.0.
Description
The application is that the application number of submitting on June 24th, 2004 is dividing an application of 200410013345.7 patent applications.
Technical field
The invention belongs to the new plant technical field.Be specifically related to a kind of isolated culture method of long-grained nonglutinous rice, more particularly, relate to the improvement and the application in long-grained nonglutinous rice isolated culture and transgenosis thereof of long-grained nonglutinous rice callus subculture and division culture medium.
Background technology
Substratum is the key of plant isolated culture method, and the foundation of any plant isolated culture method all mainly is to carry out around the screening of substratum.The plant isolated culture is the process of indoors artificial simulating plant self-sow, different plant species has different growth conditionss and environment, therefore, can imagine, designing a kind of substratum that is fit to all species tissue culture is impossible realize that every kind of substratum all has its suitableeest species even genotype scope.As everyone knows, several classical substratum of widespread use at present is because its component concentration difference.Its effect and effect are also inequality.B
5(Gamborg OL etc., 1968.Nutrient requirements of suspension culture of soybean roots cells.Exp Cell Res 50:150-158) mainly are applicable to the isolated culture of cress to substratum; N
6(.1975.Establishment of an efficientmedium for anther culture of rice through comparative experiments on the nitrogen sources.ScientiaSinica such as Chu CC 18:659-668) mainly is suitable for the tissue culture of japonica rice to substratum; MS substratum (Murashige T, Skoog is medium for rapid growth and bioassays with tobacco cultures.Plant Physiol F.1962.Arevised, 15:473-493) makes the isolated culture of plants of Solanaceae such as tobacco, tomato, potato become very easy and convenience.These substratum are called as the substratum commonly used of plant tissue culture and are used till today.
The composition of above-mentioned these classical substratum generally is made up of minimum medium and supplementary component, and wherein minimum medium mainly comprises inorganic salt (macroelement and trace element), organotrophy compositions such as VITAMIN, glycine and inositol, carbon source etc.Plant hormone and organic additive then are to adjust with different cultivation periods according to different culturing purposes, different Objects of Development.The substratum that the part composition of minimum medium is adjusted is called the minimum medium of improvement, as MS, the B of improvement of improvement
5Substratum or the like.
So far, in rice variety isolated culture process, behind not anti-subculture of callus and the subculture callus be difficult to the differentiation be still very distinct issues, this also is the major cause that long-grained nonglutinous rice becomes the difficult species of genetically modified something lost.Because be used for any maturation method of plant transgene now all is to be based upon on the plant isolated culture method basis, and needs the screening and the subculture process of the kanamycin-resistant callus tissue of a long period.At present, most all is to adopt MS or N about callus subculture in the rice variety isolated culture process and the test of differentiation report
6Substratum carries out, but uses this class substratum effect very can not be satisfactory.Subject matter be existing subculture callus growth speed very slow, and brownization of callus is serious, succeeding transfer culture continuously, thereby cause the whole test failure.Therefore, screening is fit to the substratum of long-grained nonglutinous rice callus subculture and differentiation, thereby the isolated culture method of setting up a kind of rice variety is significant for the tissue culture of long-grained nonglutinous rice and transgenic research and application thereof.
Summary of the invention
The objective of the invention is to set up a kind of method of long-grained nonglutinous rice isolated culture, screening is suitable for callus subculture and the division culture medium that the long-grained nonglutinous rice isolated culture is used, this new cultural method and substratum can be adapted to long-grained nonglutinous rice tissue culture and genetically modified application under the isolated condition widely.
