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Method for inducing loblolly pine embryonic callus, and dedicated culture medium thereof

An embryogenic callus and induction medium technology, applied in the field of plant tissue culture, can solve the problems of high cost, low efficiency, and difficulty in meeting production requirements, and achieve the effects of good embryogenicity, high efficiency, and improved yield and quality.

Active Publication Date: 2020-06-16
JIANGXI ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the research on vegetative propagation technology of Pine loblolly includes methods such as cutting, grafting, and conventional tissue culture, but the cost is high, the efficiency is low, and it is difficult to meet the production requirements.

Method used

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  • Method for inducing loblolly pine embryonic callus, and dedicated culture medium thereof
  • Method for inducing loblolly pine embryonic callus, and dedicated culture medium thereof
  • Method for inducing loblolly pine embryonic callus, and dedicated culture medium thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A method for inducing embryogenic callus of pine taeda, comprising the following steps:

[0024] 1. Collection of Explants

[0025] In mid-to-late July, take natural pollinated immature cones from the middle and upper periphery of the canopy of the taeda pine mother plant with good growth, no pests and diseases, and a large amount of fruit in the 1st generation seed garden of taeda loblolly, and bring them back to the laboratory 4 ℃ refrigerator on the day of collection save.

[0026] 2. Disinfection of explants

[0027] Take the immature taeda pine cones out of the refrigerator, peel off the immature seeds with branch shears, remove the seed wings and place them in a sterile inoculation bottle, and then put them on the ultra-clean workbench for disinfection. Disinfect with 75% alcohol for 30 seconds, rinse with sterile water for 3-5 times, then soak in 0.1% mercury liter for 7-8 minutes, shake the bottle continuously for 5-6 times, rinse with sterile water for 5-7 ti...

Embodiment 2

[0033] Example 2 Effects of different genotypes on the induction of embryogenic callus of Pine taeda

[0034] According to the operation method of Example 1, the collected explants of 6 different genotypes such as Rongshan 52 were placed on the induction medium for embryogenic callus induction, and 10 callus were inoculated in each dish. The induction results are shown in Table 1.

[0035] Table 1 Effect of different genotypes on induction of embryogenic callus

[0036]

[0037] It can be seen from Table 1 that there are significant differences in the induction rate of embryogenic callus among different genotypes. The highest induction rate of embryogenic callus is Rongshan 52, with an average induction rate of 63.33%, and the lowest is Wulin 1. The induction rate was only 10%, and there were significant differences in the induction rate of embryogenic callus among different genotypes, indicating that genotype is one of the important factors affecting embryogenic callus.

Embodiment 3

[0038] Example 3 Effects of different basal mediums on the induction of embryogenic callus of Pine taeda

[0039] According to the operation method of Example 1, the explants were inoculated into 6 different basal media such as LP, DCR, mWV5, LV, B5 and WPM for induction, and 1.0 mg·L -1 2,4-D, 0.5mg·L -1 6-BA, 500mg·L -1 Inositol, 450mg·L - 1 L-Glutamine, 500mg·L -1 Hydrolyzed casein, 30g·L -1 Sucrose and 7.5g·L -1 Table 1 shows the effects of carrageenan and different basal media on the induction of embryogenic callus.

[0040] The results in Table 2 show that the induction rate of embryogenic callus in DCR medium is the highest, reaching 34.00%; followed by WPM and LP medium, the induction rate of embryogenic callus is 15.00% and 10.00% respectively; while LV, mMV5 and The induction rate in B5 medium was low, and there was a significant difference between DCR medium and other media, indicating that basal medium was very important for inducing embryogenic callus.

[...

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Abstract

The invention discloses a method for inducing a loblolly pine embryonic callus, and a dedicated culture medium thereof and belongs to the technical field of plant tissue culture. The method is characterized in that an immature loblolly pin seed is put on an inducing medium for inducing culture and an embryonic cell line with rapid growth and strong vigor is obtained, wherein the inducing culture medium takes a DCR culture medium as a basic culture medium and is added with 2,4-D, 6-BA, NAA, ABA, brassinolide, L-glutamine, casein hydrolysate, inositol, and 2-(N-morpholine) ethanesulfonic acid monohydrate. The invention provides the stable and efficient method for inducing the loblolly pin embryonic callus, wherein the yield and quality of the embryonic callus are increased, subculture multiplication of the embryonic callus is normal, good embryonic natures are kept, and important technological support is provided for further implementation of studies in somatic embryo seedling culture, genetic transformation, germplasm innovation and the like.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and more specifically relates to a method for inducing embryogenic callus of taeda pine and a special culture medium thereof. Background technique [0002] Loblolly pine (Pinus taeda L.) is native to the southeastern United States. It is the most important fast-growing coniferous timber tree species among southern pine trees. Because of its excellent characteristics such as fast growth, wide application, and strong adaptability, it has been successfully introduced in my country. And it has become an important fast-growing and high-yielding tree species in the subtropical low mountain and hilly areas of my country. In the early 1980s, the genetic improvement of Pine loblolly began in my country, and a lot of work was carried out in the aspects of introduction and cultivation, provenance testing, seed orchard construction, progeny determination, early selection, etc., and achieved certa...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 杨春霞丁伟谷振军程强强杜强黄宝祥
Owner JIANGXI ACAD OF FORESTRY
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