A method for efficient and rapid regeneration of adventitious buds from pear leaves

A leaf and fast technology, applied in plant regeneration, horticultural methods, botanical equipment and methods, etc., can solve the problems of difficult application of genetic transformation system and difficulty in regeneration of Oriental pear varieties, so as to overcome easy browning, accelerate growth, reduce The effect of condensation

Active Publication Date: 2018-05-11
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to explore the factors that affect the regeneration of snow pear leaves with snow pear leaves as materials, establish a high-efficiency propagation system for snow pear tissue culture seedlings, provide a method for efficient and rapid regeneration of adventitious buds from pear leaves and Culture medium to solve the problems of difficult regeneration of oriental pear varieties and difficult application of genetic transformation system in oriental pear varieties

Method used

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  • A method for efficient and rapid regeneration of adventitious buds from pear leaves
  • A method for efficient and rapid regeneration of adventitious buds from pear leaves
  • A method for efficient and rapid regeneration of adventitious buds from pear leaves

Examples

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Effect test

Embodiment 1

[0034] A method for efficiently and quickly regenerating adventitious buds from pear leaves, comprising: obtaining sterile tissue culture seedlings, inducing adventitious buds, primary culture and subculture of adventitious buds, etc., the specific operation process is as follows:

[0035] 1) Acquisition of aseptic tissue culture seedlings of snow pear: when the tree body just starts to germinate in spring, cut 40 cm long branches with full bud eyes on the snow pear tree, put them in 2% sucrose solution, and cultivate them at about 25°C Indoor germination; after the axillary buds germinate, cut the branches into stem segments, each stem segment has 3 buds, and rinse with running water for 40 minutes; in the ultra-clean workbench, first treat the buds with 70% ethanol for 30 seconds, and then wash them with sterile water 3 times, and then treated with 2.5% sodium hypochlorite for 4 minutes, and finally washed 4 times with sterile water, filter paper to absorb excess water, after...

Embodiment 2

[0042] The inventor investigated the influence of different ratios of hormones TDZ and IBA in the induction medium and the dark culture time on the regeneration effect of snowflake pear leaves. Other operations are the same as in Example 1, and the results are shown in Table 1. figure 2 and image 3 shown.

[0043] Table 1 Effects of different hormone ratios and dark culture time on leaf regeneration of snow pear

[0044]

[0045] The inventor investigated the influence of different leaf age snowflake pear leaves on the browning rate in the regeneration in step 2), and other operations are the same as in Example 1, and the investigation results are shown in Table 2.

[0046] Table 2 Effects of different leaf ages on regeneration browning rate of pear tree leaves

[0047] leaf age

[0048] The inventor investigated the influence on leaf culture when the petri dish was placed upside down and placed upside down during dark culture in step 3) and the operation was ...

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Abstract

The invention discloses a method for efficiently and rapidly regenerating adventitious buds from pear leaves. The method comprises steps as follows: acquiring sterile tissue culture seedlings, generating adventitious buds by inducing the leaves and performing primary culture and subculture on the adventitious buds, wherein an inducing medium is a combination of a basic culture medium NN69, auxin IBA and cytokinin TDZ, the leaf age is about 20-50 d, dark culture is performed for 21 d, the regeneration effect of leaves of Pyrus bretschneideri Rehd. is remarkable, the regeneration rate can reach 70.83%, the average number of regeneration buds of each leaf is 2.06, the callus occurrence rate reaches 100%, the browning rate is very low and is even 0, and demands for germplasm preservation of pear varieties and requirements for the regeneration rate of the Pyrus bretschneideri Rehd. serving as a genetic transformation material are met. Requirements for the leaves are low, the material sources are enriched, and the problem of high browning rate in the woody plant tissue regeneration process is solved. The problem that the regeneration rate of the oriental pear varieties is generally low is effectively solved, an effective material is expected to be provided for genetic transformation of the oriental pear varieties, and breakthrough of genetic transformation of pears is realized.

Description

technical field [0001] The invention belongs to the category of biotechnology and breeding, and relates to a method for pear leaf regeneration, in particular to an efficient and fast method for regenerating adventitious buds from pear leaves and a culture medium. Background technique [0002] Pear is a perennial woody plant that mainly reproduces asexually. It has a long generation cycle, a high degree of heterozygosity in genetics, and a long childhood period. Breeding is time-consuming, laborious, and high-input. The application of genetic transformation technology based on plant tissue culture and isolated tissue regeneration in pears has greatly promoted the germplasm innovation and new variety selection of pears, and has become an effective way for pear genetic engineering breeding. But compared with other species, the progress of genetic transformation of pear is slow, and the efficiency of genetic transformation is very low. The main reason is that the regeneration o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 吴俊闫允青张绍铃刘伦常耀军谷超殷豪齐开杰谢智华
Owner NANJING AGRICULTURAL UNIVERSITY
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