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A method for induction of somatic embryos and plant regeneration of Dutch pea
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A technology of somatic embryos and cell embryos, applied in the field of edible plant biology, to achieve good social and economic benefits
Active Publication Date: 2022-02-22
HEBEI NORMAL UNIVERSITY OF SCIENCE AND TECHNOLOGY
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However, domestic and foreign scholars' research on Dutch pea is limited to its cultivation, seed germination, cross breeding, resistant varieties and germplasm resource investigation, and the establishment of the somatic embryo regeneration system of Dutch pea has not been reported at home and abroad.
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Embodiment 1
[0035] Embodiment 1, the establishment of Dutch pea aseptic culture system
[0036] The test materials were young pods of snow peas purchased in supermarkets. Select well-grown Dutch pea pods and gently scrub the surface dirt under running water, then blot the water with filter paper and place them in a clean beaker. Soak in 75% alcohol for 30s, pour it off quickly, immerse in 0.1% mercuric chloride solution to sterilize for 10 minutes, wash 5-6 times with sterile water on the ultra-clean workbench, and cut off the edge of the pod after the surface of the inoculated material is drained Some of the immature embryos or cotyledons were taken as explants.
[0038] The immature embryos or cotyledons in Example 1 were cut into small pieces of 1.0-2.0 mm, and inoculated on the medium for inducing callus. The callus induction medium is MS+0.5mg / l BA+0.3mg / l IBA.
[0039] In addition, the medium contains 3% sucrose and 0.9% agar, and the pH is adjusted to 5.5-5.7 before adding agar, and then sterilized at 121° C. and pressure 101 kp under high temperature and high pressure conditions for 15 minutes. They were cultured for 10d, 20d, and 30d in the dark at a constant temperature of 25°C until the callus was induced.
[0040] After 15 days, the entire cotyledon gradually swelled, and wrinkles appeared at the incision. After 20 days, a loose, granular, milky yellow (milky white) callus began to be produced at the incision. After 30 days, the entire cotyledon callus produced a large amount of callus .
Embodiment 3
[0041] Embodiment 3, induction of embryogenic callus
[0042] The callus induced in Example 2 was inoculated on the embryogenic callus induction medium, and the embryogenic callus induction medium added different kinds of hormones in the MS medium, as follows:
[0043] (1) MS+4mg / l 2,4-D+2.0mg / l NAA
[0044] (2) MS+4mg / l 2,4-D+2.0mg / l BA
[0045] (3) MS+4mg / l 2,4-D+2.0mg / l KT
[0046] (4) MS+4mg / l 2,4-D+0.2mg / l NAA+0.5mg / l BA
[0047] (5) MS+4mg / l 2,4-D+0.2mg / l NAA+0.5mg / l KT
[0048] (6) MS+4mg / l 2,4-D+0.2mg / l NAA+1.0mg / l BA
[0049] (7) MS+4mg / l 2,4-D+0.2mg / l NAA+1.0mg / l KT
[0050] (8) MS+4mg / l 2,4-D+0.2mg / l NAA+2.0mg / l BA
[0051] (9) MS+4mg / l 2,4-D+0.2mg / l NAA+2.0mg / l KT
[0052] (10) MS+4mg / l 2,4-D+0.2mg / l NAA+3.0mg / l BA
[0053] (11) MS+4mg / l 2,4-D+0.2mg / l NAA+3.0mg / l KT
[0054] (12) MS 0
[0055] In addition, the medium contains 3% sucrose and 0.9% agar, and the pH is adjusted to 5.5-5.7 before adding agar, and then sterilized at 121° C. and pressure 101 kp u...
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Abstract
The invention relates to a method for inducing somatic embryos of Dutch pea and regenerating plants, and belongs to the field of edible plantbiotechnology. The present invention utilizes young embryos or cotyledons of Dutch pea to induce somatic embryos and establish regenerated plant clones. The steps include: 1. Aseptic treatment of Dutch pea pods and selection of explants; 2. Induction of callus; 3. Induction of embryogenic callus; 4. Occurrence and development of somatic embryos; 5. Proliferation and rooting of somatic embryos; 6. Transplantation into seedlings. The somatic embryo induction and plant regeneration method of the edible plant snow pea provided by the present invention lays a foundation for the accelerated industrial production of the fast propagation of the snow pea, and the research on the genetic transformation and gene improvement of the snow pea.
Description
technical field [0001] The invention relates to the field of edible plant biotechnology, and specifically provides a method for inducing somatic embryos of Dutch pea and regenerating plants. Background technique [0002] Dutch beans, also known as soft pod peas, pod-eating peas, green Dutch beans, small cold beans, Huai beans, hemp beans, green beans, Liu beans, golden beans, Huihui beans, etc., are vegetables in the legume family that are served with tender beans or tender pods . Snow pea also contains a kind of its unique plant lectin, branching element and gibberellin GA20, etc. These substances play an important role in enhancing the metabolic function of the human body. Snow pea has high economic value and will become a competitive commodity. Planned and scientific development and utilization will receive good social benefits, and its application prospects are broad. [0003] The development of plants through somatic embryos is a major advance in cellengineering. So ...
Claims
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Application Information
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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 韩金玲李向岭张敏王健杨晴杨敏
Owner HEBEI NORMAL UNIVERSITY OF SCIENCE AND TECHNOLOGY
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