CN102257958A - Medium maturity virus-free potato basic seedling culture medium and preparation method thereof - Google Patents

Medium maturity virus-free potato basic seedling culture medium and preparation method thereof Download PDF

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CN102257958A
CN102257958A CN2011101258252A CN201110125825A CN102257958A CN 102257958 A CN102257958 A CN 102257958A CN 2011101258252 A CN2011101258252 A CN 2011101258252A CN 201110125825 A CN201110125825 A CN 201110125825A CN 102257958 A CN102257958 A CN 102257958A
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mother liquor
preparation
medium
culture medium
carrageen
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CN102257958B (en
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曹君迈
焦向炜
祁金涛
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North Minzu University
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Abstract

The invention relates to a medium maturity virus-free potato basic seedling culture medium and a preparation method thereof. The method is characterized by comprising the following steps: (1) preparing basic mother liquor of the culture medium: preparing macroelement mother liquor, preparing trace element mother liquor, preparing ferric salt mother liquor and preparing No.1 organic matter mother liquor; (2) weighing 5 grams of carrageen every liter of culture medium, adding 200ml of distilled water to every gram of carrageen, boiling the carrageen until the carrageen is clear and transparent and then stopping heating; (3) adding the mixed mother liquor in a beaker and a C source to the boiled carrageen; (4) adjusting the pH value to 5.8-6.0 with sodium hydroxide or hydrochloric acid; and (5) bottling the culture medium and then carrying out high-pressure sterilization. The culture medium and the preparation method have the following beneficial effects: not only are the nutrition composition and components in the culture medium reduced, but also the plantlets grow vigorously and the period is shortened since the low-cost carrageen, instead of agar, is used as a coagulant, thus effectively reducing the seedling culture energy consumption and cost.

