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A kind of subculture method of anti-rust black poplar tissue culture seedling

A subculture, rust-resistant technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of easy lack of rust-resistant resistance, and achieve the effect of large-scale production

Inactive Publication Date: 2016-08-31
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention aims to solve the problem that the resistance to rust resistance is easily lost during the current tissue culture propagation process and after subculture, and provides a method for subculture of rust-resistant black poplar tissue culture seedlings

Method used

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  • A kind of subculture method of anti-rust black poplar tissue culture seedling

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specific Embodiment approach 1

[0011] Embodiment one: the subculture method of the anti-rust black poplar tissue culture seedling of the present embodiment, carry out according to the following steps:

[0012] 1. Screening of rust-resistant black poplar:

[0013] Separate fully expanded leaves with a leaf interval index of 5 to 9 from the dormant branches of 1-year-old black poplar in activation culture, take 8 to 12 leaves from each black poplar, inoculate with rust fungus, and then place the inoculated leaves at 16°C In the growth room of the black poplar, carry out 16h light every day, continue to cultivate for 7 days, calculate the disease index of each black poplar, select the black poplar plant with the disease index<25, clean and disinfect the black poplar leaves, inoculate into 1 black poplar containing 300mg / LAmp On the / 2MS solid medium, carry out subculturing for 20 days, screen out the black poplar plants with disease index<25 after subculturing for 20 days as rust-resistant black poplar;

[00...

specific Embodiment approach 2

[0017] Specific embodiment two: the difference between this embodiment and specific embodiment one is that: the method for inoculating rust bacteria in step one is: suspending spores with a Tween 20 aqueous solution of 0.004% with a mass percent concentration, and then spraying inoculation, wherein the concentration of spores 40000 / mL. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0018] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: the calculation method of disease index is: disease index=100 * ∑ (number of sick leaves at all levels * representative value at all levels) / (investigation total leaf number × the highest level representative value), the disease index <25 means disease resistance, 25 < disease index <50 means moderate disease resistance, 50 < disease index <75 means susceptible disease, and 75 < disease index means highly susceptible disease; among them, the disease level and The representative values ​​of each level are as follows: level 1 without erythrosporium on the leaves, with a representative value of 0; level 2 with uredospores on the area below 1 / 4 of the leaves, with a representative value of 1; 1 / 4 to 1 / 2 of the area of ​​the leaves If there are uredospores produced, it is grade 3, with a representative value of 2; if 1 / 2 to 3 / 4 of the leaf area has uredospores produced, it...

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Abstract

The invention discloses a subculture method for leaf-rust-resistant black poplar tissue culture seedlings, and relates to a subculture method for black poplar tissue culture seedlings. The subculture method is used for solving the problem that the resistance to leaf rust is likely to lose in tissue cultivation and propagation expansion processes and after subculture. The method comprises the following steps: I, separating leaves from activated cultured one-year-old black poplar dormant branches, inoculating rust funguses, putting the inoculated leaves in a growth chamber, illuminating every day, culturing for 7 days, calculating disease indexes of each black poplar, selecting the black poplars with disease indexes lower than 25, cleaning and disinfecting the black poplar leaves, inoculating the black poplar leaves into a culture medium to perform subculture, and screening out the black poplars with diseases indexes lower than 25 after sub-culturing for 20 days; and II, propagating and breeding the screened leaf-rust-resistant black poplar tissue culture seedlings under a sterile operation condition in a form of splitting multiple shoots, and inoculating the multiple shoots into the subculture culture medium to culture. The subculture method is used for the tissue culture field.

Description

technical field [0001] The invention relates to a method for subculture of black poplar tissue culture seedlings. Background technique [0002] Larch-poplar rust fungus (Melampsora larici-populina), also known as poplar leaf rust fungus, is a long-lived rust fungus with a long life history that is widely distributed and harmful. The poplar leaf rust caused by it is the Populus spp.) is an important leaf disease at the seedling and sapling stage, which can attack more than 20 kinds of poplars such as Qingyangpai, Heiyangpai and their hybrids. Physiological differentiation exists in Larch-poplar rust. So far, the physiological races that have been discovered include E1, E2, E3, and the later discovered physiological race E4. Generally, black poplar (Populus nigra) and its hybrids are resistant to rust fungi. The black poplar resistant to rust fungus was screened out through the inoculation test, but encountered problems such as browning, vitrification and loss of resistance ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 李丹蕾王峰陈俏丽张瑞芝牛春阳王博文
Owner NORTHEAST FORESTRY UNIVERSITY
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