CN108934272B - Method for promoting germination of seeds of sequoia jiangnanensis - Google Patents

Method for promoting germination of seeds of sequoia jiangnanensis Download PDF

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CN108934272B
CN108934272B CN201810857105.7A CN201810857105A CN108934272B CN 108934272 B CN108934272 B CN 108934272B CN 201810857105 A CN201810857105 A CN 201810857105A CN 108934272 B CN108934272 B CN 108934272B
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germination
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CN108934272A (en
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刘雄盛
蒋燚
肖玉菲
王勇
刘菲
姜英
黄荣林
韦铄星
陈风帆
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Guangxi Zhuang Autonomous Region Forestry Research Institute
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01CPLANTING; SOWING; FERTILISING
    • A01C1/00Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting

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Abstract

The invention discloses a method for promoting germination of seeds of sequoia jiangnanensis, which comprises the following steps: (1) seed selection: selecting seeds with the shapes of ellipses, brown and glossy front surfaces, light yellow back surfaces, wider middle parts or wider middle lower parts of the seeds, and thousand seed weights of 155g, and then floating and selecting full seeds; (2) seed disinfection: soaking the plump seeds obtained by seed selection by using a potassium permanganate solution, and then washing by using distilled water; (3) accelerating germination: soaking the sterilized seeds in gibberellin solution with the concentration of 50-300 mg/L for 12-48 h at the temperature of 25-40 ℃; (4) seed germination: and (3) placing the seeds subjected to the pregermination treatment in a culture dish paved with 2 layers of filter paper, and then placing the culture dish into an incubator for culture. The method can obviously improve the germination rate and the seedling quality of the south China fir seeds, shorten the germination time of the seeds, and provide a scientific and reliable method for the large-scale cultivation and the standardized production of the south China fir.

Description

Method for promoting germination of seeds of sequoia jiangnanensis
Technical Field
The invention belongs to the technical field of plant cultivation, and particularly relates to a method for promoting germination of seeds of sequoia jiangnanensis.
Background
South China fir (keteleria fortunei var. cyclolepis) belongs to Pinaceae (Pinaceae) fir (keteleria), is a unique local precious tree species in China, and is only sporadically scattered in provinces such as Fujian, Zhejiang, Jiangxi, Hunan, Guangdong, Guangxi, Yunnan, Guizhou and the like in China. The tree trunk of the south China fir is tall, straight, dense and green in branches and leaves, compact in material, beautiful in material color, durable, tree roots contain colloid and are commonly used for papermaking sizing materials in folk, so the south China fir is a valuable tree species using materials and landscaping ornamental tree species. Meanwhile, the south China fir has the excellent characteristics of strong stress resistance, fast growth, strong adaptability and the like, and is also an excellent mountain afforestation tree species. In recent years, due to the comprehensive factors of artificial felling, own biological characteristics, natural environment and the like, the distribution range of the south China fir is reduced, the wild quantity is sharply reduced, and the south China fir is classified as a local protection tree species by provinces.
In order to relieve the contradiction between resource shortage and development and utilization, the development of artificial cultivation is an effective way. In the previous work of investigation of planting resources and breeding and seedling of the south China fir, the seed setting rate of the south China fir is low, the seed abortion phenomenon rate is as high as 80%, the germination rate of the south China fir is low in the sowing process, the germination is irregular, and the conditions seriously restrict the standardized planting and production of the south China fir. On the other hand, the method suitable for seed germination of the south China fir is different due to the large difference of plant attributes and seed physiological habits of different plants, so that the method suitable for promoting the seed germination of the south China fir, particularly the south China fir, is not found in the prior art. Therefore, in order to promote the scale cultivation and the standardized production of the south China fir, researches on methods for shortening the germination time of the south China fir seeds and improving the germination rate and the seedling quality of the seeds are urgent.
Disclosure of Invention
Aiming at the defects of low germination rate, long germination time and the like of the south China fir seeds in the prior art, and aiming at the characteristics of hard seed coat, poor water permeability and rich oil of the south China fir seeds, the invention provides a method for promoting the germination of the south China fir seeds, which promotes the germination of the south China fir seeds, shortens the germination time of the seeds, and improves the germination rate of the seeds and the quality of nursery stocks.
