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Method and culture medium for carrying out induction culture on regenerated plants by adopting pulsatilla chinensis anthers

A technology for inducing culture medium and regenerating plants, which is applied in the field of inducing and cultivating regenerated plants of Pulsatilla anthers, and achieves the effects of high induction rate and improved culture efficiency.

Active Publication Date: 2018-02-23
FORESTRY RES INST OF HEILONGJIANG PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are few reports on the in vitro cultivation of Pulsatillae, mainly focusing on explants such as seeds and leaves for in vitro culture. There is no report on how to use Pulsatilla pollen as explants to successfully cultivate Pulsatilla regenerated plants in vitro

Method used

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  • Method and culture medium for carrying out induction culture on regenerated plants by adopting pulsatilla chinensis anthers
  • Method and culture medium for carrying out induction culture on regenerated plants by adopting pulsatilla chinensis anthers
  • Method and culture medium for carrying out induction culture on regenerated plants by adopting pulsatilla chinensis anthers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] The basal medium used for Pulsatilla anther-induced regeneration plants is MS modified medium, which consists of:

[0079] Macroelements: ammonium nitrate 1300mg / L, potassium nitrate 1500mg / L, potassium dihydrogen phosphate 210mg / L, magnesium sulfate 370mg / L and calcium chloride 400mg / L;

[0080] Trace elements: potassium iodide 0.86mg / L, boric acid 6.2mg / L, manganese sulfate (MnSO 4 4H 2 O) 22.3mg / L, zinc sulfate (ZnSO 4 ·7H 2 O) 8.6mg / L, sodium molybdate (Na 2 MoO 4 2H 2 O) 0.25mg / L, copper sulfate (CuSO 4 ·5H 2 O) 0.025mg / L and cobalt chloride (CoCl 2 ·6H 2 O) 0.025mg / L;

[0081] Iron salt: disodium edetate (Na 2 EDTA) 37.3mg / L and ferrous sulfate (FeSO 4 ·7H 2 O) 27.8mg / L;

[0082] Organic Matter: Inositol (C 6 h 12 o 6 2H 2 0) 100mg / L, Niacin (NC 5 h 4 COOH) 0.5mg / L, thiamine hydrochloride (C 12 h 17 ClN 4 OS·HCl) 0.1mg / L, pyridoxine hydrochloride (C 8 h 11 o 3 N·HCl) 0.5mg / L and glycine (NH 2 ·CN 2 · COOH) 2mg / L.

Embodiment 2

[0083] Example 2 Pulsatilla pollen induced culture regeneration plant

[0084] 1. Material collection and processing

[0085] From April to May, the fine wild varieties of Pulsatilla pulsatillae in Maoer Mountain, Harbin City, Heilongjiang Province were selected. At this time, the pollen is in the vigorous growth period, and the collection time is selected from 9 to 10 am in sunny weather. During the period from budding (emergence of flower buds) to flowering, flower buds were collected every 2 days. The collected flower buds were placed in a refrigerator at 4°C for 48 hours at low temperature and set aside.

[0086] Before inoculation, the stage of pollen development was identified by microscopic examination, and the pollen of Pulsatilla grandis in the middle stage of mononuclei was selected.

[0087] The microscopic examination method for determining the pollen development period is as follows: tear off the corolla with tweezers and take out the anthers, place the anthers ...

Embodiment 3

[0093] The culture medium and steps for inducing callus were the same as in Example 2, except that the developmental stage of the anthers was selected as the uninucleate marginal stage.

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Abstract

The invention provides a basic culture medium for carrying out induction culture on regenerated plants by adopting pulsatilla chinensis anthers, and belongs to the technical field of culturing of plant tissues. Using amounts of macroelements and a part of trace elements are improved based on an MS culture medium. Based on the basic culture medium provided by the invention, induction culture on theregenerated plants by adopting the pulsatilla chinensis anthers is carried out, and induction is achieved successfully so as to obtain the regenerated plants of the pulsatilla chinensis anthers. Theinvention also provides a method for carrying out induction culture on the regenerated plants by adopting the pulsatilla chinensis anthers. The method comprises the following steps: selecting and inoculating the pulsatilla chinensis anthers in monocyte periods into the induced culture medium; performing dark culture until green and bright shoots are formed on the surfaces of wound callus, transferring the shoots into a proliferation culture medium for proliferation culture, and carrying out induction to form clustered shoots; transferring the clustered shoots into a rooting culture medium forculturing to obtain the regenerated plants of the pulsatilla chinensis. The pulsatilla chinensis anthers are cultured by adopting the method provided by the invention, so that an induction ratio, a proliferation ratio and a rooting ratio are high, and culture time is short, and the culturing efficiency of the regenerated plants of the pulsatilla chinensis is effectively improved.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a basal medium, an induction medium, a proliferation medium and a rooting medium for inducing and cultivating regenerated plants of Pulsatilla pulsatillae anthers, and a method for inducing and cultivating regenerated plants of Pulsatilla chinensis anthers. Background technique [0002] Pulsatilla chinensis (Bunge) Regel, also known as sheep's beard flower, old crown flower, general grass, big bowl flower, husband flower, old sister-in-law flower and Maoguduo flower, is a plant of the genus Pulsatilla in the family Ranunculaceae. Plant height 15-35 cm, rhizome 0.8-1.5 cm thick, basal leaves 4-5, leaves broadly ovate, 3-lobed, middle segment stalked or nearly sessile, scape 1(-2), Plumber, erect flowers, sepals blue-purple, oblong-ovate, aggregate fruit 9-12 cm in diameter, achenes spindle-shaped, flat, flowering from April to May. Pulsatilla mostly grows in areas wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 舒钰李晶王丹赵学丽王承义王颖
Owner FORESTRY RES INST OF HEILONGJIANG PROVINCE
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