藏红T在种子染色标记示踪方面的应用技术领域 本发明涉及藏红T的一种新用途，具体涉及藏红T应用于种子染色标记示踪。 TECHNICAL FIELD safranine T in terms of dye labeled tracer seeds  The present invention relates to a new use of safranine T, T applied particularly to seed safranine dye labeled tracer. 背景技术 种子传播、扩散等将母株生殖周期的末端与它们后代种群的建立连结了起来，现在已广泛认为其对植被结构具有深刻的影响。  seed dispersal and diffusion mother plant reproductive cycle will end with the establishment of their offspring populations link up, is now widely considered to have a profound impact on vegetation structure. 种子传播对植物种群、群落和生态系统生物学具有很多意义，它不仅影响种群的动态和持续生长，遗传区化，影响生物间的相互作用， 还能改变物种的丰富度和分布，进而对群落的结构产生影响。 Seed dispersal has many implications for plant population, community and ecosystem biology, it not only affects the dynamic and sustained growth of population, the area of genetics, affect the interaction between biological, but also changes in abundance and distribution of species, and thus on the community the structure of an impact. 而且，种子传播对农业生态系统中的杂草区系也具有十分重要的意义。 Moreover, seed dispersal is also of great importance to agricultural weed flora in the ecosystem. 研究各种媒介对植物种子的传播、扩散的影响一般采用植物调查种子库数量和空间分布动态或用小球模拟的方法。 Research various media to spread plant seeds, diffusion of general or dynamic simulation method with a small ball investigate seed bank number and spatial distribution of plants. 这些方法能够定性地揭示各种传播途径对植物种子的传播作用和对植物种子库空间动态的影响，不能确定杂草植物种子的传播比例，也难以将植物种子传播与各个生态系统中的群落连结起来。 These methods can be qualitatively reveal the effects of various transmission on Seed Dispersal of seeds and plant space dynamic libraries can not be propagated to determine the proportion of weed seeds, it is difficult to link the community plant seed propagation respective ecosystems stand up. 首要的原因是，种子传播循环是复杂的：在种子产生和新一代成株形成之间有很多中间步骤和过程； 另一个原因是，种子传播难以跟踪，研究者很难跟踪母株产生的种子传播到新的地方，因而也难以确定它们的命运。 The primary reason is that the seed propagation cycle is complex: and in the seed production process and there are many steps between the intermediate generation adult form; Another reason is that it is difficult to track the spread of seeds, seed researchers difficult to track parent strain produced spread to new places, and therefore it is difficult to determine their fate. 近年来，一些新技术和方法的应用，将传播的种子进行标记示踪， 在将种子和幼苗与母株对应方面显示了很好的前景。 In recent years, new techniques and methods, seed propagation is labeled tracer, shows great promise in the seeds and seedlings and the corresponding terms of the mother plant. 这些标记示踪技术包括：放射性同位素标记法、荧光染料标记法、稳定同位素分析和分子遗传标记等。 These labeled tracer techniques comprising: a radioisotope labeling, fluorescent labeling, stable isotope analysis and molecular markers like. 使用这些方法并成功标记往往需要较大粒的、含有丰富胚乳的种子，且这些被标记种子的检测需要特殊的仪器和较为复杂的操作，使用不方便，费时费工，其中荧光染料标记法和放射性同位素更是存在安全性和环境污染问题，另外，这些方法的花费都较昂贵，因而它们的应用范围受到限制。 And indicia of success using these methods often require a large grain, endosperm rich seeds, and these seeds were labeled detection requires special equipment and complicated operations, inconvenient, time consuming, and wherein the fluorescent labeling of radioactive isotopes more security and environmental pollution problems, in addition, take these methods are more expensive, and thus the scope of their application is limited. 