Rosa Vietnamese for promoting porphyra haitanensis filamentous algal filaments to develop into shell sporangia branches

Abstract

The invention provides a Rosa Vietnamese for promoting porphyra haitanensis filamentous algal filaments to develop into shell sporangia branches, and the provided strain can promote formation of the shell sporangia branches of porphyra haitanensis filaments so as to shorten the breeding cycle of the porphyra haitanensis filaments. The preservation number of the rose Vietnamese provided by the invention is CGMCC (China General Microbiological Culture Collection Center) NO.34643. According to the method, the screened rosa Vietnamese strain bacterial liquid is added into the solution for culturing the protonema, so that the development time of the conchosporangium branches can be effectively shortened, the formation rate of the conchosporangium branches can be increased, and the problems that the formation rate of the conchosporangium branches is insufficient, the formation period is too long and the like are solved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of algae breeding, and particularly relates to a Vietnamese rose fungus for promoting the development of Porphyra haitanensis filamentous algae filament into a cystocarpic branch. BACKGROUND

[0002] Porphyra haitanensis is a large marine red alga with important economic and ecological value in China, and its yield accounts for about 75% of the total yield of Porphyra in China. Its life history includes two important stages of filamentous body (sporophyte) and leafy body (gametophyte). The formation of cystospores is a key period for the transformation from the filamentous body stage to the leafy body stage, and the development of filamentous body into cystocarpic branch is a necessary basis for the production of cystospores. The speed of cystocarpic branch formation directly determines the efficiency of artificial breeding and the yield of seedlings of Porphyra haitanensis. However, the formation period of cystocarpic branch usually takes 2-3 months at present, and the slow formation of cystocarpic branch leads to long breeding cycle, high cost and great difficulty in management, which seriously restricts the high-quality development of Porphyra haitanensis breeding industry. Therefore, shortening the time of cystocarpic branch formation and realizing the centralized large-scale production of seedlings are the hot and difficult problems in the process of Porphyra haitanensis seedling breeding.

[0003] At present, methods such as optimization of cultivation conditions, addition of exogenous hormones and selection of varieties with short filamentous body development time are tried to shorten the formation time of cystocarpic branch and improve the production efficiency of seedlings. Under laboratory and factory breeding conditions, even if the cultivation conditions are optimized by increasing temperature, reducing light intensity, shortening light period and increasing phosphorus fertilizer, the formation time of cystocarpic branch is still not significantly shortened. The long development time makes the seedlings susceptible to external conditions, which further affects the whole seedling production process. It is considered that the selection of cystocarpic branch formation varieties is a potential solution, but the selection of corresponding new varieties takes a long period, consumes a lot of manpower and material resources, and is difficult to select. In addition, the addition of physiologically active organic matters such as seaweed oligosaccharide, plant hormone abscisic acid and methyl jasmonic acid can promote the formation of cystocarpic branch, but it is seriously limited in practical production application due to safety and cost problems. Therefore, it is urgent to explore a new method of promoting the formation of cystocarpic branch in a green, high-quality and efficient way, and effectively promote the optimization and upgrading of Porphyra haitanensis breeding industry. SUMMARY

[0004] The present application aims to provide a Vietnamese rose fungus for promoting the development of Porphyra haitanensis filamentous algae filament into a cystocarpic branch, which can promote the formation of cystocarpic branch of Porphyra haitanensis filamentous body, so as to shorten the breeding cycle of Porphyra haitanensis filamentous body.

[0005] The Vietnamese rose fungus provided by the present application can promote the formation of cystocarpic branch of Porphyra haitanensis filamentous body, so as to shorten the breeding cycle of Porphyra haitanensis filamentous body. Rossellomorea vietnamensis) Ph5-1 strain, the Vietnamese rose fungus was deposited in the China General Microbiology Culture Collection Center (CGMCC) on May 22, 2025, with the deposit number CGMCC NO.34643, and the deposit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0006] The present invention also provides a use of the screened Ph5-1 strain, which is the use in preparing a product for promoting the development of filamentous algae filaments of Porphyra haitanensis into conchosporangia branches; The product, as a specific example, is a bacterial liquid preparation, which is a mixture of the selected Vietnamese rose fungus ( Rossellomorea vietnamensis , Ph5-1) strain was inoculated into the culture medium and cultured, and then the centrifugal precipitate was prepared and washed with sterile seawater; The culture medium, as a specific record of the embodiment, is 2216E culture medium; The culture preparation is to culture the strain to an OD value of 0.5-0.8, and then centrifuge the culture solution to obtain a strain precipitate; The centrifugal speed is 8000-10,000 rpm.

