CN104737919A - Tissue culture rapid seedlings growing method used for cherry stock M9 - Google Patents

Abstract

The invention discloses a tissue culture rapid seedling growing method used for cherry stock M9. The tissue culture rapid seedling growing method comprises the following steps: (1) a sterile material is obtained; (2) primary culture of cherry is carried out; (3) enrichment culture of the cherry is carried out; (4) strong plantlet and rootage culture is carried out; (5) domestication and transplantation are carried out. The tissue culture rapid seedling growing method used for cherry stock M9 has the beneficial effects that since the method is realized on the basis of a M9 culture medium, the kinds and of concentrations of a mass of elements are adjusted, the concentration of agar is regulated and controlled by optimizing hormone, a balanced and reasonable nutritional requirement can be met for subculture proliferation of cherry tissue culture seedlings, the rapid, efficient and robust growth of the cherry tissue culture seedlings can be ensued, the internode elongation is effectively promoted, and the rooting coefficient is high.

Description

The group training fast seedling-cultivating method of a kind of cherry rootstock M9

Technical field

The present invention relates to the group training fast seedling-cultivating method of a kind of cherry rootstock M9.

Background technology

At present, cherry belongs to rose family deciduous tree fruit tree, and time ripe, color is scarlet, exquisitely carved, delicious shape is spoilt, tree performance is graceful, is rich in sugar and irony, nutritious, important fruit tree and Evergreen garden plant, have again very high edible and medical value, extensively cultivate in China in recent years, development potentiality is huge.

At present, mainly adopt cuttage, press strip and point root turion breeding in the production of cherry, not only reproduction coefficient is low, and causes deterioration of variety, damage by disease and insect its spread in china.Adopt tissue culture technique can Fast-propagation New Crop Varieties, accelerate the promotion rate of new varieties, the plantlet in vitro that Plant Tissue Breeding is simultaneously cultivated, gene are highly consistent, administration measure, nursery stock quality parameter is good, can produce the nursery stock providing high-quality for industrialization.

At present, the technical problem of cherry group training factorial seedling growth does not also solve completely, when adopting MS medium cherry plantlet in vitro shoot proliferation to cultivate, often there is following problem: group training seedling base portion callus of taking root expands, the phenomenons etc. such as plantlet in vitro growth potential is weak, blade is large, stem is short and small, internode elongation growth is slow, growth coefficient is low.

Summary of the invention

The object of this invention is to provide the group training fast seedling-cultivating method of a kind of cherry rootstock M9, solve kind of the problem that the growth of cherry group training stage internode elongation is slow, growth coefficient is low.

The technical solution adopted for the present invention to solve the technical problems is:

A group training fast seedling-cultivating method of cherry rootstock M9, is characterized in that, comprise the following steps:

(1) sterilizable material is obtained:

The budlet that clip cherry is newly sprouted spring, after the alcohol immersion 20-30s cleaning material of 75%, soaks after 15-20 minute on superclean bench in pure water, then inhales budlet soma surface moisture with blotting paper;

(2) Initial culture of cherry:

Get budlet base portion, be cut into the segment that lcm is long, be inoculated on budlet Fiber differentiation, the composition of described budlet inducing culture is: MN+2.0 mg/L 6-BA+0.5 mg/L 2,4-D+0.2 mg/L NAA+ 30 g/L white sugar+6g/L agar powder, budlet was inoculated in budlet inducing culture after 19 days, carried out shoot proliferation cultivation;

(3) Multiplying culture of cherry:

Bud with callus is inoculated on the adventitious bud proliferation medium that pH value is 5.8-6.2 and carries out Multiplying culture, the composition of described proliferated culture medium is potassium nitrate 0.3mg/L+ potassium dihydrogen phosphate 0.1mg/L+ magnesium sulfate 0.2mg/L+ potassium iodide 0.2mg/L zinc sulphate 0.3mg/L+ sodium molybdate 0.1mg/L+ ferrous sulfate 0.5mg/L+ puridoxine hydrochloride 0.2mg/L+ nicotinic acid 0.1mg/L+ heteroauxin 0.2mg/L+6-BA1.0-3.0mg/L+ sucrose 30g/L+ agar 6g/L, wherein, first carry out dark treatment 1 week, give illumination every day 16h again, the cultivation temperature of Multiplying culture is 24-26 DEG C,

(4) Rooting and hardening-off culture:

Induce Multiple Buds in adventitious bud proliferation medium after, Multiple Buds being divided into little Cong or simple bud and being inoculated in pH value is in the Rooting and hardening-off culture base of 5.8-6.2, the composition of described Rooting and hardening-off culture base is 0.2mg/LIBA+NAA0.5-1.0mg/L+IBA2.0-6.0mg/L+ sucrose 30g/L+ agar 6g/L, carries out taking root and strong seedling culture;

(5) rooting culture:

When root length is for 2-4cm, carry out rooting culture, blake bottle is moved to hardening 6-10 days in booth, wash away medium, root being soaked 5% liquor potassic permanganate carries out disinfection, and is transplanted in the dish of cave, removes film, give full exposure, carry out rich water quality management after 1 week.

Beneficial effect: the present invention is owing to adopting on the basis of M9 medium, by adjusting kind and the concentration of macroelement and optimizing hormone regulating and controlling agar concentration etc., for cherry peach plantlet in vitro shoot proliferation provides the nutritional need of balanced and reasonable, ensure that it is quick, efficient, robust growth, coefficient of taking root is high, effectively facilitate internode elongation, improve growth coefficient.

