A kind of method for in vitro regeneration of immature embryos of hemp bamboo

An immature embryo, in vitro regeneration technology, applied in the field of plant tissue culture, can solve the problems of difficulty in disinfection and sterilization of shoot tip explants, low efficiency of embryogenic callus induction, low frequency of adventitious bud differentiation, etc. Effects of induction frequency and adventitious bud differentiation frequency, increasing adventitious bud differentiation frequency, and expanding explant types

Active Publication Date: 2022-07-29
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problem of this technology is that it is difficult to sterilize the shoot tip explants, and there is serious explant pollution, which is time-consuming and labor-intensive.
Secondly, the induction efficiency of embryogenic callus is low, the frequency of adventitious bud differentiation is low, and the cycle is long

Method used

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  • A kind of method for in vitro regeneration of immature embryos of hemp bamboo

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Embodiment 1

[0029] like figure 1 As shown, the present embodiment provides a method for the in vitro regeneration of immature embryos of hemp bamboo. The immature hemp seed (30 days after flowering) is surface sterilized with 75% alcohol for 60s, rinsed twice with sterile water, and then soaked in 0.1% mercuric chloride solution for 20 min, rinsed with sterile water for 3 times, the embryos were stripped and placed on callus induction medium with scutellum upward, and cultured at 25°C for 21 d in the dark to obtain callus. The induced callus was transferred to embryogenic callus induction medium, cultured at 25°C in the dark, and subcultured once every 21 d, and subcultured 3-4 times. The embryogenic callus was transferred to the differentiation medium, and the culture conditions were 25°C and 16 hours of light per day. Robust adventitious shoots were selected and transplanted into rooting medium. The culture temperature was 25°C, and the light was 16 hours a day. After 2 weeks, the ro...

Embodiment 2

[0036] Using immature embryos of hemp as explants, callus induction and embryogenic callus induction can be combined into one, and the efficiency of embryogenic callus induction can be increased from 17-38% to 47.3-61.7%. , the callus induction period was shortened from 2-4 months to 2 months, and the adventitious bud differentiation frequency also increased significantly, from 5.81-13.78% to 26.4-36.9%. In addition, since the immature embryo is still wrapped in the complete immature seed, the seed coat has a good protective effect on the embryo during sterilization, and can be completely sterilized without worrying about the survival of the embryo, which can effectively reduce pollution.

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Abstract

The invention discloses a method for in vitro regeneration of immature embryos of Bamboo rubra, comprising the following steps: 1) induction of callus of immature embryos of Bamboo vulgaris; 2) induction and proliferation of embryogenic callus of Bamboo vulgaris; 3) The differentiation of the embryogenic callus of Hempaba; 4) the rooting of the differentiated seedlings of Hemp; 5) the transplanting of the regenerated plants of Hemp. The invention uses immature embryos as explants, which solves the problem that in the past, it is difficult to judge the anther development period when the anthers are used as explants, the shoot tips are explants but not easy to sterilize, and embryogenic callus exists in both. Induce low frequency problems. Secondly, the callus induction medium and adventitious bud differentiation medium suitable for the immature embryos of Ma bamboo were established. It breaks through the callus induction and adventitious bud differentiation technology of different explant types, improves the frequency of adventitious bud differentiation, and expands the types of explants.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for in vitro regeneration of immature embryos of hemp bamboo. Background technique [0002] Hemp bamboo (Dendrocalamus latiflorus Munro) belongs to the Poaceae Bamboo family and is native to China. It is naturally distributed in Fujian, Taiwan, Guangdong, Guangxi, Guizhou, Yunnan and other provinces and regions. It is the southern and southeastern coastal areas of my country. It is one of the important high-quality, fast-growing and dual-purpose large-scale cluster bamboo species. The breeding of bamboo plant varieties is mainly based on the breeding of excellent clones in nature, and the genetic improvement is limited, and there is a lack of excellent new varieties that are urgently needed in production. At present, callus can be induced and regenerated plants can be obtained by using anthers and noded stem segments as explants, but there are st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001
Inventor 乔桂荣卓仁英黄碧芸邱文敏王雨珺
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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