[0007] Another aspect of the present invention is to provide a method for promoting the maturation of conchosporangium branches of Porphyra haitanensis conifers, which comprises adding the above-mentioned strain to a conifer culture medium and co-culturing it with the free conifers of Porphyra haitanensis. As a preference, the Ph5-1 strain is 10 -8 The volume ratio of CFU / mL to culture medium was 1:100 for addition; The above-mentioned free filamentous body culture medium is seawater with added nutrients; The preferred culture conditions are as follows: temperature 29 ± 0.5 °C, light intensity 30 μmol photons m -2 s -1 , photoperiod 9 h:15 h L / D; The present invention adds the screened Vietnamese rose fungus ( Rossellomorea vietnamensis , Ph5-1) can effectively shorten the development time of conchosporangium branches and increase the formation rate of conchosporangium branches, alleviating the problems of insufficient conchosporangium branch formation rate and too long formation period. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 : Phylogenetic clustering diagram of 16s rDNA fragments of strains.

[0009] Figure 2Figure 1 is a photo of the formation of the conchocelis gametophyte of Porphyra haitanensis by the bacterial agent. DETAILED DESCRIPTION

[0010] The present application isolates a strain of bacteria from the filamentous stage of Porphyra haitanensis, which has a high-efficiency effect of promoting the formation of conchospores, thereby providing a feasible and controllable method for efficient and healthy seedling culture.

[0011] The filamentous body culture solution in the present application selects filtered seawater, and 48.4 mg / L FeCl3·6H2O, 1 g / L CoCl2·6H2O, 12.096 g / L NaEDTA, 4 mg / L H3BO3, 143 mg / L ZnCl2, 840 mg / L MnCl2·4H2O, 42.5 g / L KI, 6 g / L NaNO3, 260 mg / L Na2HPO4, 0.5 mg / L FeC6H5O7, 20.56 g / L VB1, 1 mg / L Tris and 1 mg / L VB2 nutritional elements are added to the seawater. -1 -1 -1 -1 -1 -1 -1 -1 -1 -1 -1 -1 -1 The seawater is sterilized by high pressure before use. However, other culture solutions used for culturing Porphyra filamentous bodies can also be used.

[0012] The present application will be further described in detail in combination with specific embodiments.

[0013] Example 1: Isolation of the strain In 2023, the free filamentous bodies of "Gao Liang No. 1" Porphyra haitanensis collected from Fujian Province Pingtan County Aquatic Breeding Experimental Co., Ltd. in Fuzhou City, Fujian Province were cultured for propagation. The filamentous bodies were crushed to 200-500 µm / section by a cell wall breaking machine, 1 mL of culture solution was diluted to 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 times in turn.

[0014] ​​​​​​​​​​​​Bacteria that promote conchosporangial branch formation were isolated from conchosomes using the dilution spread plate method. 10 μL of each dilution solution was spread onto 2216E seawater solid medium and incubated at 30°C in a shaker at 180 rpm for 1-7 days. The isolated strains were purified using the plate streak method. After incubation at 30°C for 24-48 hours, bacterial morphology was observed. Single colonies with distinct morphological characteristics were selected for isolation and purification. The fastest-growing colony was selected for verification of conchosporangial branch formation. The resulting bacteria were numbered and designated strain Ph5-1. The purified strains were stored in 40% glycerol at -80°C.

