CN101554133A - Liquid medium pollen, preparing method and application thereof - Google Patents
Liquid medium pollen, preparing method and application thereof Download PDFInfo
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- CN101554133A CN101554133A CNA2009100944445A CN200910094444A CN101554133A CN 101554133 A CN101554133 A CN 101554133A CN A2009100944445 A CNA2009100944445 A CN A2009100944445A CN 200910094444 A CN200910094444 A CN 200910094444A CN 101554133 A CN101554133 A CN 101554133A
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Abstract
The invention discloses liquid medium pollen, a preparing method and applications thereof. The invention aims at improving the activity and the quality of pollen, reducing the use level of pure pollen and reducing the cost of seed production. Tobacco pure pollen and 0.2 to 0.5 mM polyamine solution are mixed, the mixture ratio of the pollen to the solution is 50 to 80 mg/ml, and the liquid medium pollen is obtained after uniform fixing. The polyamine solution is selected form one of putrescine, spermidine or spermine. Simultaneously the invention provides the application of the liquid medium pollen in tobacco artificial pollination.
Description
Technical field
The invention belongs to the artificial pollination technical field, more particularly, the present invention relates to the liquid medium pollen of tobacco hybrid seeding.Simultaneously, the invention still further relates to the preparation method and the application of described liquid medium pollen.
Background technology
Be the intellectual property of protecting breeding man and the purpose that reaches control tobacco leaf planting area, China's large tracts of land on leaf tobacco production is promoted the use of tobacco male sterile kind at present, and cultivated area is enlarging year by year, and sowing quantity is also increasing year by year.Through incomplete statistics, the area of popularizing planting tobacco male sterile line has surpassed 530,000 hectares (see the Zhejiang agricultural science for details, 2009,1:22～25) on the national leaf tobacco production in 2007.The popularizing planting of tobacco male sterile kind has become one of main direction of China's leaf tobacco production.The seed of sterile variety needs to obtain by artificial supplementary pollination, and high vigor pollen is an essential condition guaranteeing production of hybrid seeds success.The tobacco pollen amount is few, and the pollen collection is a job of wasting time and energy, and pollen devitalization easily in collection and storage.
Prior art uses the pure pollen of tobacco to pollinate, but consumption is big during pollination, and the pollen effective rate of utilization is low, and especially when running into strong wind weather, effective utilization ratio of pollen is lower, thereby causes breeding cost too high.Therefore, under the prerequisite that guarantees seed quality and output, how to improve the pollen vigor, reduce the pollen consumption, improve the effective rate of utilization of pollen, reduce breeding cost, just become tobacco hybrid seeding personnel technical barrier anxious to be solved.
In the process of artificial supplementary pollination, being mixed with liquid medium pollen with boric acid, the sucrose solution of pollen and suitable concentration carries out supplementary pollination and can promote pollen germination and pollen tube growth, can save the pollen consumption, improve percentage of fertile fruit, this is more existing researchs on crops such as fruit tree, pear tree, peach.For example with pollen with after the BAS of suitable concentration mixes, to the pollination of spraying of peach and sweet cherry, can improve percentage of fertile fruit (Agricultural University Of He'nan's journal, 2005,39 (4): 394～397; The China fruit tree, 2006, (6): 23～25).When air was dry, the mucus of secretion stigma was easily done, and pollen is difficult for adhering to, and is unfavorable for pollen germination.Low concentration sucrose can be used as the spreader-sticker in the liquid medium pollen, can keep the pollen vigor of cabbage type rape, grape etc. to a certain extent, improves the column cap environment, help pollen and adhere to sprouting, save consumption (Chinese oil crop journal, 2004,26 (2): 15～18 of pollen greatly; Bulletinof the Fukuoka Agricultural Research Center, 2006,25:89～94).
Polyamines, common have putrescine (Putrescine, Put), spermidine (Spermidine, Spd), (Spermine Spm) etc., is the low-molecular-weight fat nitrogenous base of the biologically active that produces in the organism metabolic process to spermine.Polyamines is ubiquity in plant corpus, and is especially higher at the floral organ intensive amount, about the more existing researchs of the relation of polyamines and pollen vigor.There are many researchs to show that the polyamines solution of suitable concentration has the crop pollen of raising vigor, promotes function (fruit tree journal, 2002,19 (6): 377～380 of pollen germination and pollen tube growth, raising percentage of fertile fruit; The Hebei agricultural science, 2005,9 (3): 99～102).
