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CN-115806930-B - Culture medium and culture method of small anti-sphagnum gametophyte

CN115806930BCN 115806930 BCN115806930 BCN 115806930BCN-115806930-B

Abstract

The invention relates to the technical field of bioengineering, and provides a culture medium and a culture method of a gametophyte of small anti-torsion moss. The spore culture medium of the small anti-torque gametophyte consists of ：MgSO 4 ·7H 2 O 250mg/L、KH 2 PO 4 250mg/L、FeSO 4 ·4H 2 O 12.5mg/L、KNO 3 1010mg/L、 ammonium tartrate 921mg/L, caCl 2 ·2H 2 O147 mg/L and glucose 5000mg/L. Through the technical scheme, the formation and propagation of the small anti-sphagnum gametophyte can be realized in a short time, a feasible way is provided for the cultivation and the subculture of the small anti-sphagnum, and seedling materials and propagation technology are provided for environmental restoration and garden landscaping by using moss.

Inventors

  • WANG YUTING
  • Dun Yanting
  • CUI XIAOJING

Assignees

  • 河北师范大学

Dates

Publication Date
20260505
Application Date
20221220

Claims (9)

  1. 1. The spore culture medium of the gametophyte of the small anti-torsion moss is characterized by comprising the following components: MgSO 4 ·7H 2 O 250mg/L、KH 2 PO 4 250mg/L、FeSO 4 ·4H 2 O 12.5mg/L、KNO 3 1010mg/L、 Ammonium tartrate 921mg/L, caCl 2 ·2H 2 O147 mg/L and glucose 5000mg/L.
  2. 2. The spore culture medium of the gametophyte of small anti-torque according to claim 1, characterized in that the pH of KH 2 PO 4 is 6.5.
  3. 3. The proliferation culture medium of the small anti-torque gametophyte is characterized by comprising the components of any one of claims 1-2, 4-D0.5 mg/L and 8g/L of agar powder.
  4. 4. The culture method of the small anti-torque gametophyte is characterized by comprising the following steps: S1, inoculating a spore suspension of the small anti-torsion moss into the spore culture medium according to any one of claims 1-2, and culturing to obtain protonema and gametophyte; S2, inoculating the protonema and gametophyte into the proliferation culture medium of claim 3, and culturing to obtain a new gametophyte; s3, inoculating the new gametophyte into a humid humus culture medium, and culturing to obtain the small anti-torsion gametophyte.
  5. 5. The method according to claim 4, wherein the culturing in S1, S2, S3 is performed at a temperature of 25.+ -. 2 ℃ and an illumination intensity of 2500 lx, and an illumination time of 16 h/day.
  6. 6. The method for culturing gametophyte of small anti-torsion moss according to claim 4, wherein the spore suspension in S1 is prepared by crushing the sterilized sporocysts to prepare a spore suspension; the capsule sterilization is specifically carried out by sequentially using water washing, sterilizing with ethanol solution with volume fraction of 75% for 5min, and water washing.
  7. 7. The method of claim 4, wherein the culturing time in S1 is 30 days.
  8. 8. The method of claim 4, wherein the culturing time in S2 is 40 days.
  9. 9. The method of claim 4, wherein the culturing time in S3 is 10 days.

Description

Culture medium and culture method of small anti-sphagnum gametophyte Technical Field The invention relates to the technical field of bioengineering, in particular to a culture medium and a culture method of a small anti-sphagnum gametophyte. Background The small anti-physcomitrella belongs to Cong Xianke and anti-physcomitrella, the plant body is densely clustered, a 'moss blanket' can be formed, the plant body is dark green, the plant body is slightly brown-green in the later growth period, the plant height is about 2-3 cm, and the stem is single or is branched in a bundle shape. She Congsheng in the top of the stem, the tip of the leaf is often curled into a cylinder shape when dried, the leaf edge is multi-toothed when wet, the middle rib is thick and strong under the leaf tip, and the basal cell of the leaf is long rectangle. The capsule is upright, cylindrical, has obvious bench, is yellow green and is brown in old time. The capsule is thin, about 1cm long, and often twists when dried, and is hermaphroditic. Is a special species in China, and is widely distributed in places such as Heilongjiang, hebei, gansu, yunnan and Tibet in China. The small anti-sphagnum is widely distributed in cities, is easy to collect, has double-layer cells, has good drought resistance and cold resistance, can complete life history in the field, and is easy to produce sporocysts. Meanwhile, the water-absorbing and water-holding material also has good water-absorbing and water-holding capacity, and has great development potential in the aspects of water-absorbing and water-holding material manufacturing, outdoor and indoor greening, environment indication, habitat reconstruction, repair and maintenance and the like. The bryophyte plants are short and have little development and utilization in ecological and economic aspects, so that the research on the bryophyte tissue culture at home and abroad is relatively little, the content is concentrated on spore germination or protonema development, and the research on the propagation and secondary culture of the bryophyte is little. At present, no related report is about propagation of small anti-sphagnum gametophytes in culture flasks by using Boea sporangia, and no research report is found about soil colonization of small anti-sphagnum. Disclosure of Invention The invention provides a culture medium and a culture method of small anti-torsion gametophytes, which can realize the formation and propagation of a large number of small anti-torsion gametophytes in a short time and the gametophytes of small anti-torsion gametophytes planted in soil. The technical scheme of the invention is as follows: spore culture medium of the gametophyte of the small anti-torque moss comprises MgSO4·7H2O 200～300mg/L、KH2PO4 200～300mg/L、FeSO4·4H2O 10～15mg/L、KNO3 1000～1500mg/L、 ammonium tartrate 900-1000 mg/L, caCl 2·2H2 O100-150 mg/L and glucose 4800-5200 mg/L. As a further technical scheme, the method comprises MgSO4·7H2O 250mg/L、KH2PO4 250mg/L、FeSO4·4H2O12.5mg/L、KNO3 1010mg/L、 ammonium tartrate 921mg/L, caCl 2·2H2 O147 mg/L and glucose 5000mg/L. As a further technical scheme, the pH of KH 2PO4 is 6.5. The proliferation culture medium of the small anti-torque gametophyte comprises the components of a spore culture medium, 2, 4-D0.5 mg/L and agar powder 8g/L. The culture method of the small anti-sphagnum gametophyte comprises the following steps: S1, inoculating spore suspension of small anti-torsion moss into the spore culture medium, and culturing to obtain protonema and gametophyte; s2, inoculating the protonema and the gametophyte into the proliferation culture medium, and culturing to obtain a new gametophyte; S3, inoculating the new gametophyte into a field planting culture medium for culture to obtain the small anti-torque gametophyte. As a further technical scheme, when the culture is carried out in the S1, the S2 and the S3, the temperature is 25+/-2 ℃, the illumination intensity is 2500lx, and the illumination time is 16 h/day. As a further technical scheme, the spore suspension in the S1 is prepared by crushing the sterilized sporocysts to prepare the spore suspension; the capsule sterilization is specifically carried out by sequentially using water washing, sterilizing with ethanol solution with volume fraction of 75% for 5min, and water washing. As a further technical scheme, the culture time in the S1 is 30 days. As a further technical scheme, the culture time in the S2 is 40 days. As a further technical scheme, the planting medium in the step S3 is a moist humus soil medium, and the culture time is 10 days. The subculture method of the small anti-sphagnum gametophyte comprises the following steps: S1, inoculating spore suspension of small anti-torsion moss into the spore culture medium, and culturing to obtain protonema and gametophyte; s2, smashing the protonema and the gametophyte, and inoculating the smashed protonema and gametophyte into a secondary culture medium to obtain a secondary s