CN105660414A - Lagarosolen leave tissue culture two-step seedling culture method - Google Patents

Abstract

The invention provides a lagarosolen leave tissue culture two-step seedling culture method, which comprises the following steps of taking tender lagarosolen leaves as explants to be subjected to sterilization treatment; then, cutting the tender lagarosolen leaves and inoculating the cut tender lagarosolen leaves onto an induction and differential culture medium; after 1 to 1.5 months, selecting sterile buds and seedlings which have more than two leaves, are 1.5 to 3cm in height and are differentiated from the tender leaves to be transplanted onto a strong seedling rooting culture medium; after each strain of the sterile seedling grows more than 4 roots and the root length reaches 2.5 to 4cm, opening a bottle cap; after the indoor seedling domestication is performed for 3 to 4 days, taking out tissue culture seedlings from a culture bottle; washing out the culture medium; transplanting the seedlings into a sterilized culture substrate. The seedlings obtained by using the method are health; the survival rate is high; a great number of lagarosolen high-quality seedlings can be provided in a short period; the steps of preparing a culture medium and transferring bottles are reduced; a great amount of labor and materials are saved, so that simplicity is realized; the implementation is easy; the production cost is reduced; the lagarosolen scaled seedling and seed resource storage problems are effectively solved.

Description

A kind of Lu Shi 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 Lu Shi thin cylinder lettuce tongue vane group trains two step seedling methods.

Background technology

The thin cylinder lettuce tongue of Lu Shi is Gesneriaceae Chiritopsis plant, is the novel species found for 2008, and the thin cylinder lettuce tongue of Lu Shi is perennial herb, root stock cylinder, blade kidney shape or circle, peduncle, inflorescence and florescence are outer by wire pubescence and purple body of gland, the month at florescence 5-6, really June phase. China's Endemic and rare species, special product Guangxi Pingyue County, it to be born on Limestone Mountain rock, height above sea level is about 200-250m.

Lu Shi thin cylinder lettuce tongue distributed areas are extremely narrow, only on the limestone cliff of area, Guangxi Pingyue County, 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 Lu Shi 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 Lu Shi 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 Lu Shi 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 Lu Shi thin cylinder lettuce tongue seedling of cultivation.

To achieve these goals, the technical scheme is that

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

(1) blade induction differentiation Adventitious bud culture: take Lu Shi thin cylinder lettuce tongue young leaflet tablet, surface sterilization 30 ~ 45s in 75% ethanol, sterilized water 2 ~ 3 times, put into 0.1%Hcl₂5min, sterilized water 2 ~ 3 times, 0.1%Hcl₂3min, 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 in temperature, intensity of illumination 1800 ~ 2600lux, 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 Lu Shi 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; a large amount of Lu Shi high quality seedling of thin cylinder lettuce tongue 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 Lu Shi 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.

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

(1) blade induction differentiation Adventitious bud culture: take Lu Shi thin cylinder lettuce tongue young leaflet tablet, surface sterilization 30 ~ 45s in 75% ethanol, sterilized water 2 ~ 3 times, put into 0.1%Hcl₂5min, sterilized water 2 ~ 3 times, 0.1%Hcl₂3min, 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 Lu Shi 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

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

Claims (1)

1. a Lu Shi thin cylinder lettuce tongue vane group trains two step seedling methods, it is characterised in that:

(1) blade induction differentiation Adventitious bud culture: take Lu Shi thin cylinder lettuce tongue young leaflet tablet, surface sterilization 30 ~ 45s in 75% ethanol, sterilized water 2 ~ 3 times, put into 0.1%Hcl₂5min, sterilized water 2 ~ 3 times, 0.1%Hcl₂3min, 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 Lu Shi 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.