CN105062950A - Nutrient solution, olive pollen nutrient mixing solution and artificial pollination method - Google Patents
Nutrient solution, olive pollen nutrient mixing solution and artificial pollination method Download PDFInfo
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Abstract
The invention discloses a nutrient solution, an olive pollen nutrient mixing solution, and an artificial pollination method. The nutrient solution is prepared by adding the following components in water: cane sugar, boric acid and monopotassium phosphate, wherein the pH of the nutrient solution is 6-6.5; the olive pollen nutrient mixing solution is prepared by adding olive pollen into the nutrient solution; the artificial pollination method of olive comprises the step of spraying the olive pollen nutrient mixing solution on blooming olive flowers at twice, wherein the time interval between the twice of pollination does not exceed 72h. Proved by a contrast experiment, compared with the effect obtained by spraying a solution composed of cane sugar and boric acid only, the inventor finds that the germination rate of olive pollen is improved by 8.36-11.33%, and the difference is very obvious. The olive pollen nutrient mixing solution is easy to prepare, convenient to use, and suitable for popularization and application. According to the artificial pollination method of the olive, the germination activity of pollen in artificial pollination can be obviously improved, so that the pollination success rate is improved, and finally, the percentage of fertile fruit of the olive is improved.
Description
Technical field
The invention belongs to plant sexual propagation field, relate to a kind of nutritive medium, and Fructus oleae europaeae pollens nutrition mixed solution, also relate to the method using this Fructus oleae europaeae pollens nutrition mixed solution to carry out Fructus oleae europaeae artificial pollination simultaneously.
Background technology
Fructus oleae europaeae (
oleaeuropaeal.) primary in Mediterranean country, China started introduction and acclimatization in 1964, by the screening of 50 years, had now formed multiple excellent main breed: mainly plant No. 8 with Lay star, Fo Ao, Hubei Province, skin cuts profit, A Si, No. 32, Chenggu etc. for representative.In Gansu, Yunnan, Sichuan, Shaanxi, the ground such as Guizhou successfully promotes and plants.
Research shows that the potentiality to be exploited of Fructus oleae europaeae oil is huge; be proved to be can stimulate circulation, improve digestive system function, protection skin, improve endocrine system function, benefit to Skeletal system, anti-cancer, radioprotective, the effect such as anti-ageing, prevention cardiovascular and cerebrovascular diseases obviously, be high-grade, green, healthy natural edible oil.The sweet oil that Chinese festiva import is more than 40,000 tons, current domestic sweet oil annual production is only 3% ~ 5% of year import total amount, needs raising output badly.
Contriver finds, can Fructus oleae europaeae pollen germination rate and germination of pollen tube length be the essential condition determining complete fertilization process, during pollination, pollen starts to sprout after contacting with pistil stigma is affine, pollen tube constantly extends and enters blastular and embryo of silkworms has merged fertilization, affect the factor of normal fertilization except outside temperature, humidity and wind, also have column cap activity and Pollen Activity.But the nutritive substance needed for pollen germination almost comes from pollen itself entirely, if when pollen germination, supplementing of the nutrition such as boric acid, sucrose, can promote the sprouting of pollen greatly.
Fructus oleae europaeae belongs to hermaphrodite flower, and setting percentage of pollinating between the tree body with kind is extremely low or shaky, and Olive Fruit rate is relatively low, and generally 3% ~ 8%, improve Olive Fruit rate is one of important channel of improving output always.And improve pollination efficiency by artificial hybridization pollination technique and improve percentage of fertile fruit the most effective approach.
Realizing in process of the present invention, contriver finds to study and promotes that the raising of ratio of nutrient solution to artificial pollination efficiency of Fructus oleae europaeae pollen germination is significant.Contriver, through test of many times design and multiple batches of test, progressively finds out the ratio of nutrient solution of optimum Fructus oleae europaeae pollen germination and pollen tube growth.And apply this proportioning and add a certain amount of Fructus oleae europaeae pollen, the form sprayed with water smoke carries out palmification to the Fructus oleae europaeae in the florescence, promotes Fructus oleae europaeae artificial pollination efficiency with this technology, and the method can improve percentage of fertile fruit and output preferably.
Summary of the invention
Given this, the object of the invention is to provide a kind of nutritive medium that can promote Fructus oleae europaeae pollen germination, and this nutritive medium can significantly improve pollination efficiency and percentage of fertile fruit compared to prior art.Meanwhile, present invention also offers the Fructus oleae europaeae pollens nutrition mixed solution for Fructus oleae europaeae artificial pollination.Present invention also offers the method using this Fructus oleae europaeae pollens nutrition mixed solution to carry out Fructus oleae europaeae artificial pollination.
