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Genetic Transformation Method of Dreb Gene Transformation Anthurium

A method of genetic transformation, a technology of genetic transformation, applied in the field of molecular breeding

Inactive Publication Date: 2011-12-21
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, the introduction of DREB gene to improve the stress resistance of crops has been successful in crops such as rapeseed, tobacco, tomato, wheat and chrysanthemum, but the research on using DREB gene to transform anthurium has not been reported yet.

Method used

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  • Genetic Transformation Method of Dreb Gene Transformation Anthurium
  • Genetic Transformation Method of Dreb Gene Transformation Anthurium
  • Genetic Transformation Method of Dreb Gene Transformation Anthurium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Obtaining aseptic callus:

[0041] 1) Explant selection

[0042] The stems, leaves, and petioles were used to induce callus on the subculture medium 1 / 2MS+6-BA1.0mg / L+IBA0.1mg / L, 40 days later for experiment. The culture conditions are: temperature 24~26℃, light intensity 40μmol·m -2 ·S -1 , 12h / 12h light and dark alternately. Results: The stem segment was used as an explant for tissue culture and bud induction rate was the highest and the most successful (Table 1, figure 1 ).

[0043] Table 1: Callus induction of Anthurium andraeanum by different explants

[0044] Explant

[0045] 2) The effect of different hormone conditions on the callus of the stem of Anthurium andraeanum

[0046] Take the tissue cultured seedlings of Anthurium andraeanum, cut the stem tip of Anthurium andraeanum with a pair of leaves, add 0.1mg / l IBA and 0.1mg / l NAA to the medium 1 / 2MS+6-BA1.0mg / L to induce callus Tissues, and use 1 / 2MS+6-BA1.0mg / L without hormones to induce callus as a cont...

Embodiment 2

[0054] Example 2 Agrobacterium genetic transformation

[0055] 1) The effect of hygromycin (Hgy) concentration on the callus differentiation and bud growth of Anthurium andraeanum

[0056] Select callus and buds of the same size and inoculate them in induction medium containing different concentrations of hygromycin, the concentrations of which are 0, 5mg / L, 10mg / L, 20mg / L, 30mg / L, 50mg / L, Change the fresh medium once every two weeks, and observe the statistical results after 45 days, see Table 5. Table 5 shows that at a very low concentration, the inhibitory effect of hygromycin on the callus differentiation and bud growth of Anthurium andraeanum is not obvious, the differentiation rate of callus reaches 92%, and the bud growth is robust; but when the concentration reaches 30mg At 50 mg / L, there is browning around the callus, and bud growth is slow; with the increase of hygromycin concentration, the inhibitory effect is proportional to its concentration. At 50 mg / L, the callus is...

Embodiment 3

[0080] Example 3 The effect of IBA concentration on root differentiation

[0081] The callus blocks with adventitious buds obtained by resistance screening were transferred to regeneration medium MS+6-BA1.0mg / L+IBA 1.0mg / L without antibiotics at a temperature of 26±2℃, Cultivate under the condition of light time of 16h / d. When the adventitious buds grow to 2.0-3.0cm, cut the adventitious buds and inoculate with the IBA concentration of 0mg / L, 0.1mg / L, 0.5mg / L, 1.0mg / L, 1.5mg / L 1 / 2MS medium. The results showed that when the concentration of IBA was 0.1mg / L, rooting was faster, and roots took place in about two weeks. The roots were thick and abundant; as the concentration of hormones increased, the time for roots to occur was prolonged, especially when the concentration of IBA increased. When it exceeds 1.0 mg / L, the root is not only difficult to occur, but also very easy to heal, with the root tip swelling and yellowing. Therefore, the optimum concentration is when the IBA con...

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Abstract

The invention belongs to the field of molecular breeding and discloses a genetic transformation method for transforming anthurium with DREB gene. The method is to incise the aseptic callus of Anthurium anthurium, infect with the Agrobacterium liquid containing the recombinant plasmid pCAMBIA1301-DREBa for 10 minutes, co-cultivate for 3 days, perform antibacterial culture for 5 days, and screen to obtain hygromycin-resistant rooted shoots. . It was confirmed by PCR detection that the exogenous gene had been transferred into the genome DNA of Anthurium anthurium. The present invention establishes its genetic transformation system for Anthurium for the first time, provides theoretical basis and experimental basis for Anthurium molecular breeding, and will effectively promote its molecular breeding process.

Description

Technical field [0001] The invention belongs to the field of molecular breeding, and relates to a genetic transformation method for transforming DREB gene into Anthurium andraeanum. Background technique [0002] Anthurium is a perennial herb of the anthurium of the Araceae family. It is native to southern Colombia and is a rare flower in the world. Anthurium has gorgeous buddha buds, long flowering period, annual flowering, and waxy leaves. It is an indoor ornamental plant with excellent foliage and flowers. The optimum temperature for growth of Anthurium is 18-20℃ at night and around 25℃ during the day. In summer, if the room temperature at night exceeds 25℃ or the room temperature during the day exceeds 30℃, and in winter, if the room temperature at night is below 12℃ or the room temperature during the day is below 18℃, the leaves will lose luster and yellow , Foyan buds have shorter flowering period, are easy to wither, and can also cause leaf rot, which seriously affects it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C12N15/66A01H5/00
Inventor 滕年军陶清良陈发棣陈琳房伟民蒋甲福陈素梅刘兆磊
Owner NANJING AGRICULTURAL UNIVERSITY
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