CN103396981A - In vitro culture and vitality determination method for water lily pollen - Google Patents
In vitro culture and vitality determination method for water lily pollen Download PDFInfo
- Publication number
- CN103396981A CN103396981A CN2013102690647A CN201310269064A CN103396981A CN 103396981 A CN103396981 A CN 103396981A CN 2013102690647 A CN2013102690647 A CN 2013102690647A CN 201310269064 A CN201310269064 A CN 201310269064A CN 103396981 A CN103396981 A CN 103396981A
- Authority
- CN
- China
- Prior art keywords
- pollen
- water lily
- slide glass
- cultivated
- vitro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses an in vitro culture and vitality determination method for water lily pollen. The method comprises the following steps: 1) a preparing a mixed liquid medium containing cane sugar, boric acid, potassium nitrate, calcium chloride, polyethylene glycol, hormone and water lily flower stigma disc; 2) acquiring pollen of blooming water lily last ten-days of July in blooming period; 3) dripping the liquid medium on a piece of slide glass, dipping a small amount of pollen into the liquid medium and culturing the pollen at 25 DEG C in a dark environment; and 4) culturing the slide glass for 6 h and observing germination status under an optical microscope, thereby determining the vitality of water lily pollen. The in vitro culture method for water lily in vitro provided by the present invention is scientific and reasonable; the vitality determination method for water lily pollen is simple, has strong operationality and reliable data, and can completely realize quantitative determination. Therefore, the invention has high actual application value for raising breeding efficiency of water lily.
Description
Technical field
The present invention relates to a kind of method that in-vitro pollen is cultivated, a kind of particularly isolated culture method, also relate to the vigour-testing method of water lily pollen, belongs to the plant hybridization breeding technical field.
Background technology
Water lily be the Nymphaeceae Nymphaea (
Nymphaea) general term of plant, with its abundant pattern, holy and pure implied meaning and the cauline leaf strong adsorptive power to nutrient rich in water and objectionable impurities, very popular, be widely used in water feature design.
The water lily Pollen Activity of different varieties is measured the surviving rate that can distinguish tested kind water lily pollen, to instructing the water lily breeding, has most important theories and realistic meaning.Have several different methods can measure the vigor of pollen, wherein in-vitro pollen germination can be observed intuitively death or lack blodynamic pollen, and the data science of mensuration is reliable, and fully quantitatively.For many plants, in-vitro pollen germination all has dependency with solid and Seed Development, therefore in-vitro pollen germination is measured, is still the prefered method of Pollen viability in present cross-breeding.The sprouting method that exsomatizes is measured Pollen Activity needs specific substratum and culture condition, and the main component of substratum generally comprises Ca
2+, boron and carbon source etc., some plants also need other materials such as plant hormone 6-BA or PEG etc.At present, have no the substratum of water lily in-vitro pollen germination and the report of culture condition.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of water lily in-vitro pollen cultural method is provided, realize water lily in-vitro pollen germination mensuration.
The present invention is achieved by the following technical programs:
A kind of water lily in-vitro pollen is cultivated and vigour-testing method, comprises the following steps:
1) obtaining liq substratum
Press following component obtaining liq substratum: the boric acid of the sucrose of 300 g/L, 10 mg/L, the sal epsom of 350 mg/L, 900 mg/L saltpetre, 800 mg/L calcium chloride and 100 g/L plant hormone PEG1500, all the other are the sweet liquid on water lily style cephalic disc;
2) water lily pollen gathers
In water lily the flowers are in blossom time last ten-days period in July, morning, 10:00 gathered water lily pollen in full bloom, and water lily pollen is packed in centrifuge tube, and lucifuge is standby;
3) water lily pollen is cultivated
Liquid nutrient medium is dropped on slide glass, with writing brush, a small amount of water lily pollen is dipped in liquid nutrient medium, do not cover slide glass, after planting slide glass is placed in the culture dish that is lined with moistening filter paper, under dark surrounds, cultivate;
4) the water lily Pollen Activity is measured
Slide glass was cultivated after 6 hours, was placed in optical microphotograph Microscopic observation germination situation, can observe and sprout water lily pollen, and the described pollen tube length of water lily pollen of having sprouted is greater than the pollen diameter.
Aforesaid water lily in-vitro pollen is cultivated and vigour-testing method, and the culture temperature of wherein said step 3) is 24～26 ℃.
The present invention has following beneficial effect with respect to prior art:
Water lily in-vitro pollen cultural method is scientific and reasonable, and at first, sucrose, boric acid and calcium are essential for pollen germination and pollen tube growth, are the main components of in-vitro pollen germination substratum.Sucrose mainly plays in pollen germination provide the energy and regulate the osmotic pressure effect, and a certain amount of boric acid and calcium ion can make pollen tube look straight and thick, unlikely long.Secondly, plant hormone PEG1500(polyoxyethylene glycol) membrane structure in pollen is changed, change the film surface charge, the soft degree of film and permeability are improved, thereby promote pollen germination and pollen tube growth.In addition, the sweet liquid on water lily column cap dish contains the material favourable to pollen germination.Obtaining liq substratum of the present invention is conducive to the water lily pollen germination, has more than sowing pollen and uniform advantage.Water lily pollen viability measuring method of the present invention is simple, workable, and data are reliable, and can be completely achieved detection by quantitative, for improving the water lily breeding efficiency, has higher actual application value.
