High-temperature detoxification method based on lily scales and leaf tips and application of high-temperature detoxification method

A lily and scale technology, applied in the field of high-temperature detoxification, can solve the problems of difficulty in obtaining stem tip materials, long heat treatment time, high cost, etc., and achieve the effects of high reproduction coefficient, increased detoxification rate, and low cost

Active Publication Date: 2021-07-27
YANGTZE NORMAL UNIVERSITY
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a high-temperature detoxification method based on lily scale leaf tips and its application, so as to solve the problems of difficulty in obtaining stem tip materials, high cost and long heat treatment time in the existing lily detoxification method. long question

Method used

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  • High-temperature detoxification method based on lily scales and leaf tips and application of high-temperature detoxification method
  • High-temperature detoxification method based on lily scales and leaf tips and application of high-temperature detoxification method
  • High-temperature detoxification method based on lily scales and leaf tips and application of high-temperature detoxification method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 A kind of lily high temperature detoxification method based on scale leaf tip

[0033] 1) First-generation culture: take the virus-carrying 'Siberia' lily corm as material, wash it with washing powder, rinse it in running water for more than 30 minutes, and then put the bulbil in a sterile container on an ultra-clean workbench, wash it with 75% alcohol Soak for 40s, rinse with sterile water and then add 0.1% HgCl 2 Disinfect for 12 minutes, rinse with sterile water 6-8 times for use. Then the cleaned scales are placed on the primary culture medium with the inner side facing up to carry out induction culture for 2-3 weeks to obtain sterile buds.

[0034] 2) Subculture: use aseptic lily buds as explants, inoculate them in the subculture medium and culture them for 3 to 4 weeks to obtain normal-growing sterile bulblets.

[0035] 3) Thermal exercise treatment: place the lily tissue culture seedlings in a light incubator, treat them at a high temperature of 37...

Embodiment 2

[0047] The high temperature treatment temperature is 85° C., and other steps are the same as in Example 1.

[0048] After the high temperature treatment at 85℃, the growth of small scale leaf tip of 'Siberia' lily is shown in Table 3, and the growth of leaf tip of surviving 'Siberia' lily is as follows Figure 4 As shown, when the leaf tip was inserted into the medium, the leaf tip swelled and began to form adventitious buds in about 20 days; the adventitious buds grew leaves and developed roots in about 40 days, and developed into test-tube plantlets in 60 days.

[0049] Table 3 The growth of small scale leaf tips of 'Siberia' lily treated at 85℃

[0050]

[0051]

[0052] Survival rate = normal survival number / total number of inoculations

Embodiment 3 100

[0053] The detection of virus in the lily material of embodiment 3

[0054] Lily RNA extraction: In order to detect the detoxification effect, a certain number of seedlings were randomly selected from the lily test-tube seedlings prepared in Examples 1-2, and the total amount of leaf total was extracted using the Tiangen Plant RNA Small Extraction Kit (R4151). RNA was stored at -20°C for later use.

[0055] cDNA synthesis: The total volume of each cDNA synthesis reaction is 20 μl, and the first-strand cDNA is synthesized using the plant cDNA rapid reverse transcription kit (KR116) from TIANGEN Company. The reaction reagents include 2 μl of total RNA, 2 μl of 5×gDNA Buffer, and RNase-Free ddH 2 O 6μl, incubate at 42°C for 5min, place on ice; add 10×King RT Buffer 2, FastKing RTEnzyme Mix 1μl, FQ-RT Primer Mix 2μl, RNase-Free ddH 2 O 5 μl, the reaction program was incubated at 42°C for 30 minutes and inactivated at 70°C for 5 minutes.

[0056] PCR amplification: The PCR reacti...

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Abstract

The invention provides a high-temperature detoxification method based on lily scales and leaf tips and application of the high-temperature detoxification method. According to the method, the lily scales or bulbils are used as materials, sterile bulblets are cultured, 37-DEG C and 23-25-DEG C variable-temperature alternate treatment is adopted, then 95-DEG C high-temperature treatment and accurate control over the treatment time are combined, the detoxification efficiency of the scale leaf tips on lily viruses CMV, LSV and LMoV are greatly improved, the length of the stripped leaf tips is 1.0-2.0 mm, the leaf tips with the length do not need instruments and equipment such as an anatomical lens, generally, one bulblet has 6-10 or even more leaf tips, the materials are easier to obtain, operation is easy and convenient, and cost is low. Compared with traditional heat treatment, the heat treatment time is short, and the detoxification rate is high. The method breaks through the situation that stem tip culture is mainly adopted in the prior art, provides a new way for culturing lily non-toxic seedlings, has a good application prospect, and lays a foundation for industrialized seedling production.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture and detoxification, and specifically relates to a high-temperature detoxification method based on lily scale leaf tips and an application thereof. Background technique [0002] Lily is a perennial bulbous flower with many types and strains, a long flowering period, huge flowers, rich colors, and beautiful flowers. It has not only excellent ornamental value, but also excellent medicinal and edible value. Siberia (Lilium ‘siberia’) is one of the main cut lilies that sell well in the market. It belongs to the oriental line of lilies. It has large flowers and is pure white. The current market price is about 15 yuan a piece. In recent years, with the expansion of lily production scale, the self-propagation of bulbs and the rapid circulation have made the accumulation of lily virus diseases more and more serious. In the production of lilies, bulbs or scales are often used for asexual repro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 符勇耀张豪夏萍刘虹伶徐文姬杨利平
Owner YANGTZE NORMAL UNIVERSITY
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