The invention is realized by adopting the following technical scheme:
a method for promoting germination of seeds of Sequoia junior comprises the following steps:
(1) seed selection: selecting seeds which are oval in shape, brown and glossy in front, light yellow in back, wide in middle or middle and lower parts of the seeds, and 130-fold-weight seeds or 155 g-weight seeds, mixing the seeds with clear water, fully stirring the mixture, removing floating empty seeds and shrivelled seeds, adding sodium hydroxide into water, soaking, rubbing and washing the mixture to obtain full seeds;
(2) seed disinfection: soaking the plump seeds obtained by seed selection in 0.2 percent potassium permanganate solution for 10-15 min, and then washing with distilled water for 3-5 times;
(3) accelerating germination: soaking the sterilized seeds in gibberellin solution with the concentration of 50-300 mg/L for 12-48 h at the temperature of 25-40 ℃;
(4) seed germination: and (3) placing the seeds subjected to the pregermination treatment in a culture dish paved with 2 layers of filter paper, then placing the culture dish in an incubator, and continuously culturing for 24 h as a period under the conditions that the temperature is 25 ℃, the humidity is 90%, the illumination time is 12h, the illumination is dark, and the illumination intensity is 1000 LX.
Further optimizing, in the seed selection in the step (1), 0.2 g of sodium hydroxide and 1.5U of alkaline lipase are added into each 100 ml of clear water, the seeds are rubbed after being soaked for 12 hours, and the settled seeds are fished out to obtain full seeds.
Further optimizing, the germination accelerating treatment in the step (3) is carried out, and the sterilized seeds are soaked for 48 hours at the temperature of 30 ℃ by using gibberellin solution with the concentration of 200 mg/L.
Compared with the prior art, the technical scheme has the following beneficial effects:
1. in the seed selection step of the method, seeds with oval shapes, brown and glossy front surfaces, light yellow back surfaces and wider middle parts or wider middle and lower parts of the seed wings are selected, and the thousand seed weight of the seeds is required to reach 155 g. The thousand kernel weight is an important index for reflecting the size and the fullness of the seeds and is an important basis for measuring the quality of the seeds. The method screens the seeds according to the standards of the shape, the color, the wing shape, the thousand seed weight and the like of the seeds, can effectively ensure that the screened seeds with good plumpness and excellent quality are obtained, and improves the germination rate of the seeds; the sodium hydroxide and the alkaline lipase are added to soak and scrub the seeds, so that a large amount of grease of the seed coats can be effectively removed, the seed coats are softened, the dosage of the sodium hydroxide is reduced by adding the alkaline lipase, and the safety of the method is improved.
2. In the germination accelerating treatment step of the method, aiming at the characteristics of hard seed coat, poor water permeability, rich oil, slow germination and low germination rate of the south China fir seeds, the sterilized seeds are soaked by gibberellin solution with proper concentration, and the soaking temperature and time are scientifically controlled, so that the germination of the south China fir seeds is promoted, and the germination time of the seeds can be shortened.
3. In the seed germination step of the method, the seeds are cultured under the condition of 90 percent of humidity, so that the south China fir seeds are ensured to fully absorb moisture during germination, the swelling and softening of seed coats are promoted, the penetration of oxygen is increased, various physiological activities of the seeds are enhanced, the absorption and utilization of nutrient substances stored in the seeds are improved, the germination rate of the seeds is improved, and the germination time of the seeds is shortened.
4. The experimental data prove that the method of the invention can not only improve the germination rate of the seeds and shorten the germination time of the seeds, but also improve the germination vigor, the germ length and the germ root length of the south China fir seeds compared with the prior art. Therefore, the method can comprehensively improve the vitality of the seeds and improve the quality of the seedlings.
5. The method is suitable for promoting the germination of the seeds of the south China fir, is simple to operate, low in cost and easy to popularize, and meets the requirements of large-scale cultivation and standardized production of the south China fir.
Detailed Description
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto. The specific experimental conditions and methods not indicated in the following examples are generally conventional means well known to those skilled in the art.