For solving above technical problem, technical scheme provided by the invention is, provides a kind of nutritive medium, and this nutritive medium can promote Fructus oleae europaeae pollen germination, and this nutritive medium is in water, add following component and content thereof:
Sucrose: 12% ~ 13%wt;
Boric acid: 50mg/L ~ 60mg/L;
Potassium primary phosphate: 0.8g/L ~ 1.2g/L;
The pH of nutritive medium is 6 ~ 6.5.
According to a preferred implementation of nutritive medium of the present invention, described nutritive medium is in water, add following component and content thereof:
Sucrose: 12%wt;
Boric acid: 50mg/L;
Potassium primary phosphate: 1g/L;
The pH of nutritive medium is 6 ~ 6.5.
According to an embodiment of nutritive medium of the present invention, this nutritive medium also comprises following component and content thereof:
Component one: vitamins B
1: 20mg/L ~ 25mg/L;
Or/and
Component two: growth hormone: 4.5mg/L ~ 5.5mg/L;
Plant hormones regulators,gibberellins: 0.8mg/L ~ 1.2mg/L.
According to a preferred implementation of nutritive medium of the present invention, each component of described nutritive medium and content thereof are:
Component one: vitamins B
1: 20mg/L;
Or/and
Component two: growth hormone: 5mg/L;
Plant hormones regulators,gibberellins: 1mg/L.
The present invention also provides a kind of Fructus oleae europaeae pollens nutrition mixed solution based on aforementioned nutritive medium, adds Fructus oleae europaeae pollen in described nutritive medium, and the addition of pollen is 0.2g/L ~ 0.5g/L.
According to an embodiment of Fructus oleae europaeae pollens nutrition mixed solution of the present invention, the described Fructus oleae europaeae pollens nutrition mixed solution shelf time is no more than 4h.
According to an embodiment of Fructus oleae europaeae pollens nutrition mixed solution of the present invention, described Fructus oleae europaeae pollens nutrition mixed solution is prepared by following steps:
1) with pure tap water as solvent, first add sucrose, boric acid successively, then add all the other components, stir;
2) be 6 ~ 6.5 with sodium hydroxide or salt acid for adjusting pH value;
3) add subzero treatment to cross and the Fructus oleae europaeae pollen of the 1h ~ 2h that at room temperature rises again, it is fully stirred and Homogeneous phase mixing in the solution.
Present invention also offers one utilizes aforementioned Fructus oleae europaeae pollens nutrition mixed solution to carry out artificial pollination method, is applicable to Fructus oleae europaeae, Fructus oleae europaeae pollens nutrition mixed solution is sprayed to open Fructus oleae europaeae and takes; By Phenological Observation, carry out first time artificial pollination when setting body and having 40 ~ 50% the flowers are in blossom and put, carry out second time when setting body 80% ~ 90% the flowers are in blossom and putting and pollinate, twice pollination time interval is no more than 72h.As preferably, twice pollination time interval is no more than 48h.
According to an embodiment of artificial pollination method of the present invention, the collection also comprising pollen with preserve step: before Fructus oleae europaeae is bloomed, with sulfuric acid paper bag to inflorescence bagging, full-bloom stage is collected the pollen that sheds and is removed foreign material; Or gather compared with prematurity, branch of blooming that flower density is larger, in the loft drier of 30 DEG C, place 12h, below loft drier, lay the template collecting pollen, collect pollen and removal of impurities; Pollen is placed in vial is airtight, half-light, cryopreservation, be used in 4 DEG C ~-20 DEG C preservations then, next year is used in-20 DEG C of preservations.
According to an embodiment of artificial pollination method of the present invention, at room temperature rise again before the pollen that cryopreservation is crossed uses 1h ~ 2h.
Compared with prior art, a technical scheme tool in technique scheme has the following advantages:
1, nutrient solution prescription of the present invention: sucrose+boric acid+potassium primary phosphate, or sucrose+boric acid+potassium primary phosphate+vitamins B
1, or sucrose+boric acid+potassium primary phosphate+growth hormone+Plant hormones regulators,gibberellins, or sucrose+boric acid+potassium primary phosphate+vitamins B
1+ growth hormone+Plant hormones regulators,gibberellins can significantly improve the germination rate of Fructus oleae europaeae pollen.Contriver is proved by contrast experiment, and compared to sucrose+boric acid, the germination rate of Fructus oleae europaeae pollen improves 8.36% ~ 11.33%, and level of difference reaches extremely significantly (P
0.05< 0.01).
2, Fructus oleae europaeae pollens nutrition mixed solution preparation of the present invention is simple, easy to use, is suitable for applying.