[0015] Example 2: 16s rRNA gene sequence analysis of strains Bacterial genomic DNA was extracted and 16S rDNA was amplified by PCR using the following primers (sequences: 799F: 5'-GAGT TTGATCCTGGCTCAG-3'; 1193R: 5'-GGTTACCTTGTTACGACTT-3'). Figure 1 ), identified the bacteria as Rossellomorea genus, named Rosa vietnamese ( Rossellomorea vietnamensis ) Ph5-1 strain, deposited in the China General Microbiology Culture Collection Center (CGMCC) on May 22, 2025, with the deposit number CGMCC NO.: 34643, and the deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0016] Example 3: Analysis of functional properties of strains After the strain was activated, 10 μL of bacterial solution was spread on starch hydrolysis medium, Asubei medium and phosphate-solubilizing bacterial medium (organic phosphorus bacterial medium and inorganic phosphorus bacterial medium) for characteristic analysis, and 100 μL of activated bacterial solution was added to 100 mL of DF liquid medium and ADF liquid medium, and the OD was measured by spectrophotometer. 600 Assays were performed to examine the ACC deaminase activity of the strains. The strain Ph5-1 was found to have nitrogen fixation, solubilization of organic and inorganic phosphorus, and starch hydrolysis, but lacked ACC deaminase activity. These results indicate that the strain Ph5-1 screened by the present invention exhibits growth-promoting properties but does not reduce ethylene levels in the filaments.

[0017] Example 4: Effect of strains in promoting the formation of free conchosporangium branches After the strain was activated, 100 μL of bacterial solution was inoculated into 100 mL of 2216E liquid medium and cultured at 30 °C until the OD 600When the concentration reaches 0.5-0.8, the bacterial solution is centrifuged at 9000 rpm for 5 min, the strain precipitate is collected, and it is resuspended in sterile seawater to obtain the strain solvent for later use.

[0018] Take 0.01 g of free filaments in good growth state and add them to 100 mL of sterilized seawater with nutrient solution. -8 The volume ratio of CFU / mL to culture medium was 1:100 and added to seawater. The culture was placed in a light incubator and cultured under the following conditions: 29 ± 0.5 °C, light intensity of 30 μmol photons m −2 s −1 , photoperiod 9 h:15 h L / D. After 20 days, observe the formation of conchosporangium branches and record the maturity rate. Figure 2 As shown in the results, after 20 days of culture, the formation rate of conchosporangium branches in the culture without the addition of the strain solvent was 55%, while the formation rate of conchosporangium branches in the culture with the strain solvent was 87%. The results show that the strain screened by the present invention can significantly promote the formation of conchosporangium branches in Porphyra haitanensis.

[0019] In summary, the Rosobacter vietnamese strain screened by the present invention ( Rossellomorea vietnamensis , Ph5-1) strain contains components that can effectively promote the formation of conchosporangium branches, effectively alleviate the problems of long seedling cycle and inconsistent formation rate during the formation of conchosporangium branches, increase the formation rate of conchosporangium branches, and provide an important prerequisite for the release of conchospores.

Claims

1. A Vietnamese rose fungus, characterized in that The preservation number of the Vietnamese rose fungus is CGMCC NO.: 34643.

2. Use of the Rosa vietnamese according to claim 1 in the preparation of a product for promoting the development of filamentous algae filaments of Porphyra haitanensis into conchosporangium branches.

3. The use according to claim 2, characterized in that The product is a bacterial liquid preparation.

4. A bacterial liquid preparation, characterized in that: The bacterial liquid preparation is prepared by inoculating the Vietnamese rose fungus described in claim 1 into a culture medium for cultivation, preparing a centrifugal precipitate, and suspending it in sterilized seawater.

5. The bacterial liquid preparation according to claim 4, characterized in that The culture medium is 2216E culture medium.

6. The bacterial liquid preparation according to claim 4, wherein The bacterial liquid preparation is obtained by centrifuging the culture liquid when the bacterial strain is cultured to an OD value of 0.5-0.8 to obtain a bacterial strain precipitate.

7. A method for promoting the maturation of conchosporangium branches of Porphyra haitanensis, characterized in that: The method comprises adding the Rosa vietnamese described in claim 1 to a conical culture medium and co-culturing the culture medium with the free conical conical bodies of Porphyra haitanensis.

8. The method according to claim 7, wherein The filamentous body culture medium is seawater with added nutrient salts.

9. The method according to claim 7, wherein The co-culture temperature was 29 ± 0.5 °C and the light intensity was 30 μmol photons m -2 s -1 , photoperiod 9 h:15 h L / D.

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