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防风种子休眠生理与栽培技术研究

Study on Seed Dormancy Physiology and Culture Technique of Saposhnikovia Divaticata

【作者】 周艳玲

【导师】 赵雨森; 赵敏;

【作者基本信息】 东北林业大学 , 水土保持与荒漠化防治, 2009, 博士

【摘要】 防风(Saposhnikovia divaricata)是一种重要的药用植物,同时也是一种固沙植物。随着防风野生资源的日益枯竭,人工种植防风研究逐渐成为热点。而防风人工栽培中最大的问题是由于种子休眠特性加上东北地区春旱的气候特点造成的出苗缓慢、出苗不齐。本文以东北和内蒙古东部地区防风为研究对象,开展防风群落学特征、种子休眠生理和栽培技术研究,对人工优质栽培防风具有重要的意义。东北地区防风多分布于下列群落中:狼针草、白扦云杉林中;矮丛苔草、花木蓝、辽东栎林中;荆条、黄背草灌草丛中;麻黄、冰草灌草丛中;黄囊苔草、多裂叶荆芥、狼针草草原中;寸草、羊草、大针茅草原中;线叶菊、羊茅草原中;羊草、羊茅草原中;小白蒿、冰草、羊茅草原中;阿尔泰针茅、糙隐子草草原中。从我国防风的主产区东北及内蒙古东部地区共采集36份防风种子样品,通过测定这些种子样品的净度、千粒重、含水量、生活力与发芽率,对测定结果进行统计分析后制定了防风种子的质量分级标准。防风种子共分4级:Ⅰ级防风种子,发芽率≥80%、生活力≥92%,净度≥93%,千粒重≥3.7g、含水量≤9%;Ⅱ级防风种子,60%≤发芽率<80%、82%≤生活力<92%、84%≤净度<93%、3.2 g≤千粒重<3.7 g、含水量<9%;Ⅲ级防风种子,40%≤发芽率<60%、72%≤生活力<82%、75%≤净度<84%、2.7 g≤千粒重<3.2 g、含水量≤9%;Ⅳ级(不合格种子)防风种子发芽率≤40%、生活力≤72%、净度≤75%、千粒重≤2.7g、含水量≥9%。对防风种子的休眠原因进行了研究,结果表明防风种子去皮后的种仁发芽率、吸水率和呼吸强度均未见提高,说明种皮障碍不是导致防风种子休眠的原因;通过制作石蜡切片观察防风种子胚在贮藏期间的发育动态,发现防风种子胚存在形态后熟现象,但贮藏至翌年春季,绝大多数种子的胚已发育完全,因此不是造成田间出苗率低、发芽缓慢的主要原因;本文用首创的原位作用研究方法,直接证实了防风种子中内源抑制物质是抑制其种子萌发、导致种子休眠的主要原因。通过定期观测6种不同贮藏条件下防风种子的生活力、电导率、粗提物抑制活性与发芽率等生理生化指标,确立能够保持防风种子生活力、提高种子发芽率的适宜贮种方式。结果表明:冷藏沙藏、冷冻沙藏不适用于防风的贮种;室温干藏、冷藏干藏与冷冻干藏适用于防风种子的长期贮种,其中又以冷藏干藏最好;室温沙藏不适用于长期贮种,建议防风种子收获后采取干藏的方式,播种前一个月室温条件下层积,可以加快防风种子的发芽速率。分别采用GA溶液浸种,细沙摩擦,有机溶剂浸种,流水冲洗,杨、柳枝浸出液浸种等方式处理防风种子。结果表明有效解除防风种子休眠的方法:始温为45℃的温水浸种24 h,防风种子发芽率可提高至67.5%;流水冲洗48 h,防风种子发芽率可提高至75%;首先用始温为45℃的温水浸种24 h,然后加入柳枝浸出液,防风种子发芽率可提高至75%。采用有机溶剂提取法对防风种子中的内源抑制物质进行初步提取与分离,获得乙醚相Ⅰ、乙醚相Ⅱ、乙醚相Ⅲ、乙醚相Ⅳ、水相Ⅱ、水相Ⅲ、水相Ⅳ共7个提取分离物。生物测定结果表明,3个水相提取物具有较高的抑制活性。用硅胶柱层析对3个水相提取物进一步分离与纯化,根据生物测定结果将有活性的过柱收集液浓缩后进行HPLC-MS分析鉴定,并查阅相关文献推测防风种子中的内源抑制物质有:亚麻酸、脱落酸、β-古甾醇阿魏酸酯、齐墩果酸、5-O-甲基阿米醇、升麻酸、β-桉叶醇、6-O-乙酰基熊果苷、绿原酸。防风大规模栽培成功的关键技术在于如何提高田间出苗率,种子经温水浸泡后掺沙播种,播前与覆土后踩格子或木石磙镇压,可使防风种子田间出苗率提高至52%,效果显著。