3, Fructus oleae europaeae artificial pollination method of the present invention, obviously can promote the germination of pollen in artificial pollination, thus improves pollination success ratio, finally can improve Olive Fruit rate.
4, can Fructus oleae europaeae pollen germination rate and germination of pollen tube length be the essential condition determining complete fertilization process.The present invention can make the pollen 24h germination rate of Fructus oleae europaeae reach 10% ~ 20%, and pollen tube length obviously extends, and greatly can improve Fructus oleae europaeae pollen fertilization effect.
Embodiment
Specific embodiment is described below.
Embodiment 1
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 12%wt, boric acid 50mg/L, potassium primary phosphate 0.8g/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 2
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 13%wt; Boric acid 60mg/L; Potassium primary phosphate 1.2g/L; The pH of nutritive medium is 6 ~ 6.5.
Embodiment 3
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 12%wt; Boric acid 50mg/L; Potassium primary phosphate 1g/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 4
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 12%wt, boric acid 50mg/L, potassium primary phosphate 0.8g/L, vitamins B
120mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 5
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 13%wt, boric acid 60mg/L, potassium primary phosphate 1.2g/L, vitamins B
125mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 6
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 12%wt, boric acid 50mg/L, potassium primary phosphate 1g/L, vitamins B
120mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 7
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 12%wt, boric acid 50mg/L, potassium primary phosphate 0.8g/L, growth hormone 4.5mg/L, Plant hormones regulators,gibberellins 0.8mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 8
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 13%wt, boric acid 60mg/L, potassium primary phosphate 1.2g/L, growth hormone 5.5mg/L, Plant hormones regulators,gibberellins 1.2mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 9
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 12%wt, boric acid 50mg/L, potassium primary phosphate 1g/L, growth hormone 5mg/L, Plant hormones regulators,gibberellins 1mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 10
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 12%wt, boric acid 50mg/L, potassium primary phosphate 0.8g/L, vitamins B
120mg/L, growth hormone 4.5mg/L, Plant hormones regulators,gibberellins 0.8mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 11
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 13%wt, boric acid 60mg/L, potassium primary phosphate 1.2g/L, vitamins B
125mg/L, growth hormone 5.5mg/L, Plant hormones regulators,gibberellins 1.2mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 12
Nutritive medium described in the present embodiment adds each component by following standard in distilled water or pure tap water: sucrose 12%wt, boric acid 50mg/L, potassium primary phosphate 1g/L, vitamins B
120mg/L, growth hormone 5mg/L, Plant hormones regulators,gibberellins 1mg/L.The pH of nutritive medium is 6 ~ 6.5.
Embodiment 13
What the present embodiment said description is a kind of Fructus oleae europaeae pollens nutrition mixed solution, is the Fructus oleae europaeae pollen adding 0.2g/L ~ 0.5g/L on the basis of embodiment 1 ~ 12 any embodiment Middle nutrition liquid.Concrete interpolation standard can be 0.2g/L, 0.3g/L, 0.4g/L or 0.5g/L, and any amount in 0.2 ~ 0.5 scope, and such as, interpolation standard is 0.2g/L, namely adds 0.2g Fructus oleae europaeae pollen in every 1L nutritive medium.
The Fructus oleae europaeae pollens nutrition mixed solution shelf time is no more than 4h, namely need join and namely use.Through experimental observation, the shelf time is greater than 4h, and pollen tube will sprout elongation rapidly, can reduce the effect using the artificial pollination of Fructus oleae europaeae pollens nutrition mixed solution, finally can reduce Pollination Effect, reduce Fructus oleae europaeae output.
Fructus oleae europaeae pollens nutrition mixed solution preparation process is as follows:
1) with pure tap water as solvent, first add sucrose, boric acid successively, then add all the other components, stir; The addition of respective components is in following scope: sucrose: 12% ~ 13%wt; Boric acid: 50mg/L ~ 60mg/L; Potassium primary phosphate: 0.8g/L ~ 1.2g/L; Vitamins B
1: 20mg/L ~ 25mg/L; Growth hormone: 4.5mg/L ~ 5.5mg/L; Plant hormones regulators,gibberellins: 0.8mg/L ~ 1.2mg/L.Such as, with the solvent of pure tap water 1L as nutritive medium, 120 ~ 130g sucrose can be added, 50 ~ 60mg boric acid, 0.8 ~ 1.2g potassium primary phosphate, 20 ~ 25mg vitamins B
1, 4.5 ~ 5.5mg growth hormone, 0.8 ~ 1.2mg Plant hormones regulators,gibberellins.
2) be 6 ~ 6.5 with 0.1mol/L sodium hydroxide or salt acid for adjusting pH value.
