Method of removing virus for Jiangxi Qianshan red-bud taro through vitrification cryopreservation

A technology of ultra-low temperature preservation and vitrification, which is applied in horticultural methods, botanical equipment and methods, horticulture, etc., and can solve problems such as virus accumulation, production loss of red bud taro, and species degradation.

Inactive Publication Date: 2013-08-28
SHANGRAO NORMAL UNIV
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Problems solved by technology

However, due to the long-term asexual reproduction in production, the virus has accumulated more and more, the species has d

Method used

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Embodiment Construction

[0004] The present invention will be further described in conjunction with following embodiment:

[0005] (1) Cutting the stem tip of red bud taro in Qianshan, Jiangxi

[0006] The shoot tip of a single bud of Jiangxi Qianshan red bud taro with a length of about 1 mm was cut under a stereo microscope.

[0007] (2) Pre-cultivation of stem tip of red bud taro in Qianshan, Jiangxi

[0008] The red bud taro stem tip is pre-cultured on MS+KT2mg / L+NAA0.5mg / L solid medium with 0.75M sucrose for 3 days. The culture conditions are: light time 14h / d, light intensity about 1500-2000lx, culture temperature 25± 2°C.

[0009] (3) Dehydration and liquid nitrogen preservation of stem tip of red bud taro in Yanshan, Jiangxi

[0010] After pre-cultivation of the shoot tip of red bud taro, 60% cryoprotectant PVS2 (30% (w / v) glycerol+15% (w / v) ethylene glycol+15% (w / v) dimethyl sulfoxide+ 0.4M sucrose, pH5.8) was dehydrated at 25±2°C for 60 minutes, and then dehydrated with 100% PVS2 at 0°C f...

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Abstract

The invention provides a method of removing virus for Jiangxi Qianshan red-bud taro through vitrification cryopreservation. The method comprises: (1) cutting shoot tips with a length being about 1 mm of single buds of the red-bud taro under a microscope; (2) pre-culturing the shoot tips on a solid medium of MS + KT 2 mg/L + NAA 0.5 mg/L +0.75 M sucrose for three days; (3) after the shoot tips are pre-cultured, using a 60 % PVS2 to dehydrate for 60 minutes under 25 DEG C, then using a 100 % PVS2 to dehydrate for 90 minutes under 0 DEG C, replacing with a new PVS2, and then putting into liquid nitrogen; (4) after preservation, defrosting the shoot tips in a 37 DEG C water bath, washing with a liquid medium of MS + KT 2 mg/L + NAA 0.5 mg/L +1.2 M sucrose for 3 times, 10 minutes for each time, culturing the washed shoot tips in dark for 5 days under 25 DEG C, and then culturing under a normal photoperiod with an illumination time being 14 hours per day, a light intensity being about 1500-2000lx, and a culturing temperature being 25 DEG C, wherein the shoot tips can grow into normal buds after about 15 days; and (5) culturing the germinated buds in a rooting medium of 1/2 MS + NAA 0.1-0.5 mg/L, thereby obtaining the virus-free seedlings of the red-bud taro. Compared with prior art, the tube-test plantlets of the red-bud taro are high in virus removing rate, the shoot tips can be taken in a large amount and the operation is simple and convenient.

Description

Technical field: [0001] The invention relates to a method for breeding plant seedlings, in particular to a method for detoxification of Jiangxi Yanshan red bud taro by ultra-low temperature preservation through vitrification. Background technique: [0002] Red bud taro is a traditional superior product in Qianshan County, Jiangxi Province, with a long history of planting. In the past ten years, this advantageous industry has developed rapidly and has become the leading industry for farmers in Yanshan County to increase their income and become rich. Qianshan County has also become the largest red bud taro production base in Jiangxi Province. In March 2007, it was approved by China Green Food Committee and The Ministry of Agriculture named it "National Green Food Standardized Production Base County of Red Bud Taro". However, due to the long-term asexual reproduction in production, the virus is increasingly accumulated, the species is degraded, and the yield and quality have d...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 洪森荣尹明华
Owner SHANGRAO NORMAL UNIV
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