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A method that can significantly increase the microspore callus yield of highland barley variety Himala 22

A technology of microspore and microspore culture, applied in the field of bioengineering, can solve the problems affecting the regeneration frequency and difference of highland barley microspores

Active Publication Date: 2022-03-11
SHANGHAI ACAD OF AGRI SCI
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

We found that when the barley microspore induction culture is actually operated, there is still a significant difference in the callus yield induced by the highland barley microspore and the high-frequency microspore regeneration barley material, which directly affects the late regeneration frequency of the highland barley microspore

Method used

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  • A method that can significantly increase the microspore callus yield of highland barley variety Himala 22
  • A method that can significantly increase the microspore callus yield of highland barley variety Himala 22
  • A method that can significantly increase the microspore callus yield of highland barley variety Himala 22

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: Low temperature treatment of highland barley, barley material seedlings

[0019] The barley varieties "Hua 30" and "Hua 11" were bred by the Cell Engineering Research Office of the Shanghai Academy of Agricultural Sciences. Companies can buy. "Himala 22", "Hua 30" and "Hua 11" were soaked in a 15cm petri dish with filter paper until they turned white, and the germinated seeds were moved to a refrigerator at 4°C for 30 days at low temperature. After being transplanted into an artificial climate chamber at a low temperature, the plant culture conditions are: temperature 18°C ​​at night, 20°C during the day; humidity 65%; 12 hours of light and 12 hours of darkness.

Embodiment 2

[0020] Embodiment 2: Highland barley, barley material microspore culture

[0021] Select "Himala 22" and "Hua 30" ears whose floret microspore development in the middle part of the young panicle of the plant is in the early-middle stage of uninucleation, cut off the leaves (keep about 1.5cm from the base of the upper two leaves), and put them in a fresh-keeping bag to keep moist , indicating the date and quantity of materials collected, etc., and put them in a 4°C refrigerator for 2 to 4 weeks of low-temperature pretreatment. Free microspores from flower buds of "Himala 22" and "Hua 30" treated at low temperature were obtained according to the method of Lu Ruiju et al. Effects on seedlings. Acta Plant Physiology, 2001, 27(2): 135-140). Each test tube connects the flower buds of 4 spikes, and adds 12ml extract (mannitol 60g / L+CaCl 2 1.1g / L+MES 0.976g / L+colchicine 20mg / L, pH 5.8), using a high-speed disperser for ultra-speed rotary cutting. Filter the spin-cut suspension wit...

Embodiment 3

[0022] Example 3: "Himala 22" microspore inoculation of "Hua 30" and "Hua 11" ovaries

[0023] The ovaries of barley varieties "Hua 30" and "Hua 11" were taken after artificial detasseling for 2 days, and the microspores in each plate of "Himala 22" were cultured according to 0, 0.4 / cm 2 and 0.8 pieces / cm 2 The ovaries were placed in each dish, and 3 replicates were set for each treatment. When cultured in dark until 20 days, weigh the microspore calli of the two materials with different treatments, and the average weight of the calli of "Himala 22" that was not inoculated with the ovary of "Hua 30" was 4.7mg, and the weight was 0.4 / cm 2 The average callus weight of "Himala 22" with added ovary is 46.7 mg, at 0.8 callus / cm 2 The average callus weight of "Himala 22" added with ovary was 25.6 mg, and the microspore-induced callus yield of "Himala 22" increased significantly after adding "Hua 30" ovary. There is a significant difference under the condition of p2 Add the "Flowe...

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Abstract

The invention relates to the technical field of bioengineering, in particular to a method capable of significantly increasing the callus yield of highland barley variety "Himala 22" microspore culture, which is to add barley variety "Hua 30" ovary. In the present invention, by adding a certain amount of ovaries of the barley variety "Hua 30" with excellent tissue culture characteristics to the microspore induction medium of "Himala 22", the final callus yield is significantly higher than that of barley microspores without adding ovaries. Significant improvement; the invention provides a practical and effective method for the high-efficiency regeneration of microspores of "Himala 22" and the haploid breeding of highland barley.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method capable of significantly increasing the yield of microspore-cultured calluses of the highland barley variety "Himala 22". Background technique [0002] Highland barley is an important food crop in the Qinghai-Tibet Plateau region of my country, and is also the staple food of compatriots in Tibetan areas. It is also an excellent feed and raw material for wine making, and plays an important role in my country's agricultural production. The selection and breeding of fine barley varieties is related to the food security in my country's frontier areas and the improvement of the living standards of Tibetan compatriots, and has important political significance and social and economic benefits. With people's emphasis on health and the diversification of dietary structure, highland barley is changing from a regional ration crop to a global healthy food source crop due to i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00A01H6/46
CPCA01H4/001A01H4/005A01H4/008A01H6/4624
Inventor 郭桂梅李颖波刘成洪宗营杰何婷陆瑞菊陈志伟
Owner SHANGHAI ACAD OF AGRI SCI
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