3) add subzero treatment to cross and the Fructus oleae europaeae pollen of the 1h ~ 2h that at room temperature rises again, it is fully stirred and Homogeneous phase mixing in the solution.The pollen gathered is placed in that vial is airtight, half-light, cryopreservation, and when using then, pollen is preserved under 4 DEG C ~-20 DEG C conditions; Next year, when using, preserves under-20 DEG C of conditions.
Embodiment 14
Described by the present embodiment is the artificial pollination method of Fructus oleae europaeae.Comprise and gather and preserve the steps such as pollen, Fructus oleae europaeae pollens nutrition Compound mixed solution and spraying pollination.Wherein, Fructus oleae europaeae pollens nutrition Compound mixed solution carries out according to embodiment 13, and the present embodiment repeats no more this.
The collection of pollen and preservation: before Fructus oleae europaeae is bloomed, with sulfuric acid paper bag to inflorescence bagging, full-bloom stage is collected the pollen that sheds and is removed foreign material; Or gather compared with prematurity, branch of blooming that flower density is larger, in the loft drier of 30 DEG C, place 12h, below loft drier, lay the template collecting pollen, collect pollen and removal of impurities; Pollen is placed in vial is airtight, half-light, cryopreservation, be used in 4 DEG C ~-20 DEG C preservations then, next year is used in-20 DEG C of preservations; At room temperature rise again before the pollen that cryopreservation is crossed uses 1h ~ 2h.
Fructus oleae europaeae pollens nutrition mixed solution carries out artificial pollination: Fructus oleae europaeae pollens nutrition mixed solution is sprayed to open Fructus oleae europaeae and takes; By Phenological Observation, carry out first time artificial pollination when setting body and having 40 ~ 50% the flowers are in blossom and put, carry out second time when setting body 80% ~ 90% the flowers are in blossom and putting and pollinate, twice pollination time is no more than 72h, preferably more than 48h.Orchard worker accurately can not control spray amount usually when field spray, and spraying should be even, should not spray too much, on flower slightly water droplet time spray and can stop.
For further illustrating beneficial effect of the present invention, be described below by way of specific experiment.For convenience of more scientifically observing and the germination rate of Fructus oleae europaeae pollen and the length of pollen tube being described, this experiment is carried out in the lab.Nutritive medium is made into as substratum, and this substratum is on the basis of nutritive medium, with the addition of the agar of 0.5%wt, and agar does not then produce the impact of biological chemistry aspect on the germination of pollen and the elongation of pollen tube.Therefore, this substratum truly can reflect the function and efficacy of nutritive medium.
Specific experiment step is as follows:
(1) collection of pollen and preservation: the Fructus oleae europaeae branch that random selecting stamen is vigorous, collects the pollen half-light shed and is kept in the refrigerator of-20 DEG C.
(2) sterilizing before inoculation: before inoculation pollen, clean the utensil such as culture dish, tweezers with distilled water, and dry at 80 DEG C, more all utensils are placed in high pressure steam sterilization 30mim under 121 DEG C of conditions.
(3) making of substratum: the Fructus oleae europaeae pollens nutrition mixed solution of Fructus oleae europaeae artificial pollination produces utilization in the wild, and the tap water that choice for use is pure is more convenient as solvent, when conditions permit, also can use distilled water.For absolutely proving the promoter action of each component to pollen germination power, the factor beyond experimental design under laboratory condition, should be avoided the impact of experiment effect as far as possible, therefore, selecting distilled water as solvent.
