Method for embryo rescue at different developmental stages of early phase of lilium formolongi distant hybridization

A new iron cannon lily and developmental stage technology is applied in the field of embryo rescue at different developmental stages in the early stage of distant hybridization of new iron cannon lily, and can solve problems such as single flower color and pattern, obstacles after fertilization, etc.

Active Publication Date: 2015-12-09
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the new iron gun lily has a single flower color and flower shape, which are white and trumpet-shaped respectively. In order to breed new lily varieties with a short growth period and rich flower colors and flower shapes, it is necessary to carry out distant hybridization with other lily hybrid varieties. This distant hybridization has serious post-fertilization obstacles, and effective early embryo rescue measures before embryo abortion 40 days after pollination can greatly increase the success rate of hybridization
At present, there are no relevant reports at home and abroad

Method used

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  • Method for embryo rescue at different developmental stages of early phase of lilium formolongi distant hybridization
  • Method for embryo rescue at different developmental stages of early phase of lilium formolongi distant hybridization
  • Method for embryo rescue at different developmental stages of early phase of lilium formolongi distant hybridization

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Experimental program
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Effect test

Embodiment 1

[0033] Example 1 Screening of ovary slice culture conditions in different periods of early pollination 5d-25d

[0034] 1. Materials and methods

[0035] (1) Extract the ovaries of the new Iron Pao Lily 'Augusta' as the female parent and the Oriental Lily 'Sorbonne' as the male parent, 5d, 10d, 15d, 20d, 25d after pollination; (2) the extracted ovaries Use a cotton ball dipped in alcohol with a volume percentage of 75% to wipe the surface for disinfection, soak in sodium hypochlorite with 1% available chlorine for 20 minutes, shake the container while soaking to make it fully contact; then rinse with sterile water 3 times, 2 minutes each time (3) cut the ovary into 2-3mm thin slices with a sharp scalpel and place them horizontally on the medium; (4) the medium formula for screening is basic medium B 5 , 1 / 2B 5 , MS, 1 / 2MS (pH=5.8, agar mass percentage concentration 0.8%) were added 3%, 6%, 9% or 12% sucrose with different mass percentage concentrations respectively to form a ...

Embodiment 2

[0041] Example 2 Ovary Slice Culture Obtained Embryo Ovule Culture Condition Screening in Vitro

[0042] 1. Materials and methods

[0043] (1) Inoculate the embryonic ovules obtained under the optimal conditions of ovary culture in different periods in Example 1 into corresponding and consistent medium to continue to cultivate; (2) The medium formula is consistent with the previous ovary slice culture, that is, basically Medium B 5 , 1 / 2B 5 , MS, 1 / 2MS (pH=5.8, agar mass percentage concentration 0.8%) were added with 3%, 6%, 9% or 12% sucrose with different mass percentage concentrations respectively to form a total of 12 treatment combinations for screening suitable for different The best medium formula for further culture of embryonic ovules obtained during this period; (3) The culture conditions are 25±1°C, dark culture, and the seedling formation of ovules is observed and counted.

[0044] 2. Results and Analysis

[0045] Table 2. Screening results of culture condition...

Embodiment 3

[0050] 35d-40d ovule direct in vitro culture condition screening after pollination of embodiment 3

[0051] 1. Materials and methods

[0052] (1) Extract the new iron gun lily 'Augusta' as the female parent, and the Oriental lily 'Sorbonne' as the male parent, and the ovary of 35d-40d after pollination; (2) the extracted ovary under sterile conditions Wipe the surface with 75% volume percent alcohol cotton ball for 3 times, soak in sodium hypochlorite with 1% available chlorine for 20 minutes, and rinse with sterile water for 3 times; Screening medium. (4) The formula of the screening medium is MS as the basic medium, and the mass percentage concentration of agar is 0.8%. Different concentrations of NAA, sucrose and medium pH are screened, and the results of different treatments are analyzed by range analysis and variance analysis . (5) The culture temperature is 25±1° C., dark culture, and each treatment is repeated 3 times, with 20-30 ovules per repetition.

[0053] 2. R...

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Abstract

The invention belongs to the field of tissue culture of plants and particularly provides a method for embryo rescue at different developmental stages of the early phase of lilium formolongi distant hybridization. The method comprises steps as follows: 1), after lilium formolongi distant hybridization, ovaries are taken to be sterilized and disinfected; 2), embryo rescue at different developmental stages is performed in 5-25 days after pollination as follows: the ovaries are sliced and cultured, ovules are obtained and subjected to tissue culture, and hybridized tissue culture seedlings are obtained; embryo rescue at different developmental stages is performed in 35-40 days after pollination as follows: in vitro culture of ovules or in vitro culture of embryos is performed, and the hybridized tissue culture seedlings are obtained. The invention further provides an application of the method for embryo rescue at different developmental stages of the early phase of lilium formolongi distant hybridization in lily breeding. According to the method for embryo rescue at different developmental stages of the early phase of lilium formolongi distant hybridization, the barrier of incompatibility after distant hybridization fertilization of lilium formolongi and lilium oriental hybrid lines can be overcome, and the ovules with the embryos and hybrid seedlings are obtained.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and in particular relates to a method for rescuing embryos of different developmental stages in the early stage of distant hybridization of Lilium chinensis. Background technique [0002] Lily hybrid breeding technology includes intra-species hybridization and inter-species hybridization. Inter-species hybridization is distant hybridization. There are not only obstacles before fertilization, such as pollen tubes cannot germinate normally in the style, but also serious obstacles after fertilization. Obtain hybrid seedlings. Obstacles after fertilization are mainly due to the inconsistency of the embryo or endosperm and the mother's physiological functions, resulting in the abnormal growth and development of the embryo after fertilization, and the abortion usually begins at about 40 days of development, resulting in the failure of hybridization. Embryo rescue technology of immature embryos is t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 贾桂霞李玉帆韩秀丽田晓明周艳萍张铭芳张锡庆汪莲娟崔祺高雪
Owner BEIJING FORESTRY UNIVERSITY
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