Somatic embryogenesis and plant regeneration method for Picea asperata Mast

A technology for spruce spruce and embryogenesis, which is applied in plant regeneration, botanical equipment and methods, horticultural methods, etc., can solve problems such as no literature reports, achieve optimization of culture conditions, maintain embryogenic ability, and reduce loss of embryogenicity. The effect of risk on ability

Inactive Publication Date: 2013-12-25
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are no literature reports at home and abroad on the methods of somatic embryogenesis and plant regeneration of Picea twigs

Method used

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  • Somatic embryogenesis and plant regeneration method for Picea asperata Mast
  • Somatic embryogenesis and plant regeneration method for Picea asperata Mast
  • Somatic embryogenesis and plant regeneration method for Picea asperata Mast

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Isolation of immature zygotic embryos: collect immature cones produced by free pollination around May 1, and start collecting immature cones in early June, collecting once every 7 to 10 days, a total of 15 collections, 12 genotypes , Place the harvested cones at a low temperature at 4°C. The cones were treated with absolute ethanol for 15 minutes, and washed twice with sterile water. Under sterile conditions, remove the seeds and peel off the seed coat and endosperm, and immature zygotic embryos (such as figure 1 shown) to induce embryogenic callus.

Embodiment 2

[0060] Embryogenic callus induction stage: The immature zygotic embryos obtained in Example 1 are subjected to embryogenic callus induction, using 1 / 2LM basic medium, and the concentrations of additional hormones are shown in the following table:

[0061] Processing number

2,4-D / μM

6-BA / μM

1

5

2.5

2

5

5

3

5

10

4

10

2.5

5

10

5

6

10

10

7

15

2.5

8

15

5

9

15

10

[0062] And, add 1g·L -1 The natural complex enzymatically hydrolyzes casein, the medium contains 1% sucrose, 500mg·L -1 glutamine and 0.2% plant gel (gellan gum, Sigma). Adjust the pH to 5.8 before adding the gel, and then sterilize for 20 minutes at 121° C., pressure 101 kp, high temperature, and high pressure. About 35ml of semi-solid medium is contained in a glass Petri dish with a diameter of 90mm. The culture temperature is 24±1°C, cultured in dark condition...

Embodiment 3

[0066] Embryogenic callus induction stage: The immature zygotic embryos obtained in Example 1 were subjected to embryogenic callus induction, using 1 / 2LM basic medium, and the concentrations of additional hormones were 2, 4-D10 μM and 6-BA5 μM, respectively, adding 1g·L -1 The natural complex enzymatically hydrolyzes casein, the medium contains 1% sucrose, 500mg·L -1 of glutamine and 0.2% vegetable gel. Before adding the gel, adjust the pH to 5.8, then sterilize at 121°C, pressure 101kp, high temperature, and high pressure for 20 minutes, culture at 24±1°C, and culture in the dark until embryogenic callus is induced. Under the best collection date, the average induction rate of multiple genotypes reached over 64%.

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Abstract

The invention relates to a somatic embryogenesis and plant regeneration method for Picea asperata Mast. The method comprises the following steps: collecting freely-pollinated clonal cones of the Picea asperata Mast, and isolating immature zygotic embryos for somatic embryogenesis. The somatic embryogenesis comprises four phases: an embryonic callus induction phase, an embryonic callus proliferation phase, a somatic embryo mature phase and a somatic embryo germination phase. Compared with the conventional international research on the same or similar tree species, the frequency of embryonic callus induction is greatly increased, and the differentiation capacity and germination capacity of somatic embryos are also greatly increased. Meanwhile, a long-time keeping and proliferation method for embryonic callus is obtained, thus greatly reducing the risk of embryonic loss.

Description

technical field [0001] The invention relates to the field of cell engineering seedling propagation in forestry, in particular to a method for somatic cell embryogenesis and plant regeneration of spruce twigs. Background technique [0002] Picea asperata Mast (Picea asperata Mast) is a species of Picea in the family Pinaceae. It is commonly known as spruce in China, also known as Picea large fruit, and it is the most widely distributed species of Picea in my country. Evergreen tree, up to 45m, DBH up to 1m. Bark light grayish brown, cracked into irregular scales or slightly thick fast flakes off; branchlets have sparse or dense pubescence, or glabrous, light brownish yellow, brownish yellow or light reddish brown when annual, leaf pillows with White powder, or white powder is not obvious; winter buds are conical, resinous, and swollen at the base. Coniferous quadrangular strips, 1-2cm long, 1-1.5mm wide, slightly curved, slightly pointed or acute at the apex, quadrangular i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 王军辉张建伟张守攻
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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