In tobacco hybrid seeding process, carry out artificial supplementary pollination if the polyamines solution of suitable concentration and the pure pollen of tobacco are mixed with liquid medium pollen by a certain percentage, improving the pollen vigor, reducing pure pollen consumption and reduce and to obtain effect preferably aspect the seed production cost.At present, the correlative study of polyamines liquid medium pollen on crop and application also do not appear in the newspapers as yet.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, a kind of pollen of raising vigor that has is provided, reduce the polyamines liquid medium pollen of pure pollen consumption function,, thereby reduce breeding cost with instead of pure pollen.
Another object of the present invention provides a kind of method for preparing described liquid medium pollen.
The present invention also aims to provide the application of described liquid medium pollen in the tobacco hybrid seed production and pollination.
Purpose of the present invention is achieved by the following technical programs.
Except as otherwise noted, the percentage that is adopted among the present invention is mass percent.
A. the invention provides a kind of liquid medium pollen, it is characterized in that: with the pure pollen of tobacco is that putrescine, spermidine or the spermine solution of 0.2～0.5mM mixes with concentration respectively, the proportioning of pollen and solution is 50～80mg/ml, mixes and promptly obtains required polyamines liquid medium pollen.
B. the invention provides a kind of preparation method of described liquid medium pollen, this method adopts following steps:
1. the collection of pollen---treating that the tobacco male parent flower reaches begins to gather pollen when picking flowers standard, and the pollen after the collection is preserved standby down at 4 ℃;
2. the preparation of polyamines solution---putrescine, spermidine or spermine are made into the solution that concentration is 0.2～0.5mM respectively, and the solution for preparing is preserved standby in 4 ℃ of lower seals;
3. the preparation of polyamines liquid medium pollen---in the ratio of 50～80mg/ml, be that the polyamines solution of 0.2～0.5mM evenly mixes, promptly obtain required liquid medium pollen pure pollen of tobacco and concentration;
C. the invention provides the application method of a kind of described liquid medium pollen in tobacco artificial pollination, specific practice is as follows: dip in writing brush and get described liquid medium pollen and evenly be applied in maternal fireworks column cap, carry out the hybrid seeding artificial pollination.
Compared with prior art, the present invention has following outstanding advantage:
1. prove through continuous 3 years field experiment that the appropriate liquid medium pollen can improve the pollen vigor, improve the percentage of fertile fruit of field cigarette strain, improve seed production;
2. the use of liquid medium pollen does not make significant difference to the germination of seed, thereby has guaranteed seed production quality;
3. improve the effective rate of utilization of pure pollen, reduced the consumption of pure pollen, reduced breeding cost.Of the present invention applying has good economy and society benefit.
Embodiment
The present invention is described in further detail below in conjunction with embodiment and Application Example, but they are not the qualification to protection domain of the present invention.
Embodiment 1
Selected model field is positioned at Xishuangbanna, Yunnan China Tobacco Seed Yuxi Co., Ltd. production of hybrid seeds base.The tobacco seedling mode adopts floating seedlings, and Cheng Miaohou transplants the land for growing field crops.Field planting density is: line-spacing 1.2m, spacing in the rows 0.5m.
Carried out the field on 10 mu of ground implements in production of hybrid seeds base.2 mu of male parents that are used to plant cloud and mist 87 wherein, all the other 8 mu are used to plant cloud and mist 87 female parents.Treating that cloud and mist 87 male parent flowers reach begins to adopt powder when picking flowers standard, the flower pesticide of gathering cloud and mist 87 male parents dries and makes its loose powder, sifts out pollen with 50 mesh sieve, and the pollen of collection is preserved standby at 4 ℃ of lower seals.In the 50mg/ml ratio, take by weighing pure pollen and join in the putrescine solution that concentration is 0.2mM and mix, be mixed with required liquid medium pollen.
Embodiment 2
Repeat embodiment 1, have the putrescine solution of following difference: 0.2mM to change the spermidine solution that concentration is 0.2mM into.
Embodiment 3
Repeat embodiment 1, have the putrescine solution of following difference: 0.2mM to change the spermine solution that concentration is 0.2mM into.
Embodiment 4～6
Repeat embodiment 1, have the putrescine solution of following difference: 0.2mM to change putrescine, spermine or the spermidine solution that concentration is 0.5mM respectively into.
Embodiment 7～12
Repeat embodiment 1～6, following difference is arranged: the ratio of pollen and polyamines solution is 80mg/ml in the liquid medium pollen.