Example 1:
a method for promoting germination of seeds of Sequoia junior comprises the following steps:
(1) seed selection: selecting seeds with the shape of an ellipse, brown and glossy front surface, light yellow back surface, wider middle part or wider middle lower part of a seed wing, and the weight of a thousand grains of 130 g, mixing the seeds with clear water, fully stirring, removing floating empty grains and shriveled grains, then adding 0.2 g of sodium hydroxide and 1.5U of alkaline lipase into per 100 ml of clear water, soaking for 12 hours, rubbing and washing the seeds, and fishing out the settled seeds to obtain full seeds;
(2) seed disinfection: soaking the plump seeds obtained by seed selection for 12 min by using a potassium permanganate solution with the mass fraction of 0.2 percent, and then washing the seeds for 4 times by using distilled water;
(3) accelerating germination: soaking the sterilized seeds for 48 hours at the temperature of 30 ℃ by using gibberellin solution with the concentration of 200 mg/L;
(4) seed germination: and (3) placing the seeds subjected to the pregermination treatment in a culture dish paved with 2 layers of filter paper, then placing the culture dish in an incubator, and continuously culturing for 24 h as a period under the conditions that the temperature is 25 ℃, the humidity is 90%, the illumination time is 12h, the illumination is dark, and the illumination intensity is 1000 LX.
Example 2:
a method for promoting germination of seeds of Sequoia junior comprises the following steps:
(1) seed selection: selecting seeds with the shape of an ellipse, brown and glossy front surface, light yellow back surface, wider middle part or wider middle lower part of a seed wing, and heavy thousand seeds of 135 g, mixing the seeds with clear water, fully stirring the mixture, removing floating empty particles and shriveled particles, then adding 0.2 g of sodium hydroxide and 1.5U of alkaline lipase into 100 ml of clear water, soaking the seeds for 12 hours, rubbing the seeds, and fishing out the settled seeds to obtain full seeds;
(2) seed disinfection: soaking the plump seeds obtained by seed selection for 10 min by using a potassium permanganate solution with the mass fraction of 0.2 percent, and then washing the seeds for 3 times by using distilled water;
(3) accelerating germination: soaking the sterilized seeds for 36 h at the temperature of 25 ℃ by using gibberellin solution with the concentration of 200 mg/L;
(4) seed germination: and (3) placing the seeds subjected to the pregermination treatment in a culture dish paved with 2 layers of filter paper, then placing the culture dish in an incubator, and continuously culturing for 24 h as a period under the conditions that the temperature is 25 ℃, the humidity is 90%, the illumination time is 12h, the illumination is dark, and the illumination intensity is 1000 LX.
Example 3:
a method for promoting germination of seeds of Sequoia junior comprises the following steps:
(1) seed selection: selecting seeds with the shape of an ellipse, brown and glossy front surface, light yellow back surface, wider middle part or wider middle lower part of a seed wing and the thousand-grain weight of 155g, mixing the seeds with clear water, fully stirring the mixture, removing floating empty grains and shriveled grains, then adding 0.2 g of sodium hydroxide and 1.5U of alkaline lipase into each 100 ml of clear water, soaking the seeds for 12 hours, rubbing the seeds, and fishing out the settled seeds to obtain full seeds;
(2) seed disinfection: soaking the plump seeds obtained by seed selection for 15 min by using a potassium permanganate solution with the mass fraction of 0.2 percent, and then washing the seeds for 5 times by using distilled water;
(3) accelerating germination: soaking the sterilized seeds for 48 hours at the temperature of 35 ℃ by using gibberellin solution with the concentration of 100 mg/L;
(4) seed germination: and (3) placing the seeds subjected to the pregermination treatment in a culture dish paved with 2 layers of filter paper, then placing the culture dish in an incubator, and continuously culturing for 24 h as a period under the conditions that the temperature is 25 ℃, the humidity is 90%, the illumination time is 12h, the illumination is dark, and the illumination intensity is 1000 LX.
Example 4:
a method for promoting germination of seeds of Sequoia junior comprises the following steps:
(1) seed selection: selecting seeds with the shape of an ellipse, brown and glossy front surface, light yellow back surface, wider middle part or wider middle lower part of a seed wing, and 145 g thousand-grain weight, mixing the seeds with clear water, fully stirring the mixture, removing floating empty grains and shrunken grains, then adding 0.2 g of sodium hydroxide and 1.5U of alkaline lipase into each 100 ml of clear water, soaking the seeds for 12 hours, rubbing the seeds, and fishing out the settled seeds to obtain full seeds;
(2) seed disinfection: soaking the plump seeds obtained by seed selection for 13 min by using a potassium permanganate solution with the mass fraction of 0.2 percent, and then washing the seeds for 4 times by using distilled water;
(3) accelerating germination: soaking the sterilized seeds for 48 hours at the temperature of 40 ℃ by using a gibberellin solution with the concentration of 50 mg/L;
(4) seed germination: and (3) placing the seeds subjected to the pregermination treatment in a culture dish paved with 2 layers of filter paper, then placing the culture dish in an incubator, and continuously culturing for 24 h as a period under the conditions that the temperature is 25 ℃, the humidity is 90%, the illumination time is 12h, the illumination is dark, and the illumination intensity is 1000 LX.