Description

In ripe potato detoxicating basis seedling medium and preparation method thereof
Technical field
The present invention relates to ripe potato detoxicating basis seedling medium and preparation method thereof in a kind of.
Background technology
It is to produce potato detoxicating basis seedling by group culturation rapid propagating technology that China's virus-free potato primary stock industrialization is at present produced, and it is transplanted hardening again and produces micro potato.And the MS medium is the medium of the numerous extensive employing of present potato detoxicating basis seedling industrialization the expansion, and its agar consumption is too high, have in addition to 1.7%, and the agar cost is higher, and basic seedling is grown under the matrix environment really up to the mark, and crisp flexibility of stem is relatively poor, cause basic seedling growing way poor, a basis seedling and propagating generation needs about 25 days, before the transplanting hardening, takes out basic seedling from bottle, pour in the warm water, agar around the root is washed off, and the root easy damaged influenced transplanting survival rate when agar was washed root more firmly.Do not select medium for use, strengthened industrial production cost, make basic seedling growth potential relatively poor by the kind maturing stage.Increased cost of drugs in the training period and used electrical loss, and basic seedling growth potential is relatively poor, the poor quality, unfavorable to suitability for industrialized production.
Summary of the invention
One of purpose of the present invention provides ripe potato detoxicating basis seedling medium in a kind of, adopts the detoxification basis seedling of this medium with short production cycle, growth is vigorous, production cost is low.
Two of purpose of the present invention provides a kind of preparation method of ripe potato detoxicating basis seedling medium in above-mentioned.
For achieving the above object, the technical solution used in the present invention is.
Ripe potato detoxicating basis seedling medium is characterized in that in a kind of, calculates with the 1L medium and contains: the KNO of 1890～1910mg 3, 169～171mg KH 2PO 4, 369～371mg MgSO 47H 2The CaCl of O, 439～441mg 22H 2The MnSO of O, 22.27～22.23mg 44H 2The KI of O, 0.827～0.833mg, the H of 6.17～6.23mg 3BO 3, 8.57～8.63mg ZnSO 47H 2The Na of O, 0.247～0.253mg 2MoO 4H 2The CuSO of O, 0.0247～0.0253mg 45H 2The CoCl of O, 0.0247～0.0253mg 26H 2The Na of O, 37.0～37.3mg 2The FeSO of EDTA and 26～28mg 47H 2The VB of O, 1.99～2.01mg glycine, 0.09～0.11mg 1, 0.49～0.51mg VB 6, and the VB of 0.49～0.51mg 5Or VPP; Surplus is a water.
The NH that wherein also contains 402.5～422.5mg 4NO;
The C source sucrose that wherein also contains 24900～25100mg.
The preparation method of ripe potato detoxicating basis seedling medium in a kind of, its special feature is, comprises the steps:
(1) preparation of the basic mother liquor of medium
A, preparation macroelement mother liquor
Get the KNO of 18900～19100mg 3, 1690～1710mg KH 2PO 4, 3690～3710mg MgSO 47H 2The CaCl of O and 4390～4410mg 22H 2O, more than all kinds of materials fully dissolve with distilled water respectively, put in order by it and pour constant volume in the 1L volumetric flask successively into, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
B, prepare micro-mother liquor
Get the MnSO of 2227～2223mg 44H 2The KI of O, 82.7～83.3mg, the H of 617～623mg 3BO 3, 857～863mg ZnSO 47H 2The Na of O, 24.7～25.3mg 2MoO 4H 2The CuSO of O, 2.47～2.53mg 45H 2The CoCl of O and 2.47～2.53mg 26H 2O, more than all kinds of materials fully dissolve with distilled water respectively, constant volume is in the 1L volumetric flask, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
C, preparation mother liquid of iron salt
Get the Na of 370～373mg 2The FeSO of EDTA and 260～280mg 47H 2O, more than all kinds of materials fully dissolve with distilled water respectively, constant volume is in the 1L volumetric flask, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
D, No. 1 organic matter mother liquor of preparation
Get the VB of 199～201mg glycine, 9～11mg 1, 49～51mg VB 6, and the VB of 49～51mg 5Or VPP, more than all kinds of materials fully dissolve with distilled water respectively, constant volume is in the 1L volumetric flask, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
Measure 100mL macroelement mother liquor, mother liquid of iron salt respectively by every liter of medium, and 10mL trace element mother liquor and No. 1 organic matter mother liquor, it is standby to put into beaker;