Embodiment 13～24
Repeat embodiment 1～12, following difference is arranged: used pollen is the pollen of K 326 male parents.
The present invention by a large amount of field trials with determine suitable putrescine, spermidine, spermine solution concentration and with the ratio of pollen.Ratio in 50mg～80mg/ml, add to the pure pollen of tobacco in putrescine, spermidine or the spermine solution that concentration is 0.2～0.5mM and evenly mixing, be prepared into different liquid medium pollens, and be contrast with pure pollen, in cloud and mist 87 and two large tracts of land recommended varieties of K326, carry out the hybrid seeding test.Studied liquid medium pollen to field cigarette strain percentage of fertile fruit and every mu of seed average yield, the influence of indexs such as indoor 25 ℃ of potentialities of seed of sprouting down, germination rate.Discover, after using suitable polyamines liquid medium pollen to carry out the field pollination, the percentage of fertile fruit and the seed production of cigarette strain are higher than contrast, the germination vigor of seed and germination rate and contrast ratio do not have significant difference, use the polyamines liquid medium pollen to reach and improve the pollen vigor, save pure pollen consumption, reduce the purpose of breeding cost.
Now, further specify technique effect of the invention process by the field demonstration example.
Application Example 1
Dip in cloud and mist 87 liquid medium pollens (pollen concentration is 50mg/ml, and putrescine concentration is 0.2mM) of getting embodiment 1 system with writing brush, be applied in maternal fireworks column cap, carry out artificial pollination.The field capsule is brown gathers after ripe, dries (water content is less than 6.0%) after the seed cleaning, 15 ℃ of low temperature storages.Add up every mu of seed average yield, carry out seed germination experiment under indoor 25 ℃, statistics germination vigor and germination rate.Result of the test shows, the 1ml liquid medium pollen can be to a flower surplus the cloud and mist 87 maternal pollinations 80, field, solid back percentage of fertile fruit reaches 87.6%, average every mu of seed production reaches 6.34kg, potentiality of seed reaches 92.3%, germination rate reaches 95.3%, (adopt cloud and mist 87 pure pollen pollinations with contrast ratio, the pure pollen of the 50mg flower surplus in the of 60 of can pollinating, the field percentage of fertile fruit is 82.2%, and average every mu of seed production is 5.60kg, and germination vigor reaches 92.0%, germination rate reaches 95.0%) seed production significantly increases, and seed sprouting does not then have significant difference.
Application Example 2
Dip in cloud and mist 87 liquid medium pollens (pollen concentration is 50mg/ml, and spermidine concentration is 0.2mM) of getting embodiment 2 systems with writing brush, be applied in maternal fireworks column cap, carry out artificial pollination.Capsule is brown gathers after ripe, dries (water content is less than 6.0%) after the seed cleaning, 15 ℃ of low temperature storages.Add up every mu of seed average yield, carry out seed germination experiment under indoor 25 ℃, statistics germination vigor and germination rate.The result shows that the 1ml liquid medium pollen can be to a flower surplus the cloud and mist 87 maternal pollinations 80, and solid back percentage of fertile fruit reaches 89.3%, and average every mu of seed production reaches 6.57kg, and potentiality of seed all reaches more than 93.3%, and germination rate reaches more than 95.3%.With contrast ratio (adopt cloud and mist 87 pure pollen pollinations, the pure pollen of the 50mg flower surplus in the of 60 of can pollinating, the field percentage of fertile fruit is 82.2%, average every mu of seed production is 5.60kg, germination vigor reaches 92.0%, and germination rate reaches 95.0%) seed production significantly increases, and seed sprouting does not then have significant difference.
Application Example 3
Dip in cloud and mist 87 liquid medium pollens (pollen concentration is 50mg/ml, and spermine concentration is 0.2mM) of getting embodiment 3 systems with writing brush, be applied in maternal fireworks column cap, carry out artificial pollination.Capsule is brown gathers after ripe, dries (water content is less than 6.0%) after the seed cleaning, 15 ℃ of low temperature storages.Add up every mu of seed average yield, carry out seed germination experiment under indoor 25 ℃, statistics germination vigor and germination rate.Result of the test shows, the 1ml liquid medium pollen can be to a flower surplus the cloud and mist 87 maternal pollinations 80, solid back percentage of fertile fruit reaches 88.5%, average every mu of seed production reaches 7.16kg, potentiality of seed reaches 92.0%, germination rate reaches 95.3%, (adopt cloud and mist 87 pure pollen pollinations with contrast ratio, the pure pollen of the 50mg flower surplus in the of 60 of can pollinating, the field percentage of fertile fruit is 82.2%, and average every mu of seed production is 5.60kg, and germination vigor reaches 92.0%, germination rate reaches 95.0%) seed production significantly increases, and seed sprouting does not then have significant difference.Illustrate that the polyamines liquid medium pollen helps to improve cloud and mist 87 pollen vigor, improve percentage of fertile fruit and seed production, save the pollen consumption, but seed quality is uninfluenced.