The present invention is achieved through the following technical solutions:
A kind of method of long-grained nonglutinous rice isolated culture, its step comprises:
1) with the long-grained nonglutinous rice explant of the mature seed, rataria or the flower pesticide that shell, place 75% alcohol to clean 1 minute, 0.15% mercuric chloride soaked 10-15 minute, rinsed with sterile water inserts MS+2, on the substratum of 4-D 2.5mg/L+ glutamine 300mg/L+ proline(Pro) 500mg/L+3-5% sucrose+0.3% vegetable jelly (Phytagel), the pH of this substratum is 5.9, culture condition is 26 ± 1 ℃, and dark the cultivation goes out callus with described explant induction;
2) the resulting callus of step 1) is inserted subculture medium (being numbered J3) and cultivate, the component of this substratum is: KNO
31900-2000mg/L, NH
4NO
31500-1650mg/L, KH
2PO
4150-170mg/L, MgSO
47H
2O350-370mg/L, CaCl
22H
2O 350-400mg/L, FeSO47H
2O 35-42mg/L, Na
2EDTA 50-56mg/L, MnSO
44H
2O 80-100mg/L, ZnSO
47H
2O 15-25mg/L, H
3BO
325-30mg/L, KI 6.0-8.0mg/L, CuSO
45H
2O 0.2-0.3mg/L, Na
2MoO
42H
2O 2.0-3.0mg/L, CoCl
26H
2O 0.2-0.3mg/L, glycine 2.0-3.0mg/L, V
B10.5-1.0mg/L, V
B61.0-1.5mg/L, nicotinic acid 1.0-1.5mg/L, inositol 100-150mg/L, glutamine 300-500mg/L, proline(Pro) 500-800mg/L, 2,4-D 2.0-3.0mg/L, maltose 30g/L, vegetable jelly 3.0g/L, pH is 5.9, and culture condition is 26 ± 1 ℃, the dark cultivation, every 20 days subcultures once;
3) get step 2) resulting subculture callus inserts division culture medium and (is numbered DL
3), the component of this substratum is as follows: KNO
32800-3000mg/L, (NH
4)
2SO
4350-450mg/L, KH
2PO
4300-400mg/L, MgSO
47H
2O180-200mg/L, CaCl
22H
2O 170-200mg/L, FeSO47H
2O 45-55mg/L, 60-74mg/L Na
2EDTA, MnSO
44H
2O 80-100mg/L, ZnSO
47H
2O 15-25mg/L, H
3BO
325-30mg/L, KI 6.0-8.0mg/L, CuSO
45H
2O 0.2-0.3mg/L, Na
2MoO
42H
2O 2.0-3.0mg/L, CoCl
26H
2O 0.2-0.3mg/L, glycine 2.0-3.0mg/L, V
B10.5-1.0mg/L, V
B61.0-1.5mg/L, nicotinic acid 1.0-1.5mg/L, inositol 100-150mg/L, glutamine 300-500mg/L, proline(Pro) 500-800mg/L, maltose 30-50g/L, the fast cry of certain animals of 6-benzylamino 2mg/L, kinetin 2mg/L, indolylacetic acid 0.2mg/L, naphthylacetic acid 0.2mg/L, vegetable jelly 5g/L, pH are 6.0, and culture condition is 26 ± 1 ℃, illumination cultivation (16 little time/8 hour dark), plant regenerates.
A kind of long-grained nonglutinous rice callus subculture medium that is used for isolated culture (is numbered J
3), its component is as follows:
KNO
31900-2000mg/L, NH
4NO
31500-1650mg/L, KH
2PO
4150-170mg/L, MgSO
47H
2O350-370mg/L, CaCl
22H
2O 350-400mg/L, FeSO47H
2O 35-42mg/L, Na
2EDTA 50-56mg/L, MnSO
44H
2O 80-100mg/L, ZnSO
47H
2O 15-25mg/L, H
3BO
325-30mg/L, KI 6.0-8.0mg/L, CuSO
45H
2O 0.2-0.3mg/L, Na
2MoO
42H
2O 2.0-3.0mg/L, CoCl
26H
2O 0.2-0.3mg/L, glycine 2.0-3.0mg/L, V
B10.5-1.0mg/L, V
B61.0-1.5mg/L, nicotinic acid 1.0-1.5mg/L, inositol 100-150mg/L, glutamine 300-500mg/L, proline(Pro) 500-800mg/L, 2,4-D 2.0-3.0mg/L, maltose 30g/L, vegetable jelly 3.0g/L.
A kind of long-grained nonglutinous rice callus division culture medium that is used for isolated culture (is numbered DL
3), its component is as follows:
KNO
32800-3000mg/L, (NH
4)
2SO
4350-450mg/L, KH
2PO
4300-400mg/L, MgSO
47H
2O180-200mg/L, CaCl
22H
2O 170-200mg/L, FeSO47H
2O 45-55mg/L, 60-74mg/L Na
2EDTA, MnSO
44H
2O 80-100mg/L, ZnSO
47H
2O 15-25mg/L, H
3BO
325-30mg/L, KI 6.0-8.0mg/L, CuSO
45H
2O 0.2-0.3mg/L, Na
2MoO
42H
2O 2.0-3.0mg/L, CoCl
26H
2O 0.2-0.3mg/L, glycine 2.0-3.0mg/L, V
B10.5-1.0mg/L, V
B61.0-1.5mg/L, nicotinic acid 1.0-1.5mg/L, inositol 100-150mg/L, glutamine 300-500mg/L, proline(Pro) 500-800mg/L, maltose 30-50g/L, 6-benzylaminopurine 2mg/L, kinetin 2mg/L, indolylacetic acid 0.2mg/L, naphthylacetic acid 0.2mg/L, vegetable jelly 5g/L.