Example 5:
a method for promoting germination of seeds of Sequoia junior comprises the following steps:
(1) seed selection: selecting seeds with the shape of an ellipse, brown and glossy front surface, light yellow back surface, wider middle part or wider middle lower part of a seed wing and the weight of thousand seeds of 140 g, mixing the seeds with clear water, fully stirring the mixture, removing floating empty grains and shriveled grains, then adding 0.2 g of sodium hydroxide and 1.5U of alkaline lipase into 100 ml of clear water, soaking the seeds for 12 hours, rubbing the seeds, and fishing out the settled seeds to obtain full seeds;
(2) seed disinfection: soaking the plump seeds obtained by seed selection for 10 min by using a potassium permanganate solution with the mass fraction of 0.2 percent, and then washing the seeds for 5 times by using distilled water;
(3) accelerating germination: soaking the sterilized seeds for 12 hours at the temperature of 35 ℃ by using gibberellin solution with the concentration of 200 mg/L;
(4) seed germination: and (3) placing the seeds subjected to the pregermination treatment in a culture dish paved with 2 layers of filter paper, then placing the culture dish in an incubator, and continuously culturing for 24 h as a period under the conditions that the temperature is 25 ℃, the humidity is 90%, the illumination time is 12h, the illumination is dark, and the illumination intensity is 1000 LX.
Example 6:
a method for promoting germination of seeds of Sequoia junior comprises the following steps:
(1) seed selection: selecting seeds with the shape of an ellipse, brown and glossy front surface, light yellow back surface, wider middle part or wider middle lower part of a seed wing, and the weight of a thousand grains of 150g, mixing the seeds with clear water, fully stirring, removing floating empty grains and shriveled grains, then adding 0.2 g of sodium hydroxide and 1.5U of alkaline lipase into per 100 ml of clear water, soaking for 12 hours, rubbing and washing the seeds, and fishing out the settled seeds to obtain full seeds;
(2) seed disinfection: soaking the plump seeds obtained by seed selection for 15 min by using a potassium permanganate solution with the mass fraction of 0.2%, and then washing the seeds for 3 times by using distilled water;
(3) accelerating germination: soaking the sterilized seeds for 24 hours at the temperature of 35 ℃ by using a gibberellin solution with the concentration of 300 mg/L;
(4) seed germination: and (3) placing the seeds subjected to the pregermination treatment in a culture dish paved with 2 layers of filter paper, then placing the culture dish in an incubator, and continuously culturing for 24 h as a period under the conditions that the temperature is 25 ℃, the humidity is 90%, the illumination time is 12h, the illumination is dark, and the illumination intensity is 1000 LX.
Test example:
designing an orthogonal test, and taking gibberellin solution concentration, soaking time and soaking temperature as factors to test and determine the germination rate, germination vigor, germination index, germination time lag, germ length and germ root length of the south China fir seeds under different germination accelerating treatments. The test comprises the following specific steps:
(1) seed source: the tested seeds are from the Jiangnan oil fir forest with good growth in six-beat separation of the Guishan forest farm in Hezhou city. Collecting cones in 11 months of 2015, then stacking the cones indoors for several days, spreading the cones under the sun after seeds in the cones are fully mature, naturally releasing the seeds, airing, and then putting the cones into a refrigerator at 4 ℃ for low-temperature storage;
(2) seed selection: selecting filled seeds for testing according to the seed selection method described in the above example 1;
(3) seed disinfection: seed disinfection was performed according to the seed disinfection method described in example 1 above;
(4) accelerating germination: adopting gibberellin solution, soaking temperature and soaking time to perform pregermination treatment on the south China fir seeds. Taking gibberellin solution concentration, soaking time and soaking temperature as factors, arranging tests (table 1) for 4 levels of each factor, totally treating 16 treatments (table 2), and taking clean water for soaking for 12h at room temperature as a Control (CK);
TABLE 1 Germination test factor level table
Figure DEST_PATH_IMAGE001
TABLE 2 Germination accelerating treatment test combinations
Figure DEST_PATH_IMAGE002
(5) Seed germination: placing the seeds after germination accelerating treatment in culture dishes paved with 2 layers of filter paper, placing 30 seeds in each culture dish, repeating each treatment for 3 times, and then placing the seeds in an incubator with the temperature of 25 ℃, the humidity of 90%, the illumination intensity of 1000 LX and the alternate illumination time of 12h illumination/12 h darkness for culture;