Application Example 4
Dip in the K326 liquid medium pollen (pollen concentration is 80mg/ml, and putrescine concentration is 0.5mM) of getting embodiment 22 systems with writing brush, be applied in maternal fireworks column cap, carry out artificial pollination.Capsule is brown gathers after ripe, dries (water content is less than 6.0%) after the seed cleaning, 15 ℃ of low temperature storages.Add up every mu of seed average yield, carry out seed germination experiment under indoor 25 ℃, statistics germination vigor and germination rate.Result of the test shows, the 1ml liquid medium pollen can be to a flower surplus the maternal pollination 130 of K326, solid back percentage of fertile fruit reaches 90.5%, average every mu of seed production reaches 8.96kg, potentiality of seed reaches 93.0%, germination rate reaches 96.0%, (adopt the pure pollen pollination of K326 with contrast ratio, the pure pollen of the 80mg flower surplus in the of 100 of can pollinating, the field percentage of fertile fruit is 84.6%, and average every mu of seed production is 7.23kg, and germination vigor reaches 91.3%, germination rate reaches 96.0%) seed production significantly increases, and seed sprouting does not then have significant difference.
Application Example 5
Dip in the K326 liquid medium pollen (pollen concentration is 80mg/ml, and spermidine concentration is 0.5mM) of getting embodiment 23 systems with writing brush, be applied in maternal fireworks column cap, carry out artificial pollination.Capsule is brown gathers after ripe, dries (water content is less than 6.0%) after the seed cleaning, 15 ℃ of low temperature storages.Add up every mu of seed average yield, carry out seed germination experiment under indoor 25 ℃, statistics germination vigor and germination rate.The result shows that the 1ml liquid medium pollen can be to a flower surplus the maternal pollination 130 of K326, and solid back percentage of fertile fruit reaches 91.0%, and average every mu of seed production reaches 8.92kg, and potentiality of seed all reaches 93.3%, and germination rate reaches 96.0%.With contrast ratio (adopt the pure pollen pollination of K326, the pure pollen of the 80mg flower surplus in the of 100 of can pollinating, the field percentage of fertile fruit is 84.6%, average every mu of seed production is 7.23kg, germination vigor reaches 91.3%, and germination rate reaches 96.0%) seed production significantly increases, and seed sprouting does not then have significant difference.
Application Example 6
Dip in the K326 liquid medium pollen (pollen concentration is 80mg/ml, and spermine concentration is 0.5mM) of getting embodiment 24 systems with writing brush, be applied in maternal fireworks column cap, carry out artificial pollination.Capsule is brown gathers after ripe, dries (water content is less than 6.0%) after the seed cleaning, 15 ℃ of low temperature storages.Add up every mu of seed average yield, carry out seed germination experiment under indoor 25 ℃, statistics germination vigor and germination rate.Result of the test shows, the 1ml liquid medium pollen can be to a flower surplus the maternal pollination 130 of K326, solid back percentage of fertile fruit reaches more than 85.5%, average every mu of seed production reaches 8.74kg, potentiality of seed reaches 93.3%, germination rate reaches 96.0%, (adopt the pure pollen pollination of K326 with contrast ratio, the pure pollen of the 80mg flower surplus in the of 100 of can pollinating, the field percentage of fertile fruit is 84.6%, and average every mu of seed production is 7.23kg, and germination vigor reaches 91.3%, germination rate reaches 96.3%) seed production significantly increases, and seed sprouting does not then have significant difference.Illustrate that the polyamines liquid medium pollen helps to improve the pollen vigor of K326, improve percentage of fertile fruit and seed production, save the pollen consumption, but seed quality is uninfluenced.
Claims (4)
1. liquid medium pollen is characterized in that: with pure pollen of tobacco and concentration is that the polyamines solution of 0.2～0.5mM mixes, and the proportioning of pollen and solution is 50～80mg/ml, mixes and promptly obtains required liquid medium pollen.
