Super-low step-by-step freezing preservation method for kelp swarm spore

Abstract

The present invention discloses an ultra-low temperature step-by-step freeze storage method of sea-tangle swarm (embryo) spore. Said method includes the following steps: material treatment, equilibration, step-by-step cooling freeze-storing and anabiosis. It can store lots of germplasm material and is survival rate is high.

Description

1. Technical field [0001] The invention belongs to the technical field of marine organisms, and relates to a germplasm preservation technology of large-scale economic seaweed-kelp. 2. Background technology [0002] Kelp (Laminaria japonica Aresch) is an important economic algae in China. Its breeding output and scale have ranked first in the world for many years. The rich germplasm resources are an important material basis for fishery production. The protection and utilization of kelp germplasm resources will improve the level of kelp varieties. , to provide an important guarantee for the realization of improved kelp production. The life history of kelp is divided into two distinct generational stages: the sporophyte generation and the gametophyte generation. The gametophyte generation is the haploid stage of kelp. The zoospore is the beginning stage of the gametophyte generation. Afterwards, the zoospores are released, and the zoospores swim for about 2 hours, or attach or ...

AI Technical Summary

Problems solved by technology

However, this traditional preservation technique has obvious deficiencies: (1) liquid culture is the only way of preservation, and the method is single, which limits the scale and efficiency of preservation; (2) liquid culture preservation generally needs to update the culture medium within half a month, The workload is heavy, the operation is cumbersome, and frequent operations also increase the chance of germplasm contamination and mixing; (3) the kelp species cannot be aseptically treated before the spores are collected, and the entire preservation process is under bacterium-containing conditions, which cannot completely eliminate microorganisms, especially Pollution by miscellaneous algae; (4) Abnormal cell morphology and parthenogenesis occur from time to time during culture, and the culture factors need to be further optimized

The method of ultra-low temperature cryopreservation of algae is mainly two-step freezing method (two-step freezing). In this method, an antifreeze protective agent is added to the preservation material for pre-freezing, and then the pre-frozen material is put into liquid nitrogen for quick freezing. At present, it has become a routine method for the cryopreservation of algae germplasm; the main reason for the limitation of the application of ultra-low temperature cryopreservation technology is that the establishment of freezing operation procedures mainly depends on empirical experimental results, and the preservation technology suitable for one kind of algae may affect another kind of algae. not applicable at all