Ascendens mucronatum somatic embryogenesis method
An embryogenesis and somatic cell technology, applied in the field of forest cell engineering, can solve the problems of long-term continuous production difficulties, long breeding cycle, low seedling emergence rate, etc., and achieve the effect of realizing mass reproduction, meeting market demand and improving induction rate.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0034] (1) Preparation of immature zygotic embryos of Zhongshan fir
[0035] From late August to mid-September, the immature cones of Zhongshan fir were collected every 7 days, and quickly stored in a refrigerator at 4°C. Take out the zygotic embryo and observe it under a dissecting microscope. Take the seeds that have developed to the early cotyledon embryo, soak them in 75% ethanol for 45-60 seconds, then sterilize them with 2.5% sodium hypochlorite solution for 18-20 minutes, and finally rinse them with sterile water for 3-60 seconds. 4 times, then under sterile conditions, peel off the hard seed coat outside the seeds with a sterile scalpel and tweezers under a dissecting microscope, take out the immature zygotic embryos, and inoculate them in solid embryogenic callus induction culture Inoculate 8-10 grains per culture dish. Zhongshan fir immature zygotic embryos such as figure 1 shown.
[0036] (2) Induction of embryogenic callus
[0037] Under sterile conditions, imm...
Embodiment 2
[0057] It is basically the same as Example 1, except that the induction solid medium in step (2) is: improved DCR basic medium+NAA 4.0mg / L+6-BA 1.5mg / L+L-Glu 0.3g / L, Pro 0.2g / L+Inositol 0.2g / L+Maltose25g / L+agar 8.5g / L, pH 5.6.
[0058] The maintenance and proliferation medium in step (3) is: 3 / 4 improved DCR basic medium+NAA2.0mg / L+6-BA0.5mg / L+KT 1.0mg / L+Pro 0.2g / L+Inositol 0.2g / L+CH 0.5g / L+Maltose 30g / L+agar 8.5g / L, pH 5.8.
[0059] The somatic embryo induction medium in step (4) is: the improved DCR medium is basic medium+ABA 2.0mg / L+6-furfuryl aminopurine (KT) 2.0mg / L+methyl jasmonate 0.2g / L+polyethylene Diol 8000 (PEG8000) 100g / L+Inositol 0.5g / L+LH 1g / L+Vc 15mg / L+Maltose 20g / L+agar 8.5g / L), pH 6.0. Other each step and control condition are all identical with embodiment 1, and the result shows, also can reach the induction effect of embodiment 1.
Embodiment 3
[0061] Basically the same as Example 1, the difference is that the somatic embryo induction medium in step (4) is: the improved DCR medium is basic medium+ABA 3.0mg / L+6-furfuryl aminopurine (KT) 2.0mg / L+ Methyl jasmonate 0.2g / L+ polyethylene glycol 8000 (PEG8000) 100g / L+Inositol 0.5g / L+LH 1g / L+Vc15mg / L+Maltose 20g / L+agar 8.5g / L), pH 6.0. Other each step and control condition are all identical with embodiment 1, and the result shows, also can reach the induction effect of embodiment 1.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 


