CN103651134A

CN103651134A - Tissue culture method for chirita wentsaii

Info

Publication number: CN103651134A
Application number: CN201310656572.0A
Authority: CN
Inventors: 杨铁顺; 赵亮; 柯盛发; 刘青; 吴建平; 闵宇; 刘阳
Original assignee: Tianjin Binhai International Flower Technology Park Co Ltd
Current assignee: Tianjin Binhai International Flower Technology Park Co Ltd
Priority date: 2013-12-05
Filing date: 2013-12-05
Publication date: 2014-03-26

Abstract

The invention belongs to the field of plant tissue culture, and especially relates to a tissue culture method for chirita wentsaii. The method comprises: removing tender leaves on petioles, flushing with running water for 30 min, draining off water, immersing with alcohol with a concentration of 70% for 30 s on an ultra-clean workbench, then disinfecting for 7-8 min with a mercuric chloride solution with a concentration of 0.1%, flushing with sterile water for 5-6 times to fully remove residual mercuric chloride; cutting the disinfected leaves into small slices with an area of 0.5-1 square centimeter, inoculating to a bud induction medium, and putting in a culture room with a temperature of 23 DEG C for culture; and gradually performing subculture, wherein the illumination time in every day is 10-12 h, the illumination intensity is 1600-2000 Lx and the humidity is 85%-90%. By employing the tissue culture method provided by the invention, chirita wentsaii can rapidly grow, the leaves show bottle green and are thick, straight and abundant in luster, the plant type is tidy, bracts are pure and gorgeous in color, and chirita wentsaii is high in ornamental value.

Description

The tissue culture method of literary grace lip post lettuce tongue

Technical field

The invention belongs to Plant Tissue Breeding field, especially relate to the tissue culture method of a kind of literary grace lip post lettuce tongue.

Background technology

Literary grace Chirita, in Gesneriaceae (Gesneriaceae), has and views and admires and medical value, and leaf is kidney shape, and it is heart-shaped that base portion is, actinodrome, and the oblique mitriform of corolla, flower purple, sliver is circular, and stamen and staminodium and are born in the nearly base portion of corolla tube.Can be cultivated and be bred by sowing, cuttage, plant division and tissue, wherein front 3 kinds of modes be the simplest and conventional.

Summary of the invention

The problem to be solved in the present invention is to provide the tissue culture method of a kind of literary grace lip post lettuce tongue, to meet the needs of extensive ectogenesis literary grace lip post lettuce tongue.

For solving the problems of the technologies described above, the technical solution used in the present invention is: the tissue culture method of a kind of literary grace lip post lettuce tongue, remove the tender leaf blade of petiole, after flowing water rinses 30min, drop water purification part, the alcohol with 75% on superclean bench infiltrates 30s, then the mercuric chloride solution that is 0.1% by concentration sterilization 7-8min, through aseptic water washing 5-6 time, fully remove residual mercuric chloride.The fritter that the blade of the poison that disappeared is cut into 0.5-1 square centimeter is seeded on bud inducing culture, is placed on temperature and is in the culturing room of 23 ℃ to cultivate; And progressively subculture is cultivated.Illumination every day 10-12h, intensity of illumination 1600-2000Lx, humidity 85%-90%.

Used medium: inducing culture MS+2mg/L6-BA+0.2mg/L NAA, differential medium MS+1mg/L6-BA+0.1mg/L NAA, root media 1/2MS+0.2mg/L IBA.

Advantage and good effect that the present invention has are: adopt tissue culture method of the present invention can make literary grace lip lettuce tongue Fast Growth, and its blade is bottle green, blade is thick tall and straight, rich gloss; Plant type is neat; Bract color is pure gorgeous, and appreciative value is strong.

Embodiment

In order to understand the present invention, below by specific embodiment, the invention will be further described.Be divided into experimental group and control group, the tender leaf blade that 30 of each processing selecting are removed petiole is test material.