the agar-agar soln of the distilled water preparation 0.5%wt boiled with 1L, then dissolve in 120g sucrose, 50mg boric acid successively, steam sterilizing 20min at 121 DEG C;
when culture medium solution temperature is down to 40 DEG C, according to the proportioning of following i ~ vi substratum, in agar-agar soln, add the potassium primary phosphate of corresponding content, vitamins B
1, growth hormone, Plant hormones regulators,gibberellins bacteriological filtration solution, stir:
I, 0.5%wt agar+12%wt sucrose+50mg/L boric acid+1g/L potassium primary phosphate.Increase parallel laboratory test group, the concentration arranging potassium primary phosphate is 3g/L, and all the other conditions are constant simultaneously;
II, 0.5%wt agar+12%wt sucrose+50mg/L boric acid+1g/L potassium primary phosphate+20mg/L vitamins B
1+ 5mg/L growth hormone+1mg/L Plant hormones regulators,gibberellins.Increase parallel laboratory test group, setting up the concentration of putting potassium primary phosphate is 3g/L, vitamins B simultaneously
125mg/L, set up growth hormone 7.5mg/L-Plant hormones regulators,gibberellins 3mg/L and combine, all the other conditions are constant;
III, 0.5%wt agar+12%wt sucrose+50mg/L boric acid+1g/L potassium primary phosphate+20mg/L vitamins B
1.Increase parallel laboratory test group, setting up the concentration of putting potassium primary phosphate is 3g/L, vitamins B simultaneously
125mg/L; All the other conditions are constant;
IV, 0.5%wt agar+12%wt sucrose+50mg/L boric acid+1g/L potassium primary phosphate+5mg/L growth hormone+1mg/L Plant hormones regulators,gibberellins.Increase parallel laboratory test group, setting up the concentration of putting potassium primary phosphate is 3g/L, and set up growth hormone 7.5mg/L-Plant hormones regulators,gibberellins 3mg/L and combine, all the other conditions are constant simultaneously;
V, 0.5%wt agar+12%wt sucrose+50mg/L boric acid+20mg/L vitamins B
1.Increase parallel laboratory test group simultaneously, set up and put vitamins B
125mg/L; All the other conditions are constant;
Vi, 0.5%wt agar+12% sucrose+50mg/L boric acid,
Wherein, vi, as the control experiment of i, II, III, IV, v, passes behind experimental study potassium primary phosphate, vitamins B
1, growth hormone, Plant hormones regulators,gibberellins is to the effect of Fructus oleae europaeae pollen germination;
be finally 6 ~ 6.5 by the sodium hydroxide of 0.1mol/L or salt acid for adjusting pH scope.
(4) pollen is inoculated: the bottom different culture media prepared being evenly distributed in respectively sterile petri dish, thickness 0.3 ~ 0.5cm.With tweezers or writing brush, the Fructus oleae europaeae pollen of the 1h ~ 2h that rises again is seeded on substratum equably, after covering the upper flap of culture dish, slightly shakes culture dish, make 1/3 ~ 1/2 of inoculation pollen surface to contact with substratum.
(5) pollen cultures and observation: inoculation is had the substratum of Fructus oleae europaeae pollen puts into 25 DEG C, the incubator of 3000Lx light intensity cultivates.After cultivation 4h, 24h and 48h, random observation 12 ~ 30 visuals field under the object lens of × 10, repeat 3 times, record and calculate germination rate and the germination of pollen tube length of Fructus oleae europaeae pollen, the results are shown in Table 1 to table 3.
Situation is sprouted during the pollen 4h of table 1 different culture media process
| Process | Germination rate (%) | Length (μm) |
| A1B1C1 | 0.92±0.22 JK | 101.57±15.79 GHI |
| A1B1C2 | 0.89±0.12 JK | 91.82±39.95 HI |
| A1B1C3 | 1.52±0.32 FGHIJK | 171.92±22.55 EFG |
| A1B2C1 | 1.18 ±0.30 IJK | 171.24±21.44 EFG |
| A1B2C2 | 2.10±0.33 EFGHIJK | 199.20±29.03 DEF |
| A1B2C3 | 0.64±0.12 K | 212.19±31.78 CDF |
| A1B3C1 | 0.85 ±0.17 JK | 190.67 ±51.74 DEF |
| A1B3C2 | 1.48±0.29 GHIJK | 142.93±32.16 FGH |
| A1B3C3 | 1.40±0.21 HIJK | 166.48±35.55 EFGH |
| A2B1C1 | 6.68±0.75 ABC | 221.84±17.11 CDEF |
| A2B1C2 | 6.91±0.78 AB | 237.09±18.25 CDE |
| A2B1C3 | 5.93±0.42 BC | 194.22±18.18 DEF |
| A2B2C1 | 7.79±0.79 A | 342.56±14.91 A |
| A2B2C2 | 5.53±1.09 BC | 265.21±18.17 BCD |
