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Hybrid liriodendron somatic embryogenesis synchronization control method

A control method, the technology of Liriodendron body, applied in the field of synchronization control of somatic embryogenesis of hybrid Liriodendron, to achieve the effects of improving synchronization, reducing difficulty, and shortening culture time

Active Publication Date: 2012-04-11
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chen Zhi et al. (2007) also preliminarily established the embryogenic callus suspension culture of hybrid Liriodendron tulipifera by studying some physical factors that affect suspension culture, such as shaking speed, initial cell density, subculture cycle, and selection of suspended matter. system, but the establishment of a high-frequency, synchronized somatic embryogenesis suspension culture system of Liriodendron chinensis has not been reported

Method used

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  • Hybrid liriodendron somatic embryogenesis synchronization control method
  • Hybrid liriodendron somatic embryogenesis synchronization control method
  • Hybrid liriodendron somatic embryogenesis synchronization control method

Examples

Experimental program
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Effect test

Embodiment 1

[0056] A kind of hybrid Liriodendron somatic embryogenesis synchronization control method, as attached figure 2 The operation route shown includes the following steps:

[0057] (1) Take about 2 g of embryogenic callus preserved on solid medium and inoculate it in a 250 ml Erlenmeyer flask, add 50 ml of induction medium I, place it on a shaker, 95 r min -1 , cultured in dark, subcultured every other week with a volume ratio of 1:9 culture and induction medium I, and continuously subcultured for 2 to 3 weeks to obtain the initial material. The initial material was fully shaken to form a uniform particle, such as image 3 As shown in A.

[0058] (2) Pour the initial material processed in step (1) into a 50ml graduated cylinder, automatically settle for 3mins, pour off the supernatant, add induction medium II with a volume ratio of 1:9 culture and medium ratio, and transfer it into a 250ml Erlenmeyer flask In the same step (1) culture conditions, continue to cultivate for 1 wee...

Embodiment 2

[0063] According to the method operation step (1), (2) of embodiment 1, step (3) can also adopt following operations:

[0064] For cell clusters of 38-150 μm, the method of 18% Ficoll density gradient centrifugation or direct centrifugation of culture medium can be continued to be used to remove tubular empty cells and loose cell clusters, and dense cell clusters should be taken to adjust the density to 500-600 μm. The cell mass / mL was transferred to the solid medium covered with filter paper for culture, and 2-10 mg·L -1 ABA promotes the maturation of somatic embryos and inhibits the generation of abnormal embryos. At the same time, the cell clusters after sieving and centrifugation can also be continuously cultured in liquid maturation medium for about two weeks, and then sieved a second time, and the embryogenic cell clusters of 150-280 μm are transferred to semi-solid maturation medium for culture. The formula of the mature medium is: MS medium, 2~10mg·L -1 ABA, 100mg·L ...

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Abstract

The invention discloses a hybrid liriodendron somatic embryogenesis synchronization control method. In the method, 2,4-D concentration is reduced gradually by utilizing a growth hormone regulating mechanism; kinetin (KT) with the dose of 0.5 mg.L<-1> is added to induce uniform embryonic cell masses; the embryonic cell masses are subjected to fractional culture by a mechanically sieving method; and different subsequent synchronization control methods are adopted according to different cell mass sizes and somatic embryogenesis modes. After the synchronization control, the somatic growth synchronization ratio of hybrid liriodendron subjected to classified screening at the stage of globular embryo reaches about 90 percent; the synchronization ratio of fractional treatment at the stages of heart-shaped embryo and torpedo-shaped embryo is about 40 percent; after a proper period of refrigeration, the synchronization ratio is increased to some extent; and the conversion rate of a mature embryo, which develops from a material at the stage of globular embryo, to a regeneration plant can reach 70 to 80 percent.

Description

technical field [0001] The invention relates to the field of biological technology of seedling propagation in forestry, in particular to a method for synchronous control of somatic embryogenesis of hybrid Liriodendron tulipifera. Background technique [0002] There are only two species of Liriodendron (Liriodendron) in the Magnoliaceae family, namely, North American Liriodendron (L.tulipifera Linn.) and Liriodendron (L.chinense (Hemse.) Sarg), and the existing resources are very scarce. It is a second-class precious and endangered tree species in my country. Liriodendron has the characteristics of rapid growth, straight stem shape, wide use of wood, and high ornamental value of leaves and flowers. In particular, Professor Ye Peizhong from Nanjing Forestry University and others successfully obtained hybrid Liriodendron ( Liriodendron chinense×L.tulipifera), has obvious heterosis, fast growth, strong resistance, beautiful tree posture, beautiful flower color, unique leaf shape...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 施季森李婷婷陈金慧龙伟
Owner NANJING FORESTRY UNIV
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