Experimental group experimental technique: remove the tender leaf blade of petiole, after flowing water rinses 30min, drop water purification part, alcohol with 75% on superclean bench infiltrates 30s, the mercuric chloride solution that is 0.1% by concentration again sterilization 7～8min, through aseptic water washing 5～6 times, fully removes residual mercuric chloride.The fritter that the blade of the poison that disappeared is cut into 0.5～1 square centimeter is seeded on bud inducing culture, is placed in culturing room and cultivates; And progressively subculture is cultivated.Illumination every day 10～12h, intensity of illumination 1600～2000Lx, temperature is (23 ± 1) ℃, humidity 85%～90%.

Example 1

Adopt inducing culture: MS+2mg/L6-BA+0.2mg/L NAA; MS+2mg/L6-BA+0.1mg/L NAA; MS+2mg/L6-BA+0.02mg/L NAA

Cultivation condition is illumination every day 10～12h, intensity of illumination 1600～2000Lx, and temperature is (23 ± 1) ℃, humidity 85%～90%.

Example 2

Adopt differential medium:

Adopt root media: 1/2MS+0.2mg/L MS+1mg/L6-BA+0.5mg/L NAA; MS+0.5mg/L6-BA+0.1mg/L NAA; MS+1mg/L6-BA+0.1mg/L NAA; Cultivation condition is illumination every day 10～12h, intensity of illumination 1600～2000Lx, and temperature is (23 ± 1) ℃, humidity 85%～90%.

Example 3IBA; 1/2MS+1.0mg/L IBA; MS+0.2mg/L IBA;

Result:

Evaluation metrics: comprise bud clump number, radical.

Analyze: as can be seen from Table 1, along with 6-BA/NAA increases, bud clump number is just larger.Work as 6-BA2mg/L, during NAA0.2mg/L, the inductivity of bud clump is the highest, and the bud of generation is maximum.Forming the optimum medium of bud clump is MS+2mg/L6-BA+0.2mg/L NAA+ agar+sugar, pH value 5.6～5.8.Young leaflet tablet is inoculated on above-mentioned appropriate media, cultivates after 30 days, blade can directly differentiate many Multiple Buds, inductivity 100%.The little Multiple Buds differentiating is cut apart, receive on differential medium MS+1mg/L6-BA+0.1mg/L NAA and continue to cultivate, within 15～20 days, get final product subculture once.

In table 2,6-BA can suppress the growth of root; IBA increases, and rooting rate increases; When medium is when MS becomes 1/2MS, rooting rate reaches 100%.

Adopt the blade of of the present invention group of Pei Wencai lip post lettuce tongue to be bottle green, blade is thick tall and straight, not sagging, glossy; Plant type is neat; Bract color is pure gorgeous, and appreciative value is strong.

The impact that table 16-BA and NAA form group training seedling bud clump

Table 26-BA and the IBA impact on group training seedling rooting

。

Claims

1. the tissue culture method of a literary grace lip post lettuce tongue, it is characterized in that: the tender leaf blade of removing petiole, after flowing water rinses 30min, drop water purification part, alcohol with 75% on superclean bench infiltrates 30s, the mercuric chloride solution that is 0.1% by concentration again sterilization 7-8min, through aseptic water washing 5-6 time, fully removes residual mercuric chloride.The fritter that the blade of the poison that disappeared is cut into 0.5-1 square centimeter is seeded on bud inducing culture, is placed on temperature and is in the culturing room of 23 ℃ to cultivate; And progressively subculture is cultivated.Illumination every day 10-12h, intensity of illumination 1600-2000Lx, humidity 85%-90%.

2. the tissue culture method of literary grace lip post lettuce tongue according to claim 1, it is characterized in that: used medium: inducing culture MS+2mg/L6-BA+0.2mg/L NAA, differential medium MS+1mg/L6-BA+0.1mg/L NAA, root media 1/2MS+0.2mg/L IBA.