(2) add 5g by every L medium and take by weighing carragheen, add 200ml distilled water, add and boil, stop heating then to as clear as crystal by every g carragheen;
(3) the mixing mother liquor in the beaker, C source are added in the well-done carragheen, add water again and make culture medium C source quality concentration reach 2.49～2.51%, the carragheen mass concentration reaches 0.5%, heats 80-121 ℃ and stirs;
(4) with sodium hydroxide or hydrochloric acid the pH value is transferred to 5.8～6.0, promptly finishes the preparation of solid culture medium;
(5) bottling back autoclaving gets final product.
Wherein the C source is a sucrose, and addition is 24900～25100mg.
The NH that wherein in the macroelement mother liquor, also adds 4025～4225mg 4NO 3
Wherein regulating the pH value is sodium hydroxide or the hydrochloric acid of using 1mol/L, splashes in the rapid prepared culture medium of previous step, stirs after dripping, and repeats this process and is transferred to 5.8～6.0 until the pH value, promptly finishes the preparation of solid culture medium.
Wherein bottling back autoclaving is the solid culture medium that will prepare, and every bottled 30～40mL seals tight bottleneck, puts into the autoclaving basket, and 121 ℃ of sterilizations are taken out after 15～19.5 minutes and got final product.
Compare with background technology, in the medium of the present invention not only nutrition composition and composition reduce, and with after carragheen replaces agar as coagulating agent cheaply, the vigorous and cycle shortening of test-tube plantlet growth, thereby effectively reduced energy consumption and cost in growing seedlings.
Embodiment
Embodiment 1:
Ripe potato test-tube plantlet medium components by weight percent raw material is produced as follows in the detoxification:
A, macroelement mother liquor (mother liquor 1) are formed: 18900～19100mg potassium nitrate (KNO 3), 4025～4225mg ammonium nitrate (NH 4NO 3), 1690～1710mg potassium dihydrogen phosphate (KH 2PO 4), 3690～3710mg magnesium sulfate (MgSO 47H 2O), 4390～4410mg calcium chloride (CaCl 22H 2O), more than all kinds of materials fully dissolve with distilled water respectively, put in order by it and pour constant volume in the 1L volumetric flask successively into, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
B, micro-mother liquor (mother liquor 2) are formed: the MnSO of 2227～2223mg 44H 2The KI of O, 82.7～83.3mg, the H of 617～623mg 3BO 3, 857～863mg ZnSO 47H 2The Na of O, 24.7～25.3mg 2MoO 4H 2The CuSO of O, 2.47～2.53mg 45H 2The CoCl of O and 2.47～2.53mg 26H 2O, more than all kinds of materials fully dissolve with distilled water respectively, constant volume is in the 1L volumetric flask, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
C, mother liquid of iron salt are formed (mother liquor 3): 370～373mg disodium ethylene diamine tetraacetate (Na 2EDTA), 260～280mg ferrous sulfate (FeSO 47H 2O), more than all kinds of materials fully dissolve with distilled water respectively, constant volume reinstalls 4 ℃ of preservations, standby in the reagent bottle that posts the concentration label in the 1L volumetric flask;
D, No. 1 organic matter mother liquor are formed (mother liquor 4): the VB of 199～201mg glycine, 9～11mg 1, 49～51mg VB 6, and the VB of 49～51mg 5Or VPP, more than all kinds of materials fully dissolve with distilled water respectively, constant volume reinstalls 4 ℃ of preservations, standby in the reagent bottle that posts the concentration label in the 1L volumetric flask;
Measure 100mL mother liquor 1, mother liquor 3 according to every liter of medium respectively, measure 10mL mother liquor 2, mother liquor 4, it is standby to put into beaker:
(2) add the 5g carragheen altogether by every L medium, weigh in the balance and get carragheen, add 200ml distilled water, add and boil, stop heating then to as clear as crystal by every g carragheen;
(3) the mixing mother liquor in the beaker, C source are added in the well-done carragheen, add water again and make culture medium C source quality concentration reach 2.49～2.51%, the carragheen mass concentration reaches 0.5%, heats 80-121 ℃ and stirs;
(4) with sodium hydroxide or the hydrochloric acid of 1mol/L, splash in the rapid prepared culture medium of previous step, stir after dripping, repeat this process and be transferred to 5.8～6.0, promptly finish the preparation of solid culture medium until the pH value;
