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Method for rapidly obtaining stable homozygous nitrogen efficient material by using hybrid F1-generation microspore culture

A technology of microspore culture and microspores, applied in the field of agriculture, can solve problems such as long time, and achieve the effect of excellent nitrogen utilization rate

Active Publication Date: 2021-01-29
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But heterosis mainly exists in F 1 Generation, due to the law of Mendelian segregation, it will obviously decline from the second generation, and nitrogen efficiency is largely controlled by recessive genes, screening needs to be carried out in high generation, which will take longer, even 6-10 Homozygous nitrogen-efficient germplasm

Method used

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  • Method for rapidly obtaining stable homozygous nitrogen efficient material by using hybrid F1-generation microspore culture
  • Method for rapidly obtaining stable homozygous nitrogen efficient material by using hybrid F1-generation microspore culture
  • Method for rapidly obtaining stable homozygous nitrogen efficient material by using hybrid F1-generation microspore culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Hybridization of Barley Low Nitrogen Tolerant Parents

[0057] Barley (Hordeum vulgare L.) is the fourth most important cereal crop in the world after wheat, rice and maize. It is considered to be an excellent model plant for studying microspore culture high-frequency regeneration technology and functional food. Take 100 seeds of two barley low-nitrogen-resistant materials BI04 and BI28 (Chen Zhiwei et al. 201030 (1): 158-162) obtained by the laboratory screening in the early stage, disinfect the surface with 10% sodium hypochlorite for 8-10 minutes, rinse with distilled water, Put them on wet gauze and accelerate germination overnight at room temperature in the dark. After 7 days of normal germination, get the seedlings with consistent growth (the seedling length is 5 cm) and transplant them to the field with a spacing of 3 cm and a row spacing of 30 cm. The soil contains 34.0g / kg of organic matter, 2.41g / kg of total nitrogen, and 37.30mg / kg of available nit...

Embodiment 2

[0058] Example 2: F 1 generation plant propagation

[0059] The hybrid seeds are planted in the field during the sowing season as above, and get F 1 Subsequent growth plants are used as culture materials for free microspores.

Embodiment 3

[0060] Example 3: Free F 1 Homozygous DH strain obtained from microspore culture

[0061] Select the spikelets whose microspore development is in the early and middle stages of uninucleate in the middle part of the field from the field, and treat them at a low temperature of 5°C for 2-3 weeks. When inoculating, the spikelets are soaked in 75% alcohol for 30 seconds, sterilized with a saturated bleach solution for 15 minutes, and sterile water Rinse 4 times, connect 10 ears in each test tube, pour into 15ml pre-cooled extract (containing 6% mannitol, 1.1g / L CaCl 2 and 0.976g / LMES), using a high-speed disperser for ultra-speed rotary cutting, 300 mesh screen filter, and the filtrate is 700r min -1 Centrifuge for 5 minutes and repeat 3 times to collect microspores.

[0062] The microspores were pretreated with the pretreatment solution at 25°C in the dark for 2 days. The pretreatment liquid contains 6% mannitol, 1.1g / L CaCl 2 and 0.976g / L MES.

[0063] Before culturing, the ...

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Abstract

The present invention relates to the technical field of agriculture, particularly to a method for quickly obtaining a genetically stable homozygous nitrogen efficient material by using hybrid F1-generation microspore culture. The method comprises the following steps: selecting two identified low-nitrogen-tolerant barley parents for hybridizing, harvesting F1-generation microspores, after in-vitroculture and through a differentiation and dedifferentiation process of the microspores, performing rapid homozygosis and immobilization on low-nitrogen-tolerant parent nitrogen efficient (dominant orrecessive) genes, performing culture to obtain regenerated doubled haploid M0-generation seeds, through multi-year multi-point multi-repetition low-nitrogen stress culture identification via water culture and field trials, and finally, rapidly performing screening to obtain the homozygous barley germplasm material whose seedling-stage biomass, mature-stage single-plant yield and nitrogen utilization efficiency are stably improved under a low-nitrogen condition and exceed those of parents.

Description

technical field [0001] The invention relates to the field of agricultural technology, in particular to a method for rapidly creating homozygous germplasm materials of barley with stable super-parent nitrogen utilization efficiency. Background technique [0002] Nitrogen is one of the essential mineral elements for plant growth, accounting for 1.5-2% of plant dry weight and 16% of protein. The use of nitrogen fertilizers has increased ten-fold in half a century to boost crop yields. A large amount of nitrogen fertilizer is used in farmland. As a result, most high-yielding crop varieties bred in recent decades are very dependent on nitrogen and other nutrients. Many barren-tolerant crop varieties are close to disappearing, which greatly increases the cost of planting. At the same time, a large amount of nitrogen fertilizer is not effectively used, and the nitrogen that can be used by crops accounts for about 30-40%. In China, more than 15 million tons of waste nitrogen are l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/02A01H1/04A01H4/00A01H6/46
CPCA01H1/02A01H1/04A01H4/001A01H4/008A01H6/4624Y02P60/21
Inventor 徐红卫高润红张述伟宗营杰李静黄琳丽陈志伟刘成洪
Owner SHANGHAI ACAD OF AGRI SCI
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