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Oil sunflower inbred line in vitro culture method

A technology of in vitro culture and inbred lines, applied in the field of agricultural biology, can solve problems such as differences in medium for adventitious bud induction and difficult regeneration of sunflowers

Pending Publication Date: 2020-05-08
INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies have reported on sunflower root tips, stem tips, immature embryos, anthers and ovaries, protoplasts, cotyledons, hypocotyls, true leaves and other explant regeneration plants, but sunflower is a plant that is difficult to regenerate, and the genotype Different, in vitro culture explant selection, and adventitious bud induction medium will also have significant differences
However, there is no relatively mature in vitro culture method for oil sunflower inbred lines.

Method used

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  • Oil sunflower inbred line in vitro culture method
  • Oil sunflower inbred line in vitro culture method
  • Oil sunflower inbred line in vitro culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: A method for in vitro cultivation of oil sunflower inbred lines, which includes the following steps:

[0026] (1) Obtaining aseptic seedlings: In this example, the seeds of the oil sunflower inbred line HT80-97, 75-33B, 0272, 0177, 0196, 0365 provided by the Sunflower Research Group of the Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences were wrapped in gauze , Rinse under running water for 0.5-1h, disinfect with 75% alcohol for 1min, and then use 0.1% HgC1 2 Sterilize for 8 minutes, rinse with sterile water for 3-4 times, then inoculate the sterilized seeds on a solid medium and culture to obtain sterile seedlings. The culture conditions are 24-26°C, illumination 16h / d, and culture for 5 days; this example The solid medium is a 1 / 2MS solid medium without any hormones and a pH of 5.8-6.2. Aseptic vaccine figure 1 Shown.

[0027] (2) Induction of adventitious buds: cut the cotyledon nodes of sterile seedlings inoculated with 5 days of age, cut 1...

Embodiment 2

[0031] Example 2: Comparative test:

[0032] 1Materials and methods

[0033] 1.1 Material

[0034] Oil sunflower inbred lines HT80-97, 75-33B, 0272, 0177, 0196, 0365 were provided by the Sunflower Group of the Institute of Crop Sciences, Inner Mongolia Academy of Agriculture and Animal Husbandry.

[0035] 1.2 method

[0036] 1.2.1 Obtaining aseptic vaccine

[0037] First select two inbred lines 75-33B and HT80-97 for experiment. Take the seeds of two inbred lines separately, wrap them with gauze, rinse under running water for 0.5-1h, disinfect with 75% alcohol for 1 min, and then use 0.1% HgC1 2 Disinfect for 8 minutes, rinse with sterile water 3-4 times, and inoculate the seeds on 1 / 2MS solid medium without any hormones. The culture conditions are 22-26°C, and the light is 16h / d.

[0038] 1.2.2 Adventitious bud induction

[0039] 1.2.2.1 The effect of different explants and hormone combinations on the induction of adventitious buds

[0040] Cut the cotyledons and true leaves of 75-33B an...

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Abstract

The invention discloses an oil sunflower inbred line in vitro culture method. The method includes the following steps: (1) performing sterile seedling obtaining, taking oil sunflower inbred line seedsto disinfect after wrapping the seeds with gauze, and inoculating the disinfected seeds to a solid medium to cultivate to obtain sterile seedlings; (2) performing adventitious bud induction, cuttingcotyledonary nodes of the inoculated 5-day-old sterile seedlings, cutting 1-2 small cuts vertical to leaf veins, inoculating the cotyledonary nodes on an adventitious bud induction culture medium to induce adventitious buds; (3) performing the root growing of the adventitious buds, cutting the buds from explants to transfer to a root-growing culture medium to induce roots to grow when the inducedadventitious buds are grown to 1-2 cm; and (4) performing seedling hardening and transplanting when the roots are grown to 3-5 cm. The method takes oil sunflower inbred lines as a test material; technical methods of obtaining regeneration plants through oil sunflower in vitro culture can be optimized; and foundations of realizing genetic transformation of oil sunflowers can be established.

Description

Technical field: [0001] The invention belongs to the field of agricultural biotechnology, and particularly relates to a method for in vitro cultivation of oil sunflower inbred lines. Background technique: [0002] Sunflower (Helianthus annuus L.) is an important oil crop in the world and the main economic crop in Inner Mongolia. Over the years, through conventional breeding, people have improved sunflower yield, oil content, resistance to diseases and insects, and bred many excellent new sunflower varieties. However, the conventional breeding procedures are cumbersome, take a long time, and it is not easy to concentrate on excellent traits. Offspring. The use of Agrobacterium-mediated plant genetic engineering technology has opened up a new way for the improvement of sunflower varieties. However, whether any target gene can be transferred to the recipient and obtain transgenic plants, one of the first prerequisites is to establish an in vitro culture regeneration plant. method....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01H6/14
CPCA01H4/001A01H4/008A01H6/1464
Inventor 孙瑞芬吴新荣韩平安聂利珍郭树春牛素清张立华李素萍于海峰李晓东聂惠牟英男常悦唐宽刚任芹勇
Owner INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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