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A method for constructing Lycoris genetic transformation system

A technology of genetic transformation system and construction method, applied in the field of construction of Lycoris genus genetic transformation system, can solve problems such as poor fertility, difficulty in new varieties, low positive rate, etc., achieve high success rate, improve transformation success rate, repeatability Good results

Active Publication Date: 2020-01-14
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because some species of this genus are of hybrid origin, the fertility is poor, and it is difficult to obtain new varieties through hybrid breeding; even if the hybridization is successful, the repetition rate is poor and the positive rate is low

Method used

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  • A method for constructing Lycoris genetic transformation system

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Establishment of GUS Transgenic System of Amaryllis Lycoris

[0068] 1 Callus induction The sterile bulblets of Amaryllis chinensis were transferred to 30g / L sucrose+7.5g / L agar+MS+6.0mg / L 6-BA+1.5mg / LNAA medium. After 2 weeks, white protrusions appeared around the small bulbs, which was the germination of callus tissue. On 20 days, pale yellow tissues appeared around the white protrusions, and after 30 days, pale yellow callus masses were seen. Subculture until callus clumps larger than 1 cm in diameter are induced. The culture temperature is 25±1°C, and the culture is dark.

[0069] 2 vector construction

[0070] (1) The cloning vector EF-3-GUS-21 μL was introduced into Agrobacterium as a recombinant plasmid.

[0071] (2) Transformation: take the Agrobacterium strain EHA105 stored at -70°C, melt it on ice, add 4 μL of the ligation product when it reaches a semi-thawed state, flick and mix well, put it in an ice bath for 20-30 minutes, heat shock at 42°C for 30 seco...

Embodiment 2

[0093] Establishment of Transgenic System of Genes Related to Synthesis of Lycoris Anthocyanins

[0094] 1 Callus induction The sterile bulblets of Lycoris were transferred to 30g / L sucrose+7.5g / L agar+MS+6.0mg / L6-BA+1.5mg / LNAA medium. After 2 weeks, white protrusions appeared around the small bulbs, which was the germination of callus tissue. On 20 days, pale yellow tissues appeared around the white protrusions, and after 30 days, pale yellow callus masses were seen. Subculture until callus clumps larger than 1 cm in diameter are induced. The culture temperature is 25±1°C, and the culture is dark.

[0095] 2 vector construction

[0096] The source of the gene and the carrier: Lycoris anthocyanin synthesis gene MYB119 was synthesized by Gene Synthesis Company according to the nucleotide sequence shown in SEQ ID No.3 in the sequence list, and the restriction site SalI was added at both ends of the nucleotide sequence and XbaI. The pCAMBIA2301 plasmid with its own GUS report...

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Abstract

The invention relates to a Lycoris herb genetic transformation system constructing method, and belongs to the technical field of genetics. According to the Lycoris herb genetic transformation system constructing method, a target exogenous gene is introduced into Lycoris herb callus by using a genetic engineering method, and Lycoris herb test-tube seedlings are obtained through differentiation culture and strong seedling culture, such that the new Lycoris herb variety having the target exogenous gene controlled biological traits is obtained. According to the present invention, the method has characteristics of good repeatability and high success rate, wherein the success rate reaches 60-80%; and the establishment of the Lycoris herb genetic transformation system develops the new way for thecultivation of new Lycoris herb varieties.

Description

technical field [0001] The invention belongs to the technical field of genetics, and in particular relates to a method for constructing a genetic transformation system of the genus Lycoris. Background technique [0002] Lycoris Herb. belongs to Amaryllidaceae (Amaryllidaceae), a perennial herb. Bulbs ovoid, leaves in spring, leaves band-shaped, umbels with 4-6 small flowers; flowers yellow, red, white; perianth lobes recurled and shrunken. The flowering period is from July to August. The flower stems are tall and straight, the flower shape is beautiful, and the flower color is gorgeous. All species of this genus have high ornamental value, and have gradually become new-type fresh-cut flowers, garden ground covers and potted bulbous flowers. [0003] But at present, the utilization of Lycoris in my country is limited to the domestication and cultivation of wild resources. The cultivation and promotion of new varieties of Lycoris is only in its infancy. At present, only t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H4/00
CPCA01H4/001A01H4/005A01H4/008C12N15/8205
Inventor 郑玉红张鹏翀庄维兵傅力刘天宇鲍淳松韩福贵
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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