(5) with the medium for preparing, every bottled 30～40mL seals tight bottleneck, puts into the autoclaving basket, sterilizes 15～19.5 minutes, and can finish the preparation of medium for 121 ℃.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling: individual plant seedling fresh weight 262mg, dry weight 12.4mg, plant height 6.77cm, the thick 1.03mm of stem, the number of blade 5.5, several 8.4 of root bar, the long 5.41cm of root.
Embodiment 2:
The difference of present embodiment and embodiment 1 is: potassium nitrate (KNO3) content is that 4750mg, ammonium nitrate (NH4NO3) are 0mg in every liter of mother liquor 1, and cane sugar content is 1.5%.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 62mg, dry weight 3.5mg, plant height 4.69cm, the thick 1.09mm of stem, the number of blade 5.1, several 5.0 of root bar, the long 3.68cm of root.
Embodiment 3:
The difference of present embodiment and embodiment 1 is: potassium nitrate (KNO3) content is 4750mg in every liter of mother liquor 1, and mother liquor 4 mysoinositol content are 2500mg, and cane sugar content is 2%.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 135mg, dry weight 6.4mg, plant height 5.59cm, the thick 0.90mm of stem, the number of blade 6.4, several 5.4 of root bar, the long 5.29cm of root.
Embodiment 4:
The difference of present embodiment and embodiment 1 is: potassium nitrate (KNO3) content is that 4750mg, ammonium nitrate (NH4NO3) are 8250mg in every liter of mother liquor 1, and mother liquor 4 mysoinositol content are 5000mg.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 204mg, dry weight 9.6mg, plant height 6.41cm, the thick 1.26mm of stem, the number of blade 6.9, several 6.8 of root bar, the long 5.55cm of root.
Embodiment 5:
The difference of present embodiment and embodiment 1 is: potassium nitrate (KNO3) content is that 9500mg, ammonium nitrate (NH4NO3) are 0mg in every liter of mother liquor 1, and mother liquor 4 mysoinositol content are 2500mg.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 129mg, dry weight 6.9mg, plant height 4.92cm, the thick 0.92mm of stem, the number of blade 5.3, several 6.0 of root bar, the long 4.42cm of root.
Embodiment 6:
The difference of present embodiment and embodiment 1 is: potassium nitrate (KNO3) content is 9500mg in every liter of mother liquor 1, and mother liquor 4 mysoinositol content are 5000mg, and cane sugar content is 1.5%.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 45mg, dry weight 3.0mg, plant height 3.81cm, the thick 0.95mm of stem, the number of blade 5.6, several 3.5 of root bar, the long 3.27cm of root.
Embodiment 7:
The difference of present embodiment and embodiment 1 is: potassium nitrate (KNO3) content is that 9500mg, ammonium nitrate (NH4NO3) are 8250mg in every liter of mother liquor 1, and cane sugar content is 2.0%.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 80mg, dry weight 4.7mg, plant height 4.43cm, the thick 1.17mm of stem, the number of blade 4.8, several 4.4 of root bar, the long 3.12cm of root.
Embodiment 8:
The difference of present embodiment and embodiment 1 is: ammonium nitrate (NH4NO3) is 0mg in every liter of mother liquor 1, and mother liquor 4 mysoinositol content are 5000mg, and cane sugar content is 2.0%.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 73mg, dry weight 4.8mg, plant height 4.21cm, the thick 0.79mm of stem, the number of blade 4.7, several 3.6 of root bar, the long 3.85cm of root.
Embodiment 9:
The difference of present embodiment and embodiment 1 is: ammonium nitrate (NH4NO3) is 8250mg in every liter of mother liquor 1, and mother liquor 4 mysoinositol content are 2500mg, and cane sugar content is 1.5%.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 133mg, dry weight 6.8mg, plant height 7.68cm, the thick 1.12mm of stem, the number of blade 6.3, several 7.3 of root bar, the long 3.71cm of root.
Embodiment 10:
Present embodiment (MS medium) is with the difference of embodiment 1: ammonium nitrate (NH4NO3) is 16500mg in every liter of mother liquor 1, and mother liquor 4 mysoinositol content are 10000mg.
On the medium of this mother liquor preparation, when cultivating 12d, measure potato detoxicating basis each economical character index of seedling and be respectively: individual plant seedling fresh weight 83mg, dry weight 8.3mg, plant height 4.6cm, the thick 1.07mm of stem, the number of blade 4.2, several 4.6 of root bar, the long 4.7cm of root.
Through evidence, culture medium cost of the present invention is lower than widely used MS (CK) 21.7% in the present production.

