Method for culturing non-heading Chinese cabbage microspore plantlets

A technique for headed cabbage and microspores is applied in the field of cultivating non-headed cabbage microspore plants, which can solve the problems of cumbersomeness, pollution, and low embryo rate.

Active Publication Date: 2019-08-06
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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  • Method for culturing non-heading Chinese cabbage microspore plantlets
  • Method for culturing non-heading Chinese cabbage microspore plantlets
  • Method for culturing non-heading Chinese cabbage microspore plantlets

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, Chimaera, and Caixin are varieties that can produce embryos; Caixin, Wutaicai, and Wuyuemanan are difficult-to-embryo varieties. 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 difficult-to-embryo varieties all produced embryos, and the easy-to-embryo variety Kuaicai reached 13.62 embryos / bud.

[0040] Such as Figure 4 to Figure 7 Respectively are 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. Image 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 a...

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Abstract

The invention discloses a method for culturing non-heading Chinese cabbage microspore plantlets. The method comprises the following specific steps of (1) flower bud selection, wherein flower buds withthe number ratio of 1:1 between petals and anthers are selected; (2) culture of microspore derived embryos, wherein the flower buds collected in the step (1) are disinfected by a sodium hypochloritesolution for 10-15 minutes, washed with sterilization water and rinsed by an NLN-13 culture medium; the rinsed flower buds are completely ground, filtered, centrifuged, diluted by the NLN-13 culture medium containing 10-30 mg/L of a boron substance and subpackaged in culture dishes, activated carbon is added, heat shock treatment is conducted, dark culture is conducted at 24-26 DEG C for 14-16 days, and the embryoids are formed; (3) germination into the plantlets from mature embryos, wherein the embryoids are subjected to shake culture, mature embryos turning to green are transferred into a solid MS culture medium for culture, and after 38-42 days of culture, acclimatization and transplant can be directly conducted. According to the method, on the premise of ensuring the embryo formation rate, the operation steps are simplified, the operation time is greatly shortened, and the plantlet production efficiency is improved.

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 Applications(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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