CN105475270A - Preservation method of Zostera marina L. seedlings - Google Patents

Abstract

The invention relates to a preservation method of Zostera marina L. seedlings. The method comprises the following steps: preserving new healthy Zostera marina L. seeds collected in July to August in natural seawater at 0-4DEG C in dark environment, cleaning the seeds with natural seawater with the temperature same to the temperature of the above natural seawater and replacing the natural seawater with the natural seawater with the same temperature every week, picking germinant seedlings from January in the next year, placing the germinant seedlings in the natural seawater at 0-4DEG C in the dark environment, and replacing the natural seawater with the natural seawater with the sample temperature every week. The method fully utilizes the physiologic phenomenon of the state of dormancy of Zostera marina L. in 0-4DEG C natural seawater environment, and allows the germinant Zostera marina L. seedlings to be preserved at a low temperature in dark and low-pressure environment for a long time and to extremely slowly grow, so the preservation time of the Zostera marina L. seedlings is effectively prolonged.

Description

A kind of store method of Zostera marina seedling

Technical field

Sea grass preserving seed field belonging to the present invention, relates to a kind of store method of Zostera marina seedling.

Background technology

Zostera marina (ZosteramarinaL.) belongs to ocean spermatophyte, be widely distributed in Northern Hemisphere Coastal temperate, the sociales of Ye Shi China Shandong, Hebei and Liaoning Coast's silt substrate plate with coast sea grass group, build the careless bed ecosystem of the Zostera marina that formed by it and all play very important effect in the self-cleanings of the multifarious maintenance of neritic organism and marine ecosystems.But recent two decades makes its population in China's slump of disastrous proportions due to factors such as ocean eutrophication, therefore one of its population conservation important content of having become northern China marine ecosystems to recover with restoration and reconstruction.Wherein, whether grass wrack seeds to survive in population be an issues that need special attention if sprouting the seedling formed, therefore, relate at present the growth of its seedling and physiology, biochemical pathway research more.Grass wrack seeds gathers in 7-8 month in China marine site, be stored in the room temperature even natural sea-water of 0-4 DEG C and just start November to sprout successively in then, January next year enters a large amount of germination period, March, namely sprouting entered coda, after germinateing, Zostera marina seedling just grows true leaf, root and stem very soon, enters fast growing period.Such germination and growth rhythm make the relevant physiological of its seedling, growth the research phase shorter, therefore, how to extend the storage life of its seed or seedling, the more sufficient time is striven in the correlative study that can be Zostera marina seedling.

Summary of the invention

The object of this invention is to provide a kind of low cost, the simple to operate and store method of effective Zostera marina seedling, the method can preserve the vigor of Zostera marina seedling for a long time.

The technical solution used in the present invention is:

A store method for Zostera marina seedling, the steps include:

(1) put into the large container that natural sea-water is housed after the healthy grass wrack seeds cleaning of being collected back from marine site in 7-8 month, in 0-4 DEG C, preserve under dark environmental condition;

(2) weekly with equitemperature natural sea-water cleaning seed, and the replacing of an equitemperature natural sea-water is carried out;

(3) from January next year, the seedling of germination is chosen, is positioned over and is equipped with in the large container of natural sea-water, in 0-4 DEG C, preserve under dark ambient conditions, water consumption is 10ml/ strain;

(4) carry out weekly the replacing of an equitemperature natural sea-water, and remove unsound seedling, clean container.

The technology of the present invention effect is: Zostera marina seedling only needs natural sea-water and common vessel in preserving, period only changes seawater, without the need to special aeration equipment, also without the need to illumination, make full use of Zostera marina under 0-4 DEG C of natural sea-water environment, enter this physiological phenomenon of resting state, the seedling that Zostera marina has just been sprouted can preserve vigor for a long time under low temperature, dark and low compression ring border, period seedling growth very slow, effectively extend the holding time of Zostera marina seedling.

Embodiment

A kind of low cost of the present invention, the simple to operate and store method of effective Zostera marina seedling, can ensure that the Zostera marina seedling vigor of 8 months is preserved.Comprise following examples, the following examples can further illustrate the present invention, but do not limit the present invention in any way.