Claims (8)

1. ripe potato detoxicating basis seedling medium in a kind is characterized in that, calculates with the 1L medium and contains: the KNO of 1890～1910mg 3, 169～171mg KH 2PO 4, 369～371mg MgSO 47H 2The CaCl of O, 439～441mg 22H 2The MnSO of O, 22.27～22.23mg 44H 2The KI of O, 0.827～0.833mg, the H of 6.17～6.23mg 3BO 3, 8.57～8.63mg ZnSO 47H 2The Na of O, 0.247～0.253mg 2MoO 4H 2The CuSO of O, 0.0247～0.0253mg 45H 2The CoCl of O, 0.0247～0.0253mg 26H 2The Na of O, 37.0～37.3mg 2The FeSO of EDTA and 26～28mg 47H 2The VB of O, 1.99～2.01mg glycine, 0.09～0.11mg 1, 0.49～0.51mg VB 6, and the VB of 0.49～0.51mg 5Or VPP; Surplus is a water.
2. ripe potato detoxicating basis seedling medium is characterized in that: the NH that wherein also contains 402.5～422.5mg in as claimed in claim 1 4NO.
3. ripe potato detoxicating basis seedling medium is characterized in that: the C source sucrose that wherein also contains 24900～25100mg in as claimed in claim 1 or 2.
4. the preparation method of ripe potato detoxicating basis seedling medium in a kind is characterized in that, comprises the steps:
(1) preparation of the basic mother liquor of medium
A, preparation macroelement mother liquor
Get the KNO of 18900～19100mg 3, 1690～1710mg KH 2PO 4, 3690～3710mg MgSO 47H 2The CaCl of O and 4390～4410mg 22H 2O, more than all kinds of materials fully dissolve with distilled water respectively, put in order by it and pour constant volume in the 1L volumetric flask successively into, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
B, prepare micro-mother liquor
Get the MnSO of 2227～2223mg 44H 2The KI of O, 82.7～83.3mg, the H of 617～623mg 3BO 3, 857～863mg ZnSO 47H 2The Na of O, 24.7～25.3mg 2MoO 4H 2The CuSO of O, 2.47～2.53mg 45H 2The CoCl of O and 2.47～2.53mg 26H 2O, more than all kinds of materials fully dissolve with distilled water respectively, constant volume is in the 1L volumetric flask, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
C, preparation mother liquid of iron salt
Get the Na of 370～373mg 2The FeSO of EDTA and 260～280mg 47H 2O, more than all kinds of materials fully dissolve with distilled water respectively, constant volume is in the 1L volumetric flask, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
D, No. 1 organic matter mother liquor of preparation
Get the VB of 199～201mg glycine, 9～11mg 1, 49～51mg VB 6, and the VB of 49～51mg 5Or VPP, more than all kinds of materials fully dissolve with distilled water respectively, constant volume is in the 1L volumetric flask, it is standby to reinstall in the reagent bottle that posts the concentration label 4 ℃ of preservations;
Measure 100mL macroelement mother liquor, mother liquid of iron salt respectively by every liter of medium, and 10mL trace element mother liquor and No. 1 organic matter mother liquor, it is standby to put into beaker;
(2) add 5g by every L medium and take by weighing carragheen, add 200ml distilled water, add and boil, stop heating then to as clear as crystal by every g carragheen;
(3) the mixing mother liquor in the beaker, C source are added in the well-done carragheen, add water again and make culture medium C source quality concentration reach 2.49～2.51%, the carragheen mass concentration reaches 0.5%, heats 80-121 ℃ and stirs;
(4) with sodium hydroxide or hydrochloric acid the pH value is transferred to 5.8～6.0, promptly finishes the preparation of solid culture medium;
(5) bottling back autoclaving gets final product.
5. the preparation method of ripe potato detoxicating basis seedling medium in as claimed in claim 4, it is characterized in that: wherein the C source is a sucrose, addition is 24900～25100mg.
6. the preparation method of ripe potato detoxicating basis seedling medium is characterized in that: the NH that wherein also adds 4025～4225mg in the macroelement mother liquor in as claimed in claim 4 4NO 3
7. the preparation method of ripe potato detoxicating basis seedling medium in as claimed in claim 4, it is characterized in that: wherein regulating the pH value is sodium hydroxide or the hydrochloric acid of using 1mol/L, splash in the rapid prepared culture medium of previous step, stir after dripping, repeat this process and be transferred to 5.8～6.0, promptly finish the preparation of solid culture medium until the pH value.
8. the preparation method of ripe potato detoxicating basis seedling medium in as claimed in claim 4, it is characterized in that: wherein bottling back autoclaving is the solid culture medium that will prepare, every bottled 30～40mL, seal tight bottleneck, put into the autoclaving basket, 121 ℃ of sterilizations are taken out after 15～19.5 minutes and are got final product.
CN 201110125825 2011-05-17 2011-05-17 Medium maturity virus-free potato basic seedling culture medium and preparation method thereof Expired - Fee Related CN102257958B (en)

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CN102674966A (en) * 2012-05-21 2012-09-19 新疆生产建设兵团农六师农业科学研究所 Special culture medium for potato virus-free seedling transplanting and preparation method thereof

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CN101849509A (en) * 2010-06-29 2010-10-06 浙江省农业科学院 Root tip detoxification and rapid propagation technology for purple potatoes

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CN101790935A (en) * 2010-03-31 2010-08-04 四川农业大学 Potato isolated culture one-step seedling culture medium and optimization method and seedling method thereof
CN101849509A (en) * 2010-06-29 2010-10-06 浙江省农业科学院 Root tip detoxification and rapid propagation technology for purple potatoes

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Publication number Priority date Publication date Assignee Title
CN102674966A (en) * 2012-05-21 2012-09-19 新疆生产建设兵团农六师农业科学研究所 Special culture medium for potato virus-free seedling transplanting and preparation method thereof

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