AU2021101773A4 - Medium combination for rapid in vitro cultivation of actinidia chinensis seedlings - Google Patents

Abstract

The present disclosure relates to the technical field of Actinidia chinensis seedling cultivation, and discloses a medium combination for rapid in vitro cultivation of Actinidia chinensis seedlings, including a virus-free scion wood induction medium, a virus-free scion wood propagation medium, 5 a virus-free scion wood rooting medium, a tissue cultivation rootstock induction medium, a tissue cultivation rootstock propagation medium, and a tissue cultivation rootstock rooting medium. The present disclosure integrates the traditional grafting technology with the rapid virus-free seedling propagation technology. The medium combination, when used for Actinidia chinensis cultivation, realizes the healthy, rapid, and large-scale cultivation of virus-free seedlings and solves the problem 10 that traditional seedlings are susceptible to pathogenic bacteria and have underdeveloped root systems.

Description

MEDIUM COMBINATION FOR RAPID IN VITRO CULTIVATION OF A CTINIDIA CHINENSIS SEEDLINGS TECHNICAL FIELD

The present disclosure relates to the technical field of Actinidia chinensis seedling cultivation,

and in particular to a medium combination for rapid in vitro cultivation of Actinidia chinensis

seedlings.

BACKGROUND

Actinidia chinensis is an ancient vine deciduous fruit tree in the family of Actinidia. It is

currently one of the most successful plant domestication examples from wild to artificial

commercial cultivation. As a center of origin, evolution and distribution for Actinidia plants, China

is very rich in natural resources of Actinidia chinensis. With high nutritional value (especially rich

in vitamin C and the like), unique flavor, and high economic value, Actinidia chinensis fruit is

known as "King of Fruits" and "King of Vc" and is very popular among people. Compared with

other fruits, Actinidia chinensis cultivation and production has high economic benefits and a

promising future, which has become a high-efficiency industry with local characteristics and an

important channel for farmers in mountainous areas to increase income.

At present, breeding methods of Actinidia chinensis seedlings still mainly include grafting,

cutting, and seedling propagation methods. These methods have made great contributions to the production of Actinidia chinensis seedlings, but also show obvious limitations. Since Actinidia chinensis is highly heterozygous, gender differentiation will occur in offspring, and the offspring obtained through seedling propagation have various traits and cannot effectively retain excellent traits. It is difficult to reproduce a large number of seedlings in a short period of time by the traditional grafting and cutting methods due to a limited number of branches and low survival rate.

In addition, with the increase in the number of Actinidia chinensis cultivation years and the

Actinidia chinensis cultivation area in China, non-standard cultivation management techniques,

frequent disease infection, and increased hazard level have become the main factors that threaten

and restrict the development of the Actinidia chinensis industry. In particular, Actinidia chinensis

canker is devastating to the Actinidia chinensis industry. At present, there are no effective chemical

agents for the prevention and treatment of Actinidia chinensis virus diseases at home and abroad.

With the characteristics of high propagation coefficient, high efficiency, annual production, etc.,

plant tissue cultivation is widely used in the preservation of superior varieties and the expansion of

high-quality non-toxic seedlings, which is the future development direction of industrialized

agricultural seedling cultivation. In recent years, a huge progress has been made in the research on

tissue cultivation of Actinidia chinensis, but there are still some urgent problems to be solved, such

as serious explant browning and abnormal seedlings during the cultivation process. Therefore, the

research on key technologies for breeding Actinidia chinensis seedlings by tissue cultivation is of

important practical significance for solving the following problems: current shortage of seedling supply, low conventional propagation coefficient, variable quality, serious disease infection, variety deterioration, and the like. Therefore, the inventors provide a medium combination for rapid in vitro cultivation of Actinidia chinensis seedlings.

SUMMARY

Based on the above problems, the present disclosure provides a medium combination for rapid

in vitro cultivation of Actinidia chinensis seedlings. The medium combination realizes the healthy,

rapid, and large-scale cultivation of virus-free seedlings and solves the problem that traditional

seedlings are vulnerable to disease and have underdeveloped root systems, so as to obtain seedlings

with strong adaptability.

To solve the above technical problem, the present disclosure provides the following technical

solution.

