CN102257958B - 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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CN102257958B
CN102257958B CN 201110125825 CN201110125825A CN102257958B CN 102257958 B CN102257958 B CN 102257958B CN 201110125825 CN201110125825 CN 201110125825 CN 201110125825 A CN201110125825 A CN 201110125825A CN 102257958 B CN102257958 B CN 102257958B
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mother liquor
culture medium
carrageen
preparation
preparing
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CN102257958A (en
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曹君迈
陈彦云
焦向炜
陈科元
祁金涛
涂江萍
张珊珊
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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 virus-free potato foundation seedlings substratum and preparation method thereof
Technical field
The present invention relates to a kind of in ripe virus-free potato foundation seedlings substratum and preparation method thereof.
Background technology
It is to produce virus-free potato foundation seedlings by group culturation rapid propagating technology that China's virus-free potato primary stock industrialization is at present produced, then it is transplanted hardening produce micro potato.And the MS substratum is the substratum that numerous extensive employing is expanded in present virus-free potato foundation seedlings industrialization, and its agar consumption is too high, have even to 1.7%, and the agar cost is higher, and basic seedling is grown under 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 transplanting hardening, takes out basic seedling from bottle, pour in warm water, agar around root is washed off, and when agar is washed root more firmly, the root easy damaged, affect transplanting survival rate.Do not select substratum by the kind ripening stage, strengthened industrial production cost, make basic seedling growth potential relatively poor.Increased in the training period cost of drugs and electricity 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 be to provide a kind of in ripe virus-free potato foundation seedlings substratum, adopt the detoxification basis seedling of this substratum with short production cycle, growth is vigorous, production cost is low.
Two of purpose of the present invention be to provide a kind of above-mentioned in the preparation method of ripe virus-free potato foundation seedlings substratum.
For achieving the above object, the technical solution used in the present invention is.
In a kind of, ripe virus-free potato foundation seedlings substratum, is characterized in that, calculates with the 1L substratum and contain: 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 water.
Wherein also contain the NH of 402.5～422.5mg 4NO;
The C source sucrose that wherein also contains 24900～25100mg.
The preparation method of ripe virus-free potato foundation seedlings substratum in a kind of, its special feature is, comprises the steps:
(1) preparation of the basic mother liquor of substratum
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, above all kinds of materials fully dissolve with distilled water respectively, put in order by it and pour successively constant volume in the 1L volumetric flask into, reinstall in the reagent bottle that posts the concentration label 4 ℃ and save backup;
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, above all kinds of materials fully dissolve with distilled water respectively, and constant volume reinstalls in the reagent bottle that posts the concentration label 4 ℃ and saves backup in the 1L volumetric flask;
C, preparation mother liquid of iron salt
Get the Na of 370～373mg 2The FeSO of EDTA and 260～280mg 47H 2O, above all kinds of materials fully dissolve with distilled water respectively, and constant volume reinstalls in the reagent bottle that posts the concentration label 4 ℃ and saves backup in the 1L volumetric flask;
D, No. 1 organism 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, above all kinds of materials fully dissolve with distilled water respectively, and constant volume reinstalls in the reagent bottle that posts the concentration label 4 ℃ and saves backup in the 1L volumetric flask;
Measure respectively 100mL macroelement mother liquor, mother liquid of iron salt by every liter of substratum, and 10mL trace element mother liquor and No. 1 organism mother liquor, put into beaker standby;
(2) add 5g by every L substratum and take carrageenin, add 200ml distilled water by every g carrageenin, add and boil to as clear as crystal, then stopped heating;
(3) the mixing mother liquor in beaker, C source is added in well-done carrageenin, then add water and make culture medium C source quality concentration reach 2.49～2.51%, the carrageenin 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, namely completes the preparation of solid medium;
(5) after the bottling, autoclaving gets final product.
Wherein the C source is sucrose, and add-on is 24900～25100mg.
Wherein also add the NH of 4025～4225mg in the macroelement mother liquor 4NO 3
Wherein regulate the pH value and be sodium hydroxide or hydrochloric acid with 1mol/L, splash in the substratum that previous step prepares, stir after dripping, repeat this process until the pH value is transferred to 5.8～6.0, namely complete the preparation of solid medium.
Wherein after the bottling, autoclaving is the solid medium that will prepare, and every bottled 30～40mL is tamping 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 substratum of the present invention not only nutrition composition and composition reduce, and with after carrageenin replaces agar as peptizer cheaply, the vigorous and cycle shortening of test-tube plantlet growth, thereby effectively reduced energy consumption and cost in growing seedlings.
Embodiment
Embodiment 1:
In detoxification, ripe potato test-tube plantlet substratum components by weight percent waste is as follows:
A, macroelement mother liquor (mother liquor 1) form: 18900～19100mg saltpetre (KNO 3), 4025～4225mg ammonium nitrate (NH 4NO 3), 1690～1710mg potassium primary phosphate (KH 2PO 4), 3690～3710mg sal epsom (MgSO 47H 2O), 4390～4410mg calcium chloride (CaCl 22H 2O), above all kinds of materials fully dissolve with distilled water respectively, put in order by it and pour successively constant volume in the 1L volumetric flask into, reinstall in the reagent bottle that posts the concentration label 4 ℃ and save backup;
