Storage method of cunninghamia lanceolata pollen

Abstract

The invention relates to the technical field of plant pollen storage, in particular to a storage method of cunninghamia lanceolata pollen. The storage method comprises the following steps: S1, collecting male flower clusters; s2, the male flower clusters are dried for 1-2 days in the environment with the temperature being 30-35 DEG C and the relative humidity being 40-50%; s3, sieving the dried male flower clusters to obtain pollen; s4, the pollen is refrigerated in a shading mode. According to the storage method, male flower clusters are stored under the warm-temperature and low-humidity conditions with the temperature being 30-35 DEG C and the relative humidity being 40-50%, compared with natural drying, the germination rate is remarkably increased, and the pollen yield can reach 10% or above.

Description

Technical Field

[0001] The invention relates to the technical field of plant pollen storage, and in particular to a storage method for fir pollen. Background Art

[0002] Chinese fir is a subtropical tree species, up to 30 meters tall, with a breast diameter of 2.5 to 3 meters. Chinese fir wood is yellow-white and easy to process, and is an important greening and construction material. The bark of Chinese fir contains tannin, which has the effects of dispelling wind and relieving pain, dispersing blood stasis and stopping bleeding. It can be used for chronic tracheitis, stomachache, rheumatic joint pain, traumatic injuries and other diseases.

[0003] Hybrid breeding is one of the important ways of crop breeding, and pollen vigor determines the effectiveness of hybridization. Before pollination in Chinese fir hybrid breeding, pollen from male flowers of Chinese fir must be collected first. The male flower branches of Chinese fir are directly collected and scattered pollen is collected in containers or sealed bags. The collected pollen is easily inactivated when stored at room temperature. The Chinese utility model patent with application number 202022508277.9 discloses the steps of drying mature male flower pollen at 40-55°C for preservation, but the dried pollen is stored at room temperature, the activity is lower and faster, and the germination rate is not high. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a storage method for fir pollen with high germination rate.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is: a storage method of fir pollen, comprising the following steps: S1, collect male flower clusters; S2, drying the male flower clusters at a temperature of 30-35°C and a relative humidity of 40-50% for 1-2 days; S3, sieving the dried male flower clusters to obtain pollen; S4. Shield the pollen from light and refrigerate.

[0006] The beneficial effect of the present invention is that the storage method of the present invention significantly improves the germination rate and the powder yield rate can reach more than 10% by placing the male flower clusters under warm temperature and low humidity conditions of 30-35°C and 40-50% relative humidity compared with natural drying. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1 This is a statistical chart of the germination rate of male flower clusters at different stages after drying under different conditions; Figure 2 This is a comparison chart of the germination rates of male flower clusters at different stages after drying under different conditions; Figure 3 This is a comparison chart of pollen germination rates under different cold storage conditions; Figure 4 This is a graph showing the change of pollen germination rate over time under sealed refrigerated storage conditions at 8°C and relative humidity of 70%; Figure 5 This is a graph showing the change of pollen germination rate over time under the conditions of unsealed refrigerated storage at a temperature of 8°C and a relative humidity of 70%. Figure 6 This is a graph showing the change of pollen germination rate over time under indoor storage conditions of 20.1℃ and 72.9% relative humidity; Figure 7 This is a graph showing the change in pollen germination rate over time under storage conditions in a climate box at a temperature of 22.1°C and a relative humidity of 45.1%. DETAILED DESCRIPTION

[0008] In order to explain the technical content, achieved objectives and effects of the present invention in detail, the following is an explanation in combination with the implementation modes and the accompanying drawings.

[0009] A method for storing fir pollen comprises the following steps: S1, collect male flower clusters; S2, drying the male flower clusters at a temperature of 30-35°C and a relative humidity of 40-50% for 1-2 days; S3, sieving the dried male flower clusters to obtain pollen; S4. Shield the pollen from light and refrigerate.

[0010] From the above description, it can be seen that the beneficial effect of the present invention is that the present invention obtains pollen by drying the male flower clusters and then sieving them, and the pollen yield can reach more than 10%. Traditional fir pollen is usually dried in a dryer or baked at below 40°C. The former has a small processing capacity and is only suitable for experimental research. The latter is difficult to control the temperature. It takes more than 3 days to process a batch, which easily inactivates the pollen, and the pollen vitality can only be maintained for about four days; and neither of them regulates the relative humidity during drying. Through research, the present invention found that the microsporangium ball is carried out under warm temperature and low humidity conditions with a temperature of 30~35°C and a relative humidity of 40~50%, which increases the germination rate by more than 26.% compared with natural drying, and the pollen yield can reach more than 10%.

