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Application of mortierella mycorrhiza fungi to culture of tissue culture seedling of dendrobium officinale

A technology of mycorrhizal fungi and Mortierella, applied in the field of microorganisms and tissue culture, can solve the problems of low survival rate of transplanting and domestication, slow growth of tissue culture seedlings, etc.

Inactive Publication Date: 2014-06-25
ZHEJIANG FORESTRY UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

People began to seek artificial cultivation methods to cultivate Dendrobium officinale, and the development of tissue culture technology has realiz

Method used

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  • Application of mortierella mycorrhiza fungi to culture of tissue culture seedling of dendrobium officinale
  • Application of mortierella mycorrhiza fungi to culture of tissue culture seedling of dendrobium officinale
  • Application of mortierella mycorrhiza fungi to culture of tissue culture seedling of dendrobium officinale

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Experimental program
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Embodiment 1

[0019] Embodiment 1: Mortierella mycorrhizal fungus in the present invention Mortierella Isolation and identification of FL9 strain

[0020] Collected from the fresh nutrient roots of Chunlan in the wild state of Daming Mountain, Zhejiang, rinse the sediment on the roots with water, sterilize the surface of the material, rinse it with sterile water, cut it into a block with a growth of about 5mm under sterile conditions, and place it on a PDA In the culture medium, the cut surface faces the culture medium, and cultured at a constant temperature of 25°C and protected from light. After 7 days of culture, mycelia grew out from the tissue block to form colonies of a certain size, and the mycelium was picked from the edge and moved to another PDA plate to repeat until purification to obtain strain FL9.

[0021] Morphological observation of strain FL9 and staining of lactophenol Meslin, combined with Figure 1-3 , the biological characteristics are expressed as: growing on PDA med...

Embodiment 2

[0024] Embodiment 2: the rooted strong seedling of dendrobium candidum tissue culture seedling

[0025] Select seedlings with basically the same growth state, with a weight of about 0.28±0.4g and a plant height of 2.5±0.5cm, and inoculate them into the medium for rooting and strong seedlings. The basic medium is 1 / 2MS, with an additional 0.7mg L -1 IBA+1.0mg·L -1 NAA sucrose 30mg·L -1 , agar 6.8mg·L -1 , activated carbon 1mg·L -1 , with a pH of 5.8, the effects of different organic additives such as coconut milk, banana puree, and potatoes on the rooting and strengthening of Dendrobium candidum tissue culture seedlings, 5 plants were inoculated in each bottle, and repeated three times. After two months of cultivation, the plant height, fresh weight and root number of the seedlings were investigated. The results showed that adding coconut milk was beneficial to the differentiation of Dendrobium officinale seedlings and the generation of new roots.

Embodiment 3

[0026] Embodiment 3: Screening of bacterin symbiotic medium

[0027] A: DE1 medium, formula: CaCl 2 1.0mmol·L –1 , MgSO 4 0.5mmol·L –1 , K 2 SO 4 1.0mmol·L –1 , KH 2 PO 4 0.4mmol·L –1 , FeSO 4 100μmol L –1 , H 3 BO 4 25μmol·L –1 , MnCl 2 33μmol·L –1 , ZnSO 4 2.8μmol·L –1 , NaMoO 4 1.0μmol L –1 , Na 2 EDTA140μmol·L –1 , yeast juice 1.0g·L –1 , agar 8.0g·L –1 .

[0028] B: Choose Dijk and Eck's improved medium for symbiotic culture of orchids and fungi, referred to as DE2 medium, and its formula is: CaCl 2 1.0μmol·L –1 , MgSO 4 0.5μmol·L –1 , K 2 SO 4 1.0μmol·L –1 , KH 2 PO4 0.4μmol·L –1 , FeSO 4 100μmol·L –1 , H 3 BO 4 25μmol·L –1 , MnCl 2 33μmol·L –1 , ZnSO 4 2.8μmol·L –1 , NaMoO 4 1.0μmol·L –1 , Na 2 EDTA140μmol·L –1 , yeast juice 1.0g·L –1 , agar 8.0g·L –1 , Soluble starch 9.0g / L.

[0029] C: MS medium, sucrose 10g·L -1 .

[0030] D: MS medium, sucrose 15g L -1 .

[0031] E: MS medium, sucrose 20g L -1 .

[0032]...

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Abstract

The invention relates to application of mortierella mycorrhiza fungi to culture of a tissue culture seedling of dendrobium officinale, belonging to a microorganism and tissue culture technology, and particularly relating to separation and identification of mycorrhiza fungi, tissue culture of the dendrobium officinale, and a function of the mycorrhiza fungi to the increase of the biomass of the dendrobium officinale and mineral elements. The mortierella mycorrhiza fungi is applied to rooting and strong seedling culture of the tissue culture seedling of the dendrobium officinale, including three steps: isolated culture of the mycorrhiza fungi, rooting and strong seedling culture of the tissue culture seedling of the dendrobium officinale, and mycorrhization of the dendrobium officinale. By adopting the invention, the tissue culture seedling of the dendrobium officinale is remarkably increased in terms of fresh weight growth rate, dry weight, plant height growth rate, tillering, rooting and the like, and is remarkably increased in absorbing speed for a mineral substance, especially a large quantity of elements; and fungi (i) Mortierella (/i)FL9 and the dendrobium officinale have a good symbiotic relationship.

Description

technical field [0001] The invention relates to the application of a mycorrhizal fungus of the genus Mortierella in the cultivation of dendrobium candidum tissue culture seedlings, and belongs to the technical field of microorganisms and tissue culture. Background technique [0002] Most orchids in nature have symbiotic mycorrhizal fungi in different growth states. It is difficult for orchid seeds to germinate. Under natural conditions, orchids grow well because orchids and fungi form a good symbiotic relationship, which can promote the growth of orchids. [0003] Dendrobium officinale is a perennial herb of the Orchidaceae Dendrobium genus, which is a traditional Chinese medicinal material and ranks first among the nine immortal grasses in my country. In recent years, with the enhancement of people's awareness of health care, the market demand for Dendrobium officinale has increased, and wild Dendrobium resources have been severely harvested. People began to seek artifici...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01H4/00C12R1/645
Inventor 崔永一王芬钱仁卷
Owner ZHEJIANG FORESTRY UNIVERSITY
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