CN106035084A

CN106035084A - Subculture medium for blueberries and preparation method of medium

Info

Publication number: CN106035084A
Application number: CN201610393651.0A
Authority: CN
Inventors: 王媛花; 颜志明; 冯英娜; 王全智
Original assignee: Jiangsu Polytechnic College of Agriculture and Forestry
Current assignee: Jiangsu Polytechnic College of Agriculture and Forestry
Priority date: 2016-06-03
Filing date: 2016-06-03
Publication date: 2016-10-26

Abstract

The invention discloses a subculture medium for blueberries. The medium contains macroelement mother liquor, microelement mother liquor, iron salt mother liquor, calcium salt mother liquor, organic matter mother liquor and hormone mother liquor, wherein the organic matter mother liquor in every one liter of the subculture medium for the blueberries is prepared from 100 mg of inositol, 1 mg of niacin, 1 mg of vitamin B1, 2mg of vitamin B6 and 2 mg of vitamin C, and the hormone mother liquor in every one liter of the subculture medium for the blueberries is prepared from 4 mg of zeatin. A plant growth hormone, namely, the zeatin, is added to the subculture medium, so that blueberry tissue cultured plants can grow well, have many ramets and root easily, transplantation of the blueberries into fields after culture is facilitated, and the survival rate of the blueberries in the fields is increased; meanwhile, under the assistance of a matched amount of the vitamin C, the vitrification effect of the zeatin on the blueberries can be eliminated, better growth of the blueberry plants can be promoted, and cultured seedlings grow stronger, are not browned to death easily and have greener and flat leaves.

Description

A kind of blue berry subculture medium and compound method thereof

Technical field

The present invention relates to a kind of culture medium, be specifically related to a kind of subculture medium.

Background technology

Blue berry is the fruit that a kind of nutritive value is the highest, rich in abundant vitamin, protein, meals fibre in sarcocarp Dimension, fat and the nutrient such as mineral, far above other fruit rich in nutrition, wherein unique in blue berry anthocyanin pigments In many fruit, proportion is unusual height.And in the cultivation of blue berry, the requirement to soil is higher, soil organic matter content and PH value is whether blue berry cultivates the key factor survived, and typically requires the content of organic matter 5%, and the soil of pH value 4.0～5.5. Blue berry seedling breeds the method all using tissue culture in a large number at present, equally in tissue culture, and the blue berry requirement to culture medium Also it is that comparison is high, it is desirable to high organic content and weakly acidic pH value.

Culture medium conventional in plant tissue culture is MS culture medium, and MS culture medium feature is inorganic salt and ion concentration Higher, it is more stable ionic equilibrium solution.WPM culture medium is in the cultivation the most commonly used when xylophyta is cultivated Base.But in the tissue culture of blue berry, no matter by MS or WPM culture medium, effect is not the most fine.Although reporting at present Many blue berries are cultivated and are used WPM culture medium, but for being both for some blue berry kind, therefore, the training reported Support based formulas the most general in most blue berry kinds.

Zeatin is generally used in tissue culture procedures promoting cell division, promotes that plant carries out plant division, but zeatin The excessive concentration used, or during subculture, the peace of zeatin access times is numerous, easily causes the glass of blue berry plant Changing, present vitrified blue berry Seedling, its stem, leaf surface are without waxiness, and internal polar compound level is higher, cell retention ability Difference, transpiration is strong, it is impossible to normally transplant, and causes the significant wastage of human and material resources, financial resources, is the training of current blueberry tissue The biggest obstacle that in supporting, tissue cultured seedling produces.And do not use zeatin or zeatin concentration too low in the case of, blue berry is planted Strain can not normal growth, blue berry plant can be made time more serious dead.Therefore by reducing zeatin concentration or being not added with It is infeasible that Semen Maydis usually solves the vitrification phenomenon of blue berry.

Summary of the invention

Goal of the invention: it is an object of the invention to overcome the deficiencies in the prior art, it is provided that a kind of blue berry successive transfer culture Base and compound method thereof, to adapt to the cultivation needs of different blue berry kind, it is ensured that well-grown.