| A2B2C3 | 5.38±0.51 BCD | 169.46±22.31 EFG |
| A2B3C1 | 6.70±0.98 ABC | 331.83±20.18 AB |
| A2B3C2 | 5.08±0.52 CD | 256.61±19.28 CD |
| A2B3C3 | 5.34±0.59 BCD | 165.03±17.89 EFGH |
| A3B1C1 | 3.81±0.67 DE | 150.96±15.59 FGH |
| A3B1C2 | 2.89±0.38 EFGH | 66.66±15.52 I |
| A3B1C3 | 2.85±0.18 EFGHI | 67.86±4.82 I |
| A3B2C1 | 3.80±0.16 DE | 202.92±12.47 DEF |
| A3B2C2 | 3.14±0.30 EFG | 99.60±3.36 GHI |
| A3B2C3 | 3.74±0.18 DE | 144.63±11.98 FGH |
| A3B3C1 | 5.33±0.51 BCD | 282.75±20.30 ABC |
| A3B3C2 | 3.17±0.24 EF | 102.98±8.27 GHI |
| A3B3C3 | 2.49±0.26 EFGHIJ | 66.66±3.41 I |
Situation is sprouted during the pollen 24h of table 2 different culture media process
| Process | Germination rate (%) | Length (μm) |
| A1B1C1 | 4.45±0.54 DEFGH | 395.34±46.77 ABCD |
| A1B1C2 | 2.59±0.53 FGH | 175.39±25.79 HIJKLM |
| A1B1C3 | 4.41±0.59 DEFGH | 307.71±33.12 CDEFG |
| A1B2C1 | 8.34±0.67 ABC | 430.73 ±37.21 ABC |
| A1B2C2 | 2.43±0.30 GH | 367.99±59.53 ABCDE |
| A1B2C3 | 3.21±0.43 FGH | 322.08±36.59 BCDEF |
| A1B3C1 | 4.43±0.46 DEFH | 484.64 ±39.39 A |
| A1B3C2 | 3.30±0.51 EFGH | 317.38±34.91 BCDEF |
| A1B3C3 | 2.07±0.27 H | 212.42±19.83 GHIJKL |
| A2B1C1 | 10.31±1.35 AB | 444.39±39.74 AB |
| A2B1C2 | 8.47±1.07 ABC | 251.30±27.20 FGHIJ |
| A2B1C3 | 7.77±2.27 BCD | 197.87±29.72 GHIJKM |
| A2B2C1 | 8.36±0.95 ABC | 458.50±36.83 A |
| A2B2C2 | 11.33±3.46 A | 285.24±23.83 EFGHI |
| A2B2C3 | 7.71±1.50 BCD | 240.78±7.72 FGHIJ |
| A2B3C1 | 8.35±1.79 ABC | 416.75±34.12 ABCD |
| A2B3C2 | 6.80±0.91 CDE | 232.06±49.68 FGHIJK |
| A2B3C3 | 6.03±1.40 CDEF | 217.43±41.71 GHIJKL |
| A3B1C1 | 7.42±1.74 BCD | 253.87±18.28 FGHIJ |
| A3B1C2 | 5.98±1.03 CDEG | 84.23±13.28 LM |
| A3B1C3 | 2.81±0.23 FGH | 71.81±22.99 M |
| A3B2C1 | 6.12±0.68 CDEF | 168.64±31.28 IJKLM |
| A3B2C2 | 4.33±0.78 DEFH | 103.03±27.95 KLM |
| A3B2C3 | 4.20±0.72 DEFH | 144.17±34.21 JKLM |
| A3B3C1 | 7.48±0.56 BCD | 394.32±36.79 ABCD |
| A3B3C2 | 4.76±0.69 DEFH | 119.27±17.61 JKLM |
| A3B3C3 | 2.68±0.41 FGH | 69.96±7.84 M |
Situation is sprouted during the pollen 48h of table 3 different culture media process
| Process | Germination rate (%) | Length (μm) |
| A1B1C1 | 5.01±0.26 DEF | 370.36±43.77 CDEF |
| A1B1C2 | 3.08±1.01 F | 211.45±20.36 GHIJK |
| A1B1C3 | 5.00±1.04 DEF | 360.99±42.62 CDEF |
| A1B2C1 | 9.77 ±1.25 ABC | 610,268±34.86 AB |
| A1B2C2 | 3.61±0.20 EF | 333.61 ±39.34 EFG |
| A1B2C3 | 3.09±0.70 F | 286.55±43.66 EFH |
| A1B3C1 | 5.12 ±1.43 DEF | 549.94 ±55.32 A |
| A1B3C2 | 3.45±0.83 F | 330.50±32.09 DEFG |
| A1B3C3 | 2.90±0.25 F | 234.18 ±36.25 GHIJ |
| A2B1C1 | 11.89±2.46 A | 456.11 ±47.52 ABC |
| A2B1C2 | 10.29±2.42 AB | 313.72 ±34.44 DEFG |
| A2B1C3 | 7.93±0.48 ABCD | 251.23 ±38.46 FGHI |
| A2B2C1 | 9.81±0.46 ABC | 451.78 ±53.24 ABC |
| A2B2C2 | 11.89±1.28 A | 287.11 ±3.70 EFGH |
| A2B2C3 | 8.49±1.20 ABCD | 274.98 ±39.50 EFGH |
| A2B3C1 | 9.76±1.49 ABC | 426.07 ±59.24 BCD |
| A2B3C2 | 5.51±1.77 DEF | 268.85 ±25.42 EFGH |
| A2B3C3 | 6.46±0.50 BCDEF | 183.81 ±16.14 HIJKL |
| A3B1C1 | 8.51±1.73 ABCD | 275.29 ±27.47 EFGH |
| A3B1C2 | 6.21±1.02 BCDEF | 86.64 ±7.10 L |
| A3B1C3 | 3.33±0.72 F | 110.78 ±17.72 KL |
| A3B2C1 | 7.85±0.89 ABCD | 215.35 ±27.64 GHIJK |
| A3B2C2 | 5.70±1.20 CDEF | 106.67 ±9.77 KL |
| A3B2C3 | 5.89±0.76 CDEF | 148.53 ±31.01 IJKL |
| A3B3C1 | 7.70±1.35 BCDE | 388.85 ±24.79 CDE |
| A3B3C2 | 5.25±0.58 DEF | 121.20 ±16.06 JKL |
| A3B3C3 | 4.37±0.85 DEF | 90.15 ±22.57 L |
Note: in table 1, table 2 and table 3, data are mean value ± standard error, and An represents potassium primary phosphate, and A1 represents that its concentration is that 0g/L, A2 represent that its concentration is that 1g/L, A3 represent that its concentration is 3g/L; Bn represents vitamins B
1, B1 represents that its concentration is that 0mg/L, B2 represent that its concentration is that 20mg/L, B3 represent that its concentration is 25mg/L; Cn represents growth hormone-Plant hormones regulators,gibberellins combination, and C1 represents that its concentration combination is that 0mg/L-0mg/L, C2 represent that its concentration combination is that 5mg/L-1mg/L, C3 represent that its concentration combination is 7.5mg/L-3mg/L.Adopt F inspection, Duncan method inspection multiple comparisons P
