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Method for inducing embryogenic cells by taking african agapanthus leaves as explant

A technology of embryogenic cells and explants, which is applied in botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of shortening the induction period of embryogenic cells and the difficulty of callus induction, and reduce the incidence of deformed seedlings occurrence, avoid the long-term subculture process, and induce the effect of rapid seedling growth

Active Publication Date: 2019-03-19
XINYANG AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

According to the distribution characteristics of the regenerative potential cells of monocotyledonous plants, the present invention utilizes leaf basal meristem tissue as explants to induce callus, breaking through the current domestic limitation that callus can only be induced by using small pedicels; the present invention Using monocotyledonous plant-specific auxin substance PIC as the main inducing hormone, it overcomes the problem of difficult callus induction of monocot plant leaves; Features: the present invention uses sucrose, glucose and maltose as carbon sources, and through the switching of types and concentration ratios, it changes the method of using only sucrose as a carbon source in traditional tissue culture and somatic embryo pathways, and improves callus and embryogenic callus. The induction rate of injured tissue shortened the induction cycle of embryogenic cells

Method used

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  • Method for inducing embryogenic cells by taking african agapanthus leaves as explant
  • Method for inducing embryogenic cells by taking african agapanthus leaves as explant
  • Method for inducing embryogenic cells by taking african agapanthus leaves as explant

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Embodiment 1

[0043] The present embodiment provides a method for inducing embryogenic cells using Agapanthus leaves as explants, and the specific steps are as follows:

[0044] Extraction of explants: the leaves of 2-year-old Agapanthus agapanthus at a distance of 0-2.0 cm from the junction of the rhizome were cut with a scalpel. Rinse the leaves of Agapanthus with running water for 90-150 minutes, place them on a clean bench, treat them with 75% (v / v) ethanol for 50-70 seconds, and wash them with ddH 2 Rinse with O for 3 to 5 times, then disinfect with 5% sodium hypochlorite for 5 to 7 minutes, then ddH 2 Rinse with O for 3 to 5 times, then treat with 75% ethanol for 50 to 70 seconds, and wash with ddH 2 Rinse 3 to 5 times, dry the water on the surface of the leaves with sterile filter paper, cut off the peripheral tissues of the leaves (including the tissues at the upper and lower ends and 2 to 3 wrapped old leaves) with a scalpel, and cut off the obtained leaves with a scalpel. The ba...

Embodiment 2

[0052] The present embodiment provides a method for inducing embryogenic cells using Agapanthus leaves as explants, and the specific steps are as follows:

[0053] Extraction of explants: the leaves of 2-year-old Agapanthus agapanthus at a distance of 0-2.0 cm from the junction of the rhizome were cut with a scalpel. Rinse the leaves of Agapanthus with running water for 90-150 minutes, place them on a clean bench, treat them with 75% (v / v) ethanol for 50-70 seconds, and wash them with ddH 2 Rinse with O for 3 to 5 times, then disinfect with 5% sodium hypochlorite for 5 to 7 minutes, then ddH 2 Rinse with O for 3 to 5 times, then treat with 75% ethanol for 50 to 70 seconds, and wash with ddH 2 Rinse 3 to 5 times, dry the water on the surface of the leaves with sterile filter paper, cut off the peripheral tissues of the leaves (including the tissues at the upper and lower ends and 2 to 3 wrapped old leaves) with a scalpel, and cut off the obtained leaves with a scalpel. The ba...

Embodiment 3

[0061] The present embodiment provides a method for inducing embryogenic cells using Agapanthus leaves as explants, and the specific steps are as follows:

[0062] Extraction of explants: the leaves of 2-year-old Agapanthus agapanthus at a distance of 0-2.0 cm from the junction of the rhizome were cut with a scalpel. Rinse the leaves of Agapanthus with running water for 90-150 minutes, place them on a clean bench, treat them with 75% (v / v) ethanol for 50-70 seconds, and wash them with ddH 2 Rinse with O for 3 to 5 times, then disinfect with 5% sodium hypochlorite for 5 to 7 minutes, then ddH 2 Rinse with O for 3 to 5 times, then treat with 75% ethanol for 50 to 70 seconds, and wash with ddH 2 Rinse 3 to 5 times, dry the water on the surface of the leaves with sterile filter paper, cut off the peripheral tissues of the leaves (including the tissues at the upper and lower ends and 2 to 3 wrapped old leaves) with a scalpel, and cut off the obtained leaves with a scalpel. The ba...

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Abstract

The invention provides a method for inducing embryogenic cells by taking african agapanthus leaves as explant, comprising the following steps: A, taking the leaf stalk bases of african agapanthus, washing and disinfecting, removing old leaves wrapping outside, and cutting off tissues at the upper and lower ends, so as to obtain leaf stalk base tissues; B, dicing the leaf stalk base tissues and then inoculating into a callus induction culture medium for culture; C, putting callus blocks obtained after culture in step B onto a callus subculture culture medium for subculture; and D, putting the callus blocks obtained after culture in step C onto an embryogenic callus induction culture medium for culture, so as to obtain the callus blocks containing mbryogenic cells. According to the invention, induction of the mbryogenic cells is successfully performed on the leaf stalk base tissues, the callus induction rate reaches 92.60%, the mbryogenic cell induction rate reaches 76.36%, and the induction cycle is only 6-7 months. The mbryogenic cell inducing seedling formation is rapid, a long-time subculture process is avoided, and occurrence of abnormal seedlings is reduced.

Description

technical field [0001] The invention relates to a method for inducing embryogenic cells using leaves of Agapanthus lilifolia as explants. Background technique [0002] Agapanthus 'Big Blue' (Agapanthus praecox ssp. orientalis 'Big Blue'), also known as "Blue Lily" and "African Lily", is native to southern Africa. It is a monocot perennial herb flower with strong ornamental value. For nearly half a century, Agapanthus has stood out in the development of the international flower industry and has become a popular fresh-cut flower, potted plant and ground cover flower all over the world, and it has shown excellent ornamental value. In addition, Agapanthus is extremely resistant, and has huge room for development in the fields of road greening and soil restoration. Currently, the supply of seedlings in the market exceeds demand. [0003] Agapanthus is commonly propagated by seeds or divisions in South Africa, the original place of origin, but after being introduced into China, t...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 岳建华董艳李文杨李露露刘文杰魏真史亚冰
Owner XINYANG AGRI & FORESTRY UNIV
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