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Seedling tissue culture method of Camellia chekiangoleosa Hu immature embryo

A technology for safflower camellia and immature embryos, which is applied in the field of plant bioengineering, can solve the problems of low natural reproduction rate, unfavorable mining and protection of safflower camellia resources, low induction rate and differentiation rate, etc., achieves reasonable hormone matching and improves rooting. rate and transplanting survival rate, and the effect of shortening the seedling cycle

Active Publication Date: 2019-12-24
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the low natural reproduction rate, which is not conducive to the excavation and protection of safflower camellia resources, there is an urgent need for a new cultivation method that can quickly cultivate Zhejiang safflower camellia in large quantities
However, when the young embryos of safflower camellia oleifera are cultivated by the existing combined seedling raising method, the induction rate and differentiation rate are relatively low.

Method used

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  • Seedling tissue culture method of Camellia chekiangoleosa Hu immature embryo
  • Seedling tissue culture method of Camellia chekiangoleosa Hu immature embryo
  • Seedling tissue culture method of Camellia chekiangoleosa Hu immature embryo

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Embodiment 1

[0036] The present embodiment provides a method for group seedling cultivation of immature embryos of safflower camellia oleifera, the steps are as follows:

[0037] 1) Explant selection and disinfection

[0038] From June to July, the fruits of Zhejiang safflower camellia oleifera 120-150 days after flowering were collected and brought back to the laboratory, and the young embryos were split open with a kitchen knife. Then select the seed coat color as white or white with rust red, soft or slightly hard immature embryos and put them into a beaker with sterilized water.

[0039] In the ultra-clean workbench, the immature embryos were sterilized with 75% (v / v) ethanol for 30 s, then washed 3 times with sterile water, then sterilized with 0.1% (m / m) HgCl2 surface for 8 min, and then rinsed with sterile water Wash 6-8 times.

[0040] 2) Callus induction

[0041] Use sterilized tweezers and a scalpel to peel off the seed coat of the immature embryo, cut the cotyledon into small...

Embodiment 2

[0054] This embodiment provides a method for group seedling cultivation of safflower camellia immature embryos. The steps and conditions are the same as in Example 1, except that the culture medium used is different. The culture medium used in this embodiment is specifically referred to as follows:

[0055] Callus induction medium includes MS medium, 0.2mg / L NAA, 1.5mg / L KT, 2.0mg / L TDZ, 70g / L sucrose, 8g / L agar, 500mg / L hydrolyzed casein , 500mg / L hydrolyzed milk protein, 100mg / L proline, 100mg / L glycine, 100mg / L glutamine, 10mg / L silver nitrate, 100mL / L coconut water, pH 5.6-5.8 .

[0056] Proliferation and differentiation medium include MS medium with macroelements reduced to 1 / 2, 0.05mg / L IBA, 2.0mg / L 6-BA, 0.005mg / LTDZ, 30g / L sucrose, 7g / L agar, 500mg / L hydrolyzed casein, 500mg / L hydrolyzed milk protein, 200mg / L proline, 200mg / L glycine, 200mg / L glutamine, 10.0mg / L silver nitrate, 100mL / L coconut water , pH is 5.8.

[0057] The rooting medium of cotyledon type somatic ...

Embodiment 3

[0059] This embodiment provides a method for group-cultivating seedlings of young embryos of safflower camellia oleifera, the steps of which are the same as in Example 1, except that the conditions of individual steps are different, and the culture medium used is different. The method used in this embodiment The different conditions and culture medium are specifically referred to as follows, and the experimental steps and conditions that are not clearly stated can be found in Example 1:

[0060] 1) Explant selection and disinfection

[0061] In the ultra-clean workbench, the immature embryos were sterilized with 75% (v / v) ethanol for 45 s, then washed 4 times with sterile water, then sterilized with 0.1% (m / m) HgCl2 surface for 10 min, and then rinsed with sterile water Wash 8 times.

[0062] 2) Callus induction

[0063] Callus induction medium includes MS medium, 0.2mg / L NAA, 0.8mg / L KT, 1.2mg / L TDZ, 50g / L sucrose, 8g / L agar, 500mg / L hydrolyzed casein , 500mg / L hydrolyzed ...

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Abstract

The invention relates to the field of plant bioengineering, in particular to a seedling tissue culture method of a Camellia chekiangoleosa Hu immature embryo. The method includes the steps: callus induction culture of fruit immature embryo cotyledons of Camellia chekiangoleosa Hu; multiplication and differentiation culture; rooting and habituated culture. Callus induction culture is implemented bythe aid of a dark culture mode, and a callus induction culture medium used by callus induction culture comprises a hormone combination of NAA, Kt and TDZ, amino acid, organic substances, anti-browning agents and 30-70g / L sucrose. According to the method, the induction rate and the differentiation rate of the Camellia chekiangoleosa Hu immature embryo are obviously increased, a lot of Camellia chekiangoleosa Hu seedlings can be rapidly cultured, and receptor materials are provided for transgenosis of the Camellia chekiangoleosa Hu.

Description

technical field [0001] The invention relates to the field of plant bioengineering, in particular to a method for group seedling cultivation of young embryos of safflower camellia oleifera. Background technique [0002] Safflower Camellia (Camellia Chekiangoleosa Hu), a plant of the genus Camellia in the family Theaceae. The oil content of the seed kernel of safflower camellia is as high as 61%, and the content of oleic acid in camellia oil is as high as 85%, which is significantly higher than that of ordinary camellia oleifera, Youxian camellia, small fruit camellia, Vietnam camellia and other white flower camellia species. [0003] Camellia oil is a unique woody edible oil plant in my country. It is rich in unsaturated fatty acids such as oleic acid and linoleic acid. It has the reputation of "Oriental olive oil". A functional edible oil with multiple health effects. Oleic acid is called "safe fatty acid" and is an important indicator for evaluating the quality of edible o...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01G31/00A01G24/28A01G24/15
CPCA01G31/00A01H4/00A01H4/001A01H4/008A01G24/15A01G24/28
Inventor 叶思诚卓仁英姚小华王开良
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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