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Method for Inducing Callus Formation from Phyllostachys pubescens Seed Embryo

A technology of callus and moso bamboo, which is applied in the field of forestry biotechnology and tissue culture, can solve the problems of difficult material collection, high pollution rate, browning, etc., and achieve the effect of ensuring disinfection effect, reducing pollution rate and good growth state

Active Publication Date: 2017-03-22
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Regarding the research on the tissue culture of moso bamboo, according to the existing reports, the browning phenomenon is more serious when the new moso bamboo shoots are used as explants
Moreover, it is mostly limited by seasons and plant development periods, and it is difficult to obtain materials.
Relatively speaking, mature seeds have the advantages of easy storage, simple operation, and are not limited by factors such as seasons and plant development periods. The research is relatively convenient, but the callus induction rate of mature seeds is low (less than 50%) and the pollution rate is higher ( higher than 30%), become the bottleneck of moso bamboo cultivation technology

Method used

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  • Method for Inducing Callus Formation from Phyllostachys pubescens Seed Embryo

Examples

Experimental program
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Effect test

Embodiment 1

[0023] The method for moso bamboo seed embryo induction callus formation comprises the following steps:

[0024] (1) Screening of seeds

[0025] Get the bamboo seeds, peel off the lemma, use the TTC staining method to measure the vitality of the bamboo seeds, and screen the bamboo seeds with 80% vigor;

[0026] (2) Disinfection of seed embryos

[0027] Take out the seed embryo of the Phyllostachys pubescens seed that screens out, excise 70% endosperm, wash 2 hours with running water, then place 75% (v / v) ethanol and soak 2 minutes, rinse with sterile water again until the surface is smooth, the back groove There is no obvious foreign matter in the tank, then under aseptic conditions, soak and shake in 2% (v / v) sodium hypochlorite solution for 15 minutes, then rinse with sterile water until the surface is smooth and there is no obvious foreign matter in the groove on the back, and finally in the Under aseptic conditions, place in 0.1% (w / v) mercuric chloride with 4 drops of T...

Embodiment 2

[0033] The method for moso bamboo seed embryo induction callus formation comprises the following steps:

[0034] (1) Screening of seeds

[0035] Get the moso bamboo seeds, peel off the lemma, adopt the TTC staining method to carry out the viability measurement to the moso bamboo seeds, and screen the moso bamboo seeds whose birth vigor is 85%;

[0036] (2) Disinfection of seed embryos

[0037] Take out the seed embryo of the Phyllostachys pubescens seed that screens out, excise 50% endosperm, wash 3 hours with running water, then place 70% (v / v) ethanol and soak for 4 minutes, rinse with sterile water again until the surface is smooth, the back groove There is no obvious foreign matter in the tank, then under aseptic conditions, soak and shake in 1.5% (v / v) sodium hypochlorite solution for 18 minutes, then rinse with sterile water until the surface is smooth and there is no obvious foreign matter in the groove on the back, and finally in the Under aseptic conditions, soak an...

Embodiment 3

[0043] The method for moso bamboo seed embryo induction callus formation comprises the following steps:

[0044] (1) Screening of seeds

[0045] Get the moso bamboo seeds, peel off the lemma, and adopt the TTC staining method to measure the viability of the moso bamboo seeds, and screen the moso bamboo seeds whose vigor is 90%;

[0046] (2) Disinfection of seed embryos

[0047]Take out the germ embryo of the Phyllostachys pubescens seed that screens out, excise 60% endosperm, wash 4 hours with running water, place 72% (v / v) ethanol then to soak 3 minutes, rinse with sterile water again until the surface is smooth, the back groove There is no obvious foreign matter in the tank, then under aseptic conditions, soak and shake in 1% (v / v) sodium hypochlorite solution for 20 minutes, then rinse with sterile water until the surface is smooth and there is no obvious foreign matter in the groove on the back, and finally in the Under aseptic conditions, place in 0.08% (w / v) mercuric c...

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Abstract

A phyllostachys pubescens embryo induced callus formation method comprises the following steps: (1) screening out phyllostachys pubescens seeds with the vitality higher than 75%; (2) taking out embryos of the screened phyllostachys pubescens seeds, cutting to remove 50% or more than 50% of endosperm, conducting treatment by use of running water, ethanol, bacteria-free water, a sodium hypochlorite solution, bacteria-free water, mercury bichloride added with Tween-80 in sequence, and then removing surface moisture for later use; (3) inoculating disinfected embryos onto an induction medium, conducting dark culture for 7-10 days to form calluses, then separating the calluses out, inoculating the separated calluses onto a propagation medium for propagation culture for 20-30 days. According to the phyllostachys pubescens embryo induced callus formation method, as the embryos of the phyllostachys pubescens seeds are selected as explants, dirt, clay, bacteria and other pollutants, hidden in gaps between the testa and internal embryos of the phyllostachys pubescens seeds can be directly removed, and accordingly, the pollution rate is effectively reduced. The phyllostachys pubescens embryo induced callus formation method has the advantages that the callus inductivity can reach 64.3%, the pollution rate can be controlled at 20% or below, and the calluses are favorable in condition and excellent in growth situation.

Description

technical field [0001] The invention relates to the fields of tree biotechnology and tissue culture, in particular to a method for inducing callus formation by moso bamboo seed embryos. Background technique [0002] Moso bamboo (Phyllostachys Pubescens), also known as Nanzhu, Mengzong bamboo, Jiangnan bamboo, belongs to Gramineae (Gramineae), Bambusoideae (Bambusoideae), Phyllostachys (Phyllostachys) plants, perennial evergreen scattered bamboo plants, is a traditional Chinese Bamboo species are sold, which can be used for wood, food, ornamental and many other purposes. It also has the function of fixing soil and wind. It is widely used in building materials, construction, green food, medical care, furniture and agricultural tools, daily necessities, tourism handicrafts, cultural, sports and cultural equipment and the environment. Landscaping and other fields. [0003] The national moso bamboo forest is about 3 million ha 2 , accounting for about 65% of the total area of ​...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 项艳赵康李媛陈丹梅王依依
Owner ANHUI AGRICULTURAL UNIVERSITY
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