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Sampling method of barley microspores

A technology of microspores and barley, which is applied in the field of agricultural biology, can solve the problems that cannot be popularized and applied, the difficulty and difference of the judgment standard of the proportion of microspore development period, and the technical requirements of the people who draw materials are high, so as to achieve the ability of differentiating green seedlings and protecting them. Bacterial environment, the effect of inducing a high rate of embryo formation

Active Publication Date: 2019-10-11
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] However, the development of microspores is continuous and complex, and there are polymorphisms, which need to be determined by the proportion of appropriate developmental stages. However, there are still difficulties and differences in the criteria for judging the proportion of microspore developmental stages
[0005] It has been reported in the previous research that the period of material collection is determined according to the color of the anthers, but the identification of the color of the anthers will vary from person to person, and the technical requirements for the material collectors are relatively high, so it cannot be popularized and applied.

Method used

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  • Sampling method of barley microspores
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  • Sampling method of barley microspores

Examples

Experimental program
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Embodiment

[0032] 1. Materials

[0033] The barley line Hua 94-30 (called "Hua 30") was planted in Qingpu Base in November 2017, and harvesting began in late March 2018.

[0034] 2. Method

[0035] 2.1 Material collection and ear pretreatment

[0036] After the flag leaf of the "Hua 30" plant starts to be pulled out until the distance between the flag leaf and the ligule is drawn out within 4cm, all materials are collected, and samples are taken according to the distance from the flag leaf to the ligule, see figure 1 and Table 1, figure 1 Among them, No. 2 is Example 3, No. 1 is Comparative Example 1, No. 3 is Comparative Example 2, and No. 4 is Comparative Example 3.

[0037] The base of the upper two leaves has a length of 1.0-1.5 cm. Cut off the leaves, put them in a fresh-keeping bag, and pretreat them at a low temperature of 5°C for 21 days.

[0038] 2.2 Collection and purification of microspores

[0039] On a sterile ultra-clean workbench, sterilize with 10% sanitin for 10 min...

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Abstract

The invention provides a sampling method of barley microspores. The method comprises the steps of (1) sampling and pretreatment of young ears and (2) collection and purification of microspores. The sampling method of the barley microspores has the advantages that when the young ears are sampled, by controlling the spacing of 1.1-2.0 centimeters from flag leaves to ligules, the induced differentiation difference in microspore culture is shortened, after the microspores obtained through separation are cultured, the inducted embryo formation rate is high, and the green seedling differentiation capacity is strong, the culture and regeneration efficiency of the microspores is improved by 3 times, the requirements for labor and equipment during culture of the microspores are greatly reduced, thequantized sampling standards can be achieved, and the sampling efficiency and the practicability of the standards can be improved.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a method for extracting barley microspores. Background technique [0002] Microspores have the characteristics of single cells and haploids, and are ideal recipients for cell engineering genetic breeding and genetic transformation, and are also ideal objects for studying the mechanism of embryogenesis. Free microspore culture is an effective way to obtain homozygous doubled haploids. The free microspore culture technology is one of the hotspots in current research. To make the free microspore culture go smoothly, it is very important to obtain materials. [0003] Microspore culture and collection of barley microspores mostly adopt the method of microscopic examination. The obtained young ears are peeled off, anthers are picked, crushed, examined under a microscope, and the developmental stage of microspores is observed to determine the appropriate period of m...

Claims

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Application Information

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IPC IPC(8): C12N5/04
CPCC12N5/04C12N2509/00
Inventor 郭桂梅何婷李颖波陈志伟陆瑞菊刘成洪
Owner SHANGHAI ACAD OF AGRI SCI
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