Method for preparing corn type II embryogenic calli

A technology of embryogenic callus and corn, applied in the field of preparation of type II corn embryogenic callus, to achieve the effects of excellent agronomic properties, shortened cultivation, and easy regeneration

Inactive Publication Date: 2011-10-19
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is commonly used in the world to distinguish between type I callus and type II callus, and the two are easy to distinguish

Method used

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  • Method for preparing corn type II embryogenic calli
  • Method for preparing corn type II embryogenic calli
  • Method for preparing corn type II embryogenic calli

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1. Preparation and verification of a maize inbred line-NGY302 capable of producing type II embryogenic callus

[0040] 1. Preparation and verification

[0041] (1) Preparation

[0042] 1. Selfing the high-oil inbred line GY302 to obtain F1 generation seeds;

[0043] In mid-August 2009, the high-oil inbred line GY302 was self-crossed at the experimental station of China Agricultural University in Changping, Beijing, and the self-bred F1 generation seeds were obtained.

[0044] 2. Type II embryogenic callus induction:

[0045] 100 immature embryos of F1 seeds were taken for embryogenic callus induction, and 100 embryogenic callus were obtained, from which type II embryogenic callus was selected, and the induction rate of type II embryogenic callus was counted.

[0046] Embryogenic callus induction method: 10 days after field self-pollination of the high-oil material GY302, corn ears were sterilized with 10% sodium hypochlorite aqueous solution for 10 minutes, t...

Embodiment 2

[0106] Example 2, Preparation of type II embryogenic callus with NGY302

[0107] 1. Method I

[0108] The immature embryos of NGY302 were inoculated on the embryogenic callus medium, and cultured in a dark environment at 28°C for 2 weeks to obtain type II embryogenic callus;

[0109] The embryogenic callus culture medium is composed of N6 medium, 2,4-D, inositol, proline, sucrose, hydrolyzed casein and phytogel;

[0110] 2, the concentration of 4-D in the described induction medium is 2.2mg / l, the concentration of inositol in the described induction medium is 0.15g / l, the concentration of proline in the described induction medium is 2.8g / l, the concentration of sucrose in the induction medium is 30g / l, the concentration of hydrolyzed casein in the induction medium is 0.15g / l, the concentration of phytogel in the induction medium 8g / l.

[0111] N6 medium was purchased from Sigma, the product catalog number is C1416.

[0112] Results All the immature embryos used for culture...

Embodiment 3

[0125] Embodiment 3, contrast

[0126] A188 was disclosed in the document "Maize Somatic Cell Culture and Screening of Disease-Resistant Variants, 1993", and the public can obtain it from China Agricultural University.

[0127] In the summer of 2010, the seeds of A188 and NGY302 were planted in Beijing Shangzhuang Experimental Station respectively, and their agronomic traits were observed. Results The plants of A188 were short, the pollination and seed setting rate was about 80%, and the male and female flowering stages were uncoordinated, separated by 2 days; while the plants of NGY302 were strong, medium in height, and the pollination and seed setting rate was high, about 100%.

[0128] The immature grain embryos of A188 and NGY302 were planted on N6 medium respectively, placed in a dark environment at 28°C for 2 weeks, and observed whether the obtained callus was type I or type II.

[0129] In the same N6 culture, A188 induced 95% callus between type I and type II; NGY302 ...

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Abstract

The invention discloses a method for preparing corn type II embryogenic calli. With the method, a corn inbred line which can produce type II embryogenic calli can be prepared. the method comprises the flowing steps: carrying out selfing among an initial corn inbred line to obtain seeds of an F1 generation; carrying out embryogenic callus induction upon embryos of the F1 generation seeds, such that embryogenic calli are obtained; collecting type II embryogenic calli; carrying out regenerating and cultivating upon the type II embryogenic calli until corn plants are obtained. The obtained corn plants form a target corn inbred line. With type II callus phenotype, the corn material provided by the present invention is easy to regenerate, and the regenerated plants have excellent agronomic properties. According to the present invention, a novel material for present transgenic acceptors is provided, a limitation that presently only non-inbred line Hill are utilized is solved, and the period for cultivating new species of corn by using transgenic technologies is shortened.

Description

technical field [0001] The invention relates to a method for preparing type II embryogenic callus of maize. Background technique [0002] Most maize inbred lines can form dense type I callus, which is poor in embryonic quality and difficult to regenerate plants after subculture; crisp type II callus grows rapidly and can be subcultured and maintained for a long time embryonic. Although the induction of maize type II callus is greatly restricted by genotype, it is still one of the most widely used recipient materials because of its relatively easy to obtain, strong regeneration ability, and relatively short cycle of obtaining transformed plants. one. [0003] It is now clear that the regeneration ability of explants is controlled by genetic factors, and the difference in the induction rate of embryogenic callus of different genotypes of immature embryos has been reported in a large number of papers. At present, most of the maize materials for Agrobacterium-mediated genetic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 陈绍江孟玉杰
Owner CHINA AGRI UNIV
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