Method for detecting varietal complexity in kelp summer seedling refrigeration water process

Abstract

The invention relates to the field of economical brown kelp seaweed varietal complexity, in particular to a method for detecting varietal complexity in kelp summer seedling refrigeration water process. The method comprises the steps that a seedling rope which is inoculated with a female gametophyte clone is put in a culture medium or a kelp seedling refrigeration water circulation system to be cultured, parthenogernesis sporophore is obtained when the seedling rope is put in the culture medium, and offspring sporophore is obtained when the seedling rope is put in the kelp seedling refrigeration water circulation system; morphological characteristics and genetype of the offspring sporophore on the seedling rope in the refrigeration water circulation system are detected and compared with those of the parthenogernesis sporophore so as to determine whether varieties are hybrid or not in the seedling process. According to the detection method, it is verified for the first time that in the kelp summer seedling production process, due to the fact that low temperature circulation water is used in common, the varietal complexity is definitely caused.

Description

technical field

[0001] The invention relates to the field of mixed species of economic brown algae kelp, in particular to a method for detecting mixed varieties of kelp summer seedlings in a cooling water process. Background technique

[0002] Kelp is a typical cold-water seaweed. In my country's sea waters, due to the high sea temperature in summer, it cannot complete the life history process in natural sea waters. Since the 1950s, the breakthrough in the cultivation technology of kelp summer seedlings in my country has promoted the rapid expansion of kelp cultivation scale. The summer seedling technology is to artificially cool down the high-temperature seawater in summer, and maintain the seawater temperature at 5-12°C, which is suitable for the survival of kelp, so as to complete the process of cultivating kelp seedlings indoors. In order to save cooling costs, kelp seedling enterprises recycle cooling water in production. Therefore, if more than two species are cultiv...

Examples

Embodiment 1

[0021] Long-term preservation of vegetatively grown filamentous Laminaria monoclonal female gametophytes (for the acquisition and growth of female gametophytes, please refer to LiJing, PangShaojun, ShanTifeng, LiuFeng, GaoSuqin. Zoospore-derived monoecious gametophytes in Undariapinnatifida (Phaeophyceae). Chinese Journal of Oceanology and Limnology. 2014-35( 371. Records in the literature), placed in a high-speed breaker, and the filamentous female gametophyte was broken to 3-10 cells at a speed of 2000r / min. Subsequently, the crushed female gametes and 50cm seedling ropes were aerated and cultured in a 1L beaker, placed in PES medium, at a temperature of 12°C, a light intensity of 10-20μmolphotonsm-2s-1, and a photoperiod of 12:12 (L:D) cultured in a light incubator.

[0022] When the female gametophyte is firmly attached to the seedling rope, increase the culture light to 50-60 μmolphotonsm-2s-1, so that the female gametophyte develops and forms oocysts. Subsequently, duri...

Embodiment 2

[0028] In August 2014, a kelp nursery enterprise in Rongcheng City, Shandong Province, my country carried out the detection of species mixing in the refrigeration water circulation system. The seedling ropes inoculated with female gametophyte clones were placed in the seedling ponds and water inlets of the low-temperature circulating water system, and the seedling ropes were collected later. The sporophytes of the progeny above were compared with the parthenogenetic sporophytes as a control. At the same time, using the microsatellite analysis method, four pairs of SSR primers were used to amplify, and the parental female gametophyte was used as a reference to analyze the genotype of the offspring sporophyte. It showed that the sporophytes of the offspring collected at the nursery pool and the water tank were normal in shape. In addition to the gene fragments of the parental female gametophytes, exogenous gene fragments were also detected in this sporophyte group, which proved to...

technical field

[0001] The invention relates to the field of mixed species of economic brown algae kelp, in particular to a method for detecting mixed varieties of kelp summer seedlings in a cooling water process. Background technique

[0002] Kelp is a typical cold-water seaweed. In my country's sea waters, due to the high sea temperature in summer, it cannot complete the life history process in natural sea waters. Since the 1950s, the breakthrough in the cultivation technology of kelp summer seedlings in my country has promoted the rapid expansion of kelp cultivation scale. The summer seedling technology is to artificially cool down the high-temperature seawater in summer, and maintain the seawater temperature at 5-12°C, which is suitable for the survival of kelp, so as to complete the process of cultivating kelp seedlings indoors. In order to save cooling costs, kelp seedling enterprises recycle cooling water in production. Therefore, if more than two species are cultiv...

Examples

Embodiment 1

[0021] Long-term preservation of vegetatively grown filamentous Laminaria monoclonal female gametophytes (for the acquisition and growth of female gametophytes, please refer to LiJing, PangShaojun, ShanTifeng, LiuFeng, GaoSuqin. Zoospore-derived monoecious gametophytes in Undariapinnatifida (Phaeophyceae). Chinese Journal of Oceanology and Limnology. 2014-35( 371. Records in the literature), placed in a high-speed breaker, and the filamentous female gametophyte was broken to 3-10 cells at a speed of 2000r / min. Subsequently, the crushed female gametes and 50cm seedling ropes were aerated and cultured in a 1L beaker, placed in PES medium, at a temperature of 12°C, a light intensity of 10-20μmolphotonsm-2s-1, and a photoperiod of 12:12 (L:D) cultured in a light incubator.

[0022] When the female gametophyte is firmly attached to the seedling rope, increase the culture light to 50-60 μmolphotonsm-2s-1, so that the female gametophyte develops and forms oocysts. Subsequently, duri...

Embodiment 2

[0028] In August 2014, a kelp nursery enterprise in Rongcheng City, Shandong Province, my country carried out the detection of species mixing in the refrigeration water circulation system. The seedling ropes inoculated with female gametophyte clones were placed in the seedling ponds and water inlets of the low-temperature circulating water system, and the seedling ropes were collected later. The sporophytes of the progeny above were compared with the parthenogenetic sporophytes as a control. At the same time, using the microsatellite analysis method, four pairs of SSR primers were used to amplify, and the parental female gametophyte was used as a reference to analyze the genotype of the offspring sporophyte. It showed that the sporophytes of the offspring collected at the nursery pool and the water tank were normal in shape. In addition to the gene fragments of the parental female gametophytes, exogenous gene fragments were also detected in this sporophyte group, which proved to...

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