Technical scheme: a kind of blue berry subculture medium of the present invention, comprises a great number of elements mother solution, trace element mother Liquid, mother liquid of iron salt, Calcisolution, Organic substance mother solution and hormone mother solution；Described in every 1L blue berry culture medium, Organic substance mother solution includes Inositol 100mg/L, nicotinic acid 1mg/L, vitamin B₁1mg/L, vitamin B₆2mg/L, Catergen mg/L, described hormone includes jade Rice element 4mg/L.

Further, described a great number of elements mother solution includes potassium nitrate 400mg/L, potassium sulfate 900mg/L, bitter salt 370mg/L, potassium dihydrogen phosphate 170mg/L, trace element include four anhydrous manganese 20mg/L, Zinc vitriol 8.6mg/L, Boric acid 6.2mg/L, copper sulfate pentahydrate 0.05mg/L, two molybdic acid hydrate sodium 0.25mg/L.

Further, described mother liquid of iron salt includes ethylenediamine o-dihydroxy ferric acetate 10mg/L.

Further, described Calcisolution includes calcium chloride hydrate 100mg/L, four water-calcium nitrate 600mg/L.

Further, each mother liquor concentrations being configured to concentrated solution, cold preservation is standby, and during configuration culture medium, every liter of culture medium adds With sucrose 30 grams, agar powder 6 grams, pH value regulates to 5.2.

Beneficial effect: 1, add auxin zeatin in culture medium of the present invention, it is possible to make blueberry tissue cultivate Plant grow fine, plant division is many, and easily takes root, beneficially blue berry cultivate after transplant enter field, improve blue berry in field Between survival rate；It is aided with ascorbic consumption to coordinate simultaneously, the vitrification impact that blue berry is caused by zeatin can either be eliminated, Can promote that again blue berry plant preferably grows, it is more healthy and stronger that seedlings cultivating grows, it is not easy to brownization is dead, and leaf color is greener, blade Open and flat；

2, formula does not use ammonium nitrate, uses the ammonium sulfate easily bought at present and carbamide to substitute, solves nitric acid Ammonium buys difficult problem；

3, owing to employing ammonium sulfate, therefore the concentration of sulfate radical increases, therefore by sub-for the sulphuric acid in tradition MS culture medium The iron salt of ferrum and disodiumedetate preparation changes Fe-EDDHA ethylenediamine o-dihydroxy ferric acetate into, and test effect proves to plant Thing growth is more preferably；

4, the cobaltous chloride in MS culture medium and copper sulfate are eliminated, because the two material consumption and few, process for preparation In due to bad weighing, it is often necessary to first configure the two material concentrated solution, adding in mother solution according to finite concentration, taking Time laborious, experiment finds that increasing by two materials does not affect plant growing, therefore removes, simplifies preparation process.

Detailed description of the invention

Below technical solution of the present invention is described in detail, but protection scope of the present invention is not limited to described enforcement Example.

Embodiment: as a example by the blue berry subculture medium configuring 1 liter:

The first step, configures 1 liter of a great number of elements, is configured to the concentration solution of 25 times, for mother solution one.

Weigh: KNO₃Potassium nitrate 10g, K₂SO₄Potassium sulfate 22.5g, MgSO₄·7H₂O bitter salt 9.25g, KH₂PO₄ Potassium dihydrogen phosphate 4.25g, is dissolved in 1 liter of distilled water.

Second step, configuration 200 person of outstanding talent rises trace element, is configured to the concentration solution of 1000 times, for mother solution two.

Weigh: MnSO₄·4H₂O tetra-anhydrous manganese 4g, ZnSO₄·7H₂O Zinc vitriol 1.72g, H₃BO₃Boric acid 1.24g、CuSO₄·5H₂O copper sulfate pentahydrate 0.01g, NaMoO₄·2H₂O bis-molybdic acid hydrate sodium 0.05g, is dissolved in 200 persons of outstanding talent and rises In distilled water.

3rd step, configuration 500 person of outstanding talent rises iron salt, is configured to the concentration solution of 200 times, for mother solution three.