发明内容 本发明的目的是为了解决现有技术中存在的缺陷，提供一种采用藏红T进行种子染色标记示踪的方法。 SUMMARY OF THE INVENTION  The object of the present invention is to solve the drawbacks present in the prior art, there is provided a method of using seed safranine T staining of the labeled tracer.  为了达到上述目的，本发明提供了一种藏红T在种子染色标记示踪方面的应用。  To achieve the above object, the present invention provides an application of safranine T in terms of dye labeled tracer seeds. 藏红T在用于未成熟脱落的种子染色时，包括以下步骤：(1)制备藏红T染色液：取藏红T或藏红T及乙醇，加水，混合均勻，使藏红T充分溶解， 制备藏红τ染色液；所述藏红T染色液中藏红T的重量百分浓度为0. 5〜5%，乙醇的体积百分浓度为0〜70% ；(2)种子染色标记：在植物种子成熟脱落前，取步骤（1)中制备的藏红T染色液，在植株的原位喷施或反复涂刷藏红T染色液于植物种子上，直至种子被染成红色。 When used Safranine T staining immature seed shedding, comprising the steps of: (1) preparing T safranin staining solution: Take safranine Safranine T and T or ethanol, water was added, mixed, fully dissolved so safranine T , τ stain solution prepared safranine; weight percent concentration of the dyeing liquid safranine T safranine T is 5~5 0.5%, by volume percent ethanol concentration 0~70%; (2) seed stained marker : off prior to plant seed maturation, from step (1) prepared in safranine T staining solution, spraying, or brushing repeatedly safranine T staining solution in situ on the plant seeds until the seeds are stained red.  藏红T在用于成熟的种子染色时，包括以下步骤：(1)制备藏红T染色液：取藏红T或藏红T及乙醇，加水，混合均勻，使藏红T充分溶解， 制备藏红τ染色液；所述藏红T染色液中藏红T的重量百分浓度为0. 5〜5%，乙醇的体积百分浓度为0〜70% ；(2)种子染色标记：采集成熟的植物种子，室内自然风干后，置于步骤（1)中制备的藏红T染色液中浸泡0. 5〜5小时，然后捞出浙干染色液，室内风干后植物种子即被染为红色。  When Safranine T for the mature seeds stain, comprising the steps of: (1) preparing T safranin staining solution: Take safranine Safranine T and T or ethanol, water was added, mixed uniformly, so that sufficient Safranine T It was dissolved to prepare a staining solution Safranin τ; the weight percent concentrations of the dyeing liquid safranine T safranine T is 5~5 0.5%, by volume percent ethanol concentration 0~70%; (2) seed staining Tags: collecting mature seeds, naturally the indoor air dried, placed in step (1) T safranin staining solution prepared in 0.5 immersed 5~5 hours, then remove and dry Zhejiang staining solution, i.e., the indoor air-dried plant seeds It is stained red.  其中，用于染色的种子来源于农田杂草、森林树木、沼生或水生植物，具体来源于以下植物：禾本科如日本看麦娘、看麦娘、茵草、杂草稻、野燕麦、双穗雀稗、稗、长芒稗、马唐、大狗尾草、狗尾草、狼尾草、狗牙根、老芒麦、黑麦草、结缕草，棒头草、长芒棒头草、耿氏硬草、雀麦、早熟禾、鹅观草、芒稷、荻、芦苇、荩草、剪股颖、杂草稻，石竹科如牛繁缕、粘毛卷耳，蓼科如羊蹄、齿果酸模、柳叶蓼，毛茛科如石龙芮、茴茴蒜，藜科如小藜，玄参科如通泉草、马先蒿、波斯婆婆纳、直立婆婆纳、婆婆纳、北水苦荬，柳叶菜科如丁香蓼，菊科如稻槎菜、苦苣菜、野艾蒿、泥胡菜、苍耳、加拿大一枝黄花、小蓬草、苏门白酒草、杓儿菜、一年蓬， 苋科如反枝苋、牛膝，锦葵科如苘麻，唇形科如宝盖草，茜草科如猪殃殃，十字花科如遏蓝  which is derived from the seeds used for dyeing farmland weeds, forest trees, marsh or aquatic plants, specifically from the following plants: Gramineae such as Japan foxtail, foxtail, Yan grass, weedy rice, wild oats, paspalum barnyard, barnyard, long barnyardgrass, crabgrass, giant foxtail, Setaria, Pennisetum, bermudagrass, E.sibiricus, ryegrass, Zoysia, Polypogon grass, polypogon monspeliensis, Gunn hard grass, brome, bluegrass, Roegneria, Mountain millet, Di, reeds, loyal grass, bentgrass, weedy rice, Caryophyllaceae such as cattle chickweed, sticky Maojuan ear, Polygonaceae such as hoof, dentatus, willow Polygonum, such as Ranunculaceae Ranunculus