Method for effectively increasing alkaloid content of pinellia ternate through low-phosphorus treatment

A kind of alkaloid content technology, applied in the direction of botany equipment and method, gardening method, plant regeneration, etc., to achieve remarkable effect, increase the alkaloid content of Pinellia pinelliae, and implement the simple effect

Active Publication Date: 2022-01-11
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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At present, there are no relevant patents and documents related to the improvement of pinellia alkaloid content mentioned in this patent

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  • Method for effectively increasing alkaloid content of pinellia ternate through low-phosphorus treatment
  • Method for effectively increasing alkaloid content of pinellia ternate through low-phosphorus treatment
  • Method for effectively increasing alkaloid content of pinellia ternate through low-phosphorus treatment

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Embodiment 1

[0018] 1. Configuration of hydroponic solution:

[0019] (1) Configure 1mol / LK respectively 2 HPO 4 , 1mol / L KH 2 PO 4 and 0.1M KCl solution, sterilized at 121°C for 20min.

[0020] (2) 8.5ml of the above-mentioned 1mol / L K2HPO4 solution and 91.5ml of the above-mentioned 1mol / L KH2PO4 solution were mixed to obtain a phosphate buffer (100mL).

[0021] (3) Low phosphorus (LP) culture medium (1L): Weigh 4.16g of commercially available Mruashige and Skoog withoutPhosphat (CAISSON, 02180014) medium (that is, MS medium without phosphate), add 900ml ddH 2 O, adjust pH to 5.8 with KOH and HCL, ddH 2 O was adjusted to 1L. After sterilizing at 121°C for 20 minutes, 100 μL of phosphate buffer and 100 ml of potassium chloride solution were added.

[0022] (4) High phosphorus (HP) culture medium (1 L): Add 100 mL of phosphate buffer to 900 ml of phosphate-free culture medium.

[0023] 2. Treat Pinellia tissue culture seedlings with low-phosphorus and high-phosphorus culture solutio...

Embodiment 2

[0026] Embodiment 2: Pinellia available phosphorus content determination

[0027] 1) Extraction of available phosphorus

[0028] Take 0.1 g of Pinellia fresh sample treated in Example 1 to a 2 mL centrifuge tube with glass beads, grind it into powder with liquid nitrogen, add 500 μL of 5M concentrated sulfuric acid, then transfer the sample to a 15 mL centrifuge tube, and add 5 mL of distilled water. In addition, 500 μL of 5M concentrated sulfuric acid and 5 mL of distilled water were used as two blank controls. Fold the 9mm qualitative filter paper according to the conventional method, and filter the above sample into the matching injection tube of the automatic sampler of the continuous flow analyzer (SKALAR, Holland). After filtering, put them in the corresponding position of the autosampler in order, put the 2 blanks first, and test them on the machine, refer to (Xu Qiming et al. Determination of inorganic phosphorus content in rice using SAN~(++) continuous flow analyzer...

Embodiment 3

[0034] Example 3: Fluorescent quantitative PCR detection of expression levels of genes related to alkaloid synthesis in Pinellia chinensis under low phosphorus stress

[0035] After HP and LP treatment for 3 days, Trizol method was used to separate and extract RNA from different tissues of Pinellia, and the above RNA was reverse-transcribed according to the method of HiScript1stStrand cDNA Synthesis Kit (10 μl reverse transcription system was added with 500 ng Total RNA) to obtain cDNA. Prepare the reaction solution according to the following components: HieffTMqPCR SYBR Green Master Mix 5 μl, PCR Forward Primer (10 μM) 0.2 μl, Reverse Primer (10 μM) 0.2 μl, cDNA 0.5 μl, ddH2O 4.1 μl. The amplification program was as follows: 94°C for 5min, 94°C for 10s, 58°C for 20s, 72°C for 20s, 40Cycles, fluorescence was collected at 72°C, and Pt18S was used as an internal reference.

[0036] Depend on figure 2 It can be seen that low phosphorus treatment induces the expression of phosph...

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Abstract

The invention discloses a method for effectively increasing the alkaloid content of pinellia ternate through low-phosphorus treatment. In the pinellia ternate planting process, pinellia ternate tissue culture seedlings are treated through a low-phosphorus culture solution, and the method for rapidly and effectively increasing the alkaloid content of the pinellia ternate is established. The pinellia ternate tissue culture seedlings induce the expression of related genes (PtCNL, PtCHD, PtKAT, PtPDC, PtAO4 and PtAHAS) of a pinellia ternate alkaloid synthetic pathway under the premise of not influencing the growth and development of the pinellia ternate tissue culture seedlings subjected to short-term low-phosphorus treatment, so that the alkaloid content of the tuber of the pinellia ternate is increased. The method is easy to implement and remarkable in effect, and powerful reference is provided for increasing the alkaloid content of the pinellia ternate and improving the quality of the pinellia ternate.

Description

technical field [0001] The invention relates to a treatment method for increasing the alkaloid content of Pinellia chinensis, in particular to a method for increasing the alkaloid content of Pinellia chinensis tissue-cultured seedlings through a low-phosphorus nutrient solution. Background technique [0002] Pinellia ternate, a plant of the Araceae family, is a medicinal plant, which not only has the functions of drying dampness and reducing phlegm, reducing adverse flow and relieving vomiting, dispelling lumps and resolving stagnation, but also has pharmacological activities such as antitumor, antibacterial, anti-inflammatory, and antiepileptic. Pinellia tuber is used as medicine, and its secondary metabolites include alkaloids, organic acids, volatile oils, flavonoids, etc., which are the main active ingredients and have various pharmacological effects. The total alkaloid components of pinellia include ephedrine, guanosine (guanosine, G), uridine (uridine, U), adenosine (a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/002A01H4/008
Inventor 洪高洁李林颖何宇青张雪颖
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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