B, micro-mother liquor (mother liquor 2) form: 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, above all kinds of materials fully dissolve with distilled water respectively, and constant volume reinstalls in the reagent bottle that posts the concentration label 4 ℃ and saves backup in the 1L volumetric flask;
C, mother liquid of iron salt form (mother liquor 3): 370～373mg disodium ethylene diamine tetraacetate (Na 2EDTA), 260～280mg ferrous sulfate (FeSO 47H 2O), above all kinds of materials fully dissolve with distilled water respectively, and constant volume reinstalls 4 ℃ of preservations, standby in the reagent bottle that posts the concentration label in the 1L volumetric flask;
D, No. 1 organism mother liquor form (mother liquor 4): the VB of 199～201mg glycine, 9～11mg 1, 49～51mg VB 6, and the VB of 49～51mg 5Or VPP, above all kinds of materials fully dissolve with distilled water respectively, and 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 substratum respectively, measure 10mL mother liquor 2, mother liquor 4, put into beaker standby:
(2) add altogether the 5g carrageenin by every L substratum, weigh in the balance and get carrageenin, add 200ml distilled water by every g carrageenin, add and boil to as clear as crystal, then stopped heating;
(3) the mixing mother liquor in beaker, C source is added in well-done carrageenin, then add water and make culture medium C source quality concentration reach 2.49～2.51%, the carrageenin mass concentration reaches 0.5%, heats 80-121 ℃ and stirs;
(4) with sodium hydroxide or the hydrochloric acid of 1mol/L, splash in the substratum that previous step prepares, stir after dripping, repeat this process until the pH value is transferred to 5.8～6.0, namely complete the preparation of solid medium;
(5) with the substratum for preparing, every bottled 30～40mL is tamping bottleneck, puts into the autoclaving basket, sterilizes 15～19.5 minutes, and can complete the preparation of substratum for 121 ℃.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings: individual plant seedling fresh weight 262mg, dry weight 12.4mg, plant height 6.77cm, the thick 1.03mm of stem, the number of blade 5.5,8.4 of root numbers, the long 5.41cm of root.
Embodiment 2:
The difference of the present embodiment and embodiment 1 is: in every liter of mother liquor 1, saltpetre (KNO3) content is that 4750mg, ammonium nitrate (NH4NO3) are 0mg, and sucrose content is 1.5%.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 5.1,5.0 of root numbers, the long 3.68cm of root.
Embodiment 3:
The difference of the present embodiment and embodiment 1 is: in every liter of mother liquor 1, saltpetre (KNO3) content is 4750mg, and mother liquor 4 mysoinositol content are 2500mg, and sucrose content is 2%.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 6.4,5.4 of root numbers, the long 5.29cm of root.
Embodiment 4:
The difference of the present embodiment and embodiment 1 is: in every liter of mother liquor 1, saltpetre (KNO3) content is that 4750mg, ammonium nitrate (NH4NO3) are 8250mg, and mother liquor 4 mysoinositol content are 5000mg.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 6.9,6.8 of root numbers, the long 5.55cm of root.
Embodiment 5:
The difference of the present embodiment and embodiment 1 is: in every liter of mother liquor 1, saltpetre (KNO3) content is that 9500mg, ammonium nitrate (NH4NO3) are 0mg, and mother liquor 4 mysoinositol content are 2500mg.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 5.3,6.0 of root numbers, the long 4.42cm of root.
Embodiment 6:
The difference of the present embodiment and embodiment 1 is: in every liter of mother liquor 1, saltpetre (KNO3) content is 9500mg, and mother liquor 4 mysoinositol content are 5000mg, and sucrose content is 1.5%.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 5.6,3.5 of root numbers, the long 3.27cm of root.
Embodiment 7:
The difference of the present embodiment and embodiment 1 is: in every liter of mother liquor 1, saltpetre (KNO3) content is that 9500mg, ammonium nitrate (NH4NO3) are 8250mg, and sucrose content is 2.0%.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 4.8,4.4 of root numbers, the long 3.12cm of root.
Embodiment 8:
The difference of the present embodiment and embodiment 1 is: in every liter of mother liquor 1, ammonium nitrate (NH4NO3) is 0mg, and mother liquor 4 mysoinositol content are 5000mg, and sucrose content is 2.0%.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 4.7,3.6 of root numbers, the long 3.85cm of root.
Embodiment 9:
The difference of the present embodiment and embodiment 1 is: in every liter of mother liquor 1, ammonium nitrate (NH4NO3) is 8250mg, and mother liquor 4 mysoinositol content are 2500mg, and sucrose content is 1.5%.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 6.3,7.3 of root numbers, the long 3.71cm of root.
Embodiment 10:
The present embodiment (MS substratum) is with the difference of embodiment 1: in every liter of mother liquor 1, ammonium nitrate (NH4NO3) is 16500mg, and mother liquor 4 mysoinositol content are 10000mg.
On the substratum of this mother liquor preparation, when cultivating 12d, measure each economical character index of virus-free potato foundation seedlings 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 4.2,4.6 of root numbers, the long 4.7cm of root.
Through evidence, culture medium cost of the present invention is lower than producing at present upper widely used MS (CK) 21.7%.

Claims (1)

1. ripe virus-free potato foundation seedlings substratum in a kind, is characterized in that, calculates with the 1L substratum and contain: 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, 22.23～22.27mg 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 CuS0 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 5
Wherein also contain the NH of 402.5～422.5mg 4NO 3
The C source sucrose that wherein also contains 24900～25100mg;
Wherein also contain the carrageenin of 5g;
PH value 5.8～6.0;
Surplus is water.
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

Citations (3)

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Publication number Priority date Publication date Assignee Title
JP3877800B2 (en) * 1996-04-30 2007-02-07 株式会社テクノバ Propagation method of aseptic sweet potato seedlings
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

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
JP3877800B2 (en) * 1996-04-30 2007-02-07 株式会社テクノバ Propagation method of aseptic sweet potato seedlings
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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