【作者】 邵玺文；

【导师】 杨利民；

【作者基本信息】 吉林农业大学 ， 药用植物， 2008， 博士

【摘要】 黄芩（Scutellaria baicalensis Georgi）为唇形科多年生草本植物，传统上以根入药，为我国常用大宗中药材，随着中药现代化的不断进步，黄芩药材需求量不断加大，野生资源锐减，已不能满足市场需求。黄芩已被我国列为三级保护濒危植物，栽培黄芩已成为我国黄芩药材的主要来源，但是，尚缺乏系统化的科学研究。因此，开展黄芩规范化种植技术和质量控制生理生态机制研究，对实现药材原料的安全供应与质量的稳定可控具有重要意义。试验选择吉林省具有代表性的临江、长春和洮南市3个不同生态区域进行人工种植黄芩，综合考虑播种时间、播种密度、施肥、最佳采收期等栽培措施和水分、光照、土壤等生态因子对中药材黄芩产量和品质的影响；明确黄芩生长发育规律，重点研究不同生态区黄芩产量、品质和生理生态特性；探讨不同生态区影响黄芩产量和品质的主要影响因子；确定黄芩最适宜的生态区，为黄芩GAP基地建设和优质药材规范化生产提供科学依据。黄芩试验期间定期取样，测定黄芩根茎叶形态指标、生物产量和黄芩苷、总黄酮含量，黄芩苷测定用美国产Agilent 1100高效液相色谱仪，总黄酮测定用UV-1700紫外分光光度计；黄芩光合生理生态指标测定用英国PP Systems公司生产的TPS-1便携式光合作用系统；常规方法测定黄芩保护酶活性和可溶性蛋白、可溶性糖含量。主要结果如下：1栽培措施对黄芩药材产量和外观品质的影响1．1环境条件对黄芩种子发芽率和保护酶活性的影响1．1．1环境条件对种子千粒重和发芽率的影响生境条件和采种时间影响黄芩种子千粒重，8月采收的洮南黄芩种子千粒重大；不同生境条件下，黄芩种子千粒重不同，但对黄芩种子发芽率没有影响，三地黄芩种子发芽率变化趋势一致，发芽率相差微小；种子贮存时间长，发芽率降低，贮存2年以上的种子不宜做生产用种；黄芩浸种时的适宜水温为40～50℃，浸种时间6h～12h，发芽适宜温度为20～25℃，在无光环境下黄芩发芽率显著提高。1．1．2黄芩种子发芽率与保护酶活性的关系超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性与黄芩种子发芽率呈显著或极显著正相关。黄芩种子发芽过程中，几种保护酶活性均是当年新种子＞1年种子＞2年种子，保护酶活性与黄芩种子抗逆性关系密切，黄芩种子随贮存年限增加，种子抗逆性降低。1．2播种时间对黄芩产量的影响1年生黄芩产量秋播和春播高于夏播，播期对2年生黄芩产量影响不明显。1．3播种方式对黄芩的影响1、2年生平作和垄作栽培黄芩干物质积累与分配规律基本一致：黄芩平作枝根多，垄作枝根少；土壤板结枝根多，土壤松软枝根少。1．4播种密度对黄芩的影响密植枝根少，稀植枝根多；适当密植单位面积产量高，外观品质好，但单株产量低；稀植地上部与地下均生长良好，单株干物质积累多，但单位面积产量低。2生态因子对黄芩药材产量和外观品质的影响2．1水分供给量对黄芩产量的影响在黄芩生长期内，150 mm供水量黄芩生物量降低，根茎变细，根长变短，改变了黄芩地上与地下物质分配比例，影响外观品质；450 mm供水量黄芩花、果重量增加，但黄芩根产量和总生物量较低；250 mm和350 mm供水量黄芩根、茎优势明显，外观品质好，根重、地上重、总生物量均高；黄芩具有较强的耐旱性。2．2施肥对黄芩产量的影响1年生黄芩单独施用氮、磷、钾肥时，氮肥效果明显，主茎变长变粗，产量提高，氮肥过量影响产量；与氮肥相比，磷、钾肥单独施用效果不明显，钾肥增量施用，黄芩干物质增加。2．3光对黄芩产量的影响相同环境条件下，光照强长势好，根和总生物量大。2．4土壤对黄芩产量的影响长春黑土黄芩根、茎、叶和总生物量大于临江棕色森林土和洮南沙质暗栗钙土。3不同生态区黄芩药材产量及生长规律3．1 1、2年生黄芩生长规律1年生黄芩根长、主茎长和生物量呈增加趋势，8月前以地上生长为主，8月后以根生物量积累为主；2年生黄芩主茎7月中旬以前生长较快，之后生长缓慢，根长和根生物量一直缓慢增加；地上重和总生物量呈单峰曲线变化，峰值出现在8月末，之后下降。3．2不同生态区黄芩产量差异及原因不同生态区1年生黄芩根生物产量差异较大，洮南最轻，长春生物产量前期低于临江，后期与临江生物产量持平；2年生黄芩根和地上生物产量长春高于临江和洮南。临江降水多，土壤和空气湿度大，1年生黄芩容易得叶枯病；洮南降水量少，黄芩出苗困难，黄芩生物量低，影响单位面积黄芩产量；长春降水量适宜，土壤肥沃，黄芩生物产量和黄芩有效成分含量都比较高，适合建立黄芩规范化种植基地。4黄芩光合生理生态特性4．1 1、2年生黄芩光合特性1年生黄芩和2年生黄芩上、中部叶片光合速率日变化趋势基本一致，早晨和中午较高；2年生黄芩下部叶片净光合速率早晨高，略有下降后升高，中午达到峰值，“光合午休”现象较轻。4．2黄芩不同时期光合特性2年生黄芩不同时期光合作用日变化均呈双峰或多峰曲线，峰谷出现的时间各不相同；5月份黄芩处于旺长期，6月份黄芩光合速率日变化较平稳，8月和9月黄芩光合速率日变化波动较小。5月、6月、8月和9月黄芩日均净光合速率是20.69μmolCO₂·m^-2·s^-1、9.81μmolCO₂·m^-2·s^-1、5.39μmolCO₂·m^-2·s^-1、4.37μmolCO₂·m^-2·s^-1。4．3黄芩光合生理生态的关系1年生黄芩光合速率与光合有效辐射呈极显著正相关；2年生黄芩上、中、下部叶片光合速率与光合有效辐射呈显著正相关，2年生黄芩下部叶片光合速率与水气压、空气相对湿度呈极显著负相关，与大气温度呈显著正相关；不同季节黄芩光合速率生育前期与光合有效辐射呈极显著正相关，与水气压呈极显著正相关，生育后期与光合有效辐射呈显著正相关，与其他环境因子的关系因季节不同差异较大。不同生态区黄芩净光合速率中午降低均为气孔限制，低的空气湿度是光合中午降低的重要生态因子；临江生态区高温高湿、长春生态区叶温高及大气CO₂浓度低、洮南生态区高温低湿是影响黄芩净光合速率的主要环境因子。5黄芩药材质量与生理生态因子关系5．1水分对黄芩药材质量的影响适宜供水量可以提高黄芩蛋白质和可溶性糖的含量，提高黄芩的抗逆性；黄芩生长期250 mm～350 mm供水量黄芩净光合速率高，各器官物质分配合理，生物产量、黄芩苷和总黄酮都比较高；1年生供水量处理黄芩苷和总黄酮与黄芩生物量呈正相关。5．2不同生态区和黄芩年龄对黄芩药材质量影响5．2．1不同生态区黄芩药材质量差异1年生黄芩苷含量，洮南略高于长春，长春高于临江，1年生黄芩根黄酮不同生态区差异较小；2年生黄芩苷含量长春＞临江＞洮南；单位面积黄芩苷平均含量长春高，洮南低。5．2．2不同年龄黄芩药材质量差异不同生态区1、2年生黄芩苷、黄酮积累动态在不同季节变化趋势基本一致；2年生黄芩的黄芩苷和总黄酮含量高于1年生黄芩。长春2年生黄芩在9月中上旬单位面积黄芩苷产量高。5．3黄芩药材质量与黄芩生物产量的关系综合分析不同生态区黄芩质量和产量关系表明：1、2年生黄芩苷和总黄酮均与黄芩根部生物量积累呈极显著负相关（p＜0.01），单位面积黄芩苷和总黄酮产量与黄芩生物产量尤其是根的产量呈正相关。5．4黄芩药材质量与黄芩可溶性蛋白和可溶性糖的关系1年生黄芩苷和总黄酮与黄芩叶片中可溶性蛋白和可溶性糖呈正相关；2年生黄芩苷和总黄酮与黄芩叶片中可溶性蛋白相关性较低，2年生黄芩苷和总黄酮与黄芩叶片中可溶性糖呈负相关。6黄芩最佳采收期2年生黄芩苷和总黄酮含量高于1年生黄芩；3年生黄芩苷和总黄酮含量高于2年生黄芩；2年生黄芩在9月中旬和第3年春季单位面积黄芩苷产量高，适宜采收，收获后有充足时间晾晒；综合产量和黄芩苷含量，栽培黄芩最佳采收时期为第二年的9月中旬或第3年春季。更多还原