0.05< 0.01 difference reaches pole conspicuous level, and between process, same letter represents electrodeless significant difference.
Table 1 to table 3 just gives contriver and is realizing the part in process of the present invention and the maximally related testing data of the present invention's scope required for protection, makes those skilled in the art can repeat to realize the present invention on the basis of the disclosure of invention.Contriver is also provided with except the concentration of component that the present embodiment provides, and also tests other concentration.Realizing in process of the present invention, contriver also tests other compounds except each compound listed by the present embodiment, comprises calcium chloride, urea etc., but has obvious restraining effect (P to the sprouting of Fructus oleae europaeae pollen
0.05< 0.01 difference reaches pole conspicuous level).
Different pollen is different for the suitableeest scope of hormone, nutritive substance, and contriver is to potassium primary phosphate, vitamins B
1, growth hormone, Plant hormones regulators,gibberellins selection, have passed through a large amount of exploratory experiment and carry out screening verification.Test-results shows: sucrose, boric acid, potassium primary phosphate, vitamins B
1, growth hormone, Plant hormones regulators,gibberellins is the important nutrient promoting Fructus oleae europaeae pollen germination, the material that can significantly improve Fructus oleae europaeae pollen germination rate has boric acid, potassium primary phosphate, vitamins B
1, the material that can significantly improve Fructus oleae europaeae pollen tube growth has sucrose, vitamins B
1; And growth hormone and Plant hormones regulators,gibberellins have slight promoter action as hormone regulator to the sprouting of Fructus oleae europaeae pollen and the elongation of pollen tube.
(6) artificial pollination opportunity and number of times: determine pollination opportunity in conjunction with Phenological Observation---should first time artificial pollination be carried out when setting body and having 40% ~ 50% the flowers are in blossom and put, when set body 80% ~ 90% the flowers are in blossom put time carry out second time pollinate, twice pollination time interval difference is no more than 72h, preferably more than 48h.
The composition of the promotion Fructus oleae europaeae pollen germination that contriver filters out, through experimental verification: the pollen 48h germination rate of Fructus oleae europaeae reaches more than 10%, and pollen tube length can reach 458 μm, substantially increases germination rate and the germination of pollen tube length of Fructus oleae europaeae pollen.Use this Medium Proportion manufacturing artificial pollination mixed-culture medium, carry out Fructus oleae europaeae artificial pollination in the mode of spraying, the germination obviously entering pollen in artificial pollination can be urged, thus improve pollination success ratio, finally can improve Olive Fruit rate.
Can Fructus oleae europaeae pollen germination rate and germination of pollen tube length determine complete the essential condition of fertilization process, and during pollination, pollen starts to sprout after contacting with pistil stigma is affine, and pollen tube constantly extends and enters blastular and embryo of silkworms has merged fertilization.The present embodiment is by demonstrating potassium primary phosphate, vitamins B under experimental conditions
1, growth hormone, Plant hormones regulators,gibberellins to the effect of Fructus oleae europaeae pollen germination, for Fructus oleae europaeae artificial water fog pollination nutrient solution proportioning experimental basis is provided.Use Fructus oleae europaeae pollens nutrition mixed solution of the present invention to carry out artificial water fog pollination, improve the germination rate of Fructus oleae europaeae pollen in artificial pollination and promote the sprouting of pollen tube, and then improving the success ratio of artificial pollination.
