CN105706930A - Tissue culture two-step seedling development method of non-cralted leaf lagarosolen hispidus blades - Google Patents

Abstract

The invention provides a tissue culture two-step seedling development method of non-cralted leaf lagarosolen hispidus blades. The method includes the steps that tender non-cralted leaf lagarosolen hispidus blades are taken and serve as explants for germicidal treatment, then the blades are cut and then inoculated to an induction and differentiation medium, and 1-1.5 months later, aseptic seedlings which have two or more blades, as high as 1.5-3 cm and differentiated from the tender blades are selected and transplanted to a strong seedling rooting culture medium; when the number of roots of each aseptic seedling reaches four or more, and the root length reaches 2.5-4 cm, a bottle cap is opened, hardening-seedling is conducted in a room for 3-4 days, then the tissue culture seedlings are taken out of a culture bottle, the culture media are washed out, and the tissue culture seedlings are transplanted to the sterile culture media. The seedlings obtained through the method are strong and high in survival rate, a large number of high-quality non-cralted leaf lagarosolen hispidus seedlings can be provided in short time, the steps of preparing the culture media and a spinner bottle are omitted, a large quantity of manpower is saved, a great number of materials are saved, the tissue culture two-step seedling development method is simple and easy to operate, production cost is lowered, and the problems of scaled seedling culture and germplasm resource preservation of non-cralted leaf lagarosolen hispidus are effectively solved.

Description

A kind of entire leaf thin cylinder lettuce tongue vane group trains two step seedling methods

Technical field

The present invention relates to biological technical field, in particular it relates to a kind of entire leaf thin cylinder lettuce tongue vane group trains two step seedling methods.

Background technology

Entire leaf thin cylinder lettuce tongue is that the thin cylinder lettuce tongue of Gesneriaceae belongs to herbaceos perennial, and root stock is closely cylindrical, blade herbaceous stem, base portion subcircular or shallow heart, the full edge in edge, corolla purple, June at florescence, fruit June phase.China's Endemic and rare species, special product Longzhou, it to be born on Limestone Mountain sylvan life crag, height above sea level is about 530m.

Entire leaf thin cylinder lettuce tongue distributed areas are extremely narrow, only on the limestone cliff of area, Longzhou, find a small amount of plant at present, liquid manure is badly lacked due to growing environment, in addition its seed is tiny, sprout the temperature range needed, humidity range and soil acidity or alkalinity scope are smaller, nature is difficult to sexual propagation, it is badly in need of setting up and effectively breeds and protection system, it is contemplated that entire leaf thin cylinder lettuce tongue germ plasm resource is effectively bred and is protected by tissue culture technique blade two step seedling method, it is at present that entire leaf thin cylinder lettuce tongue nursery is the most convenient, stable hands section, substantial amounts of manpower can be saved, material resources and place, it is easy to kind of mass transter and transfer, avoid the anthropochory of harmful disease pest.

Summary of the invention

Present invention aim at providing a kind of entire leaf thin cylinder lettuce tongue vane group to train two step seedling methods, it is possible in the short time, provide a large amount of high-qualitys to be suitable for the entire leaf thin cylinder lettuce tongue seedling of cultivation.

To achieve these goals, the technical scheme is that

Entire leaf thin cylinder lettuce tongue vane group trains two step seedling methods, it is characterised in that:

(1) blade induction differentiation Adventitious bud culture: take entire leaf thin cylinder lettuce tongue young leaflet tablet, surface sterilization 30 ~ 45s in 75% ethanol, sterilized water 2 ~ 3 times, put into 0.1%Hgcl₂5min, sterilized water 2 ~ 3 times, 0.1%Hgcl₂3min, sterilized water rinses dry rear cutout for 2 ~ 3 times and is divided into 0.5cm × 1cm size to be seeded on induction and division culture medium, is 22 ~ 26 ° of C, intensity of illumination 1800 ~ 2600lux in temperature, when light application time is 8 ~ 10 hour/day, within 10 ~ 15 days, blade differentiates adventitious bud；

(2) adventitious bud strengthening seedling and rooting is cultivated: select entire leaf thin cylinder lettuce tongue blade to break up 1 ~ 1.5 month, have more than 2 leaves, high 1.5 ~ 3cm adventitious bud is inoculated into strengthening seedling and rooting culture medium, it is 22 ~ 26 ° of C in temperature, intensity of illumination 2000 ~ 2800lux, light application time is cultivated when being 8 ~ 10 hour/day, the aseptic seedling of 1 ~ 2 month paramount 4 ~ 6cm of adventitious bud length.