Method of the present invention can be applicable on long-grained nonglutinous rice tissue culture or the transgenosis.
Long-grained nonglutinous rice callus subculture medium of the present invention and/or division culture medium can be applicable on long-grained nonglutinous rice tissue culture or the transgenosis.
The invention has the advantages that:
Utilize production upward extensively to plant, four rice varieties such as bright extensive 63 (a kind of long-grained nonglutinous rice three is to recover system) that genotypic difference is bigger, Zhenshan 97B (a kind of long-grained nonglutinous rice three is a maintenance line), in 419 (a kind of long-grained nonglutinous rices three be or two be to recover system) and W9864S (a kind of long-grained nonglutinous rice two-line sterile line) be test materials, do parallel simultaneous test with existing classical substratum, long-grained nonglutinous rice callus subculture medium that screening obtains and division culture medium and the long-grained nonglutinous rice isolated culture method of setting up thus, have specific aim and suitability widely, can be used for tissue culture and the genetically modified research and the application of most of rice varieties.
Embodiment
Embodiment 1: the screening of long-grained nonglutinous rice callus subculture medium
Test design is divided into two portions: 1, with MS, N
6, B
5And N
6(this substratum is N to B
6A large amount of nutritive ingredients and B
5Trace and organotrophy composition) four kinds of the most frequently used minimum mediums (ginseng is shown in Table 5) are the basis, 1 times (representing with X, down together), 2X are set, are combined into 16 kinds of substratum with sucrose, two kinds of carbon sources of maltose: 2, based on the MS macronutrient, maltose is carbon source, and MS, N are set
6, B
5Trace element, every kind of trace element is established 1X, 2X, four water of 5X, 10X half, Fe
2+If 1X, 1.5X, four gradients of 3X, 5X are combined into 48 kinds of substratum.Totally 64 kinds of substratum of design are inoculated the long-grained nonglutinous rice callus respectively carry out subculture, every kind of substratum connects 5 bottles of callus respectively.26 ± 1 ℃ of dark down cultivations 20 days.By observing the callus Growth situation (speed of growth, quality and color) on the more various substratum, filter out the subculture medium that is fit to the stripped callus of long-grained nonglutinous rice and (be numbered J
3).The component of this substratum is as follows: KNO
31900-2000mg/L, NH
4NO
31500-1650mg/L, KH
2PO
4150-170mg/L, MgSO
47H
2O 350-370mg/L, CaCl
22H
2O350-400mg/L, FeSO47H
2O 35-42mg/L, Na
2EDTA 50-56mg/L, MnSO
44H
2O 80-100mg/L, ZnSO
47H
2O 15-25mg/L, H
3BO
325-30mg/L, KI 6.0-8.0mg/L, CuSO
45H
2O 0.2-0.3mg/L, Na
2MoO
42H
2O 2.0-3.0mg/L, CoCl
26H
2O 0.2-0.3mg/L, glycine 2.0-3.0mg/L, V
B10.5-1.0mg/L, V
B61.0-1.5mg/L, nicotinic acid 1.0-1.5mg/L, inositol 100-150mg/L, glutamine 300-500mg/L, proline(Pro) 500-800mg/L, 2,4-D 2.0-3.0mg/L, maltose 30g/L, vegetable jelly 3.0g/L, pH are 5.9.
Embodiment 2: the comparison test of long-grained nonglutinous rice callus subculture effect
Test is adopted China to produce at present and is gone up extensively plantation, genotypic difference is bigger and four representative rice varieties: bright extensive 63 (a kind of long-grained nonglutinous rice three is to recover system), Zhenshan 97B (a kind of long-grained nonglutinous rice three is a maintenance line), in 419 (a kind of long-grained nonglutinous rices three be or two be to recover system) and W9864S (a kind of long-grained nonglutinous rice two-line sterile line).With J of the present invention
3Substratum and other three kinds (are numbered J by minimum medium MS commonly used
1), N
6(be numbered J
2) and B
5(be numbered J
4) subculture medium formed, supplementary component that adds in these substratum and plant hormone be as 2, and 4-D, proline(Pro), glutamine, maltose are identical with the vegetable jelly consumption, and difference only is the composition and the consumption of minimum medium, and pH is 5.9.Do the comparison test of substratum performance.
