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Method for high-efficiency cyclic regeneration of blackberry tissue culture seedling leaves

A technology of cyclic regeneration and tissue culture seedlings, which is applied in the field of plant tissue culture, and can solve problems such as the one-way one-round regeneration method

Active Publication Date: 2012-12-19
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on blackberry regeneration in vitro has achieved good results, but all of them are one-way one-round regeneration methods. There have been no reports at home and abroad that the blackberry seedlings can be regenerated permanently in the bottle by one-time collection outside the bottle.

Method used

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  • Method for high-efficiency cyclic regeneration of blackberry tissue culture seedling leaves
  • Method for high-efficiency cyclic regeneration of blackberry tissue culture seedling leaves
  • Method for high-efficiency cyclic regeneration of blackberry tissue culture seedling leaves

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Inducing Adventitious Buds from Stem Sections of Field Plants

[0043] Use the semi-lignified stems of "Kiowa" annual branches as explants, soak and rinse with concentrated laundry detergent, soak in 70% alcohol by volume for 45 seconds, rinse with sterile water for 3 to 5 times, and then rinse with 0.1% Soak in mercuric acid for 8 minutes; then cut the explants into (1.0-1.5) cm long bud stem segments, inoculate in MS+6-BA0.3mg / L+ZT0.5mg / L+sucrose 25g / L+agar 5.6g / L, on the culture medium of pH value 6.0, and at (25 ± 3) ℃, illuminance 2500lx, cultivate under the condition of illumination time 12h / d, the induction rate of adventitious buds after 30d is more than 85%, and each stem segment can produce 2 ~4 adventitious buds.

Embodiment 2

[0045] In Vitro Regeneration of Blackberry Tissue Cultured Leaves into Whole Plants

[0046] When the tissue culture seedling grows to more than 3 leaves, select the leaves with the longest side × the widest side being more than 0.5cm × 0.5cm, remove the edges, and trim them into leaf pieces of about 0.5cm × 0.5cm in size; Adhere to MS+TDZ 0.75mg / L+sucrose 25g / L+agar 5.6g / L, pH 6.2 culture medium, culture at (25±3)°C for 14 days in the dark, the edge of the leaf begins to curl Swell to produce a small amount of callus granules, then transfer to 1500 lx of light intensity, under the condition of 12 h / d of light time, after culturing for 20 days, light yellow and dense callus are formed, and the callus induction rate is 100%;

[0047] Remove the callus, cut into (0.3~0.6)cm×(0.3~0.6)cm blocks, inoculate into MS+6-BA1.0mg / L+IAA0.2mg / L+sucrose 30g / L+agar 5.6g / L, on the culture medium of pH value 5.8, cultivate under the conditions of (25±3)°C, 2000lx, light time 12h / d, the diffe...

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Abstract

The invention relates to a method for high-efficiency cyclic regeneration of blackberry tissue culture seedling leaves. The method for high-efficiency cyclic regeneration of blackberry tissue culture seedling leaves is characterized in that the leaves of a blackberry test-tube plantlet as an explant are induced to form callus tissues from which adventitious buds are induced and differentiated; the adventitious buds are subject to subculture proliferation, strong seedling culture and rooting induction; and finally hardening seedling and transplanting are carried out. Based on the method, the induction rate of callus tissues can reach 100 percent, the differentiation rate is more than 64.4 percent, the proliferation multiple of adventitious buds can reach 5.74, the rooting rate can maximally reach 100 percent, the average number of roots is 3.61, and the transplanting survival rate is 96.9 percent. Based on the method, the leaves of tissue culture seedling which are easy to obtain and have rich resources are used as the explant to establish the blackberry leaf in-vitro regeneration system, and meanwhile adventitious buds formed at different stages are used as the maternal plant to obtain leaves to form the blackberry leaf multiple cyclic in-vitro regeneration system, thereby solving the problem of difficult regeneration of blackberry tissue culture seedling leaves.

Description

technical field [0001] The invention relates to a method for repeated cycle regeneration of leaves of blackberry tissue culture seedlings, belonging to the technical field of plant tissue culture. technical background [0002] Blackberry (Rubus spp.) is a semi-shrub berry fruit tree of the Rosaceae Rubus genus. The fruit is an aggregate fruit, rich in nutrition, rich in sugar, fruit acid and multivitamins, and has the effects of anti-aging and improving human immunity. The content of superoxide dismutase (SOD) is the highest among fruits, and it is one of the third-generation fruits that have risen at home and abroad in recent years, and has high economic value. The cultivation and popularization of fine blackberry varieties has important application and economic value. At present, blackberry breeding mainly adopts conventional breeding methods such as seed selection, hybrid breeding, and radiation breeding, which have certain limitations, and the breeding process is slow, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 王小敏胡淑英吴文龙张春红李维林
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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