sceleratus, fennel fennel garlic, quinoa Chenopodiaceae such as small, Scrophulariaceae such as Mazus grass, Pedicularis, Persian speedwell, speedwell, Veronica, north water Kumai , Onagraceae such as loosestrife, Asteraceae Cha dishes such as rice, vegetables endive, Artemisia vulgaris, Hemistepta, cocklebur, Solidago canadensis, Conyza, Su liquor grass, vegetable dippers children a year Peng, Amaranthaceae, such as Amaranthus, Achyranthes, such as Malvaceae Abutilon, such as henbit the mint family, Rubiaceae such as cleavers, cruciferous such as Thlaspi 、荠菜，莎草科如薦草、牛毛毡、青绿苔草、球穗扁莎、水虱草、水蜈蚣、异型莎草，海桐花科如海桐，木兰科如含笑，黄杨科如小叶黄杨，榆科如榆树，椴树科如椴树，槭树科如三角枫、 五角枫、鸡爪械，胡桃科如青钱柳、枫杨，蔷薇科如委陵菜；朝天委陵菜，大戟科如地锦、大地锦、斑地锦，千屈菜科如节节菜，伞形科如蛇床等。 , Shepherd's purse, as recommended sedge grass, cow blankets, green grass green moss ball flat Sha Sui, water lice grass, water centipede, sedge, pittosporaceae such as Pittosporum, Magnoliaceae such as smiling, as lobular boxwood Branch boxwood, elm branch such as elm, linden tree branch as linden tree, maple tree branch, such as maple, Acer mono, mechanical chicken, walnut Branch as paliurus, maple, such as Rosaceae Potentilla; supina, Euphorbiaceae such as Jin, Jin earth, Euphorbia maculata, such as Lythraceae Rotala, Umbelliferae like a snake bed.  本发明相比现有技术具有以下优点：藏红T又叫番红0，是从藏红花柱头中提取的一种天然染色剂。 Compared with the prior art  The present invention has the following advantages: known safranine T Safranin 0, is extracted from saffron stigma a natural coloring agent. 采用藏红T进行种子染色标记，对种子无害，不影响种子的活性，且染色效果持久。 T uses saffron seed stain marks on seed harmless, does not affect the activity of seed and dyeing lasting effect. 利用本发明对种子进行染色标记操作简单，成本低，无污染。 With the present invention the seed marker staining simple operation, low cost and no pollution. 可应用于研究农田生态系统、河流湖泊生态系统、森林生态系统等多种生态系统中植物种子传播和种子库的研究。 Plant seed dispersal and seed bank of a variety of rivers and lakes ecosystem ecosystems, forest ecosystems can be used to study the ecosystem,. 具体实施方式 下面结合实例对本发明做进一步详细说明。 DETAILED DESCRIPTION  The present invention below with examples described in further detail.  实施例1在茵草子实成熟脱落前（5月底)使用藏红T染色液对麦田中茵草植株的种子进行原位染色标记，染色时使用软毛刷蘸藏红T染色液对茵草种子反复进行涂刷，直至将原本淡黄色的茵草种子染均勻染至红色。  Example 1 of the crop plants grass seed Yin situ staining using labeled safranine T before staining solution (5 end) Inn mature seeds as oxalyl off using a soft brush dipped safranine T staining of the staining solution Yin grass seeds repeated brushing, until the original yellow dye Yin grass seed evenly dyed to red.  实施例21.在夏熟杂草种子成熟期，采集小麦田杂草茵草spi^acAZ^X日本看m~%(AloPeCuruS japonicus)的成熟子实，室内自然风干后，用藏红T染色液浸泡2小时后捞出，放在室内风干。  Example 21. Cooked weed seed maturity in the summer, collecting grass weeds in wheat fields Yin spi ^ acAZ ^ X japonicus m ~% (AloPeCuruS japonicus) mature grains, the indoor dry naturally, with Safranine T staining solution soak 2 hours after the fish out, put indoor air dry. 原本淡黄色的茵草、日本看麦娘子实被染为红色。 Yin originally yellow grass, wheat to Japan to see a real lady was stained red.  2.被标记前后杂草种子的重量、萌发率比较和漂浮能力的比较种子重量的测定：称量草种子被标记前后的千粒重，重复5次。 