【Abstract】 Scutellaria baicalensis Georgi is a kind of perennial herbaceous plant in the labiatae family, and itsroot was used as Chinese Traditional Medicine （CTM） traditionally. Now, the wildlife resource of S.baicalensis can not meet the actual market demand with the developing modernization of CTM, thedemand was unceasingly increased while the wild resource was sharply reduced. Meanwhile the wild S.baicalensis was set to the protective endangered plant of third class by the authority. Today, the cultivatedS. baicalensis become the main source of medicine material. On the other hand, there was no systemicscientific research, on the cultivated of S. baicalensis. So, it is of key value to carry out the study ofstandardized cultivation techniques and eco-physiological mechanism on S. baicalensis to realize thesafety supply of raw medicinal materials and the stability quality controlling.Three different ecotopes were selected as the representative test plots including Linjiang, Changchunand Taonan in Jilin province. The factors affecting the quality of S. baicalensis were syntheticallyconsidered such as growing factors of seeding time, frequency, fertilization and best harvesting season andthe ecological factors of the moisture, illumination and soil. The yield, quality and eco-physiologicalcharacteristics were the key part of the study of growth and development law. The main effect factors thateffected the yield and quality were discussed among the different ecotopes, and the best feasible ecotopewas selected out. These results can provide scientific basis for construction of GAP cultivation bases andstandardized production of good quality for Chinese medicinal materials of S. baicalensis.The samples were periodical taken during the experiment for the morphological indices of root, stem,leaf, biomass, the content of baicalin and total anthoxanthin. The baicalin content was determined by 1100Agilent high performance liquid chromatography （HPLC） made in USA and total anthoxanthin contentwas determined by UV1700 ultraviolet spectrophotometer made in Japan. The photosynthetic ecophysiological indexes of S. baicalensis were determined by TPS-1 portable photosynthesis systemproduced by PP systems in England. The protective enzyme activity, contents of soluble protein and sugarwere determined by conventional methods.The main results were as follows:1 Effect of cultivated measures on yield and appearance quality of S. baicalensis1.1 Effect of environmental conditions on germination rate of seed and protective enzyme activity of S.baicalensis 1.1.1 Effect of environmental conditions on 1000-grain weight and germination rate of seedThe 1000-grain weight was changed in the different habitats, but the germination rate was not affected.Among the three plots, the different of germination rate was tiny and the change tendency of germinationrate was same. The germination rate decreased with the store years prolonged, and that stored over 2 yearswere not suitable to production. The feasible temperature of seed soaking was 40～50℃, and the last timewas 6h～12h, the feasible temperature of germination was 20～25℃, and the germination rate wassignificant improved with the dark..1.1.2 The relationship