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A method for obtaining pepper somatic embryos with explants as explants

A technology of explants and somatic cells, applied in botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of high pollution rate of seed germination and somatic cell differentiation rate of less than 1%

Active Publication Date: 2018-03-13
SPICE & BEVERAGE RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing study "Studies on Pepper Tissue Culture" carried out the research on the somatic cell regeneration of pepper with the shoot tips, hypocotyls, and leaves of pepper seedlings as explants, and provided seed sterilization and somatic cell regeneration culture methods, but the published documents Method The seed germination pollution rate is as high as 30%-60%, while the somatic cell differentiation rate is less than 1% (Liu Jinping, Tropical Agricultural Science, 2002,22(3):18-22.)
At present, there is no cultivation method suitable for pepper somatic embryogenesis in pepper production

Method used

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  • A method for obtaining pepper somatic embryos with explants as explants
  • A method for obtaining pepper somatic embryos with explants as explants

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Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1: method of the present invention

[0025] Get the mature fruit of Reyin No. 1 plant that is 5 years old, grows normally, and has no influence of diseases and insect pests; peel the fruit from the ear, soak it in 75% (V / V) alcohol for 2 minutes in a sterile environment, and then wash it with sterile water for 5 times; Remove the pulp in a sterile environment, soak the depulped seeds in 75% alcohol for 2 minutes, then wash them with sterile water for 5 times, then soak the seeds in 0.1g / L mercury chloride for 10 minutes to sterilize, and finally The seeds were transferred to a new aseptic container, washed 5 times with sterile water to obtain sterile seeds.

[0026] Under a sterile environment, the sterile seeds were placed in a sterile medium to germinate, and cultured at (28±2)°C in the dark for 60 days. The medium formula is 1 / 2MS+2g / LAC (activated carbon). After the pepper seedlings are fully grown, the outer seed coat is peeled off from the top of the ...

Embodiment 2

[0029] Embodiment 2: control method 1 (take blade, hypocotyl as explant and adopt the culture method in embodiment 1)

[0030] Get the mature fruit of Reyin No. 1 plant that is 5 years old, grows normally, and has no influence of diseases and insect pests; peel the fruit from the ear, soak it in 75% (V / V) alcohol for 2 minutes in a sterile environment, and then wash it with sterile water for 5 times; Remove the pulp in a sterile environment, soak the depulped seeds in 75% alcohol for 2 minutes, then wash with sterile water for 5 times, then soak the seeds in 0.1g / L mercury chloride for 10 minutes to sterilize, and finally The pepper seeds were transferred to a new sterile container, washed with sterile water 5 times to obtain sterile seeds.

[0031] Under a sterile environment, the sterile seeds were placed in a sterile medium to germinate, and cultured at (28±2)°C in the dark for 60 days. The medium formula is 1 / 2MS+2g / L AC. After the pepper seedlings have grown completely,...

Embodiment 3

[0034] Embodiment 3: Control Method 2 (MS culture medium is replaced with hormone-free SH culture medium among the embodiment 1)

[0035]Get the mature fruit of Reyin No. 1 plant that is 5 years old, grows normally, and has no influence of diseases and insect pests; peel the fruit from the ear, soak it in 75% (V / V) alcohol for 2 minutes in a sterile environment, and then wash it with sterile water for 5 times; Remove the pulp in a sterile environment, soak the depulped seeds in 75% alcohol for 2 minutes, then wash them with sterile water for 5 times, then soak the seeds in 0.1g / L mercury chloride for 10 minutes to sterilize, and finally The seeds were transferred to a new aseptic container, washed 5 times with sterile water to obtain sterile seeds.

[0036] Under a sterile environment, the sterile seeds were placed in a sterile medium to germinate, and cultured at (28±2)°C in the dark for 60 days. The medium formula is SH+2g / L AC. After the pepper seedlings are fully grown, ...

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Abstract

The invention relates to the technical field of tissue culture and discloses a method for acquiring pepper somatic embryogenesis by taking testa as explant. The method provided by the invention comprises the following steps: sterilizing mature pepper seeds; culturing into pepper seedlings; separating testa from seedling cotyledons; and performing callus induction culture and differentiation culture on the testa in turn, thereby acquiring the pepper somatic embryogenesis. According to the method provided by the invention, the testa is selected as the explant, the formula of the callus induction medium and differential medium is adjusted, and MS is taken as a basal culture medium and the ingredients, such as, activated carbon, polyvinylpyrrolidone and 2,4-D are added, so that the differentiation rate of the pepper somatic embryogenesis is as high as 20% and the method has significance in researching high throughput pepper seedling propagation and establishing transgenosis breeding system.

Description

technical field [0001] The invention relates to the technical field of tissue culture, in particular to a method for obtaining pepper somatic embryos with the outer testa as explants. Background technique [0002] Pepper (Piper nigrum L.) is a perennial woody vine of the family Piperaceae (Piperaceae). Its fruit has a spicy aroma and is known as the "king of spices". It is an important spice crop in the world and is also a favorite of people. condiment. In the medical field, pepper can be used as a stomachic agent, an antipyretic, and a bronchial mucosal stimulator. In the food industry, it can be used as an antioxidant, preservative and preservative. As a popular condiment and tropical agricultural product with many uses, pepper has spread to nearly 20 countries and regions in Asia, Africa and Latin America. Among them, my country is mainly distributed in Hainan, Yunnan, Guangdong, Fujian and other places, with a planting area of ​​nearly 3×10 4 hm 2 , the annual outpu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/006
Inventor 胡丽松范睿伍宝朵郝朝运谭乐和
Owner SPICE & BEVERAGE RES INST CHINESE ACAD OF TROPICAL AGRI SCI