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A kind of method for cultivating non-heading cabbage microspore plants

A technology for head-heading cabbage and microspores, which is applied in the field of cultivating non-heading cabbage microspores, can solve the problems of pollution, cumbersomeness, and low germination rate, and achieve the effects of reducing pollution, simplifying operation steps, and reducing operation time

Active Publication Date: 2022-03-08
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention mainly aims at improving the problems of the current technology such as cumbersomeness, low germination rate and pollution, thereby providing an efficient and simple method for cultivating non-heading Chinese cabbage microspore plants

Method used

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  • A kind of method for cultivating non-heading cabbage microspore plants
  • A kind of method for cultivating non-heading cabbage microspore plants
  • A kind of method for cultivating non-heading cabbage microspore plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Selection of flower buds: Collect samples from 9 am to 11 am, select flower buds with a ratio of petals to anthers close to 1:1, add a small amount of distilled water and store in a refrigerator at 4°C.

[0031] 2. Cultivate microspore embryos: Put the collected samples into a 2ml centrifuge tube, add 1% sodium hypochlorite directly to disinfect for 10-15 minutes, wash with sterilized water twice, and moisten with NLN-13 for the last time wash. Transfer the cleaned sample to the test tube of the sample grinding machine (Germany LKA control type test tube disperser S025), add a small amount of NLN-13, and grind the sample to make the sample fully ground. Prepare a 50mL centrifuge tube, place a 40μm cell filter plug on top of it to filter, centrifuge, pour off the supernatant, then add NLN-13 to shake well, then centrifuge, pour off the supernatant, leaving a yellow-green precipitate. Then add a small amount of NLN-13 to the test tube and dilute to 10 cells 5 per mL....

Embodiment 2

[0038] Embodiment 2: the cultivation of microspores of non-heading Chinese cabbage of different varieties

[0039] Choose 9 different varieties (Kaicai Jianshan white, Qinglan, Erzhuangbai, Qinggengbai, chicken feathers, cabbage core, Wutaicai, Wuyueman) and adopt the method described in Example 1 to carry out the microspore culture test, Count their embryonic rate, compare the impact of this invention on the microspore embryonic rate. Among them, Jianshabai, Qinglan, and Erzhuangbai are easy-to-germ varieties; Qinggengbai, chicken feathers, and cabbage core are varieties that can produce embryos; Such as figure 1 , such as comparing the results of embryo emergence, among the 9 selected varieties, the embryo emergence rate reached 100%, of which 3 varieties that were not easy to emerge embryos all emerged embryos, and the variety Kuaicai that was easy to emerge embryos reached 13.62 embryos / bud.

[0040] Such as Figure 4 to Figure 7 Respectively are the growth charts of th...

Embodiment 3

[0041] Example 3: Effects of Boron Element and Activated Carbon on Microspore Germination Rate of Non-heading Cabbage of Different Varieties

[0042] Two varieties with high embryo emergence rate were selected, and the research on the microspore embryo emergence rate of boron element and activated carbon was carried out. Figure 6 , divided into four groups, control (NLN-13), only adding activated carbon, only adding boron, and adding boron and activated carbon at the same time. The comparison results show that when nothing is added, the embryo emergence rate of the two varieties is very high. Low, after adding activated carbon and boron respectively, the germination rate has been improved to varying degrees, and after adding activated carbon and boron at the same time, compared with adding boron and activated carbon alone, both varieties have significantly improved, higher than the control out 4-5 times Figure 7 It is a graph comparing the emergence of embryos after adding ...

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Abstract

The invention discloses a method for cultivating microspore plants of non-heading Chinese cabbage, which specifically includes the following steps: (1) selecting flower buds: selecting flower buds with a ratio of petals to anthers of 1:1; (2) cultivating microspore embryos: The flower buds collected in step (1) are disinfected with sodium hypochlorite solution for 10-15 minutes, washed with sterilized water, and rinsed with NLN-13 medium; the rinsed flower buds are fully ground, filtered, and centrifuged to contain 10-30mg / L boron substances NLN‑13 culture medium was diluted, distributed into petri dishes and added with activated carbon, heat shock treatment, 24‑26 °C dark culture for 14‑16 days, to form embryoid bodies; (3) mature embryos germinated into plants: embryoid bodies Shaking culture is carried out, and the mature embryos that have turned green are transferred to solid MS medium for cultivation. After 38-42 days of cultivation, the hardened seedlings can be directly transplanted. Under the condition that the embryo emergence rate can be guaranteed, the method of the invention simplifies the operation steps, greatly reduces the operation time, and improves the plant production efficiency.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, in particular to a method for cultivating non-heading Chinese cabbage microspore plants. [0002] technical background [0003] Chinese cabbage is mainly a cross-pollinated plant. In traditional breeding work, it is necessary to obtain purified parents only through conventional selfing. Due to selfing decline, the selection of homozygous parents is a very difficult problem, and it takes a long time and consumes manpower. As well as financial resources, the breeding efficiency is very low. Microspore culture technology is the process of cultivating a single cell into a homozygous plant by utilizing the totipotency of the cell, which can effectively shorten the breeding period, obtain a large number of homozygous plants in a short period of time, and greatly improve the breeding efficiency. Lichter first discovered on Brassica napus in 1982 that embryoid bodies could be obtained by usi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001A01H4/00
Inventor 李英梁超凡侯喜林张娅刘同坤张昌伟
Owner NANJING AGRICULTURAL UNIVERSITY
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