(6) and (3) observing seed germination: the observation is carried out for 30 days every 24 h, and the germination is regarded as the germination when the radicle reaches 1/2. Counting germination indexes such as seed germination rate, germination vigor, germination index, germination time lag, germ and radicle length and the like, wherein the calculation formula and the description of each index are as follows:
germination percentage (%) = (total number of germinated seeds/number of test seeds) × 100%;
germination potential (%) = (number of normal germination seeds/number of test seeds within 10 d) × 100%;
germination index = ∑ (Gt/Dt), where Gt is the number of germinated seeds in t days, Dt is the number of germination days;
germination time lag (d): the days required for the first seed to germinate;
embryo and radicle length (mm): after the seeds germinate, 10 germinated seeds are selected from each culture dish to respectively measure the lengths of the embryo bud and the radicle (all the seeds are measured if the number of the seeds is less than 10);
(7) and (3) measuring results: the germination rate, the germination vigor, the germination index, the germination time lag, the germ length and the germ root length of the south China fir seeds have very obvious differences under different germination accelerating treatments (Table 3);
TABLE 3 analysis of variance and multiple comparisons of seed germination between different pregermination treatments
Figure DEST_PATH_IMAGE003
Note: ". indicates". sup. "indicatesPThe difference was extremely significant at the level of = 0.05; the letters in the same column are shown inPA significant difference at the level of =0.05, with the same letter indicating no significant difference.
(8) And (4) conclusion: compared with a control method, the seed treatment method has the advantages that the seed germination is greatly shortened, the germination vigor, the germination index, the germ length and the germ root length are superior to those of a control group, and particularly, the germination rate, the germination vigor, the germination index, the germ length and the germ root length of the sequoia jiangnanensis seeds are respectively 83.33 percent, 56.67 percent, 28.32 mm, 20.05 mm and 41.35 mm under the condition of 10 treatment; compared with the germination rate, the germination vigor, the germination index, the germ length and the radicle length of the Control (CK) treatment are respectively improved by 20 percent, 26.67 percent, 14.56 percent, 8.36 mm and 9.77 mm; meanwhile, the germination time lag of the seeds under the condition of the treatment 10 is shortened by 3.66 d compared with that of the control treatment. Therefore, the method can obviously improve the germination rate and the seedling quality of the Sequoia sempervirens seeds, shorten the germination time of the seeds and provide a scientific and reliable method for promoting the large-scale cultivation and the standardized production of the Sequoia sempervirens.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.

Claims (1)

1. A method for promoting the germination of seeds of Sequoia serrulata is characterized by comprising the following steps: which comprises the following steps:
(1) seed selection: selecting seeds with the shape of ellipse, brown and glossy front surface, light yellow back surface, wider middle part or wider middle lower part of the seed wing, and the thousand seed weight of 155g, mixing the seeds with clear water, fully stirring the mixture, removing floating empty grains and shriveled grains, adding sodium hydroxide and alkaline lipase into water, soaking and washing the mixture to obtain full seeds;
(2) seed disinfection: soaking the plump seeds obtained by seed selection in 0.2 percent potassium permanganate solution for 10-15 min, and then washing with distilled water for 3-5 times;
(3) accelerating germination: soaking the sterilized seeds for 48 hours at the temperature of 30 ℃ by using gibberellin solution with the concentration of 200 mg/L;
(4) seed germination: placing the seeds after germination accelerating treatment in a culture dish paved with 2 layers of filter paper, then placing the seeds in an incubator, and continuously culturing the seeds in 24 h as a period under the conditions that the temperature is 25 ℃, the humidity is 90%, the illumination time is 12h, the illumination is 12h, the darkness is 12h and the illumination intensity is 1000 LX;
in the step (1), 0.2 g of sodium hydroxide and 1.5U of alkaline lipase are added into each 100 ml of clear water, the seeds are washed by rubbing after being soaked for 12 hours, and the settled seeds are fished out to obtain full seeds.
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CN116171677A (en) * 2022-11-11 2023-05-30 广西壮族自治区林业科学研究院 Method for improving seed germination rate of huperzia serrata by changing environmental factors
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