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Inducing method of dioscorea alata transgenosis hairy roots

A hairy root and transgenic technology, which is applied in the field of plant cultivation and transgenic, can solve the problems of tissue culture of warm yam, etc., and achieve the effect of rapid and effective reproduction

Active Publication Date: 2019-06-28
HANGZHOU NORMAL UNIVERSITY
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Problems solved by technology

[0004] And do not see in the prior art that warm yam is carried out tissue culture, and establish the method that warm yam induces transgenic hairy root by Agrobacterium induction

Method used

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  • Inducing method of dioscorea alata transgenosis hairy roots

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Embodiment

[0027] 1. Materials and methods

[0028] 1. Materials

[0029] The tubers of Wen Yam were donated by Tianhe Biotechnology Co., Ltd., Wenzhou City, Zhejiang Province. The tubers were planted in the glass greenhouse of the unit to obtain seedlings. Take the leaves, stem segments and stem segments with a leaf axil, i.e., noded stem segments, of the seedlings as explants.

[0030] 2. Disinfection of explants

[0031] The explants were first soaked in diluted detergent for 5 minutes, rinsed with tap water, disinfected with 5 times diluted sodium hypochlorite for 20 minutes, rinsed with sterile water for 3 times, and finally blotted the surface moisture with sterile filter paper. Use scissors to cut the leaves into small pieces about 1 cm square, and cut the stems with or without leaf axils to a length of 0.5-1 cm.

[0032] 3. Culture of explants

[0033] The culture medium for inducing callus from explants uses MS basic medium, adding different proportions of 2,6-D and NAA; th...

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Abstract

The invention discloses an inducing method of dioscorea alata transgenosis hairy roots, and belongs to the technical field of plant culture and transgenosis. The method includes the steps: 1) explantsterilization; 2) explant culture; 3) infection liquid preparation; 4) hairy root induction and reproduction; 5) hairy root reproduction. According to the method, dioscorea alata stems with leaf axilla serve as explants, an induction culture medium, a differential culture medium and a rooting culture medium are optimized, an integrated plant is acquired through induction, differential and a rooting culture, stems of dioscorea alata aseptic seedlings are infected by agrobacterium rhizogenes K599 (with recombinant plasmid pRI101-AN-gfp) living bodies to obtain the transgenosis hairy roots, PCR (polymerase chain reaction) and fluorescence microscope observation identify whether the hairy roots are transgenosis hairy roots or not, an inducing system of the dioscorea alata transgenosis hairy roots is built, and dioscorea alata is rapidly and effectively bred.

Description

technical field [0001] The invention belongs to the technical field of plant cultivation and transgene, and in particular relates to a method for inducing transgenic hairy roots of warm Chinese yam. Background technique [0002] Yam has a history of more than 2,000 years of cultivation and application in my country. It can be used as miscellaneous grains and has medicinal value. Wenshanyao (Dioscorea alata, cultivar "Wenshanyao") is a special yam variety in Zhejiang Province. It has the medicinal effects of strengthening the spleen and stomach, benefiting the lungs and kidneys, nourishing qi and nourishing yin, and has a long history of cultivation. [0003] The traditional propagation method of warm yam is vegetative propagation, which not only consumes a large amount of medicinal parts, but also causes species degradation and yield reduction due to long-term vegetative propagation. Improving quality and increasing output has become an urgent problem to be solved in the pro...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 向太和金婷婷李亚菲章铖超王克云李蓉徐正贤贺宸靖
Owner HANGZHOU NORMAL UNIVERSITY
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