Weigh: Fe-EDDHA ethylenediamine o-dihydroxy ferric acetate 1g, be dissolved in 500 persons of outstanding talent and rise in distilled water.

4th step, configuration 500 person of outstanding talent rises calcium salt, is configured to the concentration solution of 50 times, for mother solution four.

Weigh: CaCl₂·2H₂O CALCIUM CHLORIDE DIHYDRATE 2.5g, Ca (NO₃)₂·4H₂O four water-calcium nitrate 15g, is dissolved in 500 Person of outstanding talent rises in distilled water.

5th step, configuration 500 person of outstanding talent rises organic solution, is configured to the concentration solution of 200 times, for mother solution five.

Weigh: inositol 10g, nicotinic acid 0.1g, vitamin B₁0.1g, vitamin B₆0.2g, vitamin C 0.2g are dissolved in 500 Person of outstanding talent rises in distilled water.

6th step, preparation 100mL concentration is the zeatin solution of 1mg/L, for mother solution six.

Weigh: 0.1g zeatin, be dissolved in 100mL distilled water.

Mother solution puts 4 degree of Refrigerator stores after preparing standby.During configuration culture medium, each liter of culture medium adds mother solution one 50mL, female Liquid two 1mL, mother solution three 5mL, mother solution four 20mL, mother solution five 5mL, mother solution six 4mL.

The compound method of above-mentioned culture medium is, adds 6g agar powder and the mixing of 30g sucrose, pH value by after every kind of mother solution mixing Regulation, to 5.2, is settled to one liter, is dispensed into culture medium and is suitable for containers for future use.

Following table is the growth conditions statistics after different blue berry kind grows 30 days in different culture media:

It can be seen that either MS culture medium or the blue berry of WPM culture medium culturing from table, in 6 different cultivars, Plant height, value-added coefficient and survival rate are all significantly lower than the culture medium of the present invention.Therefore, the culture medium of the present invention can promote Blue berry grows, and has good effect in multiple kinds.

It addition, the zeatin of the present embodiment and ascorbic concentration can effectively suppress the vitrification of blue berry Seedling, by this reality The concentration proportioning executing example compares with other concentration proportionings, as shown in the table, it is possible to find zeatin of the present invention and ascorbic Consumption coordinates can be stopped the vitrification phenomenon of blue berry, therefore seedlings cultivating is more healthy and stronger while promoting plant strain growth completely.

Claims

1. a blue berry subculture medium, it is characterised in that: comprise a great number of elements mother solution, trace element mother solution, mother liquid of iron salt, calcium Salt mother solution, Organic substance mother solution and hormone mother solution；Described in every 1L blue berry culture medium, Organic substance mother solution includes inositol 100 mg/L, cigarette Acid 1 mg/L, vitamin B₁1 mg/L, vitamin B₆2mg/L, Catergen mg/L, described hormone includes zeatin 4 mg/L.

Blue berry subculture medium the most according to claim 1, it is characterised in that: described a great number of elements mother solution includes potassium nitrate 400 mg/L, potassium sulfate 900 mg/L, bitter salt 370 mg/L, potassium dihydrogen phosphate 170 mg/L.

Blue berry subculture medium the most according to claim 1, it is characterised in that: described trace element mother solution includes four hydrations Manganese sulfate 20 mg/L, Zinc vitriol 8.6 mg/L, boric acid 6.2 mg/L, copper sulfate pentahydrate 0.05 mg/L, two hydrations Sodium molybdate 0.25 mg/L.

Blue berry subculture medium the most according to claim 1, it is characterised in that: described mother liquid of iron salt includes ethylenediamine neighbour two Hydroxyacetic acid ferrum 10 mg/L.

Blue berry subculture medium the most according to claim 1, it is characterised in that: described Calcisolution includes calcium chloride hydrate 100 mg/L, four water-calcium nitrate 600 mg/L.

6. the preparation method of blue berry subculture medium as claimed in claim 1, it is characterised in that: each mother liquor concentrations is joined Making concentrated solution, cold preservation is standby, and during configuration culture medium, every liter of culture medium adds sucrose 30 grams, agar powder 6 grams, and pH value regulates extremely 5.2。