Embodiment 1: a kind of store method of Zostera marina seedling, the steps include:

(1) August is put into the 300ml tissue culture bottle that natural sea-water is housed after the healthy grass wrack seeds cleaning that twin islet gulf, Weihai is collected back, wherein: 200 seed/bottles, totally 3 bottles;

(2) tissue culture bottle is positioned over dark in 4 DEG C of refrigerators preservation, weekly with 4 DEG C of natural sea-water cleaning seeds, and carries out the replacing of an equitemperature natural sea-water;

(3) from January next year at every turn for all the seedling just germinateed being chosen when seed changes water, the seedling chosen is positioned over and is equipped with in the 200ml tissue culture bottle of natural sea-water, in 4 DEG C, preserve under dark ambient conditions, water consumption is 10ml/ strain;

(4) seedling changes weekly 4 DEG C of natural sea-waters once, and removes unsound seedling, clean container, and period observes and finds that seedling growth is very slow, only has a slice true leaf slightly to expose;

(5) from April, vitality test is carried out to the seedling chosen: seedling 30 strain that every monthly random selecting is preserved progressively heats up, adds after light cultivates 30 days and add up its survival rate.Concrete grammar is: every strain seedling puts into the 50ml triangular flask that equitemperature natural sea-water is housed, then triangular flask is proceeded in illumination box, cultivation temperature rises 1 DEG C for every 2 days, be up to 15 DEG C, initial stage dark culturing, when temperature rises to 10 DEG C, open illumination switch, if illumination is 1000Lux, after be increased to 2000Lux gradually, after experiment shows to proceed to incubator, the true leaf of seedling starts slowly to turn green, its ramp after temperature reaches 10 DEG C, after 30 days, most of seedling grows 2-3 sheet true leaf, a pair young root, the survival results that each moon detects is in table 1, after 8 months, survival rate is 63%.

Table 1 each month seedling force inspecting result

Time	The survival rate of seedling after 30 days
		In April, 2014	95.3%
In May, 2014	96.4%
		In June, 2014	90.8%
In July, 2014	82.8%
		In August, 2014	76.3%
In September, 2014	70.6%
		In October, 2014	68.5%
In November, 2014	63.8%

In the present embodiment, illumination box used is Shanghai rich news SPX-250B-G type illumination box, and refrigerator used is Haier BCD-225SKHCB three-door refrigerator.

Embodiment 2: a kind of store method of Zostera marina seedling, the steps include:

(1) August is put into the 300ml tissue culture bottle that natural sea-water is housed after the healthy grass wrack seeds cleaning that twin islet gulf, Weihai is collected back, wherein: 200 seed/bottles, totally 3 bottles;

(2) tissue culture bottle is positioned over dark in 0 DEG C of refrigerator preservation, weekly with 0 DEG C of natural sea-water cleaning seed, and carries out the replacing of an equitemperature natural sea-water;

(3) from January next year at every turn for all the seedling just germinateed being chosen when seed changes water, the seedling chosen is positioned over and is equipped with in the 200ml tissue culture bottle of natural sea-water, in 0 DEG C, preserve under dark ambient conditions, water consumption is 10ml/ strain;

(4) seedling changes weekly 0 DEG C of natural sea-water once, and removes unsound seedling, clean container, and period observes and finds that seedling grows hardly;

(5) from April, vitality test is carried out to the seedling chosen: seedling 30 strain that every monthly random selecting is preserved progressively heats up, adds after light cultivates 30 days and add up its survival rate.Concrete grammar is: every strain seedling is put into the 50ml triangular flask that 0 DEG C of natural sea-water is housed, put into the preservation of 4 DEG C of refrigerator dark and proceed to illumination box again after 3 days, cultivation temperature rises 1 DEG C for every 2 days, be up to 15 DEG C, initial stage dark culturing, when temperature rises to 10 DEG C, open illumination switch, if illumination is 1000Lux, after be increased to 2000Lux gradually, after experiment shows to proceed to incubator, seedling starts to send rough leaf, its ramp after temperature reaches 10 DEG C, after 30 days, most of seedling grows 2-3 sheet true leaf, a pair young root, the survival results that each moon detects is in table 2, after 8 months, survival rate is 71%.

Table 2 each month seedling vitality test result

Time	The survival rate of seedling after 30 days
		In April, 2014	98.7%
In May, 2014	96.6%
		In June, 2014	92.6%
In July, 2014	87.4%
		In August, 2014	80.9%
In September, 2014	76.1%
		In October, 2014	72.1%
In November, 2014	71.7%

In the present embodiment, illumination box used is Shanghai rich news SPX-250B-G type illumination box, and refrigerator used is Haier BCD-225SKHCB three-door refrigerator.

Claims (1)

1. a store method for Zostera marina seedling, is characterized in that, the steps include:

(1) put into the large container that natural sea-water is housed after the healthy grass wrack seeds cleaning of being collected back from marine site in 7-8 month, in 0-4 DEG C, preserve under dark environmental condition;

(2) weekly with equitemperature natural sea-water cleaning seed, and the replacing of an equitemperature natural sea-water is carried out;

(3) from January next year, the seedling of germination is chosen, is positioned over and is equipped with in the large container of natural sea-water, in 0-4 DEG C, preserve under dark ambient conditions, water consumption is 10ml/ strain;

(4) carry out weekly the replacing of an equitemperature natural sea-water, and remove unsound seedling, clean container.