Determination  2. Compare the weight of the seed before and after the weed seeds are marked by weight, and floating rate is relatively germination ability: Seed grain weight was weighed before and after the mark is repeated five times.  种子萌发率测定：将直径10 cm的滤纸铺在相同直径的培养皿中，用水浸湿滤纸， 然后分别将100粒未标记和染色标记的杂草种子均勻放在滤纸上，加盖，5次重复，放在20士1°C，光：暗=12 ：12的植物培养箱中。  Seed germination rate of the: diameter 10 cm from the filter paper in Petri dishes plated the same diameter, the water soaked filter paper, and 100, respectively, and staining unlabeled marker weed seeds uniformly on a filter paper, stamped , 5 replicates, 20 persons in 1 ° C, light: dark = 12: 12 in a plant incubator. 整个实验期间保持滤纸湿润。 Keep the filter paper moist throughout the experiment. 每2天计数一次萌发的杂草种子数量，直至杂草种子不再萌发为止。 Counts the number of weed seed germination once every two days, until no weed seed germination up.  染色后的茵草和日本看麦娘种子比未染色的重量均稍微增加，但差异不显著(表1)，表明染色处理仅稍微增加了这两种杂草种子的重量，不会改变它们的漂浮特性。  After dyeing Yin grass and foxtail seeds were Japan slight increase in weight than the unstained, but the difference was not significant (Table 1), indicating that the dyeing process only slightly increase the weight of both weed seeds will not change their floating characteristics. 染色后的茵草、日本看麦娘种子的萌发率比未染色的稍微降低，差异不显著(表1)，表明染色对这两种杂草种子的萌发率没有显著影响，即染色不损害它们的活力，染色液安全。 Yin dyed grass, Japanese foxtail seed germination rate to see slightly lower than the unstained, the difference was not significant (Table 1), indicating that the staining did not significantly affect these weed seeds germination rate of the two, that is, without prejudice to their dyeing energy, security staining solution.  表1茵草、日本看麦娘种子染色前后重量及萌发率比较  Table 1 Yin grass, foxtail seeds to Japan to see before and after dyeing weight and germination rate comparison
注：平均值后的字母相同表示在0. 05水平上差异不显著。 Note: the average value of the same after the letters indicates no significant difference at the 0.05 level.  实施例3制备不同浓度的藏红τ染色液：分别取藏红T 0. 5g、lg、2g、3g、4g、5g加入到50%体积百分浓度的IOOml乙醇溶液中，混合均勻，使藏红T充分溶解，制备成藏红T染色液。  Example safranin stain solution τ 3 different concentrations were prepared: were taken 0. 5g, lg, 2g, 3g, 4g, 5g was added to a concentration of 50% by volume percent ethanol solution IOOml safranine T and mix well the safranine T fully dissolved to prepare a staining solution Safranine T. 分别将茵草种子浸泡于不同的藏红T染色液中，1小时后捞出，放在室内自然干燥后，原本淡色的茵草种子被染为红色，并比较茵草种子染色前后的发芽率变化。 Yin grass seeds were soaked in the different Tibetan red T staining solution, after 1 hour, remove, after natural drying on the interior, the original pale Yin grass seeds are stained red, and compare Yin grass seed germination rate before and after dyeing Variety.  制备不同乙醇浓度的1%藏红T染色液：取藏红T lg，分别加入体积百分浓度为0%、10%、20%、30%、40%、50%、60%、70%的IOOml乙醇溶液，混合均勻，使藏红T充分溶解，制备成藏红T染色液。  Preparation of different ethanol concentration of 1% Safranine T staining solution: Take safranine T LG, the volume percentage were added at concentrations of 0%, 10%, 20%, 30%, 40%, 50%, 60%, IOOml 70% ethanol solution, uniformly mixing the safranine T fully dissolved to prepare a staining solution Safranine T. 分别将茵草种子浸泡于不同的藏红T染色液中，1小时后捞出，放在室内自然干燥后，原本淡色的茵草种子被染为红色，并比较茵草种子染色前后的发芽率变化。 