The present disclosure provides a medium combination for rapid in vitro cultivation of

Actinidia chinensis seedlings, including a virus-free scion wood induction medium, a virus-free

scion wood propagation medium, a virus-free scion wood rooting medium, a tissue cultivation

rootstock induction medium, a tissue cultivation rootstock propagation medium, and a tissue

cultivation rootstock rooting medium.

The virus-free scion wood induction medium is based on an MS medium and further includes 3

mg/L 6-BA, 0.2 mg/L NAA, and 30 g/L sucrose; the virus-free scion wood propagation medium is based on anMS medium and further includes

0.5 mg/L 6-BA, 0.1 mg/L NAA, 0.5 mg/L A, and 30 g/L sucrose;

the virus-free scion wood rooting medium is based on a 1/2 MS medium and further includes

0.5 mg/L IBA, 0.3 mg/L IAA, and 20 g/L sucrose;

the tissue cultivation rootstock induction medium is based on an MS medium and further

includes 100 mg/L inositol and 20 g/L sucrose;

the tissue cultivation rootstock propagation medium is based on an MS medium and further

includes 1 mg/L 6-BA, 0.1 mg/L NAA, and 20 g/L sucrose; and

the tissue cultivation rootstock rooting medium is based on a 1/2 MS medium and further

includes 0.4 mg/L IBA and 20 g/L sucrose.

In the present disclosure, the medium combination described in the above technical solution is

used for in vitro cultivation of Actinidia chinensis, and a seedling cultivation method may

preferably be as follows:

A method for rapid in vitro cultivation of Actinidia chinensis seedlings may include the

following steps:

SI: Cultivation of virus-free scion wood

A female parent of virus-free scion wood is induced, propagated, and rooted by the virus-free

technology using shoot apical meristem (SAM) cultivation to obtain virus-free scion wood. A medium used at the induction stage is based on an MS medium and further includes 3 mg/L 6-BA,

0.2 mg/L NAA, and 30 g/L sucrose; a medium used at the propagation stage is based on an MS

medium and further includes 0.5 mg/L 6-BA, 0.1 mg/L NAA, 0.5 mg/L A, and 30 g/L sucrose; and

a medium used at the rooting stage is based on a 1/2MS medium and further includes 0.5 mg/L

IBA, 0.3 mg/L IAA, and 20 g/L sucrose.

S2: Cultivation of rootstock

Rootstock seeds are collected as explants, which are subjected to seed germination induction,

seedling stem sub-propagation cultivation, and rooting cultivation to obtain tissue cultivation

rootstock. An induction medium used at the seed germination induction stage is based on an MS

medium and further includes 100 mg/L inositol and 20 g/L sucrose; a propagation medium is based

on an MS medium and further includes 1 mg/L 6-BA, 0.1 mg/L NAA, and 20 g/L sucrose; and a

rooting medium is based on a 1/2 MS medium and further includes 0.4 mg/L IBA and 20 g/L

sucrose.

S3: Acquisition of virus-free seedlings

The virus-free scion wood obtained in step SI is grafted to the rootstock obtained in step S2 to

finally obtain virus-free seedlings.

Further, the female parent of virus-free scion wood in step S may be one or two of Actinidia

chinensis Planch. var. jinggangshanensis or Actinidia chinensis Planch.

Further, the rootstock species in step S2 may be Actinidia chinensis var. deliciosa.

Compared with the prior art, the present disclosure has the following beneficial effects: The

present disclosure integrates the traditional grafting technology with the rapid virus-free seedling

propagation technology, which realizes the healthy, rapid, and large-scale cultivation of virus-free

seedlings and solves the problem that traditional seedlings are susceptible to pathogenic bacteria

and have underdeveloped root systems. The female parent used in the present disclosure is a local

wild species distributed in Jinggang Mountain, and seedlings finally obtained have strong

adaptability.

DETAILED DESCRIPTION

In order to make the objectives, technical solutions, and advantages of the present disclosure

more apparent, the present disclosure will be further described in detail below with reference to

examples. The exemplary implementations and descriptions thereof in the present disclosure are

only used to explain the present disclosure, and are not intended to limit the present disclosure.