Embodiment

Embodiment:

A group training fast seedling-cultivating method of cherry rootstock M9, is characterized in that, comprise the following steps:

(1) sterilizable material is obtained:

The budlet that clip cherry is newly sprouted spring, after the alcohol immersion 20-30s cleaning material of 75%, soaks after 15-20 minute on superclean bench in pure water, then inhales budlet soma surface moisture with blotting paper;

(2) Initial culture of cherry:

Get budlet base portion, be cut into the segment that lcm is long, be seeded on budlet Fiber differentiation, the composition of described budlet inducing culture is: MN+2.0 mg/L 6-BA+0.5 mg/L 2,4-D+0.2 mg/L NAA+ 30 g/L white sugar+6g/L agar powder, budlet was inoculated in budlet inducing culture after 19 days, carried out shoot proliferation cultivation;

(3) Multiplying culture of cherry:

Bud with callus is inoculated on the adventitious bud proliferation medium that pH value is 5.8-6.2 and carries out Multiplying culture, the composition of described proliferated culture medium is potassium nitrate 0.3mg/L+ potassium dihydrogen phosphate 0.1mg/L+ magnesium sulfate 0.2mg/L+ potassium iodide 0.2mg/L zinc sulphate 0.3mg/L+ sodium molybdate 0.1mg/L+ ferrous sulfate 0.5mg/L+ puridoxine hydrochloride 0.2mg/L+ nicotinic acid 0.1mg/L+ heteroauxin 0.2mg/L+6-BA1.0-3.0mg/L+ sucrose 30g/L+ agar 6g/L, wherein, first carry out dark treatment 1 week, give illumination every day 16h again, the cultivation temperature of Multiplying culture is 24-26 DEG C,

(4) Rooting and hardening-off culture:

Induce Multiple Buds in adventitious bud proliferation medium after, Multiple Buds being divided into little Cong or simple bud and being inoculated in pH value is in the Rooting and hardening-off culture base of 5.8-6.2, the composition of described Rooting and hardening-off culture base is 0.2mg/LIBA+NAA0.5-1.0mg/L+IBA2.0-6.0mg/L+ sucrose 30g/L+ agar 6g/L, carries out taking root and strong seedling culture;

(5) rooting culture:

When root length is for 2-4cm, carry out rooting culture, blake bottle is moved to hardening 6-10 days in booth, wash away medium, root being soaked 5% liquor potassic permanganate carries out disinfection, and is transplanted in the dish of cave, removes film, give full exposure, carry out rich water quality management after 1 week.

Owing to adopting on the basis of M9 medium, by adjusting kind and concentration and the optimization hormone regulating and controlling agar concentration etc. of macroelement, for cherry plantlet in vitro shoot proliferation provides the nutritional need of balanced and reasonable, ensure that it is quick, efficient, robust growth, coefficient of taking root is high, effectively facilitate internode elongation, improve growth coefficient.

Above embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under not departing from the present invention and designing the prerequisite of spirit; the various distortion that the common engineers and technicians in this area make technical scheme of the present invention and improvement, all should fall in protection domain that claims of the present invention determine.

The part that the present invention does not relate to prior art that maybe can adopt all same as the prior art is realized.

Claims (1)

1. a group training fast seedling-cultivating method of cherry rootstock M9, is characterized in that, comprise the following steps:

(1) sterilizable material is obtained:

The budlet that clip cherry is newly sprouted in spring, after the alcohol immersion 20-30s cleaning material of 75%, is put in superclean bench on after soaking 15-20 minute in pure water, then inhales budlet soma surface moisture with blotting paper;

(2) Initial culture of cherry:

Get budlet base portion, be cut into the segment that lcm is long, be seeded on budlet Fiber differentiation, the composition of described budlet inducing culture is: MN+2.0 mg/L 6-BA+0.5 mg/L 2,4-D+0.2 mg/L NAA+ 30 g/L white sugar+6g/L agar powder, budlet was inoculated in budlet inducing culture after 19 days, carried out shoot proliferation cultivation;

(3) Multiplying culture of cherry:

Bud with callus is inoculated on the adventitious bud proliferation medium that pH value is 5.8-6.2 and carries out Multiplying culture, the composition of described proliferated culture medium is 0.3mg/L+ potassium dihydrogen phosphate 0.1mg/L+ magnesium sulfate 0.2mg/L+ potassium iodide 0.2mg/L zinc sulphate 0.3mg/L+ sodium molybdate 0.1mg/L+ ferrous sulfate 0.5mg/L+ puridoxine hydrochloride 0.2mg/L+ nicotinic acid 0.1mg/L+ heteroauxin 0.2mg/L+6-BA1.0-3.0mg/L+ sucrose 30g/L+ agar 6g/L, wherein, first carry out dark treatment 1 week, give illumination every day 16h again, the cultivation temperature of Multiplying culture is 24-26 DEG C,

(4) Rooting and hardening-off culture:

Induce Multiple Buds in adventitious bud proliferation medium after, Multiple Buds being divided into little Cong or simple bud and being inoculated in pH value is in the Rooting and hardening-off culture base of 5.8-6.2, the composition of described Rooting and hardening-off culture base is 0.2mg/LIBA+NAA0.5-1.0mg/L+IBA2.0-6.0mg/L+ sucrose 30g/L+ agar 6g/L, carries out taking root and strong seedling culture;

(5) rooting culture:

When root length is for 2-4cm, carry out rooting culture, blake bottle is moved to hardening 6-10 days in booth, wash away medium, root being soaked 5% liquor potassic permanganate carries out disinfection, and is transplanted in the dish of cave, removes film, give full exposure, carry out rich water quality management after 1 week.