[0011] Furthermore, S1 is specifically as follows: collection begins when more than 50% of the male cones in the male flower cluster change from yellow-green to brown, from firm to soft, and the bract gaps widen and present yellow grids.

[0012] From the above description, we can know that at this stage, the pollen extraction rate (pollen weight / male cone weight) of male flower clusters can reach more than 10%. Furthermore, S1 is specifically as follows: the male flower clusters entering the collection period are divided into five stages, and the male flower clusters of stage three and stage four are collected; Stage 1: The male flower cluster is green and compact, and the edges of the microsporophyll bracts are brown; Stage 2: The lower part of the male flower cluster is green-brown and the whole is compact, and the upper part of the microsporophyll bracts is brown; Stage 3: The lower part of the male flower cluster is brown, the microspore leaves in the lower third are more spaced and loose, and the bracts are brown; Stage 4: At least two-thirds of the male flower cluster is brown, the microspore leaves in the lower half are spaced wider and looser, the bracts are brown, and the bottom part begins to shed powder; Stage 5: The male flower clusters are all brown, the microspore leaves in the lower half are shedding pollen, and the bottom part is completely shedding pollen.

[0013] From the above description, it can be seen that the change from yellow-green to brown in the male flower cluster has a time limit. In order to ensure the maturity and germination rate of pollen, the collection is usually started after the male flower cluster begins to shed pollen obviously (i.e., stage five of the present invention), and the collection is stopped before all the male cones shed pollen, and the pollen is stored after being dried in the shade. However, the male cones shed pollen at different times, so this treatment makes the pollen scattered in the early stage inactivated, and the pollen collection amount and germination rate are not high.

[0014] The pollen of Chinese fir is wrapped in microsporangia. Three microsporangia grow side by side on the microsporophyll. The microsporophyll grow spirally on the main axis to form microsporangium. The microsporangium grows together at the top of the paper strip to form a male cone cluster. The male cone cluster is composed of an average of 20 microsporangium. The maturity time of each microsporangium on the male cone cluster is inconsistent. Generally, the middle microsporangium matures earlier; the maturity time of microsporangium on each microsporangium is inconsistent. Generally, the lower microsporangium matures earlier; the maturity time of microsporangium in each microsporophyll is inconsistent. Generally, the middle microsporangium matures earlier. At each stage of microsporangium development, there are microsporangiums with immature microsporangium and pollen. The present invention dries male flower clusters under warm temperature and low humidity conditions of 30-35°C and 40-50% relative humidity. Due to the high temperature, the male cone clusters mature quickly, and the pollen that is close to completing development and waiting to be released in stage three and stage four is released, thereby increasing the pollen collection amount and germination rate, and the pollen release rate can reach more than 10%. Under natural drying indoors (20°C, 70%, the natural temperature and relative humidity of indoors in spring in the south), the temperature is low, the male cones mature slowly, the pollen that is close to completing development cannot fully mature, and the pollen release rate and germination rate are low.

[0015] Furthermore, the male flower clusters collected in S1 were dried within 3 days.

[0016] From the above description, it can be seen that the longer the male flower cluster is left before drying, the higher the probability of pollen inactivation.

[0017] Furthermore, S2 specifically includes: drying the male flower clusters at a temperature of 30° C. and a relative humidity of 50% for 1 to 2 days.

[0018] Furthermore, S3 is screened using a 50-100 mesh screen.

[0019] Furthermore, the refrigeration temperature in S4 is 5-8°C and the relative humidity is 60-70%.

[0020] Preferably, the refrigeration temperature in S4 is 8° C. and the relative humidity is 70%.

[0021] From the above description, we can see that short-term storage of pollen in an environment with a temperature of 5~8℃ and a relative humidity of 60~70% can increase the germination rate of pollen, and the germination rate can be effectively maintained for more than 13 days.

[0022] Furthermore, the refrigeration time in S4 is 3 to 14 days.

[0023] From the above description, we can see that the average germination rate after refrigeration for 3 to 14 days is over 53%.

[0024] Further, the S4 is sealed during refrigeration.

[0025] From the above description, it can be seen that the average germination rate is higher and the pollen activity is more stable when sealed during refrigeration than when not sealed.

