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A method for direct somatic embryo induction of betel nut and its suspension culture to obtain regenerated plants

A technique of regenerating plants and suspension culture, which is applied in the biological field, can solve the problems such as the research report on direct somatic embryo induction suspension culture of betel nut, and achieve the effects of shortening the breeding cycle of varieties, convenient operation, and easy control of growth conditions

Active Publication Date: 2021-12-03
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There is no report on direct somatic embryo induction and suspension culture of betel nut in China

Method used

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  • A method for direct somatic embryo induction of betel nut and its suspension culture to obtain regenerated plants
  • A method for direct somatic embryo induction of betel nut and its suspension culture to obtain regenerated plants
  • A method for direct somatic embryo induction of betel nut and its suspension culture to obtain regenerated plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1. Explant acquisition: select a high-yielding healthy betel nut plant, take the uncracked young inflorescence of spathe as an explant, wash the surface of the inflorescence with a saturated washing powder solution and rinse it clean, and wash it with 75% alcohol in an ultra-clean workbench. After wiping the entire surface of the inflorescence evenly several times, use 0.1% HgCl 2 Soak for 5-12min, wash 5 times with sterile water, and place on sterile filter paper to blot the surface moisture.

[0027] 2. Direct somatic embryo induction: Take the above-mentioned sterilized explants and cut them into 1-3mm thick small pieces, inoculate them into somatic embryo induction medium, and culture them in dark at 25-28°C to obtain somatic embryos ( See figure 1).

[0028] Somatic embryo induction medium: NH 4 Cl 1000mg / L+H 3 BO 3 10mg / L+KNO 3 2400mg / L+Ca(NO 3 ) 2 620mg / L+NaH 2 PO 4 300mg / L+MgSO 4 ·7H 2 O 350mg / L+CuSO 4 ·5H 2 O 0.25mg / L+Na 2 -EDTA37.3mg / L+FeSO ...

Embodiment 2

[0039] 1. Explant acquisition: select a high-yielding healthy betel nut plant, take the uncracked young inflorescence of spathe as an explant, wash the surface of the inflorescence with a saturated washing powder solution and rinse it clean, and wash it with 75% alcohol in an ultra-clean workbench. After wiping the entire surface of the inflorescence evenly several times, use 0.1% HgCl 2 Soak for 5-12min, wash 3 times with sterile water, and place on sterile filter paper to blot the surface moisture.

[0040] 2. Direct somatic embryo induction: take the above-mentioned sterilized explants and cut them into 1-3mm thick small pieces, inoculate them into somatic embryo induction medium, and culture them in dark at 25-28°C to obtain somatic embryos.

[0041] Somatic embryo induction medium: NH 4 Cl 1100mg / L+H 3 BO 3 6mg / L+KNO 3 2500mg / L+Ca(NO 3 ) 2 560mg / L+NaH 2 PO 4 325mg / L+MgSO 4 ·7H 2 O 370mg / L+CuSO 4 ·5H 2 O 0.25mg / L+Na 2 -EDTA37.3mg / L+FeSO 4 ·7H 2 O 27.8mg / L...

Embodiment 3

[0052] 1. Explant acquisition: select a high-yielding healthy betel nut plant, take the uncracked young inflorescence of spathe as an explant, wash the surface of the inflorescence with a saturated washing powder solution and rinse it clean, and wash it with 75% alcohol in an ultra-clean workbench. After wiping the entire surface of the inflorescence evenly several times, use 0.1% HgCl 2 Soak for 5-12min, wash 5 times with sterile water, and place on sterile filter paper to blot the surface moisture.

[0053] 2. Direct somatic embryo induction: take the above-mentioned sterilized explants and cut them into 1-3mm thick small pieces, inoculate them into somatic embryo induction medium, and culture them in dark at 25-28°C to obtain somatic embryos.

[0054] Somatic embryo induction medium: NH 4 Cl 1300mg / L+H 3 BO 3 5mg / L+KNO 3 2600mg / L+Ca(NO 3 ) 2 480mg / L+NaH 2 PO 4 350mg / L+MgSO 4 ·7H 2 O 390mg / L+CuSO 4 ·5H 2 O 0.25mg / L+Na 2 -EDTA37.3mg / L+FeSO 4 ·7H 2 O 27.8mg / L...

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Abstract

The invention belongs to the field of biotechnology, and relates to a method for direct somatic embryo induction of areca nut and a method for obtaining regenerated plants by suspension culture. Somatic embryos are obtained by dark culture at 25-28° C., and then through suspension proliferation and differentiation culture to obtain somatic embryos; the somatic embryos are inoculated into somatic embryo germination medium and cultured to obtain regenerated plants. The method of the invention is simple in process and convenient in operation. The young inflorescence of areca nut is used as the explant material to directly induce and obtain early embryos, and the suspension culture technology is used to carry out large-scale proliferation, and then the somatic embryos are cultured into plants. The cycle is short, and the proliferation efficiency is high. The suspension germ line has the same genetic background, basically the same physiological state, and has good totipotency.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a method for obtaining regenerated plants through tissue culture, in particular to a method for directly inducing somatic embryos using areca nut inflorescences as explant materials and obtaining regenerated plants through suspension culture. Background technique [0002] Betel nut (Areca catechu L.) is an evergreen tree with a single growth point in the genus Areca of the palm family. It is native to the tropical rainforest of the Malay Peninsula. It is now widely distributed in India, Pakistan, Sri Lanka, and Malaysia. In China, it is mainly distributed in Hainan and Taiwan. , southern Yunnan, in addition, Guangdong, Guangxi, Yunnan, Fujian and other provinces (regions) also have a small amount of cultivation. Betel nut is an important medicinal plant, and it is the first of the four major southern medicines in my country. Its fruit has the functions of strengthening teeth an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 潘登浪曾宪海林位夫谢贵水刘钊李炜芳
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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