between germination and protective enzyme activityThe positive correlation between superoxide dismutase （SOD）, peroxidase isozymes （POD）, Catalase（CAT） activity and the germination rate of S. baicalensis seeds was significant or very significant. Theactivities of protective enzymes was in order of the new seeds > annual seeds > biennial seeds, theresistance decreased with the seed storing years prolonged.1.2 Effect of seeding time on rootThe yield of annual S. baicalensis that was planted in autumn or spring was higher than in summer,and the effect of seeding time on biennial was not remarkable.1.3 Effect of seeding pattern on rootThe accumulation and distribution law of dry matter was basically same between flat cultivation andridge cultivation of annual and biennial S. baicalensis. The flat cultivation had more branch roots thanthe ridge cultivation. And the condensed planting and hardened soil has more branch roots than the sparseplanting and soft soil.1.4 Effect of seeding density on rootThe branch roots were fewer in dense-planting and more in rare-planting. The suitably dense sowingwas good for the appearance quality and the higher yield per unit area, but the individual plant yield waslow. The sparse planting was good for the growth of the overground and underground and theaccumulation of dry matter, but the yield per unit area was low.2 Effect of ecological factors on yield and appearance quality of S. baicalensis2.1 Effect of water supply on yield of S. baicalensisThe biomass with 150mm water supply was decreased, the rhizome was thinner and the rhizomelength was shorten in the growing period, the distribution percentage between overground and undergroundwas changed, and the appearance quality was affected by this treatment. The flower and fruit biomass with450mm treatment was increased, but the yield of root and total biomass was smaller. The good influence of 250mm and 350mm treatments on root and stem were evident, the appearance quality was better, therhizome, overground biomass were highest. Otherwise S. baicalensis has better drought tolerance.2.2 Effect of fertilization on S. baicalensis yieldThe effect was evident that the application of nitrogenous fertilizer while the nitrogen, phosphorus andpotassium fertilizer was separately used on annual S. baicalensis, with which the main stem was longer andthicker But the yield would be affected by excessive application of it. Comparing with nitrogen, thephosphorus and potassium fertilizer had little affection on yield, but the dry matter was increased with theapplication of potassium.2.3 Effect of illumination on S. baicalensis yieldIt had good growth in the strong illumination under the same environmental condition, the root andtotal biomasses were increased.2.4 Effect of soil on S. baicalensis yieldThe root, stem, leaf and total biomasses of plant growed in the black soil of Changchun were more thanthose in the brown forest soil of Linjiang and sandy dark chestnut soil of Taonan.3 Growth rule and yield of S. baicalensis in different ecotopes3.1 Growth rule of annual and biennial S. baicalensisThe tendency of root and main stem length and biomass of annual S. baicalensis was increased, theoverground growth was dominant before August