Simplified tilia cordata mill tissue culture method

A technology for tissue culture and induction medium, applied in the field of plant tissue culture, can solve the problems of affecting the growth of seedlings, virus accumulation, and high cost of grafting reproduction, and achieve the effects of easy promotion, low cost, high reproduction efficiency and versatility

Active Publication Date: 2021-07-02
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 European Tilia small-leaved cultivars are mainly propagated by grafting, but the cost of grafting is relatively high, and the accumulation of viruses is prone to affect the growth of seedlings.

Method used

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  • Simplified tilia cordata mill tissue culture method
  • Simplified tilia cordata mill tissue culture method
  • Simplified tilia cordata mill tissue culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1, the selection of explant

[0028] On April 2nd, April 12th, April 22nd, May 2nd and May 12th, the healthy plants of Tilia microphylla 'Xinle' were collected as explants in the same year, and compared with different sampling times. Effect of implant contamination rate and induction rate.

[0029] After the above sprouts are collected, cut off the leaves and keep the petiole of about 1 cm, shake and soak the obtained stems in 2%-5% alkaline detergent solution for 20 minutes, rinse with running water for 2 hours, and use 75 % alcohol for 15 seconds, sterilized with 2% hypochlorous acid for 5 minutes, and washed with sterile water for 5 times, each time for 8 minutes.

[0030] The parietal leaves and axillary buds were respectively inoculated on the induction medium, the formula was: MS+6-BA 1.0mg / L+IBA 0.2mg / L+sucrose 30g / L+agar 5.8g / L, the pH value was 5.9; each treatment was inoculated 50 explants were used, and the contamination rate and induction rate w...

Embodiment 2

[0034] Embodiment 2, disinfection mode screening

[0035] Around April 22, collect the sprouting strips of the healthy plants of European small-leaved linden 'Xinle' in the same year, cut off the top 1-2 buds of the sprouting strips, and remove the leaves to keep about 1 cm of petiole, and the obtained stems are 2%-5% Shake and soak in alkaline detergent solution for 20 minutes, rinse with running water for 2 hours, and then use the following 4 disinfection methods to disinfect the surface of explants on the ultra-clean workbench, with 50 explants per treatment. The specific disinfection methods are as follows: (1) disinfect with 75% alcohol for 15 seconds, disinfect with 2% sodium hypochlorite for 5 minutes, wash with sterile water for 5 times, each time for 8 minutes; (2) disinfect with 75% alcohol for 15 seconds, Disinfect with 2% sodium hypochlorite for 10 minutes, wash 5 times with sterile water, 8 minutes each time; (3) Disinfect with 75% alcohol for 15 seconds, disinfec...

Embodiment 3

[0039] Embodiment 3, induction medium screening

[0040] Around April 22, the sprouting strips of the healthy plants of Tilia microphylla 'Xinle' were collected in the same year, 1-2 buds on the top of the sprouting strips were cut off, and the leaves were removed to keep about 1 cm of petiole. On the induction medium of ratio (sucrose 30g / L+agar 5.8g / L in each medium component, pH value is 5.9), every treatment explant is inoculated 100, the induction rate of adventitious bud and the multiplication coefficient of counting after 30d, The effects of different media on the induction of adventitious buds were compared, and the experimental results are shown in Table 3.

[0041] Table 3 Effects of different hormone components on the induction of adventitious buds of Tilia microphylla ‘Xinle’

[0042]

[0043]

[0044] It can be seen from Table 3 that when the hormone components are MS+6-BA 1.0mg / L+IBA 0.2mg / L, the effect of explant adventitious bud induction is the best, the ...

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Abstract

The invention discloses a simplified tilia cordata mill tissue culture method, and belongs to the technical field of plant tissue culture. The simplified tilia cordata mill tissue culture method provided by the invention comprises the following steps of (1) collecting coppice shoots with axillary buds on robust plants 20 to 40 days after spring buds germinate, carrying out surface disinfection, then cutting out stem segments with buds, and inoculating the stem segments into an induction culture medium for culture; (2) cutting off the coppice shoots with the axillary buds in the step (1), cutting out a stem segment with one bud, and inoculating the stem segment into an induction culture medium; (3) carrying out subculture multiplication culture on the adventitious buds obtained in the step (2); (4) transferring the adventitious buds growing to 2.0 to 3.0 cm in the step (3) into a rooting culture medium to obtain aseptic seedlings; and (5) transferring the aseptic seedlings obtained in the step (4) into a plug matrix, and hardening the seedlings in an artificial greenhouse. The simplified tilia cordata mill tissue culture method provided by the invention is high in universality and simple in operation step, and can effectively achieve the purposes of germplasm preservation and rapid propagation of different varieties of tilia cordata mill.

Description

technical field [0001] The invention relates to a simplified Tilia microphylla tissue culture method, which belongs to the technical field of plant tissue culture. Background technique [0002] Tilia is a characteristic species in the temperate and subtropical deciduous broad-leaved forests in my country. There are 23 species of plants in this genus, including 17 species in China, which is the center of species diversity distribution. Tilia has a long history of cultivation and is a multipurpose tree species for timber, medicine, fiber, honey source and landscaping. Tilia plants are usually propagated by sowing, cutting, grafting and tissue culture. Due to the simplicity and ease of seed propagation, sowing propagation is still the main propagation method of this genus. The differentiation coefficient of the progeny of single-seed propagation is relatively large, and it is difficult to preserve the traits of excellent lines. The exploration of asexual reproduction techno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001
Inventor 王欢利汤诗杰严灵君黄犀王仲伟朱珣之罗会婷
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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