The test design of different long-grained nonglutinous rice subculture mediums:
Numbering J
1: the inorganic salt of MS, trace element and organic composition (down together)+2,4-D2.5mg/L+ proline(Pro) 500mg/L+ glutamine 300mg/L+3% maltose (carbon source)+0.3% vegetable jelly, pH5.9;
Numbering J
2: N
6Inorganic salt, trace element and organic composition (down with)+2,4-D 2.5mg/L+ proline(Pro) 500mg/L+ glutamine 300mg/L+3% maltose+0.3% vegetable jelly, pH5.9;
Numbering J
3: J
3Inorganic salt, trace element and organic composition (down together)+2,4-D 2.5mg/L+ proline(Pro) 500mg/L+ glutamine 300mg/L+3% maltose+0.3% vegetable jelly, pH5.9;
Numbering J
4: B
5Inorganic salt, trace element and organic composition (down with)+2,4-D 2.5mg/L+ proline(Pro) 500mg/L+ glutamine 300mg/L+3% maltose+0.3% vegetable jelly, pH5.9.
In this test, the callus of four rice varieties that are used to test comes from the fresh callus of laboratory inoculation inducing culture about 30 days, to insert respectively after these callus weighings on four kinds and the solid subculture medium (adding agar in advance) by constant by standard sterilizing program preparation, each 5 bottles of the callus of every kind of culture medium inoculated experimental cultivar place 26 ℃ of dark cultivations immediately after the inoculation.Cultivate after 20 days the weight of weighing callus (in fresh weight), the rate of body weight gain of the callus under the different cultivations of calculating.Test-results is shown in Table 1.
The different substratum of table 1 are to the effect of long-grained nonglutinous rice callus succeeding transfer culture
Annotate: callus fresh weight and cultivate the weight in average that callus fresh weight after 20 days is 5 bottles of callus fresh weights of every kind during the inoculation added up in the table.
From the test and The results of analysis of variance as can be seen: the design four kinds of substratum, substratum J of the present invention
3Subculture effect to four rice variety callus is better than its excess-three kind substratum greatly; And J
3The quality cadmium yellow consolidation of the callus of substratum subculture presents very strong embryo sexual state, helps further cultivation.
Embodiment 3: long-grained nonglutinous rice callus subculture medium J
3Effect test to the continuous subculture of long-grained nonglutinous rice callus
By what embodiment provided above-mentioned four kinds of callus for the examination rice variety are inserted described J
3Substratum carries out continuous succeeding transfer culture test, and the time of each succeeding transfer culture is 20 days.Behind each subculture, weighing is inoculated each experimental cultivar callus and is done the differentiation vitality test for each 5 bottles, to estimate J of the present invention
3Substratum is to the hold facility of continuous subculture callus embryo.Test effect is shown in Table 2.
Table 2 long-grained nonglutinous rice callus subculture medium J
3Long-grained nonglutinous rice differentiation of calli vitality test to continuous subculture
Annotate: the gross weight of five bottles of callus of a inoculation: the total differentiated plants number of five bottles of callus of b
The result shows, subculture medium of the present invention, though along with the increase of subculture number, the differentiation vigor of callus is on a declining curve.But lowering speed is slow.Each callus for the examination material still keeps higher differentiation vigor behind the subculture three times.Can satisfy the requirement that transforms callus screening subculture fully,
Embodiment 4: the screening of long-grained nonglutinous rice division culture medium and comparison test
The processing of test materials and method and callus is carried out according to the step of embodiment 1, by observing the differentiation of calli situation (differentiation speed and differentiation seedling number) on the more various substratum, the callus division culture medium (DL that filters out
3) composition as follows:
KNO
32800-3000mg/L, (NH
4)
2SO
4350-450mg/L, KH
2PO
4300-400mg/L, MgSO
47H
2O180-200mg/L, CaCl
22H
2O 170-200mg/L, FeSO47H
2O 45-55mg/L, 60-74mg/L Na
2EDTA, MnSO
44H
2O 80-100mg/L, ZnSO
47H
2O 15-25mg/L, H
3BO
325-30mg/L, KI 6.0-8.0mg/L, CuSO
45H
2O 0.2-0.3mg/L, Na
2MoO
42H
2O 2.0-3.0mg/L, CoCl
26H
2O 0.2-0.3mg/L, glycine 2.0-3.0mg/L, V
B10.5-1.0mg/L, V
B61.0-1.5mg/L, nicotinic acid 1.0-1.5mg/L, inositol 100-150mg/L, glutamine 300-500mg/L, proline(Pro) 500-800mg/L, maltose 30-50g/L, 6-benzylaminopurine 2mg/L, kinetin 2mg/L, indolylacetic acid 0.2mg/L, naphthylacetic acid 0.2mg/L, vegetable jelly 5g/L; PH is 6.0.