2. liquid medium pollen according to claim 1 is characterized in that: described polyamines is to be selected from a kind of in putrescine, spermidine or the spermine.
3. the preparation method of claim 1 or 2 described liquid medium pollens, this method adopts following steps:
(1) collection of pollen---treating that the tobacco male parent flower reaches begins to gather pollen when picking flowers standard, and the pollen after the collection is preserved standby down at 4 ℃;
(2) preparation of polyamines solution---putrescine, spermidine or spermine are made into the solution that concentration is 0.2～0.5mM respectively, and the solution for preparing is preserved standby in 4 ℃ of lower seals;
(3) preparation of liquid medium pollen---in the ratio of 50～80mg/ml,, be that the amine aqueous solution of 0.2～0.5mM evenly mixes, promptly obtain required liquid medium pollen pure pollen of tobacco and concentration with the ratio of the pure pollen of tobacco in 50～80mg/ml.
4. the application of the described liquid medium pollen of claim 1 in tobacco artificial pollination.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102047845A (en) * | 2010-12-13 | 2011-05-11 | 湖北省烟草科研所 | Method for in-vitro liquid culture of tobacco pollen with high efficiency |
| CN103539507A (en) * | 2013-09-29 | 2014-01-29 | 四川省烟草公司凉山州公司 | Pollen starch culture medium and preparation method thereof |
| CN105112490A (en) * | 2015-09-08 | 2015-12-02 | 玉溪中烟种子有限责任公司 | Improved method for detecting activity of tobacco pollens |
| CN105123506A (en) * | 2015-09-08 | 2015-12-09 | 玉溪中烟种子有限责任公司 | Tobacco seed oil medium pollen and preparation method and purpose thereof. |
| CN107897218A (en) * | 2017-11-28 | 2018-04-13 | 新疆农业科学院园艺作物研究所 | A kind of insect attractant and application for improving jujube tree pollination rate and percentage of fertile fruit |
| CN108651271A (en) * | 2018-04-04 | 2018-10-16 | 胡勐鸿 | A kind of artificial pollination method of Picea seeds and application |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102047845A (en) * | 2010-12-13 | 2011-05-11 | 湖北省烟草科研所 | Method for in-vitro liquid culture of tobacco pollen with high efficiency |
| CN103539507A (en) * | 2013-09-29 | 2014-01-29 | 四川省烟草公司凉山州公司 | Pollen starch culture medium and preparation method thereof |
| CN103539507B (en) * | 2013-09-29 | 2015-04-01 | 四川省烟草公司凉山州公司 | Pollen starch culture medium and preparation method thereof |
| CN105112490B (en) * | 2015-09-08 | 2018-06-29 | 玉溪中烟种子有限责任公司 | A kind of tobacco pollen vigor testing methods of improvement |
| CN105123506A (en) * | 2015-09-08 | 2015-12-09 | 玉溪中烟种子有限责任公司 | Tobacco seed oil medium pollen and preparation method and purpose thereof. |
| CN105112490A (en) * | 2015-09-08 | 2015-12-02 | 玉溪中烟种子有限责任公司 | Improved method for detecting activity of tobacco pollens |
| EP3428140A4 (en) * | 2015-10-14 | 2019-05-01 | Adobs I Innovació Fitovegetal Hortus Natura S.L. | Production of improved fortifier from beer yeast water |
| CN107897218A (en) * | 2017-11-28 | 2018-04-13 | 新疆农业科学院园艺作物研究所 | A kind of insect attractant and application for improving jujube tree pollination rate and percentage of fertile fruit |
| CN107897218B (en) * | 2017-11-28 | 2020-10-16 | 新疆农业科学院园艺作物研究所 | Insect attractant for improving pollination rate and fruit setting rate of jujube trees and application |
| CN108651271A (en) * | 2018-04-04 | 2018-10-16 | 胡勐鸿 | A kind of artificial pollination method of Picea seeds and application |
| CN113249295A (en) * | 2021-04-02 | 2021-08-13 | 南京林业大学 | Method for measuring germination rate of arabidopsis in-vitro pollen |
| CN115517166A (en) * | 2022-10-21 | 2022-12-27 | 玉溪中烟种子有限责任公司 | Preparation and pollination application method of high-activity liquid-loaded composite pollen |
| CN115517166B (en) * | 2022-10-21 | 2023-10-10 | 玉溪中烟种子有限责任公司 | Preparation and pollination application methods of high-activity liquid-loaded compound pollen |
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