CN113179954B

CN113179954B - Method for cultivating tissue culture seedlings by stem segments of artemisia rupestris L

Info

Publication number: CN113179954B
Application number: CN202110674727.8A
Authority: CN
Inventors: 金艳
Original assignee: Guangxi Yao Autonomous County Yicao Fengmao Yao Medicine Co ltd
Current assignee: Guangxi Yao Autonomous County Yicao Fengmao Yao Medicine Co ltd
Priority date: 2021-06-18
Filing date: 2021-06-18
Publication date: 2022-12-30
Anticipated expiration: 2041-06-18
Also published as: CN113179954A

Abstract

The invention discloses a method for cultivating tissue culture seedlings by stem segments of blumea balsamifera, which comprises the steps of shearing off leaves of clustered buds Ai Zengshi of blumea balsamifera on a super clean workbench to form stem segments with 1 internode as culture materials; the cut stem segments are inoculated into a long-bud rooting culture medium, axillary buds on internodes of the stem segments germinate and continuously grow up and grow for 7 to 13 days, and the bud base part expands and adventitious roots grow out after 20 to 30 days to form a complete red stem Ai Zhizhu; the culture medium for rooting the long buds comprises: MS +6-BA 0.5-3.0 mg/L + TDZ 0.01-0.1 mg/L + NAA 0.1-0.5 mg/L + IAA 0.1-1.0 mg/L + sucrose 30g/L + agar 5g/L, and the pH value is 6.0. The yield of the blumea balsamifera tissue culture seedling produced by the method is more than 5 times of that of the tissue culture seedling cultured by buds conventionally, the production cost is greatly reduced, and the production efficiency is greatly improved. The method has the advantages of good repeatability and stability, simple operation, low cost and high yield.

Description

Method for cultivating tissue culture seedlings by stem segments of artemisia rupestris L

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a method for cultivating tissue culture seedlings by stem segments of Artemisia argyi.

Background

The blumea balsamifera is a blumea herb plant of the Compositae, and is a traditional Chinese medicine in China. "northern Chinese mugwort and southern red mugwort", the local person refers to the red-stem mugwort because the stem of the red mugwort is red. The red-stem moxa is mainly grown in two broad areas in south China, and the husband Bao Xiangu of Ge Hong commonly uses the red-stem moxa for moxibustion disease-removing and health care of patients, so the red-stem moxa is also called as 'Bao Guai' and also called as Shenai. The Yao nationality is warm and slightly bitter in taste, the whole herbs are used as medicines, and the Yao nationality is a life-prolonging nation in China and is a miracle medicine for health preservation, and the Yao nationality is recorded as a Baiyao Miao medicine through traditional Chinese medical records and is worried as a health-preserving holy medicine.

The red stems Ai Lingxing are distributed in the wild, the folk is often eaten as green vegetables and cakes, the medicine is mainly the prescription component of Chinese patent medicine and the moxa stick is prepared, the moxa stick can not only eliminate diseases and improve the immunity of human bodies, but also can be used anytime and anywhere due to convenient carrying and no special equipment, and becomes a necessary health care product in daily life of people. At present, the red-stem moxa is mainly wild, a small amount of seedlings are bred and planted in a plant division manner, the red-stem moxa in the wild is scattered, a large amount of manpower and material resources are needed for collection, the supply is unstable, and stable raw material guarantee cannot be provided for moxa stick production. At present, large-area planting of the blumea balsamifera in Guangxi is not reported, propagation of the blumea balsamifera Ai Chongmiao can be plant division propagation, propagation of tissue culture seedlings and propagation of seeds, and the plant division propagation can not meet the large-scale planting requirement due to low propagation coefficient, can damage wild natural resources and occupies medicinal resources; the seed propagation is mainly characterized in that the seed maturation time is inconsistent, the collection is difficult, and the large-scale seedling culture is difficult. The applicant utilizes modern biotechnology to carry out rapid propagation research on the red stems Ai Zupei, and although the propagation coefficient reaches more than 5.0, in order to further reduce the cost, the induction of the red stems Ai Zhizhu by utilizing stem segments of buds is carried out. Under normal conditions, the stem of the underground part of the Artemisia argyi can induce the plant, and the stem segment of the overground part is used as the photosynthesis and nutrition transportation tissue organ of the whole plant, so that the plant can survive by the nutrition supply of the root system and the leaves, and the plant is hard to induce because the stem segment is rotten and dead after being cut off by the nutrition supply chain.

Disclosure of Invention

The invention aims to provide a method for cultivating tissue culture seedlings by stem segments of Artemisia rupestris L.var.japonica Makino, which takes sterile multiplication buds of Jin Xiu Artemisia rupestris L.var.japonica Makino as materials, can induce roots while growing buds on the stem segments with internodes of the multiplication buds by applying a proper proportion of the types and concentrations of plant growth regulators, forms complete red stems Ai Zhizhu, and summarizes the method for cultivating the tissue culture seedlings by stem segments of the Artemisia rupestris L.var.japonici.

