CN103396981B - 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
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- CN103396981B CN103396981B CN201310269064.7A CN201310269064A CN103396981B CN 103396981 B CN103396981 B CN 103396981B CN 201310269064 A CN201310269064 A CN 201310269064A CN 103396981 B CN103396981 B CN 103396981B
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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, particularly a kind of in-vitro culture method, also relates to the vigour-testing method of water lily pollen, belongs to plant hybridization breeding technical field.
Background technology
Water lily be Nymphaeceae Nymphaea (
nymphaea) general term of plant, with its abundant pattern, holy and pure implied meaning and 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 vitality test of different varieties can distinguish the surviving rate of tested kind water lily pollen, has most important theories and realistic meaning to instructing water lily breeding.Have multiple method can measure the vigor of pollen, wherein in-vitro pollen germination can be observed death intuitively or lack blodynamic pollen, and the data science of mensuration is reliable, and completely quantitative.For many plants, in-vitro pollen germination all has dependency with solid and Seed Development, therefore in-vitro pollen germination mensuration is still the prefered method of Pollen viability in current cross-breeding.Germination in vitro method measures 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 as plant hormone 6-BA or PEG etc.At present, the substratum of water lily in-vitro pollen germination and the report of culture condition is had no.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of water lily pollen in-vitro culture method is provided, realize water lily in-vitro pollen germination and measure.
The present invention is achieved by the following technical programs:
A kind of water lily pollen isolated culture and vigour-testing method, comprise the following steps:
1) obtaining liq substratum
By following component obtaining liq substratum: the magnesium sulfate of the sucrose of 300 g/L, the boric acid of 10 mg/L, 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 oral shui solution column cap dish;
2) water lily pollen collection
In water lily the flowers are in blossom time last ten-days period in July, morning, 10:00 gathered oral shui solution pollen in full bloom, and loaded in centrifuge tube by water lily pollen, lucifuge is for subsequent use;
3) water lily pollen cultures
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 being lined with moistening filter paper, cultivates under dark surrounds;
4) water lily pollen vitality test
After slide glass cultivates 6 hours, be placed in optical microphotograph Microscopic observation Germinating status, can observe and sprout water lily pollen, the described pollen tube length having sprouted water lily pollen is greater than pollen diameter.
Aforesaid water lily pollen isolated culture and vigour-testing method, the culture temperature of wherein said step 3) is 24 ~ 26 DEG C.
The present invention has following beneficial effect relative to prior art:
Water lily pollen in-vitro culture method is scientific and reasonable, and first, sucrose, boric acid and calcium are required for pollen germination and pollen tube growth, is the main component of in-vitro pollen germination substratum.Sucrose mainly plays in pollen germination to be provided the energy and regulates osmotic pressure effect, and a certain amount of boric acid and calcium ion can make pollen tube look comparatively straight and slightly, unlikely long.Secondly, plant hormone PEG1500(polyoxyethylene glycol) pollen internal membranous structure can be made to change, change film surface charge, the softness of film and permeability are improved, thus promotes 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 water lily pollen germination, has the many and uniform advantage of sowing pollen.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, has higher actual application value for raising water lily breeding efficiency.
Advantage and disadvantage of the present invention, the non-limitative illustration by preferred embodiment below made an explanation, these embodiments only provide as an example.
Embodiment
Below in conjunction with the embodiment of water lily kind " Peter ", the invention will be further described.
The present embodiment comprises the following steps:
1) obtaining liq substratum
By following component obtaining liq substratum: the magnesium sulfate of the sucrose of 300 g/L, the boric acid of 10 mg/L, 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 oral shui solution column cap dish.
2) water lily pollen collection
The last ten-days period in July " Peter ", the flowers are in blossom the time, and morning, about 10:00 gathered oral shui solution pollen in full bloom, and loaded in centrifuge tube by water lily pollen, lucifuge is for subsequent use.
3) water lily pollen cultures
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 being lined with moistening filter paper, cultivates under 25 DEG C of dark surrounds.
4) water lily pollen vitality test
After slide glass cultivates 6 hours, be placed in optical microphotograph Microscopic observation Germinating status, the water lily of the sprouting pollen that pollen tube length is greater than pollen diameter can be observed.
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 application claims.
Claims (1)
1. water lily pollen isolated culture and a vigour-testing method, is characterized in that: comprise the following steps:
1) obtaining liq substratum
By following component obtaining liq substratum: the magnesium sulfate 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 oral shui solution column cap dish;
2) water lily pollen collection
In water lily the flowers are in blossom time last ten-days period in July, morning, 10:00 gathered oral shui solution pollen in full bloom, and loaded in centrifuge tube by water lily pollen, lucifuge is for subsequent use;
3) water lily pollen cultures
Dropped in by liquid nutrient medium on slide glass, dip in liquid nutrient medium, do not cover slide glass with writing brush by a small amount of water lily pollen, be after planting placed in by slide glass in the culture dish being lined with moistening filter paper, cultivate under dark surrounds, substratum temperature is 24 ~ 26 DEG C;
4) water lily pollen vitality test
After slide glass cultivates 6 hours, be placed in optical microphotograph Microscopic observation Germinating status, can observe and sprout water lily pollen, the described pollen tube length having sprouted water lily pollen is greater than pollen diameter.
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| CN201310269064.7A CN103396981B (en) | 2013-07-01 | 2013-07-01 | In vitro culture and vitality determination method for water lily pollen |
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| CN103396981B true CN103396981B (en) | 2015-04-29 |
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Families Citing this family (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 |
| 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 |
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 |
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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 |
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| 植物花粉培养研究进展;武冲 等;《中国农学通报》;20081130;第24卷(第11期);146-149 * |
| 水仙花粉离体萌发温度和培养液研究;罗凤霞 等;《辽宁林业科技》;20071030(第5期);13-14,25,57 * |
| 硼酸和蔗糖对‘柔毛齿叶’睡莲花粉萌发的影响;赵其龙;《亚热带植物科学》;20120410;第41卷(第4期);45-47 * |
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