Advantage and disadvantage of the present invention, will make an explanation by the non-limitative illustration of following preferred embodiment, and these embodiment only provide as an example.
Embodiment
The invention will be further described below in conjunction with the embodiment of water lily kind " Peter ".
The present embodiment comprises the following steps:
1) obtaining liq substratum
Press following component obtaining liq substratum: the boric acid of the sucrose of 300 g/L, 10 mg/L, the sal epsom of 350 mg/L, 900 mg/L saltpetre, 800 mg/L calcium chloride and 100 g/L plant hormone PEG1500, all the other are the sweet liquid on water lily style cephalic disc.
2) water lily pollen gathers
The last ten-days period in July " Peter ", the flowers are in blossom the time, and morning, about 10:00 gathered water lily pollen in full bloom, and water lily pollen is packed in centrifuge tube, and lucifuge is standby.
3) water lily pollen is cultivated
Liquid nutrient medium is dropped on slide glass, with writing brush, a small amount of water lily pollen is dipped in liquid nutrient medium, do not cover slide glass, after planting slide glass is placed in the culture dish that is lined with moistening filter paper, under 25 ℃ of dark surrounds, cultivate.
4) the water lily Pollen Activity is measured
Slide glass was cultivated after 6 hours, was placed in optical microphotograph Microscopic observation germination situation, can observe the sprouting water lily pollen of pollen tube length greater than the pollen diameter.
In addition to the implementation, the present invention can also have other embodiments, and all employings are equal to the technical scheme of replacement or equivalent deformation one-tenth, all drop in the protection domain of requirement of the present invention.
Claims (2)
1. a water lily in-vitro pollen is cultivated and vigour-testing method, it is characterized in that: comprise the following steps:
1) obtaining liq substratum
Press following component obtaining liq substratum: the sal epsom of the sucrose of 300g/L, the boric acid of 10mg/L, 350mg/L, 900mg/L saltpetre, 800mg/L calcium chloride and 100g/L plant hormone PEG1500, all the other are the sweet liquid on water lily style cephalic disc;
2) water lily pollen gathers
In water lily the flowers are in blossom time last ten-days period in July, morning, 10:00 gathered water lily pollen in full bloom, and water lily pollen is packed in centrifuge tube, and lucifuge is standby;
3) water lily pollen is cultivated
Liquid nutrient medium is dropped on slide glass, with writing brush, a small amount of water lily pollen is dipped in liquid nutrient medium, do not cover slide glass, after planting slide glass is placed in the culture dish that is lined with moistening filter paper, under dark surrounds, cultivate;
4) the water lily Pollen Activity is measured
Slide glass was cultivated after 6 hours, was placed in optical microphotograph Microscopic observation germination situation, can observe and sprout water lily pollen, and the described pollen tube length of water lily pollen of having sprouted is greater than the pollen diameter.
2. water lily in-vitro pollen as claimed in claim 1 is cultivated and vigour-testing method, and it is characterized in that: the culture temperature of described step 3) is 24～26 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310269064.7A CN103396981B (en) | 2013-07-01 | 2013-07-01 | In vitro culture and vitality determination method for water lily pollen |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310269064.7A CN103396981B (en) | 2013-07-01 | 2013-07-01 | In vitro culture and vitality determination method for water lily pollen |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103396981A true CN103396981A (en) | 2013-11-20 |
| CN103396981B CN103396981B (en) | 2015-04-29 |
Family
ID=49560698
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310269064.7A Expired - Fee Related CN103396981B (en) | 2013-07-01 | 2013-07-01 | In vitro culture and vitality determination method for water lily pollen |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103396981B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103983644A (en) * | 2014-03-10 | 2014-08-13 | 云南农业大学 | Determination method for tomato pollen vitality |
| CN106233877A (en) * | 2016-08-28 | 2016-12-21 | 湖北省农业科学院中药材研究所 | A kind of effective method measuring Herba Gelsemii Elegantis Pollen Activity |
| CN106995834A (en) * | 2017-04-25 | 2017-08-01 | 湖北省农业科学院中药材研究所 | A kind of assay method of panax japonicus Pollen Activity |
| CN110172494A (en) * | 2019-05-31 | 2019-08-27 | 南京林业大学 | A kind of detection method of perfume lotus flower pollen vigor |
| CN111387045A (en) * | 2020-04-20 | 2020-07-10 | 江苏丘陵地区镇江农业科学研究所 | Method for overcoming water lily interspecific hybridization obstacle |
