Method for inducing haploid callus by utilizing actinidia melanandra anther

A callus, kiwifruit technology, applied in horticultural methods, botanical equipment and methods, applications, etc., can solve the problems of long time consumption, less storage, restricting the industrialization process of black kiwifruit, etc., and achieves short use time and easy operation. Effect

Inactive Publication Date: 2019-01-15
XIAN BOTANICAL GARDEN SHAANXI PROV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the amount of kiwifruit stored in the wild is small, the fruit is rich in anthocyanins and is purple, the skin is smooth, etc. artificial cultivation, and its breeding work is relatively lacking, which restricts the industriali

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1. Select immature flower buds (approximately 2 to 3 mm in diameter) on branches of kiwifruit that grow well and are free from diseases and insect pests. After collection, place them in a low-temperature quick-freeze at -20°C for 30 minutes, and then refrigerate them at 4°C for 12 hours.

[0016] 2. Sterilize the flower buds after refrigeration treatment with 6% sodium hypochlorite aqueous solution for 6 minutes, then wash them with sterilized distilled water for 5 times, dry the water on the surface of the flower buds, place them in a plate, peel off the flower buds, and take out the anthers. The anthers were inoculated in a culture bottle filled with 25mL callus-inducing medium, and 10 anthers were inoculated in every bottle. The described callus-inducing medium was based on MS medium, adding 0.8mg / L zeatin, 4.5mg / L auxin, 15mL / L coconut milk, 30g / L sucrose, 5.5g / L agar, pH 5.8. Place the inoculated culture medium at 24±2°C under 16 / 8h light for 5 days. The callus f...

Embodiment 2

[0018] 1. Select immature flower buds (approximately 2 to 3 mm in diameter) on branches of kiwifruit that grow well and are free from diseases and insect pests. After collection, place them in a low-temperature quick-freeze at -20°C for 30 minutes, and then refrigerate them at 4°C for 12 hours.

[0019] 2. Sterilize the flower buds after refrigeration treatment with 8% sodium hypochlorite aqueous solution for 6 minutes, then wash them 5 times with sterilized distilled water, dry the surface of the flower buds, place them in a plate, peel off the flower buds, and take out the anthers. The anthers were inoculated in a culture bottle filled with 25mL callus induction medium, and 10 anthers were inoculated in each bottle. The callus induction medium was based on MS medium, and 1.0mg / L zeatin, 5.0mg / L auxin, 20mL / L coconut milk, 30g / L sucrose, 5.5g / L agar, pH 6.0. Place the inoculated culture medium at 24±2°C under 16 / 8h light for 5 days. The callus formed after induction was tak...

Embodiment 3

[0021] 1. Select immature flower buds (about 2 to 3 mm in diameter) on branches of kiwifruit that grow well and are free from diseases and insect pests. After collection, they are first placed in -20°C for 60 minutes, and then placed in 4°C for 15 hours.

[0022] 2. Sterilize the flower buds after refrigeration treatment with 8% sodium hypochlorite aqueous solution for 6 minutes, then wash them 5 times with sterilized distilled water, dry the surface of the flower buds, place them in a plate, peel off the flower buds, and take out the anthers. The anthers were inoculated in a culture bottle filled with 25mL callus-inducing medium, and 10 anthers were inoculated in each bottle. The callus-inducing medium was based on MS medium, adding 1.5mg / L zeatin, 6.0mg / L auxin, 30mL / L coconut milk, 30g / L sucrose, 5.5g / L agar, pH 6.0. Place the inoculated culture medium at 24±2°C under 16 / 8h light for 6 days. The callus formed after induction was taken out, and the haploid callus was ident...

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Abstract

The invention discloses a method for inducing haploid callus by utilizing actinidia melanandra anther. The method comprises the following steps: taking actinidia melanandra anther as a material and carrying out in vitro culture; based on an MS culture medium, adding zeatin, auxin, coconut juice or corn stigma juice, sucrose and agar; based on a B5 culture medium, adding naphthalene acetic acid, kinetin, potato juice, the sucrose and the agar as an induction callus medium; successfully obtaining callus of haploid actinidia melanandra after one week. The average induction rate of the method is 72.95 percent and can be up to 88.64 percent which is obviously higher than the haploid occurrence probability of a spontaneous generation pathway; in addition, the method has the advantages of simpleoperation and less time and is a relatively-effective way to obtain actinidia melanandra haploid callus; the method lays a foundation for rapid acquisition of homozygote as well as gene transformation, cell culture and genomic research of the actinidia melanandra.

Description

technical field [0001] The invention belongs to the technical field of kiwifruit cultivation, and in particular relates to a method for inducing haploid callus by using kiwifruit anthers. Background technique [0002] Kiwifruit is a perennial climbing deciduous vine fruit tree with a soft texture. It belongs to the family Actinidiaceae and the genus Actinidia. Kiwi fruit is rich in fat and juice, fragrant and delicious. It is rich in vitamin C, dietary fiber, various mineral nutrients and amino acids. . In recent years, purple fruit kiwifruit (actinidia nigra) has been loved by people because of its delicious fruit and characteristics. However, the small amount of kiwifruit stored in the wild, the lack of artificial cultivation such as a large amount of anthocyanin-rich fruits that turn purple and smooth skin, and the relative lack of breeding work restrict the industrialization process of kiwifruit. [0003] The invention patent application with the publication number CN...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/005
Inventor 陈尘张莹吴永鹏张燕毛祝新
Owner XIAN BOTANICAL GARDEN SHAANXI PROV
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