Yin grass seeds were soaked in the different Tibetan red T staining solution, after 1 hour, remove, after natural drying on the interior, the original pale Yin grass seeds are stained red, and compare Yin grass seed germination rate before and after dyeing Variety.  发芽率测定时，将茵草种子分散置于装有湿润滤纸的平皿中，每个平板50粒种子，5次重复，将平皿放在20士1°C，光：暗=12 ：12的植物培养箱中。  The germination rate measurement, the Yin grass seed dispersion containing moistened filter paper was placed in a petri dish, 50 seeds per plate, 5 replicates, 20 persons in the plates 1 ° C, light: dark = 12: plant incubator 12. 整个实验期间保持滤纸湿润。 Keep the filter paper moist throughout the experiment. 每2天计数一次萌发的茵草种子数量，直至种子不再萌发为止。 Yin counts the number of Seed Germination once every two days, until no more seeds germinated far.  表2使用不同浓度藏红T染色的茵草种子的萌发率比较  Table 2 different concentrations germination safranin staining Yin T Seed Comparison
注：平均值（士SE)后的字母相同表示在0. 05水平上差异不显著。 Note: The letter after the average value (Shi SE) represents the same no significant difference at the 0.05 level.  表3使用不同百分浓度乙醇配制的藏红T染色的茵草种子的萌发率比较 Comparison of Seed germination Yin  TABLE 3 T using safranin staining different concentrations of ethanol formulated percent of
注：平均值（士SE)后的字母相同表示在0. 05水平上差异不显著。 Note: The letter after the average value (Shi SE) represents the same no significant difference at the 0.05 level.  使用不同浓度的藏红T染色液染色后茵草的发芽率为没有显著性差异，说明0. 5%-5%的藏红T都可以用于茵草染色，且染色后不影响茵草的发芽(表2)。  No significant difference Yin grass germination rate after T safranin staining solution with different concentrations, indicating 0.5% to 5% safranine T may be used for dyeing grass Yin, and does not affect the staining Yin grass germination (table 2).  使用不同浓度乙醇配制的1%藏红T染色液染色后茵草的发芽率为没有显著性差异，说明水，或体积百分浓度不高于70%的乙醇都可以用于染色液的配制(表3)。 No  formulated with different concentrations of ethanol 1% safranin staining solution stained T Yin grass germination rate significantly different, indicating water concentration or the volume percentage of not higher than 70% ethanol can be used for staining solution formulation (table 3).  实施例4CN 102523785 A秋季在农田采集成熟的杂草稻种子，晒干后，将种子用藏红T染色液浸泡1小时后捞出，放在室内自然干燥后，原本淡色杂草稻种子被染为红色。  Example collected in autumn 4CN 102523785 A mature rice seeds of weeds in the fields, dried, the seeds with Safranine T remove stain solution after soaking for 1 hour in the indoor air drying, pale original rice weeds seeds are stained red. 种子干燥后比较杂草稻种子染色前后的发芽率变化。 Germination percentage change compared before and after dyeing weedy rice seeds after seed drying. 另外将染色的杂草稻种子分别埋入稻田5cm、10 cm、15cm 土壤中，1 年后观察染色杂草稻种子的颜色变化。 Further weed rice seeds were stained paddy buried 5cm, 10 cm, 15cm soil, the color change was observed after 1 year staining rice weed seeds.  发芽率测定时，将杂草稻种子分散置于装有湿润滤纸的平皿中，每个平板50粒种子，5次重复，将平皿放在20士1°C，光：暗=12:12的植物培养箱中。  for determination of germination, the weed seed dispersion rice dish with moistened filter paper was placed, each plate 50 seeds, repeated 5 times, the plate 20 is placed Disabled 1 ° C, light: dark = 12 : plant incubator 12. 整个实验期间保持滤纸湿润。 Keep the filter paper moist throughout the experiment. 每2天计数一次萌发的杂草种子数量，直至杂草稻种子不再萌发为止。 Counts the number of weed seed germination once every 2 days until germination of weeds in rice seeds are no longer far.  最终，杂草稻染色前的发芽率为76. 2士9. 5%，使用藏红T染色液染色后的发芽率为74. 8士10. 