Method for obtaining plant graft chimaera progeny by embryo rescue

A chimera and embryo rescue technology, applied in the fields of botanical equipment and methods, plant regeneration, horticultural methods, etc., can solve the problems of chimera sterility, lack of examples, hindering the research on the genetic variation mechanism of grafting, etc., and achieves strong operability. , the effect is remarkable

Inactive Publication Date: 2010-09-15
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, studies have found that heterogeneous cell layers in grafted chimeras can affect the reproductive and developmental characteristics of plants, leading to sterility in chimeras (see Plant cell graft chimeras obtained by co-culture of isolated protoplasts. Binding et al. Protoplasma, p. 141, No. 1, pp. 64-73, February 1987), thus hindering the research on the mechanism of genetic variation in grafting
Therefore, there is an urgent need for a complete artificial embryo rescue system to preserve the sexual offspring of grafted chimera plants, but before this there is no method for grafted chimera plant embryo rescue and plant regeneration, let alone successful examples

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Materials: This example uses artificially synthesized interspecific graft chimera TCC of Brassica oleracea (Brassica oleracea) and mustard (B. juncea). This intergraft chimera TCC is named after the cell origin of the three-layer structure of the shoot apical meristem: L1-L2-L3 from outside to inside, T= mustard, C=purple cabbage (see: Synthesis of interspecific chimeras between tuber mustard (Brassicajuncea) and cabbage (B. oleracea) and cytological analysis. Chen et al. Plant Cell Reports, Vol. 25, No. 9: 907-913, September 2006)

[0025] Step (1): Collect materials

[0026] The artificial self-pollination of TCC at the bud stage was carried out, and the ovaries of different periods after pollination were collected, and the development process of the immature embryo was observed by paraffin section method. It was found that abortion occurred in the late stage of spherical embryo, and the specific abortion time was the Day 17. Therefore, the ovary was removed on the ...

Embodiment 2-10

[0040] In Examples 2-10, the steps of the method for obtaining plant-grafted chimera progeny through embryo rescue in the present invention are the same as those in Example 1, and the differences are shown in Table 1.

[0041] Table 1

[0042] Example

Embodiment 11-12

[0044] In Examples 11-12, the steps of the method for obtaining plant-grafted chimera progeny through embryo rescue in the present invention are the same as those in Example 1, and the differences are shown in Table 2.

[0045] Table 2

[0046] Example

[0047] Example

[0048] Observation of botanical characteristics: The botanical characteristics of the selfed progeny plants of the grafted chimera TCC obtained in the above examples 2-12 are relatively similar to those of purple cabbage, and there is no significant morphological variation, and the number of chromosomes is 2n=18. Same with purple cabbage.

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Abstract

The invention discloses a method for obtaining a plant graft chimaera progeny by embryo rescue, which comprises the following steps of: (1) picking an ovary in 1 day before the immature embryo of a crucifer graft chimaera is aborted; (2) sterilizing and flushing the ovary; (3) placing the ovary on a culture dish paved with filter paper which is fully soaked in nutrient solution, dissecting one end of the ovary and then tearing the ovary along a ventral suture, selecting and cutting light green and glossy ovule, suspensor and ovary wall tissue connected with the suspensor and inoculating the three to a development culture medium, culturing the three under a dark condition at the temperature of 25 +/- 2 DEG C, and then culturing the three under light; (4) picking mature seeds, inoculating the seeds to a sprouting culture medium, and culturing the seeds under the dark condition at the temperature of 25 +/- 2 DEG C; (5) after the seeds are sprouted and grow main leaves, transferring the seedlings to a sub-cultured medium to perform culture, and then transferring the seedlings to a rooting culture medium to perform rooting; and (6) taking the routed seedlings out, transplanting the seedlings to a nutrition pot filled with matrix, hardening the seedlings for 6 to 12 days, and then transferring the seedlings to a field so as to obtain the plant graft chimaera progeny.

Description

technical field [0001] The invention relates to a method for obtaining offspring of plant grafted chimeras through embryo rescue, in particular to a technical method for rescuing sexual offspring of interspecies grafted chimeras of cruciferous plants. Background technique [0002] Grafting is the technique of connecting the buds or branches of a plant to the appropriate part of another plant, so that the two can be combined into a new plant. As a worldwide art, grafting has been widely practiced and actively contributed to the development of agriculture. Plant grafting is a method of asexual reproduction, but it can induce genetic variation in plants. As early as the Ming Dynasty, the "Compendium of Materia Medica" recorded the phenomenon that "plums follow peaches, but the strong ones are actually hairy, and plums follow apricots, but the strong ones are actually sweet". At the beginning of the 20th century, Michurin, a Soviet horticulturalist, proposed the principles and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01G31/00
Inventor 王燕陈利萍
Owner ZHEJIANG UNIV
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