Method for improving germination rate of paphiopedilum seeds preserved at ultra-low temperature by using sodium hypochlorite to treat

A technology of cryopreservation and sodium hypochlorite, which is applied in the field of plant protection biology and biology, can solve the problem of low germination rate of Paphiopedilum seeds, and achieve the effect of improving the germination rate of seeds

Inactive Publication Date: 2017-12-19
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cryopreservation of Paphiopedilum seeds is one of the most effective methods for germplasm preservation, but the germination rate of Paphiopedilum seeds after cryopreservation is extremely low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] (1) Collection and cryopreservation of seeds

[0014] Select robust Paphiopedilum armeniacum S.C.Chen&F.Y.Liu plants for artificial self-pollination during the flowering stage, collect fully mature and uncracked capsules within 4 months after pollination, and use the capsules on the ultra-clean workbench. Soak in alcohol with a volume fraction of 70% for 30 seconds, then sterilize with 0.1% mercuric chloride solution for 20 minutes, rinse with sterile water for 4 times, cut the capsules, put 8 grams of powdery mature seeds into the sterilized diameter Dehydrate on sterile filter paper with a diameter of 12 cm in a 15 cm petri dish, and 15 grams of dried discolored silica gel have been placed under the filter paper in each petri dish. After putting in the seeds, seal the culture dish with parafilm and dehydrate for 96 hours. After the seeds were dehydrated, put the seeds into 2mL cryovials and put them into liquid nitrogen at -196°C for storage. Some of the unpreserved ...

Embodiment 2

[0018] (1) Collection and cryopreservation of seeds

[0019] Select the vigorous Paphiopedilum emersonii Koop.&P.J.Cribb plant to carry out artificial self-pollination during the flowering stage, collect mature and uncracked capsules within 7 months after pollination, and place the capsules on the ultra-clean workbench. Soak in alcohol with a volume fraction of 75% for 45 seconds, then sterilize with a mercury solution of 0.15% mass fraction for 15 minutes, rinse with sterile water for 5 times, cut the capsules, and put 10 grams of powdery mature seeds into the sterilized Dehydration was carried out on sterile filter paper with a diameter of 12 cm in a petri dish with a diameter of 15 cm, and 20 grams of dried discolored silica gel had been placed under the filter paper in each petri dish. After placing the seeds, seal the culture dish with parafilm and dehydrate for 72 hours. After the seeds were dehydrated, put the seeds into 2mL cryovials and put them into liquid nitrogen ...

Embodiment 3

[0023] (1) Collection and cryopreservation of seeds

[0024] Select robust Paphiopedilum helenae Aver. plants to carry out artificial self-pollination at the flowering stage, collect mature and uncracked capsules within 9 months after pollination, and use a volume fraction of 80% of the capsules on the ultra-clean workbench After soaking in alcohol for 30 seconds, sterilize with 0.2% mercuric chloride solution for 10 minutes, rinse with sterile water for 6 times, cut the capsules, and put 14 grams of powdery mature seeds into a sterilized 15 cm diameter petri dish 22 grams of dried discolored silica gel have been placed under the filter paper in each Petri dish. After placing the seeds, seal the culture dish with parafilm and dehydrate for 48 hours. After the seeds were dehydrated, put the seeds into 2mL cryovials and put them into liquid nitrogen at -196°C for storage. Some of the unpreserved dehydrated seeds were used to determine the water content, and the water content of...

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Abstract

The invention discloses a method for improving the germination rate of paphiopedilum seeds preserved at ultra-low temperature by using sodium hypochlorite to treat. The method comprises the following steps: quickly putting the paphiopedilum seeds preserved at the ultra-low temperature into a water bath with the temperature of 38 to 42 DEG C and thawing for 1 to 3 minutes; putting the seeds into sterile water under aseptic conditions and then uniformly shaking; filtering the seeds with filter paper and then soaking the seeds by using 0.5 to 1.0 percent available chlorine sodium hypochlorite solution for 40 to 80 minutes, then flushing with the sterile water and sowing the seeds in a culture medium for culturing. According to the method for improving germination rate of the paphiopedilum seed preserved at the ultra-low temperature by using the sodium hypochlorite to treat, disclosed by the invention, the germination rate of the paphiopedilum seeds preserved at the ultra-low temperature for 12 to 24 months can be obviously improved, the germination rate of the seeds is improved from 0 to 6 percent to 30 to 60 percent. Therefore, effective protection of the paphiopedilum seeds is realized, and the foundation is laid for protection, development and utilization of germplasm resources of paphiopedilum.

Description

Technical field: [0001] The invention belongs to the fields of plant protection biology and biotechnology, and in particular relates to a method for improving the germination rate of Paphiopedilum seeds preserved in ultra-low temperature by treating with sodium hypochlorite. Background technique: [0002] Paphiopedilum is also called slipper orchid, cinderella, etc. There are more than 90 species in the world, mainly distributed in tropical Asia to Pacific islands. Paphiopedilum has a very high ornamental value due to its unique flower shape, gorgeous flower color, and long-lasting ornamental flowering period. It is a very popular high-end flower in the international flower market and has a large number of lovers in the world. However, due to the destructive excavation of wild resources due to people's excessive pursuit of P. Convention" (CITES) Appendix I and trade is prohibited. The protection of Paphiopedilum germplasm is currently a hotspot in plant protection biology....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01C1/00
CPCA01C1/00
Inventor 曾宋君文颖婷吴坤林郑枫
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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