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Method for constructing lotus regeneration system

A construction method and system technology, applied in the field of plant tissue culture, can solve the problems of high pollution rate and high fatality rate, and achieve the effects of low pollution rate, guaranteed source, and low browning rate

Active Publication Date: 2015-08-05
镇江市彩林生态农业观光有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Kong Dezheng et al. (2007) found that direct sterilization of seed embryos obtained by corroding lotus seed shells with concentrated sulfuric acid will lead to a high pollution rate, and prolonged time will lead to a high mortality rate.

Method used

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  • Method for constructing lotus regeneration system

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A construction method of a lotus regeneration system is carried out in the following steps:

[0032] (1) The preparation of the medium, including the basic medium and the components of the medium at each stage and the weight per liter:

[0033] 1) Basic medium: MS;

[0034] 2) Sterile seedling medium: MS+6-BA 1mg / L+NAA 0.5mg / L+sucrose 30g / L, agar 8g / L, pH=5.8;

[0035] 3) Callus induction medium: MS+TDZ 0.1-0.2mg / L+NAA 0.7-0.8mg / L+sucrose 50g / L, agar 8g / L, pH=5.6,

[0036] 4) Adventitious bud induction medium: MS+6-BA 50μM+sucrose 30g / L, agar 8g / L, pH=5.6,

[0037] 5) Proliferation medium: MS+6-BA 1mg / L+NAA 0.5mg / L+sucrose 30g / L, agar 8g / L, pH=5.6

[0038] 6) Rooting medium: MS+IBA 1-2mg / L+ sucrose 30g / L, agar 8g / L, pH=5.6

[0039] (2) Sterilization and shell breaking:

[0040] Experiment 1: Change the order of shell breaking and sterilization, and set up the following two treatment groups:

[0041] Treatment group 1: The round and plump lotus seeds of "Guangchang...

Embodiment 2

[0072] (1) The preparation of the medium, including the basic medium and the components of the medium at each stage and the weight per liter:

[0073] 1) Basic medium: MS;

[0074] 2) Sterile seedling medium: MS+6-BA 1mg / L+NAA 0.5mg / L+sucrose 30g / L, agar 8g / L, pH=5.8;

[0075] 3) Callus induction medium: MS+TDZ 0.1-0.2mg / L+NAA 0.7-0.8mg / L+sucrose 50g / L, agar 8g / L, pH=5.6,

[0076] 4) Adventitious bud induction medium: MS+6-BA 50μM+sucrose 30g / L, agar 8g / L, pH=5.6,

[0077] 5) Proliferation medium: MS+6-BA 1mg / L+NAA 0.5mg / L+sucrose 30g / L, agar 8g / L, pH=5.6

[0078] 6) Rooting medium: MS+IBA 1-2mg / L+ sucrose 30g / L, agar 8g / L, pH=5.6

[0079] (2) In March 2014, the round and plump seeds of 'Guangchang Bailian' were sterilized with 75% alcohol for 30 seconds, rinsed once with sterile water, soaked in 3% sodium hypochlorite for 8 minutes, and rinsed 3 times with sterile water. spare.

[0080] (3) Cut the lotus seeds with aseptic pruning shears, take out the lotus heart as an e...

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Abstract

The invention discloses a method for establishing a lotus regeneration system. The method comprises the following steps: breaking shells of sterilized Guangchang white lotus seeds by using pruning shears; taking out mature lotus nuts (embryos) as explants to culture aseptic seedlings; cutting 2-3mm of stem tips and folded tender leaves (with short petiole) as explants for inducing callus from the aseptic seedlings after 2 months; inducing and differentiating the callus to develop buds after 2 months; after 1 month, cutting off the black callus at the bottom, and proliferating the differentiated seedlings; after 3 months, inducing the differentiated seedlings to develop roots; and strengthening, domesticating and transplanting the rooted tissue culture seedlings after 1 month. By adopting the method, technical basis is provided for transgenic technology of lotus in China, and the inductive callus rate of lotus is increased and the callus is large and stable in size compared with a conventional technique; the propagation coefficients of adventitious buds are increased; as tender leaves of aseptic seedlings of lotus are innovatively taken as explants for inductive callus and are successfully differentiated to obtain seedlings, the material selection of lotus inductive callus is widened, and material guarantee is provided.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, and relates to a method for constructing a lotus regeneration system. Background technique [0002] The lotus (Nelumbo nucifera Gaertn.) is a perennial aquatic flower of the genus Nelumbo of the family Nelumbo. It is one of the ten traditional famous flowers in China. It has high ornamental value, economic value and unique cultural connotation, and is deeply loved by people. [0003] In May 2013, the whole genome sequence of the lotus was released, which put the transgenic research of the lotus on the work schedule. However, the genetic transformation and regeneration system of lotus, which is an important technical basis for transgenics, is still immature, which seriously restricts the development of lotus transgenic work. There are two ways of lotus regeneration reported at present. One way is to directly differentiate the shoot tip to form clustered seedlings, then proliferate, a...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 徐迎春戴增强盛少峰金奇江陈羡王彦杰
Owner 镇江市彩林生态农业观光有限公司
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