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In vitro culture method for hybrid embryos of lilies

An in vitro culture, hybrid embryo technology, applied in horticultural methods, botanical equipment and methods, horticulture and other directions, can solve the problems of easy graying or browning and rot of roots, small swelling of bulbs, low survival rate, etc., and achieve easy peeling. , the number of rooting, the effect of high germination rate

Inactive Publication Date: 2011-07-20
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are examples of young embryos surviving in many reports, the bulbs are often small and expand slowly, and the root system is easy to appear gray or brown and rot. After the seedlings are transplanted, the survival rate is very low. Thick and stronger bulbs are the prerequisite for successful transplanting of seedlings

Method used

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  • In vitro culture method for hybrid embryos of lilies
  • In vitro culture method for hybrid embryos of lilies
  • In vitro culture method for hybrid embryos of lilies

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1) Pick the ovary obtained from the hybridization of 'Siberia'×'Happy Birthday' that was expanded 60 days after pollination, wash it with running water for about 40 minutes, then soak it in 75% alcohol for 30 seconds under sterile conditions, and rinse it with sterile water for 3 times, Soak in 0.1% mercuric chloride solution for 15 minutes, rinse with sterile water for 3 times, 5 minutes each time; (2) cut the ovary along the carpel line with a scalpel on a plate covered with filter paper under aseptic conditions ( figure 1 ), select seeds with embryos therein; (3) use a scalpel to gently cut a small opening in the seed coat, and then pick out the seed embryos with a dissecting needle, paying attention to maintaining the integrity of the embryos.

[0033] 2) Inoculating the stripped hybrid embryos on the most suitable in vitro germination medium, the specific steps are: (1) preparing the medium. The formula of the in vitro germination medium is MS+NAA 0.01mg / L+sucrose ...

Embodiment 2

[0036]1) Extract the ovaries obtained from the hybridization of 'Bernini'×'Leodo' and 'Leodo'×'Siberia' which were inflated 70 days after pollination, wash them with running water for about 50 minutes, and then under aseptic conditions Soak in 75% alcohol for 20s, rinse with sterile water for 5 times, then soak in 0.1% mercuric chloride solution for 10 minutes, rinse with sterile water for 5 times, each time for 5 minutes; Use a scalpel to cut the ovary along the carpel line, and select the seeds with embryos in it; (3) use a scalpel to gently cut a small opening in the seed coat, and then pick out the embryos with a dissecting needle, paying attention to keeping the embryos intact. integrity.

[0037] 2) Inoculating the stripped hybrid embryos on the most suitable in vitro germination medium, the specific steps are: (1) preparing the medium. The formula of the isolated germination medium is MS+NAA 0.1mg / L+sucrose 3%+agar 0.6%, pH 5.8, with 150ml Erlenmeyer flask, the volume ...

Embodiment 3

[0040] The hybrid seedlings obtained in Examples 2 and 3 were cultured in vitro for 4 months and transferred to the proliferation medium. The specific steps are: (1) Prepare the medium. The formula of the proliferation medium is MS+NAA0.1mg / L+6-BA2.0mg / L, 3% sucrose, 0.6% agar, pH 5.8. Use a 250ml orchid vase, and the volume of the medium in each orchid vase is about 60ml; (2) Under sterile conditions, cut off the leaves of the seedlings and inoculate them on the proliferation medium, and inoculate 4 to 5 A small bulb, placed at 22°C, cultured under light conditions, light intensity 1500Lx, light time 12h·d -1 . During the culture period, the medium was changed every 3 months.

[0041] The results showed that the multiplication rates of hybrid seedlings of ‘Bornini’בLeodo’, ‘Leodo’בSiberia’ and ‘Siberia’בHappy Birthday’ were all more than 5 times ( Figure 4 ).

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Abstract

The invention provides an in vitro culture method for hybrid embryos of lilies. The method comprises the following steps of: peeling a hybrid embryo from an ovary swollen by fertilization for in vitro germination, and multiplying bulblets of the germinated hybrid embryo. The method can also comprise the steps of inducing the multiplied lily bulblets to swell and inducing to take root. The embryo age of the hybrid embryo is 60 to 70d after pollination. The in vitro culture method for the hybrid embryos of the lilies has the characteristics that: immature embryos have low pollution rate and high survival rate, seedlings have multiple multiplied bulbs, the bulbs have high swelling speed, and the seedlings take multiple roots; and the method can be used for high-efficiency in vitro culture ofthe hybrid embryos of the lilies.

Description

technical field [0001] The invention relates to a method for in vitro culture of lily hybrid embryos, in particular to a method for stripping off, in vitro culture, multiplication culture and rooting culture of lily hybrid embryos at a specific embryonic age. Background technique [0002] The fertilized hybrid fruit cannot develop normally, but gradually shrinks and turns yellow. The reason is that the embryo and endosperm are not compatible, and the endosperm is aborted, which leads to embryo abortion. It must be cultured in vitro before the fruit turns yellow, that is, in the case of complete abortion of the hybrid endosperm, the artificially prepared medium is used to provide hybrid immature embryos Nutrients, so that it continues to grow and develop until it becomes a seedling. Ovary slice culture, ovule and hybrid embryo culture, and endosperm care culture are effective methods to overcome post-fertilization obstacles, among which embryo culture is the most widely used...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 吕英民程堂仁张启翔吴沙沙焦雪辉吴锦娣刘文娟
Owner BEIJING FORESTRY UNIVERSITY
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