Bergapten endophyte separating and cultivation method and application thereof
A cultivation method and endophyte technology, applied in the biological field, can solve problems such as no research report on endophyte of ligustrum, and achieve the effects of promoting faster development, shortening rooting time, and improving rooting rate.
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Embodiment 1
[0036] Embodiment 1: A kind of separation and cultivation method of ligustilis endophytes
[0037] It includes the following steps:
[0038] (1) material preparation: select the fresh whole plant of Ligusticum edulis without mildew, and clean the surface silt for subsequent use;
[0039] (2) Surface sterilization: After sterilizing the whole plant of Ligusticum edulis by irradiation, soak it in 75% ethanol for 5 minutes, and rinse it 4 times with sterile water;
[0040] (3) Cultivation after crushing: use sterilized tools to cut Ligusticum edulis stems, leaves, and roots into pieces, and plant the 1:1:1 mixture of stems, leaves, roots, and stem-leaf roots in a petri dish. Cultivate in a constant temperature incubator for 5 days;
[0041] (4) Purification: Pick the hyphae at the edge of the colony and put them in the water agar medium, and cultivate them for 5 days to obtain the endophytes of Ligusticum edulis.
Embodiment 2
[0042] Embodiment 2: Effect of Ligusticum endophytes on the rooting time of Ligusticum cuttings
[0043] 2.1 Experimental materials: Cut the stems and branches of Ligusticum edulis, select the middle part of the stem node, the node is enlarged, and there are 240 parts with one mature leaf, and they are randomly divided into blank group, Ligusticum stem endophyte group, and Ligusticum leaf endophyte group. Bacteria group, Ligusticum root endophyte group, Ligusticum stem leaf root mixture endophyte group and 200mg / L naphthalene acetic acid group, 40 rats in each group;
[0044] 2.2 Experimental method:
[0045] Inoculate the isolated Ligusticum endophyte into an Erlenmeyer flask containing a liquid medium. After cultivating on a shaker for 8 days, insert the cuttings of Ligusticum into the Erlenmeyer flask. The blank group uses sterile water, and after continuing to cultivate for 2 days, Take out the cutting branches of Ligusticum edulis, wash them with sterile water for 3-5 ti...
Embodiment 3
[0050] Example 3: Effect of Ligusticum endophytes on Ligusticum seed germination
[0051] 3.1 Experimental materials
[0052] The Ligusticum edulis seeds harvested in autumn should be dried in the shade after harvesting. Before breeding, the Ligusticum edulis seeds should be cleaned with clear water. After the cleaned Ligusticum edulis seeds were soaked in 75% ethanol solution for 5 minutes, they should be washed 4 times with sterile water. grains, randomly divided into blank group, Ligusticum stem endophyte group, Ligusticum leaf endophyte group, Ligusticum root endophyte group, Ligusticum stem leaf root endophyte group, 100 grains in each group;
[0053] 3.2 Experimental method
[0054] The isolated endophytic bacteria of Ligusticum edulis were inoculated into conical flasks containing liquid medium, and after cultivating on a shaker for 8 days, sterilized Ligusticum edulis seeds were added to the conical flasks, and the blank group was added with sterile water. After cont...
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