With substratum DL of the present invention
3Basic nutrition component (comprising a large amount of compositions, trace ingredients and VITAMIN) and MS, N
6Minimum medium, other adds plant hormone: 6-benzylaminopurine 2mg/L, kinetin 2mg/L, indolylacetic acid (IAA) 0.2mg/L, naphthylacetic acid (NAA) 0.2mg/L, amino acid: proline(Pro) 500mg/L, glutamine 500mg/L, the substratum that different carbon sources with two kinds are formed carry out the differentiation efficiency test.The results are shown in Table 3.
The test design of different long-grained nonglutinous rice division culture mediums:
Numbering DL
1:: MS, 2.0mg/l 6-benzylaminopurine, the 2.0mg/l kinetin, the 0.2mg/l naphthylacetic acid, the 0.2mg/l indolylacetic acid, the 500mg/l proline(Pro), the 500mg/l glutamine, 5% maltose, 0.3% vegetable jelly, pH 6.0;
Numbering DL
2:: MS, 2.0mg/l 6-benzylaminopurine, the 2.0mg/l kinetin, the 0.2mg/l naphthylacetic acid, the 0.2mg/l indolylacetic acid, the 500mg/l proline(Pro), the 500mg/l glutamine, 5% sucrose, 0.3% vegetable jelly, pH 6.0;
Numbering DL
3: DL
3Minimum medium, 2.0mg/l 6-benzylaminopurine, the 2.0mg/l kinetin, the 0.2mg/l naphthylacetic acid, the 0.2mg/l indolylacetic acid, the 500mg/l proline(Pro), the 500mg/l glutamine, 5% maltose, 0.3% vegetable jelly, pH 6.0;
Numbering DL
4:: DL
3Minimum medium, 2.0mg/l 6-benzylaminopurine, the 2.0mg/l kinetin, the 0.2mg/l naphthylacetic acid, the 0.2mg/l indolylacetic acid, the 500mg/l proline(Pro), the 500mg/l glutamine, 5% sucrose, 0.3% vegetable jelly, pH 6.0;
Numbering DL
5:: N
6, 2.0mg/l 6-benzylaminopurine, the 2.0mg/l kinetin, the 0.2mg/l naphthylacetic acid, the 0.2mg/l indolylacetic acid, the 500mg/l proline(Pro), the 500mg/l glutamine, 5% maltose, 0.3% vegetable jelly, pH 6.0;
Numbering DL
6:: N
6, 2.0mg/l 6-benzylaminopurine, the 2.0mg/l kinetin, the 0.2mg/l naphthylacetic acid, the 0.2mg/l indolylacetic acid, the 500mg/l proline(Pro), the 500mg/l glutamine, 5% sucrose, 0.3% vegetable jelly, pH 6.0;
Illustrate:
aThe callus fresh weight of each triangular flask inoculation.
Illustrate:
bDifferentiated plants number in each triangular flask.
The different long-grained nonglutinous rice division culture mediums of table 3 are to the influence of long-grained nonglutinous rice callus differentiation efficiency
Test-results shows, DL of the present invention
3The differentiation efficiency utmost point of substratum is significantly higher than other substratum, promptly the design six kinds of division culture mediums in, DL
3Substratum is suitable for the long-grained nonglutinous rice differentiation of calli most and cultivates.
Embodiment 5: the application of long-grained nonglutinous rice isolated culture method of the present invention on agriculture bacillus mediated transgenosis
Adopt the explant of the described rice variety of embodiments of the invention 1-4, in another part application that detailed technological step proposes on the same day according to the applicant about the technological step that patent application provided of agriculture bacillus mediated long-grained nonglutinous rice transgenic method, induce callus with long-grained nonglutinous rice inducing culture provided by the invention, then with the described J that is numbered
3Substratum carry out the callus subculture, cultivate in advance, cultivate altogether and screening and culturing through callus, on screening culture medium, grow resistant calli.(employing is numbered DL to the resistant calli that screening is obtained through pre-differentiation and differentiation culture
3Substratum), bear at last complete transfer-gen plant again.Test-results is as shown in table 4.