and then changed into the root after that. The biennial S.baicalensis had bigger growth rate of main stem before the middle of July, then decreased, and it hadslower growth rate of root length and biomass. The overground and total biomass fluctuated as single-peakcurves, and the peaks appeared at the end of August and after that it became slow.3.2 The qualitative difference in different ecotopes and explanationThe root biomass of annual S. baicalensis had significant difference in different ecotopes. The rootfrom Taonan was the rightest, and the root from Changchun was lighter than those from Linjiang in theprevious period, and then became the same in the later period. The annual S. baicalensis was easy infectedleaf blight because of much precipitation, heavy soil and air humidity in Linjiang. The seedling emergencewas hard because of Iittle precipitation in Taonan, so the biomass was small, and the yield per unit areawas also affected. The biomasS and the content of effective components were both higher in Changchunbecause of right precipitation and rich soil, which suitable for construction of standardized cultivationbases of S. baicalensis.4 Research on photosynthetic physiological ecology characteristic of S. baicalensis 4.1 Research on photosynthetic physiological ecology characteristic of annual and biennial S. baicalensisThe diurnal changes of net photosynthetic rate at top and middle leaves position of annual andbiennial S. baicalensis was basically same, and the photosynthetic rate in the morning and noon was high;the net photosynthetic rate at bottom leaves of biennial S. baicalensis was high in the morning, anddecreased a little, and than increased, The maximum appeared in the noon, the midday depression ofphotosynthesis was light.4.2 Photosynthetic characteristic in different periodThe diurnal changes of net photosynthetic rate of biennial S. baicalensis in different period were alldouble peak or many peaks curves, the time of valley-peak was different. The May was the vigorousgrowing period of S. baicalensis, the changes in the June was stationary, and the change of netphotosynthetic rate was very small in the August and September. The diurnal net photosynthetic rate inMay, June, August and September were 20.69μmolCO₂·m^-2·s^-1, 9.81μmol CO₂·m^-2·s^-1, 5.39μmolCO₂·m^-2·s^-1,4.37μmolCO₂·m^-2·s^-1, respectively.4.3 Relationships between photosynthetic physiological ecology and environmental factorsThe photosynthetic rate of annual S. baicalensis had very significant positive correlation with thephotosynthetic active radiation. The photosynthetic rate at bottom leaves position of biennial hadsignificant negative correlation with water vapor pressure, air relative humidity, but a positive correlationwith air temperature; The photosynthetic rate of S. baicalensis had significant positive correlation with thephotosynthetic active radiation and water vapor pressure in early growing stage and positive correlationwith photosynthetic active radiation in later growing stage, the relationship between photosynthetic rateand other environmental factors was marked different in different season.The reduction of the photosynthetic rate in different ecotopes was all because of the stomatallimitation, the