The seedling that employing the inventive method obtains is healthy and strong, survival rate is high; the high quality seedling of the thin cylinder lettuce tongue of a large amount of entire leaf can be provided in a short time; and decrease the step of preparation culture medium and rolling bottle; a large amount of manpower and materials are saved; therefore simple, production cost reduces, and efficiently solves entire leaf thin cylinder lettuce tongue scale breeding and Germ-plasma resources protection problem.

Detailed description of the invention

Below in conjunction with embodiment, technical scheme is further illustrated.

Entire leaf thin cylinder lettuce tongue vane group trains two step seedling method methods, comprises the following steps:

(1) blade induction differentiation Adventitious bud culture: take entire leaf thin cylinder lettuce tongue young leaflet tablet, surface sterilization 30 ~ 45s in 75% ethanol, sterilized water 2 ~ 3 times, put into 0.1%Hgcl₂5min, sterilized water 2 ~ 3 times, 0.1%Hgcl₂3min, sterilized water rinses dry rear cutout for 2 ~ 3 times and is divided into 0.5cm × 1cm size to be seeded on induction and division culture medium, is 22 ~ 26 ° of C, intensity of illumination 1800 ~ 2600lux in temperature, when light application time is 8 ~ 10 hour/day, within 10 ~ 15 days, blade differentiates adventitious bud.

The impact on adventitious bud number of table 1 hormon

Note: bud number during Shoot propagation multiple=(after 30 days bud number during bud number-inoculation)/inoculation

(2) adventitious bud strengthening seedling and rooting is cultivated: select entire leaf thin cylinder lettuce tongue blade 1 ~ 1.5 month high 1.5 ~ 3cm adventitious bud of differentiation to be inoculated into strengthening seedling and rooting culture medium, it is 22 ~ 26 ° of C in temperature, intensity of illumination 2000 ~ 2800lux, light application time is cultivated when being 8 ~ 10 hour/day, the aseptic seedling of 1 ~ 2 month paramount 4 ~ 6cm of adventitious bud length.

The impact on adventitious bud rooting effect of the table 2 hormon level

When the every strain of aseptic seedling take root more than 4, root length reach 2.5 ~ 4cm time, open bottle cap, after indoor seedling exercising 3 ~ 4 days, tissue cultured seedling taken out from culture bottle, washes away culture medium, be transplanted in the cultivation matrix of sterilizing.

Remarks: 6-BA(6-benayl aminopurine), NAA(naphthalene acetic acid), KT(kinetins), IAA (heteroauxing), AC (activated carbon).

Claims (1)

1. an entire leaf thin cylinder lettuce tongue vane group trains two step seedling methods, it is characterised in that:

(1) blade induction differentiation Adventitious bud culture: take entire leaf thin cylinder lettuce tongue young leaflet tablet, surface sterilization 30 ~ 45s in 75% ethanol, sterilized water 2 ~ 3 times, put into 0.1%Hgcl₂5min, sterilized water 2 ~ 3 times, 0.1%Hgcl₂3min, sterilized water rinses dry rear cutout for 2 ~ 3 times and is divided into 0.5cm × 1cm size to be seeded on induction and division culture medium, is 22 ~ 26 ° of C, intensity of illumination 1800 ~ 2600lux in temperature, when light application time is 8 ~ 10 hour/day, within 10 ~ 15 days, blade differentiates adventitious bud；

(2) adventitious bud strengthening seedling and rooting is cultivated: select entire leaf thin cylinder lettuce tongue blade to break up 1 ~ 1.5 month, have more than 2 leaves, high 1.5 ~ 3cm adventitious bud is inoculated into strengthening seedling and rooting culture medium, it is 22 ~ 26 ° of C in temperature, intensity of illumination 2000 ~ 2800lux, light application time is cultivated when being 8 ~ 10 hour/day, the aseptic seedling of 1 ~ 2 month paramount 4 ~ 6cm of adventitious bud length.