The application of table 4 long-grained nonglutinous rice isolated culture of the present invention method on agriculture bacillus mediated transgenosis
Table 4 shows: four rice varieties for examination all obtain higher transformation efficiency, illustrate that agriculture bacillus mediated long-grained nonglutinous rice gene transformation method of the present invention is very effective.
Table 5 MS, N
6, B
5The minimum medium of substratum and composition thereof
Annotate: prescription source MS substratum (Murashige and Skoog, 1962); B
5Substratum (Gamborg etc., 1968); N
6Substratum (Chu etc., 1975); AA (Toriyama and Hinata, 1985).
Claims (2)
1, a kind of division culture medium of long-grained nonglutinous rice isolated culture, its component is as described below: KNO
32800-3000mg/L, (NH
4)
2SO
4350-450mg/L, kH
2PO
4300-400mg/L, MgSO
47H
2O180-200mg/L, CaCl
22H
2O 170-200mg/L, FeSO
47H
2O 45-55mg/L, 60-74mg/L Na
2EDTA, MnSO
44H
2O 80-100mg/L, ZnSO
47H
2O 15-25mg/L, H
3BO
325-30mg/L, KI 6.0-8.0mg/L, CuSO
45H
2O 0.2-0.3mg/L, Na
2MoO
42H
2O 2.0-3.0mg/L, CoCl
26H
2O 0.2-0.3mg/L, glycine 2.0-3.0mg/L, V
B10.5-1.0mg/L, V
B61.0-1.5mg/L, nicotinic acid 1.0-1.5mg/L, inositol 100-150mg/L, glutamine 300-500mg/L, proline(Pro) 500-800mg/L, maltose 30-50g/L, 6-benzylaminopurine 2mg/L, kinetin 2mg/L, indolylacetic acid 0.2mg/L, naphthylacetic acid 0.2mg/L, vegetable jelly 5g/L, pH are 6.0.
2, the application of the described substratum of claim 1 in the long-grained nonglutinous rice isolated culture.
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| CNB2006101417027A CN100445372C (en) | 2004-06-24 | 2004-06-24 | Differentiation culture medium adapted for indica rice isolated culture |
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| CNB2006101417027A Expired - Fee Related CN100445372C (en) | 2004-06-24 | 2004-06-24 | Differentiation culture medium adapted for indica rice isolated culture |
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| CN103710381B (en) * | 2013-12-23 | 2015-10-28 | 中国农业大学 | Agrobacterium is utilized to obtain method and the special culture media thereof of transgenic indica type rice |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1206435A (en) * | 1996-10-22 | 1999-01-27 | 日本烟草产业株式会社 | Method for transforming indica rice |
| CN1341350A (en) * | 2001-09-30 | 2002-03-27 | 湖北大学 | Method for high-efficiency breeding rice polyploid by combination of tissue culture and chemical induction |
| WO2002057407A2 (en) * | 2001-01-17 | 2002-07-25 | Avestha Gengraine Technologies Pvt. Ltd. | Novel method for transgenic plants by transformation & regeneration of indica rice plant shoot tips |
| CN1422524A (en) * | 2001-11-26 | 2003-06-11 | 中国科学院长沙农业现代化研究所 | Method for raising anther seedling greening-rate of rice |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1206435A (en) * | 1996-10-22 | 1999-01-27 | 日本烟草产业株式会社 | Method for transforming indica rice |
| WO2002057407A2 (en) * | 2001-01-17 | 2002-07-25 | Avestha Gengraine Technologies Pvt. Ltd. | Novel method for transgenic plants by transformation & regeneration of indica rice plant shoot tips |
| CN1341350A (en) * | 2001-09-30 | 2002-03-27 | 湖北大学 | Method for high-efficiency breeding rice polyploid by combination of tissue culture and chemical induction |
| CN1422524A (en) * | 2001-11-26 | 2003-06-11 | 中国科学院长沙农业现代化研究所 | Method for raising anther seedling greening-rate of rice |
Non-Patent Citations (2)
| Title |
|---|
| 通过转PSAG12_IPT基因培育延缓叶片衰老水稻. 林拥军，曹孟良，徐才国等.植物学报，第44卷第11期. 2002 |
| 通过转PSAG12_IPT基因培育延缓叶片衰老水稻. 林拥军，曹孟良，徐才国等.植物学报，第44卷第11期. 2002 * |
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| CN1924008A (en) | 2007-03-07 |
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