Example

A method for rapid in vitro cultivation of Actinidia chinensis seedlings included the following

steps:

SI: Cultivation of virus-free scion wood

A female parent of virus-free scion wood was induced, propagated, and rooted by the virus-free technology using SAM cultivation to obtain virus-free scion wood. In this example, the female parent of virus-free scion wood refers to one or two of Actinidia chinensis Planch. var.

jinggangshanensis and Actinidia chinensis Planch. These two female parents of virus-free scion

wood are wild Actinidia species in Jinggang Mountain, which are naturally distributed in the bushes

on Jinggang Mountain at an altitude of 200 m to 2,100 m and have strong adaptability and

high-quality fruits. A medium used at the induction stage is based on anMS medium and further

includes 3 mg/L 6-BA, 0.2 mg/L NAA, and 30 g/L sucrose; a medium used at the propagation stage

is based on an MS medium and further includes 0.5 mg/L 6-BA, 0.1 mg/L NAA, 0.5 mg/L A (A,

Adenine), and 30 g/L sucrose; and a medium used at the rooting stage is based on a 1/2MS medium

and further includes 0.5 mg/L IBA, 0.3 mg/L IAA, and 20 g/L sucrose.

S2: Cultivation of rootstock

Rootstock (Actinidiachinensis var. deliciosa (A.Chev.)) seeds were collected as explants, which

were subjected to seed germination induction, seedling stem sub-propagation cultivation, and

rooting cultivation to obtain tissue cultivation rootstock. An induction medium used at the seed

germination induction stage is based on an MS medium and further includes 100 mg/L inositol and

20 g/L sucrose; a propagation medium is based on an MS medium and further includes 1 mg/L

6-BA, 0.1 mg/L NAA, and 20 g/L sucrose; and a rooting medium is based on a 1/2 MS medium and

further includes 0.4 mg/L IBA and 20 g/L sucrose.

S3: Acquisition of virus-free seedlings

The virus-free scion wood obtained in step Si was grafted to the rootstock obtained in step S2

to finally obtain virus-free seedlings.

This example also achieves the collaborative operation of facilities and techniques such as

virus-free substrates, root control containers, fertilizer-water coupling, and pest control to cultivate

large root-controlled seedlings, with a transplantation survival rate up to 98% and a robust seedling

rate up to 95%. The virus-free seedlings cultivated in the present disclosure have developed root

systems and grow strong and healthy, which solves the prominent problem that traditional seedlings

are susceptible to pathogenic bacteria, have uncontrollable roots, and show a low survival rate in

field transplantation. Moreover, the present disclosure reduces a seedling cultivation cost by 1

yuan/plant and achieves the healthy, rapid and large-scale cultivation of commercial virus-free

seedlings.

The above is an example of the present disclosure. The above example and the specific

parameters in the example are provided merely for clearly expressing a verification process of the

present disclosure, not for limiting the protection scope of the present disclosure. The protection

scope of the present disclosure is still subject to the claims. Any equivalent structural changes made

from the content of the specification of the present disclosure should be included in the protection

scope of the present disclosure.

Claims (1)

CLAIMS:

1. A medium combination for rapid in vitro cultivation of Actinidia chinensis seedlings,

comprising a virus-free scion wood induction medium, a virus-free scion wood propagation

medium, a virus-free scion wood rooting medium, a tissue cultivation rootstock induction medium,

a tissue cultivation rootstock propagation medium, and a tissue cultivation rootstock rooting

medium; wherein,

the virus-free scion wood induction medium is based on an MS medium and further

comprises 3 mg/L 6-BA, 0.2 mg/L NAA, and 30 g/L sucrose;

the virus-free scion wood propagation medium is based on an MS medium and further

comprises 0.5 mg/L 6-BA, 0.1 mg/L NAA, 0.5 mg/L A, and 30 g/L sucrose;

the virus-free scion wood rooting medium is based on a 1/2 MS medium and further

comprises 0.5 mg/L IBA, 0.3 mg/L IAA, and 20 g/L sucrose;

the tissue cultivation rootstock induction medium is based on an MS medium and further

comprises 100 mg/L inositol and 20 g/L sucrose;

the tissue cultivation rootstock propagation medium is based on an MS medium and further

comprises 1 mg/L 6-BA, 0.1 mg/L NAA, and 20 g/L sucrose; and

the tissue cultivation rootstock rooting medium is based on a 1/2 MS medium and further

comprises 0.4 mg/L IBA and 20 g/L sucrose.