[0026] Embodiment 1 of the present invention is: a method for storing fir pollen, comprising the following steps: S1. The male flower clusters entering the collection period are divided into five stages, and the male flower clusters in stage three and stage four are collected; Stage 1: The male flower cluster is green and compact, and the edges of the microsporophyll bracts are brown; Stage 2: The lower part of the male flower cluster is green-brown and the whole is compact, and the upper part of the microsporophyll bracts is brown; Stage 3: The lower part of the male flower cluster is brown, the microspore leaves in the lower third are more spaced and loose, and the bracts are brown; Stage 4: At least two-thirds of the male flower cluster is brown, the microspore leaves in the lower half are spaced wider and looser, the bracts are brown, and the bottom part begins to shed powder; Stage 5: The male flower clusters are all brown, the microspore leaves in the lower half are shed, and the bottom part is completely shed; S2, drying the collected male flower clusters for 2 days at a temperature of 30°C and a relative humidity of 50% within 3 days; S3, sieving the dried male flower clusters through an 80-mesh sieve to obtain pollen; S4. The pollen is sealed and refrigerated in a dark place for 14 days at a temperature of 8°C and a relative humidity of 70%.

[0027] 1. Using the storage method of Example 1, the germination rates of male flower clusters at different stages after drying under different conditions were tested. The test results are shown in Figure 1 and Figure 2 The germination rate test method is as follows: the dried, unrefrigerated pollen is placed in a culture medium and cultured in a dark artificial climate box at a temperature of 25°C and a relative humidity of 70% for 5 days. The culture medium includes 1.0wt% agar, 15.0wt% sucrose and 10mg / L boric acid.

[0028] Depend on Figure 1 and Figure 2 It can be seen that: (1) When the temperature was 20℃ and the relative humidity was 70% and the pollen was naturally dried indoors for 30 hours, the germination rates of pollen collected from microsporophyll at different development stages were significantly different, with a total mean of 33.01%, which was 5.82 percentage points lower than that of pollen collected from naturally scattered pollen ( ); (2) When dried in a climate box at 30°C and 50% relative humidity for 30 h, the germination rates of pollen collected from microsporophyll at different developmental stages were not significantly different, with a total mean of 41.86%, which was 27.35 percentage points higher than that of naturally scattered pollen ( ); (3) The germination rate of pollen dried at warm temperature and low humidity was significantly higher than that dried naturally indoors by 26.81%; (4) Compared with natural drying at 20℃ and 70% relative humidity indoors, drying in a climate box at 30℃ and 50% relative humidity significantly increased the germination rate of stage 1, stage 2, stage 3 and stage 5 microsporangium pollen.

[0029] 2. Using the storage method of Example 1, the effect of refrigeration time on the germination rate of pollen under different refrigeration conditions was tested. The test results are shown in Figure 3~Figure 7 The germination rate test method is as follows: the refrigerated pollen is placed in a culture medium, and cultured in an artificial climate box in a dark environment at a temperature of 25°C and a relative humidity of 70% for 5 days. The culture medium includes 1.0wt% agar, 15.0wt% sucrose and 10mg / L boric acid.

[0030] Depend on Figure 3~Figure 7 It can be seen that: (1) After 4 days of indoor storage at a temperature of 20.1°C and a relative humidity of 72.9%, the pollen germination rate dropped sharply to around 0; (2) When stored in a refrigerated environment at 8°C and 70% relative humidity, or in a climate box at 22.1°C and 45.1% relative humidity, the pollen germination rate can be effectively maintained for more than 13 days. (3) When the temperature is 8°C and the relative humidity is 70%, the sealed refrigerated storage is 3 to 14 days, and the average germination rate is 56.69%, with little fluctuation; when the temperature is 8°C and the relative humidity is 70%, the unsealed refrigerated storage is 3 to 14 days, and the average germination rate is 53.88%; (4) After storage in a climate box at a temperature of 22.1°C and a relative humidity of 45.1% for 2 to 14 days, the average germination rate was 49.61%.

[0031] Embodiment 2 of the present invention is: a method for storing fir pollen, comprising the following steps: S1. The male flower clusters entering the collection period are divided into five stages, and the male flower clusters in stage three and stage four are collected; Stage 1: The male flower cluster is green and compact, and the edges of the microsporophyll bracts are brown; Stage 2: The lower part of the male flower cluster is green-brown and the whole is compact, and the upper part of the microsporophyll bracts is brown; Stage 3: The lower part of the male flower cluster is brown, the microspore leaves in the lower third are more spaced and loose, and the bracts are brown; Stage 4: At least two-thirds of the male flower cluster is brown, the microspore leaves in the lower half are spaced wider and looser, the bracts are brown, and the bottom part begins to shed powder; Stage 5: The male flower clusters are all brown, the microspore leaves in the lower half are shed, and the bottom part is completely shed; S2, drying the collected male flower clusters for 1 day at a temperature of 35°C and a relative humidity of 40% within 3 days; S3, sieving the dried male flower clusters through a 50-mesh sieve to obtain pollen; S4. The pollen is sealed and refrigerated in a dark place for 14 days at a temperature of 5°C and a relative humidity of 60%.