The technical scheme for realizing the purpose of the invention is as follows:

the method for cultivating the tissue culture seedlings of the blumea balsamifera by stem segments comprises the following steps:

(1) On a clean bench, cutting off leaves of cluster buds Ai Zengshi of red stalks to stem sections with 1 internode as a culture material;

(2) The cut stem segments are inoculated into a long-bud rooting culture medium, axillary buds on internodes of the stem segments germinate and continuously grow up and grow for 7 to 13 days, and the bud base part slightly expands and grows adventitious roots after 20 to 30 days to form a complete red stem Ai Zhizhu;

the culture medium for rooting the long buds comprises: MS +6-BA 0.5-3.0 mg/L + TDZ 0.01-0.1 mg/L + NAA 0.1-0.5 mg/L + IAA 0.1-1.0 mg/L + sucrose 30g/L + agar 5g/L, pH value is 6.0;

the culture conditions in the culture stage are as follows: the culture temperature is 28 +/-1 ℃, the illumination time is 10 h/d, and the illumination intensity is 5000lx.

The preferred selection of the culture medium for rooting the long buds is as follows: MS +6-BA 1.5 mg/L + TDZ 0.05mg/L + NAA0.2 mg/L + IAA0.5 mg/L + sucrose 30g/L + agar 5g/L, and the pH value is 6.0;

in the culture medium, MS is a basic culture medium, 6-BA is 6-benzylaminopurine, TDZ is thidiazuron, NAA is alpha-naphthylacetic acid, IAA is indole-3-acetic acid, the reagents are analytically pure, and water is distilled water. 6-BA has physiological effects of promoting cell division, differentiation of non-differentiated tissues, accumulation of substances in organism, lateral bud generation and aging prevention; NAA has effects of promoting cell division and expansion, and inducing to form adventitious root; TDZ can stimulate the synthesis of in vivo auxin substances, induce the generation of adventitious buds and the regeneration of root tissues; IAA can stimulate branch cell elongation, promote cambium cell division, promote xylem and phloem cell differentiation, and promote hairy root. Under the combined action of 6-BA, TDZ, NAA and IAA, the stem segment with internodes of the red stem moxa completes the processes of budding and root induction in the culture medium, and finally forms complete red stem Ai Zhizhu.

The invention has the advantages that: because each bud in the red-stem moxa propagation bud has more than 5 internodes, and each internode is cut into a stem section as the material for cultivating the red-stem moxa tissue culture seedling, the red-stem moxa tissue culture seedling produced by the method has more than 5 times of the production capacity of the tissue culture seedling cultivated by buds conventionally, the production cost is greatly reduced, and the production efficiency is greatly improved. The method has the advantages of good repeatability and stability, simple operation, low cost and high yield.

Detailed Description

The present invention will be further described with reference to the following examples, but the present invention is not limited thereto.

Example 1

(2) The cut stem segments are inoculated into a rooting medium for growing buds, axillary buds on internodes of the stem segments germinate and have slender buds after 13 days, 1-2 adventitious roots grow on the base parts of partial buds after 30 days, the base parts of partial buds do not grow roots, and the red stems Ai Zhizhu are thin and weak;

the culture medium for rooting the long buds comprises: MS +6-BA 0.5 mg/L + TDZ 0.01mg/L + NAA0.1 mg/L + IAA0.1 mg/L + sucrose 30g/L + agar 5g/L, and the pH value is 6.0;

Example 2

The method for cultivating the tissue culture seedlings by the stem segments of the artemisia rupestris L comprises the following steps:

(2) The cut stem segments are inoculated into a long bud rooting culture medium, axillary buds on the internodes of the stem segments start to germinate and continuously grow up and grow up, buds are thick and strong, but partial buds have vitrification phenomenon, the base parts of the buds expand to form callus tissues after 23 days, adventitious roots grow on the callus tissues, the roots are thick and short, leaves curl, and the red stems Ai Zhizhu grow abnormally because the concentration of a plant growth regulator in the culture medium is overhigh;

the culture medium for rooting the long buds comprises: MS +6-BA 3.0 mg/L + TDZ0.1mg/L + NAA0.5 mg/L + IAA1.0 mg/L + sucrose 30g/L + agar 5g/L, and the pH value is 6.0;

Example 3

(1) On a clean bench, cutting off leaf blades of cluster buds Ai Zengshi of red stalks into stem sections with 1 internode, and using the stem sections as culture materials;

(2) The cut stem segments are inoculated into a long-bud rooting culture medium, axillary buds on internodes of the stem segments start to germinate after 7 days, the buds grow continuously and grow to be thick and strong, the bud base part slightly expands after 20 days, 2-5 adventitious roots grow, and the stem of the red stem Ai Zhizhu is red and well grown;

the culture medium for rooting the long buds comprises: MS +6-BA 1.5 mg/L + TDZ 0.05mg/L + NAA0.2 mg/L + IAA0.5 mg/L + sucrose 30g/L + agar 5g/L, and the pH value is 6.0;

Claims

1. The method for cultivating the tissue culture seedlings by the stem segments of the artemisia rupestris L is characterized by comprising the following steps of:

(2) The cut stem segments are inoculated into a long bud rooting culture medium, axillary buds on internodes of the stem segments germinate and continuously grow up and grow for 7 days, and the bud base part slightly expands and grows 2-5 adventitious roots per plant to form complete red stems Ai Zhizhu;