4%，染色前后的杂草稻发芽率无显著性差异，说明藏红T染色液对杂草稻种子安全性好，不影响其发芽出苗。  Finally, the germination rate of rice weeds before dyeing 76.2 Disabled 9.5%, after use T safranin stain for germination rate 10.4% 74.8 disabilities, rice weeds before and after dyeing There was no significant difference in germination rate, indicating saffron T staining solution to weedy rice seed security is good, it does not affect the germination emergence. 实验发现，藏红T染色的杂草稻种子在埋入稻田5cm、10 cm、 15cm 土壤中12个月后所染红色均清晰可辨，说明此染色能持久保持。 It was found that weedy rice seeds hidden in red T-stained rice fields buried 5cm, 10 cm, 15cm soil after 12 months dyed red are clearly visible, indicating that this stain can endure.  实施例5收集在秋季成熟脱落后掉入事先设置的网兜中的椴树翅果，将翅果用藏红T染色液浸泡3小时后捞出，放在室内自然干燥后，榆树种子和果翅均被染为红色，并比较榆树种子染色前后的发芽率变化。 After  Example 5 fall into the collection linden Samara net bag set in advance in the premature fall after the fall, the Samara remove stain solution with Safranine T soaked for 3 hours in a natural drying chamber, elm seeds and fruit wings are stained red, and the germination rate change before and after comparison of elm seed staining.  发芽率测定时，将榆树染色前后的种子分别分散置于装有湿润滤纸的平皿中，每皿25粒，4次重复，将平皿放在25士1°C，光：暗=12:12的植物培养箱中。  for determination of germination, the seeds were stained before and after the dispersion elm plate equipped with moistened filter paper was placed in each dish 25, 4 replications, 25 persons in the plates 1 ° C, light: dark = 12 : plant incubator 12. 整个实验期间保持滤纸湿润。 Keep the filter paper moist throughout the experiment. 以胚根突破种皮Imm作为种子发芽标准，发芽过程中每天观察记录，注意保持滤纸湿润，直到连续2 d没有新种子发芽时视为发芽结束，统计染色前后的发芽种子数， 计算发芽率、发芽指数。 Imm radicle to break the seed coat as standard seed germination, sprouting process observed and recorded every day to keep the paper moist until germination consecutive 2 d is not seen as the end of the germination of new seeds, germinating seeds before and after dyeing number of statistics to calculate the germination rate, germination index. 计算公式如下：发芽率（G )=种子总发芽数/供试种子数XlOO %发芽指数（Gi) = Σ (G t / D t )其中：Gt —在时间t的发芽率；Dt —相应的发芽天数染色前后榆树种子发芽高峰均在2-4天时，整个发芽过程持续了约15天，染色前的发芽率为89 士9. 5%，发芽指数6. 2 士1. 4% ；使用藏红T染色液染色后的发芽率为84 士3. 3%，发芽指数5.7士0. 6%，染色前后的榆树种子发芽率、发芽指数均无显著性差异，说明藏红T染色液对榆树种子安全性好，不影响其发芽出苗。 Is calculated as follows: germination rate (G) = seed Total / test seeds XlOO% germination index (Gi) = Σ (G t / D t) Germination wherein: Gt - germination rate at time t in; Dt - corresponding germination days before and after the peak were stained elm seed germination in 2-4 days, the whole sprouting process lasted about 15 days, the germination rate of 89 persons before dyeing 9.5%, 6.2 germination index disabilities 1.4%; using reservoirs red staining solution after germination rate T 84 3.3% disabilities, persons germination index 5.7 0.6%, elm seed germination rate before and after dyeing, germination index no significant difference, indicating safranine T staining solution elm seed security is good, it does not affect the germination emergence.  实施例6在秋季采集成熟的沼生马先蒿种子，自然风干后将种子用藏红T染色液浸泡1小时后捞出，放在室内自然干燥后种子即被染为红色。  6 in the fall collected mature seed Pedicularis marsh, natural air-dry seed will remove stain solution with Safranine T soak 1 hour, i.e., on the interior naturally dried seeds embodiment stained red. 