A method for somatic embryogenesis and plant regeneration of Pinus tabulaeformis

A technology of somatic embryos and pine tabulaeformis, applied in the directions of plant regeneration, botanical equipment and methods, horticultural methods, etc., can solve the difficulty of cutting, the seedling propagation technology cannot meet the needs of large-scale afforestation, and the natural seed-setting rate of tabuliformis is low. and other problems, to achieve the effect of improving survival rate and growth, maintaining rapid proliferation ability, and promoting division

Active Publication Date: 2016-06-29
BEIJING FORESTRY UNIVERSITY
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Problems solved by technology

[0008] The main purpose of the present invention is to find a technology for somatic embryogenesis of Pinus tabulaeformis, which is a large-scale asexual of Pinus tabulaeformis, aiming at the current situation that the natural seed setting rate of Pinus tabulaeformis is low, cutting is difficult, and seedling propagation technology cannot meet the needs of large-scale afforestation. Breeding seedlings provides a method with short cycle, high reproduction rate and low cost

Method used

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  • A method for somatic embryogenesis and plant regeneration of Pinus tabulaeformis
  • A method for somatic embryogenesis and plant regeneration of Pinus tabulaeformis
  • A method for somatic embryogenesis and plant regeneration of Pinus tabulaeformis

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Embodiment 1

[0059] 1. Test materials

[0060] 1. Immature zygotic embryos of Pinus tabulaeformis

[0061] The present invention uses Beijing Forestry University forest farm (Haidian District, Beijing, longitude: 116.34, latitude: 40.00) good, mature, healthy, without pests, and 10 Chinese pine trees with high seed setting rate as material resources, from early May to June 2013 During the last ten days of the month, the cones of Pinus tabulaeformis in the 4-11th week after collecting loose powder were collected every 7 days. 30 cones were collected from each tree, and a total of 300 cones were collected each time. The cones collected on the day were stored in an ice box at low temperature and brought back to the laboratory, and then refrigerated in a refrigerator at 0‐4°C for 1-2 hours. month, spare.

[0062] 2. Plant growth regulator

[0063] The plant growth regulating substance used in the present invention adopts domestic 6-benzylaminoadenine (6-BA), 2,4-dichlorophenoxyacetic acid (...

Embodiment 2

[0086] 1. Sterilization of explants

[0087] The 30th, 37th, 44th, 51st, 58th, 65th, 72nd, and 79th days after the loose powder was stored at 0-4°C for 1 month, the cones of Pinus tabulaeformis were first washed with detergent to remove the oil on the surface of the cones, and then After washing with tap water, the cones were cut open, and the seeds were taken out; then on the ultra-clean workbench, the pine cone seeds were soaked in alcohol with a volume percentage concentration of 75% for 1 min, and then the seeds were soaked with 0.1% HgCl 2 Soak the pine cone seeds in the solution for 3-10min (preferably 5min), then rinse several times (3-6 times) with sterile water; then put the pine cone seeds on sterile dry filter paper to absorb the surface moisture Then peel off the seed coat in a sterile state to obtain immature zygote embryos, and obtain surface-sterilized immature zygote of Pinus tabulaeformis on the 30th, 37th, 44th, 51st, 58th, 65th, 72nd, and 79th days after flo...

Embodiment 3

[0100] 1. Sterilization of explants

[0101] The pinus tabulaeformis cones stored at 0-4°C for 1 month on the 51st day after loose powder were treated according to the explant sterilization method in Example 2 to obtain sterilized immature zygotic embryos of Pinus tabulaeformis;

[0102] 2. Embryogenic callus induction culture

[0103] In the ultra-clean workbench, inoculate the whole immature zygotic embryos of Pinus tabulaeformis 51 days after flowering and powder sterilized on the surface as explants in the embryogenic callus induction medium, and conduct embryogenic callus under dark conditions Induction culture, subculture once every 20 days, that is, take out the sterilized immature zygotic embryos after culture for 20 days, and place them in another fresh embryogenic callus induction medium to continue the induction of embryogenic callus In order to maintain sufficient nutrients and water in the medium, the culture temperature is (25±2)°C, the 2,4‐D used in the embryog...

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Abstract

The invention discloses a somatic embryogenesis and plant regeneration method for pinus tabulaeformis. The somatic embryogenesis and plant regeneration method comprises the following steps: 1) collecting pinus tabulaeformis fruits and performing surface sterilization to obtain sterile zygote embryos; 2) inoculating an embryonic callus induction culture medium with the sterile zygote embryos for callus induction culture; 3) inoculating a somatic embryo maturing culture medium with the embryonic calli for somatic embryogenesis culture; 4) inoculating a germination culture medium with mature somatic embryos for germination culture to obtain somatic embryo seedlings; 5) training and transplanting the somatic embryo seedlings to obtain pinus tabulaeformis plants. By an inducing method, a large number of stably growing pinus tabulaeformis somatic embryos can be obtained by inducing the embryonic calli for the first time, and complete small plants are obtained. In the method, the callus induction rate, the somatic embryogenesis rate and the germination rate are high; a large number of excellent pinus tabulaeformis test-tube seedlings can be formed within a short time; large-scale and industrial production can be carried out.

Description

technical field [0001] The invention relates to a method for plant tissue culture, in particular to a method for in vitro culture and plant regeneration of Pinus tabulaeformis, and belongs to the technical field of cell engineering seedling breeding in the forestry industry. Background technique [0002] Chinese pine (Pinustabulaeformis C.), also known as red bark pine, jack pine, etc., is a coniferous evergreen tree of the genus Pinus in the family Pinaceae. Pinus tabulaeformis is a unique conifer species in China and one of the most important afforestation tree species in North China. Its distribution area spans 14 provinces. Pinus tabulaeformis has a graceful posture, with jagged trunks and rugged sides. It is always spring in four seasons and is not afraid of wind, snow and severe cold. It has been praised as a vigorous, upright and unyielding sentiment since ancient times. Due to the well-developed root system, Pinus tabulaeformis has the advantages of resistance to dr...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 张金凤李慧赵健王晓琪付双彬康向阳罗晓芳张俊琦胡冬梅
Owner BEIJING FORESTRY UNIVERSITY
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