| CN113310767A (en) * | 2021-06-01 | 2021-08-27 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Microscopic method for pollen tube and ovule after pollination of water lily and optical microscopic tabletting manufacturing method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101292629A (en) * | 2008-06-18 | 2008-10-29 | 黑龙江省科学院亚麻综合利用研究所 | Flax free pollen culture medium and flax free pollen culture method |
| CN101633902A (en) * | 2009-08-25 | 2010-01-27 | 商洛学院 | In-vitro pollen germination liquid medium of balloon flower and method using same to determine pollen viability thereof |
-
2013
- 2013-07-01 CN CN201310269064.7A patent/CN103396981B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101292629A (en) * | 2008-06-18 | 2008-10-29 | 黑龙江省科学院亚麻综合利用研究所 | Flax free pollen culture medium and flax free pollen culture method |
| CN101633902A (en) * | 2009-08-25 | 2010-01-27 | 商洛学院 | In-vitro pollen germination liquid medium of balloon flower and method using same to determine pollen viability thereof |
Non-Patent Citations (3)
| Title |
|---|
| 武冲 等: "植物花粉培养研究进展", 《中国农学通报》 * |
| 罗凤霞 等: "水仙花粉离体萌发温度和培养液研究", 《辽宁林业科技》 * |
| 赵其龙: "硼酸和蔗糖对‘柔毛齿叶’睡莲花粉萌发的影响", 《亚热带植物科学》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103983644A (en) * | 2014-03-10 | 2014-08-13 | 云南农业大学 | Determination method for tomato pollen vitality |
| CN106233877A (en) * | 2016-08-28 | 2016-12-21 | 湖北省农业科学院中药材研究所 | A kind of effective method measuring Herba Gelsemii Elegantis Pollen Activity |
| CN106995834A (en) * | 2017-04-25 | 2017-08-01 | 湖北省农业科学院中药材研究所 | A kind of assay method of panax japonicus Pollen Activity |
| CN110172494A (en) * | 2019-05-31 | 2019-08-27 | 南京林业大学 | A kind of detection method of perfume lotus flower pollen vigor |
| CN111387045A (en) * | 2020-04-20 | 2020-07-10 | 江苏丘陵地区镇江农业科学研究所 | Method for overcoming water lily interspecific hybridization obstacle |
| CN113310767A (en) * | 2021-06-01 | 2021-08-27 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Microscopic method for pollen tube and ovule after pollination of water lily and optical microscopic tabletting manufacturing method |
| CN113310767B (en) * | 2021-06-01 | 2024-05-03 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Microscopic method for pollen tube and ovule after pollination of water lily and optical microscopic tablet manufacturing method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103396981B (en) | 2015-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103396981B (en) | In vitro culture and vitality determination method for water lily pollen | |
| Radoglou et al. | The effects of CO2 enrichment and nutrient supply on growth morphology and anatomy of Phaseolus vulgaris L. seedlings | |
| Jo et al. | In vitro sucrose concentration affects growth and acclimatization of Alocasia amazonica plantlets | |
| CN103430659B (en) | Method for determining lotus pollen viability | |
| CN102119594A (en) | Method for promoting rhododendron parvifolium seed germination | |
| CN104604672B (en) | A kind of method of rapid screening high-nicotine tobacco mutant body | |
| CN103087975A (en) | Apocarya pollen in-vitro germination liquid culture medium and application thereof in measuring pollen activity | |
| CN103999594A (en) | Method for economically and quickly breaking dormancy of sand rice seeds and accelerating sand rice seeds to germinate | |
| CN101194594B (en) | Breeding method for alsophila spinulosa | |
| CN105766893A (en) | Storage method of eucalyptus pollen and vitality measuring method of pollen stored by storage method | |
| CN104498426A (en) | Collection, storage, in-vitro culture and viability detection technology for acacia auriculiformis pollen | |
| CN102972296B (en) | Acacia rachii pollen in-vitro culture activity detection technology | |
| CN104178448A (en) | Method for determining pollen viability of watermelon | |
| CN111480563A (en) | Water culture method for experimental rice cultivated in greenhouse | |
| CN107460229A (en) | Spontaneum pollen vigor rapid identification method | |
| Moraes et al. | Activated charcoal for in vitro propagation of Brazilian orchids | |
| CN117530173A (en) | Method for creating new germplasm of pepper tetraploid by utilizing stem tip induction | |
| CN105601386B (en) | A kind of Microspore of Brassica napus seedling water planting culture solution and the method for more summer culture | |
| CN103421737B (en) | Culture medium for promoting germination of Amygdalus L. pollen, culture method and application thereof | |
| CN110172494A (en) | A kind of detection method of perfume lotus flower pollen vigor | |
| CN114766353A (en) | Rapid screening method and application of male pollination plants in actinidia arguta orchard | |
| CN103141368A (en) | Transparent device and method for directly observing relation between plant root growth and microorganism | |
| CN111690708A (en) | Actinidia chinensis in-vitro pollen activity determination method | |
| Tyagi et al. | Plant regeneration from tissue cultures initiated from immature inflorescences of a grass, Echinochloa colonum (L.) Link | |
| Nomizu et al. | Haploid plant regeneration via embryogenesis from anther cultures of Hepatica nobilis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150429 Termination date: 20160701 |