Below be only the preferred embodiment of the present invention, it should be pointed out that above-mentioned preferred implementation should not be considered as limitation of the present invention, protection scope of the present invention should be as the criterion with claim limited range.For those skilled in the art, without departing from the spirit and scope of the present invention, can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. a nutritive medium, this nutritive medium can promote Fructus oleae europaeae pollen germination, it is characterized in that, this nutritive medium is in water, add following component and content thereof:
Sucrose: 12% ~ 13%wt;
Boric acid: 50mg/L ~ 60mg/L;
Potassium primary phosphate: 0.8g/L ~ 1.2g/L;
The pH of nutritive medium is 6 ~ 6.5.
2. nutritive medium according to claim 1, is characterized in that, described nutritive medium is in water, add following component and content thereof:
Sucrose: 12%wt;
Boric acid: 50mg/L;
Potassium primary phosphate: 1g/L;
The pH of nutritive medium is 6 ~ 6.5.
3. nutritive medium according to claim 1, is characterized in that, this nutritive medium also comprises following component and content thereof:
Component one: vitamins B
1: 20mg/L ~ 25mg/L;
Or/and
Component two: growth hormone: 4.5mg/L ~ 5.5mg/L;
Plant hormones regulators,gibberellins: 0.8mg/L ~ 1.2mg/L.
4. nutritive medium according to claim 3, is characterized in that, each component of described nutritive medium and content thereof are:
Component one: vitamins B
1: 20mg/L;
Or/and
Component two: growth hormone: 5mg/L;
Plant hormones regulators,gibberellins: 1mg/L.
5. based on a Fructus oleae europaeae pollens nutrition mixed solution for nutritive medium described in claim 1,2,3 or 4, it is characterized in that, add Fructus oleae europaeae pollen in described nutritive medium, the addition of pollen is 0.2g/L ~ 0.5g/L.
6. Fructus oleae europaeae pollens nutrition mixed solution according to claim 5, it is characterized in that, the described Fructus oleae europaeae pollens nutrition mixed solution shelf time is no more than 4h.
7. Fructus oleae europaeae pollens nutrition mixed solution according to claim 5, it is characterized in that, described Fructus oleae europaeae pollens nutrition mixed solution is prepared by following steps:
With pure tap water as solvent, first add sucrose, boric acid successively, then add all the other components, stir;
Be 6 ~ 6.5 with sodium hydroxide or salt acid for adjusting pH value;
Add subzero treatment to cross and the Fructus oleae europaeae pollen of the 1h ~ 2h that at room temperature rises again, it is fully stirred and Homogeneous phase mixing in the solution.
8. utilize Fructus oleae europaeae pollens nutrition mixed solution described in claim 5,6 or 7 to carry out an artificial pollination method, be applicable to Fructus oleae europaeae, it is characterized in that, Fructus oleae europaeae pollens nutrition mixed solution is sprayed to open Fructus oleae europaeae and takes; By Phenological Observation, carry out first time artificial pollination when setting body and having 40 ~ 50% the flowers are in blossom and put, carry out second time when setting body 80% ~ 90% the flowers are in blossom and putting and pollinate, twice pollination time interval is no more than 72h.
9. method according to claim 8, is characterized in that, the collection also comprising pollen with preserve step: before Fructus oleae europaeae is bloomed, with sulfuric acid paper bag to inflorescence bagging, full-bloom stage is collected the pollen that sheds and is removed foreign material; Or gather compared with prematurity, branch of blooming that flower density is larger, in the loft drier of 30 DEG C, place 12h, below loft drier, lay the template collecting pollen, collect pollen and removal of impurities; Pollen is placed in vial is airtight, half-light, cryopreservation, be used in 4 DEG C ~-20 DEG C preservations then, next year is used in-20 DEG C of preservations.
10. method according to claim 9, is characterized in that, at room temperature rise again before the pollen that cryopreservation is crossed uses 1h ~ 2h.
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| CN106305408A (en) * | 2016-08-22 | 2017-01-11 | 文山苗乡三七科技有限公司 | Method for artificially pollinating Vietnamese pseudo-ginsengs |
| CN106342678A (en) * | 2016-08-22 | 2017-01-25 | 文山苗乡三七科技有限公司 | Medium solution for artificial pollination of Vietnam notoginseng |
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| CN106342678A (en) * | 2016-08-22 | 2017-01-25 | 文山苗乡三七科技有限公司 | Medium solution for artificial pollination of Vietnam notoginseng |
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