important ecological factor influencing the reduction of the photosynthetic rate was the lowair humidity. The high temperature and humidity in Linjiang ecotope, the high leaf temperature and low airCO₂ concentration in Changchun ecotope, and the high temperature and low humidity in Taonan ecotopewere the main environmental factors influencing the net photosynthetic rate.5 Relationship between quality of S. baicalensis herb and physiological ecology factors5.1 Effect of water on quality of S. baicalensis herbThe content of protein and soluble sugar was increased with right water supply, with which theresistance increased. The net photosynthetic rate of S. baicalensis was high with 250mm～350mm watersupply in the growth period, the distribution of dry material in every organ was reasonable. The yield, thecontent of baicalin and total anthoxanthin were increased as the result. The baicalin and total anthoxanthin contents had positive correlation with the biomass yield of annual S. baicalensis by all the water supplytreatment.5.2 Effect of age and ecotopes on quality of S. baicalensis herb5.2.1 The difference of herb in different ecotopesThe baicalin content of annual S. baicalensis in Taonan was little higher than that in Changchun, andthe content in Linjiang was evident lower than others. The difference of anthoxanthin content in the annualroot among these plots was small. The mean baicalin content per unit area in Changchun was highest andthe Taonan was the last one. The baicalin content of biennial was in the order ofChangchun>Linjiang>Taonan.5.2.2 Quality difference of S. baicalensis herb in different ageThe change tendency of dynamic accumulation of anthoxanthin and baicalin in different seasons wasalmost the same, and its content in biennial was higher than that in annual. The baicalin content per unitarea of biennial was higher in middle ten days of September of Changchun.5.3 Relationships between quality and biomass of S. baicalensis herbThe relationships between quality and yield of S. baicalensis in different ecotopes werecomprehensive analyzed, the results showed: the content of baicalin and total anthoxanthin in annual andbiennial had very significant negative correlation with biomass accumulation of the root （p<0.01）, thebaicalin and total anthoxanthin yield per unit area had positive correlation with the biomass yield of plantespecially the yield of root.5.4 Relationship between quality of S. baicalensis herb and soluble protein and sugarThe baicalin and total anthoxanthin contents of annual S. baicalensis had positive correlation with thesoluble protein and sugar in the leaf, and these contents of biennial had very little correlation with thesoluble protein and negative correlation with the soluble sugar.6 The optimal harvest time of S. baicalensisThe content of baicalin and total anthoxanthin in triennial was the highest, the biennial was the secondone, and the annual was the last. The mid-September or in the third spring is the optimal harvest time forthe biennial because of the highest yield of baicalin in unit area. And there would have enough time for theopen-air drying. So the best harvest time of S. baicalensis was the mid-September of the second year or thethird spring.更多还原