[0032] Embodiment 3 of the present invention is: a method for storing fir pollen, comprising the following steps: S1. The male flower clusters entering the collection period are divided into five stages, and the male flower clusters in stage three and stage four are collected; Stage 1: The male flower cluster is green and compact, and the edges of the microsporophyll bracts are brown; Stage 2: The lower part of the male flower cluster is green-brown and the whole is compact, and the upper part of the microsporophyll bracts is brown; Stage 3: The lower part of the male flower cluster is brown, the microspore leaves in the lower third are more spaced and loose, and the bracts are brown; Stage 4: At least two-thirds of the male flower cluster is brown, the microspore leaves in the lower half are spaced wider and looser, the bracts are brown, and the bottom part begins to shed powder; Stage 5: The male flower clusters are all brown, the microspore leaves in the lower half are shed, and the bottom part is completely shed; S2, drying the collected male flower clusters within 3 days at a temperature of 33°C and a relative humidity of 45% for 1.5 days; S3, sieving the dried male flower clusters through a 100-mesh sieve to obtain pollen; S4. The pollen was sealed and refrigerated in a dark place for 14 days at a temperature of 6°C and a relative humidity of 65%.

[0033] In summary, the storage method of fir pollen provided by the present invention, by adjusting the drying conditions and the storage environment, firstly proposes that collection should begin after more than 50% of the male cones on the male cone cluster change from yellow-green to brown as a whole, change from tight to soft, and the bract gaps widen to present yellow grids (in stage three and stage four), so that the collection time is advanced by two development stages, the pollen collection amount, pollination rate and germination rate are improved, and the maximum time limit for short-term storage of pollen is pointed out, which provides an important reference for artificial assisted pollination.

[0034] The above descriptions are merely embodiments of the present invention and are not intended to limit the patent scope of the present invention. Any equivalent transformations made using the contents of the present invention's specification and drawings, or directly or indirectly applied in related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A method for storing Chinese fir pollen, characterized in that: The following steps are involved: S1, collect male flower clusters; S2, drying the male flower clusters at a temperature of 30-35°C and a relative humidity of 40-50% for 1-2 days; S3, sieving the dried male flower clusters to obtain pollen; S4. Shield the pollen from light and refrigerate.

2. The method for storing Chinese fir pollen according to claim 1, characterized in that: Specifically, S1 is as follows: when more than 50% of the male cones in the male flower cluster change from yellow-green to brown, from tight to soft, and the bract gaps widen and appear as yellow grids, collection begins.

3. The method for storing Chinese fir pollen according to claim 1, characterized in that: Specifically, S1 includes: dividing the male flower clusters entering the collection period into five stages, and collecting the male flower clusters in stage three and stage four; Stage 1: The male flower cluster is green and compact, and the edges of the microsporophyll bracts are brown; Stage 2: The lower part of the male flower cluster is green-brown and the whole is compact, and the upper part of the microsporophyll bracts is brown; Stage 3: The lower part of the male flower cluster is brown, the microspore leaves in the lower third are more spaced and loose, and the bracts are brown; Stage 4: At least two-thirds of the male flower cluster is brown, the microspore leaves in the lower half are spaced wider and looser, the bracts are brown, and the bottom part begins to shed powder; Stage 5: The male flower clusters are all brown, the microspore leaves in the lower half are shedding pollen, and the bottom part is completely shedding pollen.

4. The method for storing Chinese fir pollen according to claim 1, characterized in that: The male flower clusters collected in S1 were dried within 3 days.

5. The method for storing Chinese fir pollen according to claim 1, characterized in that: Specifically, S2 is: drying the male flower clusters at a temperature of 30° C. and a relative humidity of 50% for 1 to 2 days.

6. The method for storing Chinese fir pollen according to claim 1, characterized in that: In the S3, a 50-100 mesh screen is used for sieving.

7. The method for storing Chinese fir pollen according to claim 1, characterized in that: The refrigeration temperature in S4 is 5-8° C., and the relative humidity is 60-70%.

8. The method for storing Chinese fir pollen according to claim 7, characterized in that: The refrigeration temperature in S4 is 8° C. and the relative humidity is 70%.

9. The method for storing Chinese fir pollen according to claim 1, characterized in that: The refrigeration time in S4 is 3 to 14 days.

10. The method for storing Chinese fir pollen according to claim 1, characterized in that: The S4 is sealed during refrigeration.

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