【Abstract】 Saposhnikovia divaricata is an important medicinal plant,and also a kind of dune-fixing plants.Research about Saposhnikovia divaticata culture became hotspot for the wide resource increasingly dry up.The main question in culture was low germination and sprouting slowly after seeding because of seed dormancy and spring drought in Northeast China.So we began to study coenology charactrristics of Saposhnikovia divaticata,seed dormancy physiology and culture techniques in Northeast China,which was important for good culture.Saposhnikovia divaricata often distributed in the follow communities.Stipa baicalensis, Picea meyeri forest.Carex humilis,Indigofera kirilowii,Quercus liaotungensis forest.Vitex negundo,Themeda japonica tussock.Ephedra sinica,Agropyron cristatum tussock.Carex korshinskyi,Schizonepeta multifida,Stipa baicalensis grassland.Carex duriuscula,Leymus chinense,Stipa grandis grassland.Filifolium sibiricum,Festuca ovina grassland.Leymus chinense,Festuca ovina grassland.Artemisia frigida,Agopyrum cristatum,Festuca ovina grassland.Stipa krylovii,Cleistogenes squarrosa grassland.We collected 36 seed samples of Saposhnikovia divaricata from main producing area, Northeast China and Eastern inner Monglia.Then investigated cleanliness,thousand-grain weight,moisture content,viability and germination percentage of these seed samples.We determined seed quality grading standard of Saposhnikovia divaricata by statistical analysis. Saposhnikovia divaricata seeds were divided into 4 grades.GradeⅠ:germination percentage≥80%,viability≥92%,cleanliness≥93%,thousand-grain weight≥3.7 g,seed moisture content≤9%.GradeⅡ:germination percentage is 60-80%,viability is 82-92%,cleanliness is 84-93%, thousand-grain weight is 3.2-3.7 g,seed moisture content≤9%.GradeⅢ:germination percentage is 40-60%,viability is 72-82%,cleanliness is 75-84%,thousand-grain weight is 2.7-3.2 g,seed moisture content≤9%.GradeⅣ:germination percentage≤40%,viability≤72%, cleanliness≤75%,thousand-grain weight≤2.7g,seed moisture content≥9%.We studied the dormancy cause of Saposhnikovia divaticata seed.Results showed that germination percentage,absorbing water capacity and respiration intensity of seeds without seed capsule were not increased.That is,seed capsule is not the cause for seed dormancy.We observed embryo growth in the storage period by paraffin section,found embryo post-ripening. But it is not the main cause for low germination and sprouting slowly in the field.We also studied the effect of endogenous inhibitory substance on seed germination in situ.Results showed that,endogenous inhibitory substance is the main factor that results in seed dormancy.To develop the suitable seed storage condition which could maintain seed vitality and increase germination percentage,we investigated seed vitality,conductivity,inhibitory activity of extracting solution and germination percentage of Saposhnikovia divaticata during the storage period.Results showed:stored in the sand at 4℃and -20℃were not suit for seed storage of Saposhnikovia divaticata,stored dry seed at room temperature,4℃and -20℃were suit for seed storage in long time,especially stored at 4℃was best,stored in the sand at room temperature was not suit for seed storage in long time.So we suggest store dry seed after harvest,store in the wet sand at room temperature one month before seeding,which could increase germination rate of Saposhnikovia divaticata.To increase seed germination percentage of Saposhnikovia divaticata,we soaked the seeds in GA solutions,rubbed the seeds with sand,soaked the seeds in organic solvent, rinsed the seeds with running water,soaked the seeds in Iixivium of willow branch and poplar branch separately.Results showed:germination percentage of seeds soaked in warm water was 67.5%,germination percentage of seeds rinsed with rushing water was 75%,germination percentage of seeds soaked in warm water and then soaked in lixivium of willow branch was 75%,these three methods increased seed germination percentage of Saposhnikovia divaticata.We extracted and separated endogenous inhibitory substance in Saposhnikovia divaticata seeds by organic solvent.Then gained 7 extractors:aetherⅠ,aetherⅡ,aetherⅢ,aetherⅣ, waterⅡ,waterⅢ,waterⅣ.Results of bioassay showed that,all of water extractors had higher inhibitory activity.Separated and purified water extractors by column chromatography.Based on the result of bioassay,we concentrated the drips separated by column chromatography and analyzed by HPLC-MS.Then consulted references,we concluded the endogenous inhibitory substance likely as follows:linolenic acid,abscisic acid,β-sitosteryl ferulate,oleanolic acid,5-O -methyl a-mminol,cimicifugc acid,β-eudesmol,6-O-acetylarbutin,Chlorogenic Acid.Critical techniques of culture Saposhnikovia divaticata were how to increase sprouting percentage in the field.This paper soaked the seeds in warm water 24 h,then mixed with sand and seeded them in the field,pressed the soil before seeding and after casing soil,which could increased the sprouting percentage to 52%.

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