比较沼生马先蒿种子染色前后的发芽率变化。 Germination rate change before and after comparison of marsh Pedicularis seed staining.  发芽率测定时，将沼生马先蒿种子分散置于装有湿润滤纸的平皿中，每个平板50 粒种子，5次重复，将平皿放在20士1°C，光：暗=12 :12的植物培养箱中。  The germination rate measurement, the marsh pedicularis seed dispersion plate containing moistened filter paper was placed, each plate 50 seeds, repeated 5 times, the plate 20 is placed Disabled 1 ° C, light: dark = 12 : plant incubator 12. 整个实验期间保持滤纸湿润。 Keep the filter paper moist throughout the experiment. 每2天计数一次萌发的杂草种子数量，直至沼生马先蒿种子不再萌发为止分别统计染色前后的发芽种子数，计算发芽率。 Count the number of weed seeds germinate once every two days, until the marsh is no longer pedicularis seed germination until the seeds are germinated statistics before and after dyeing number, germination rate is calculated.  染色前沼生马先蒿种子的发芽率为69士7. 8%，使用藏红T染色液染色后的发芽率为65士8. 2%，染色前后的榆树沼生马先蒿种子发芽率无显著性差异，说明藏红T染色液对沼生马先蒿种子安全性好，不影响其发芽出苗。  Prior to staining Numano Pedicularis germination rate of 7.8% 69 persons, the use of T safranin stain for 65 persons germination rate 8.2%, elm Numano pedicularis germination rate before and after dyeing None significant differences, indicating saffron T staining solution for security marsh Pedicularis seed, does not affect the germination emergence.  实施例6在春夏之交或秋季采集成熟的下表子实(果实和种子)，自然风干后将子实用藏红T染色液浸泡1小时后捞出，放在室内自然干燥后种子即被染为红色。 After  Example 6 collected in the spring and summer or autumn mature seeds as in the following table (fruits and seeds), naturally dried after sub utility safranine T soak 1 hour, remove the stain solution, on the interior natural drying ie seed stained red. [0031 ] 将染色和没有染色的种子分别埋藏于水稻田IOcm深的土壤中，稻田中保持5cm水深，在埋藏30天、90天、150天后，每种类染色和非染色的种子分别挖出20粒以上观察记录种子颜色的变化情况。  The stained and unstained seeds were buried in deep soil IOcm rice fields, the rice paddies to maintain water depth of 5cm, buried in 30 days, 90 days, 150 days, every kind of stain and non-staining seeds were dug 20 the granulated observe the changes in the recording seed color. 种子挖出清洗后在30 士2 qC干燥箱内干燥10-15天后，若肉眼能分别出染色和非染色的种子的区别，则每种取5粒种子在同一条件下对这些种子进行拍照，然后使用Adobe Photoshop 6.0软件对种子的颜色进行分析判别。 Seeds 10-15 days after dug washed and dried in a drying oven 30 persons 2 qC, when the eye can distinguish non-staining and staining seeds respectively, then each take 5 seeds seeds were photographed under the same conditions, then use Adobe Photoshop 6.0 software to analyze the color of the seeds of discrimination.  由表4可知，埋藏30天时所有表中的染色的杂草种子都清晰可辨，埋藏90天时只有极少数的杂草种子褪色，埋藏150天后仍然有一半以上的种子染色和非染色的可以清楚的区别开。  From Table 4, all buried stained table 30 days of weed seeds are clearly visible, buried 90 days only a handful of weed seeds fade, 150 days after the burial of the seed is still more than half staining and non-staining It can be clearly distinguished.  表4.染色的植物种子的颜色随埋藏时间的变化  Table 4. Seed dyed color variation over time of burial
注：杂草种子埋藏于水稻田中，埋藏深度10cm，且稻田中保持5厘米的水深。 NOTE: buried in the paddy weed seeds, burial depth 10cm, and held in paddy water depth of 5 cm. 平均值(士1SD)后的小写字母不同表示在0. 05水平上差异显著。 Lower case letters after the average value (Shi 1SD) different from the 0.05 level indicates a significant difference.