【作者】 王惠珍；

【导师】 张恩和；

【作者基本信息】 甘肃农业大学 ， 药用植物资源与利用， 2011， 博士

【摘要】 当归为甘肃道地药材,以质优物美的“岷归”享誉海内外。但由于现有适宜生态区有限,并且药农习惯于种植在比较平坦的低海拔地带,导致当归栽培面积有限,产品供不应求。为了探寻当归适宜栽培区,在国家自然科学基金项目“当归连作障碍的根际作用机制及生态扰动对策”（编号:31060182）和甘肃省农业生物技术研究于应用开发项目“当归连作障碍的生态扰动效应及根际生物及化学抑制剂研制”的资助下,于2009～2010年,在甘肃省岷县茶埠乡2300～2800m海拔范围内进行生态适应性试验,探索不同海拔下当归产量和品质差异,从叶片保护酶、渗透调节物质和光合特性等角度揭示海拔影响当归产量和品质的机理,研究影响当归产量和品质的关键因子及其影响大小,得到如下主要结论:1.海拔梯度通过改变光合产物分配格局和干物质积累速率而影响产量形成。随海拔升高,光合产物向根中分配提前,干物质积累速率增加。海拔导致成药期当归光合产物分配格局发生明显变化,海拔2780m处理根分配比例（≥30%）较其他两个海拔提前10d,并且后期根分配比例（约54%）明显高于其他两个海拔（49.8%～50.9%）,这为采收期当归产量形成奠定基础;生物积累量随海拔升高而呈增加趋势,分别为33.1%、43.7%和46.7%,分析海拔间生物量积累的变化幅度得出:海拔越高,变化幅度越小,在海拔2360m处理基础上,海拔2570m处理增加32%,而在海拔2570m处理基础上,海拔2780m仅增加7%;药材产量依次为海拔2570m处理（28.4g/plant）>海拔2780m处理（26.6g/plant）>海拔2360m处理（21.8g/plant）,低海拔2360m处理比其他两个海拔分别低22.6%和20.8%,差异达显著水平（P<0.05）。药材产量在海拔梯度间的变化幅度与干物质积累速率一样,随海拔升高而降低,依次为30.3%和6.3%。2.海拔梯度对当归光合色素和光合参数具有显著影响。海拔升高导致类胡萝卜素（Car）、净光合速率（Pn）、气孔导度（Gs）和胞间CO₂浓度（Ci）降低而叶绿素含量增加,而蒸腾速率（Tr）出现先增加后降低的趋势,进而影响当归光合生理生态。海拔2570m和2780m处理与海拔2360m处理相比,Car分别降低14.3%和55.4%,Pn日平均值分别降低10.0%和18.4%,Gs日平均值分别降低5.3%和16.0%,Ci日平均值分别降低7.6%和14.0%,叶绿素a分别增加7.9%和70.5%,叶绿素b分别增加21.1%和75.8%,Tr日平均值海拔2570m处理显著高于海拔2360m处理（增加13.3%）,而海拔2780m处理显著低于2360m处理（降低13.3%）。通过分析影响净光合速率环境因子表明,空气相对湿度为关键因子,其次为光合有效辐射（PAR）和空气CO₂浓度。3.由于低海拔温度较高致使当归体内活性氧过量积累,形成膜质过氧化产物丙二醛（MDA）,对细胞造成毒害,当归则通过增加细胞渗透调节物质含量降低MDA的毒害,这与当归性喜冷凉环境条件特性一致。MDA含量随海拔升高而降低,海拔2360m处理比其他两个海拔处理分别高52.4%和38.1%,处理间差异显著（P<0.05）,三个海拔处理间MDA降低幅度随海拔升高而降低,依次为52.4%和30.0%;游离脯氨酸（Pro）含量随海拔升高呈现“高-低-高”的海拔效应,海拔2360m和2780m处理间差异不显著,但均显著高于海拔2570m处理,分别高出12.4%和13.6%,而Pro在三个海拔间变化幅度随海拔升高而增加,依次为12.4%和13.6%;海拔2360m和2570m处理可溶性糖含量均显著高于海拔2780m处理（P<0.05）,分别高出19.7%和14.7%,而海拔2360m和2570m处理间差异不显著（P>0.05）,三个海拔处理间可溶性糖含量变化幅度随海拔升高而增加,依次为7.7%和14.7%;本研究还分析了渗透调节物质与膜质过氧化产物MDA间的关系,表明渗透调节物质与MDA含量变化一致,并且渗透调节物质之间具有互补关系。4.当归通过增加保护酶活性来分解过氧化物,保护细胞不受过氧化物MDA的伤害。SOD活性随海拔升高而降低,海拔2780m处理比海拔2360m和2570m处理分别降低40.8%和39.3%,差异达显著水平（P>0.05）,三个海拔处理间SOD活性降低幅度随海拔升高而增加,在海拔2360m基础上,海拔2570m处理降低2.5%,而在海拔2570m基础上海拔2780m处理降低39.3%;CAT活性随海拔升高而降低,海拔2780m处理比海拔2360m和2570m低13.6%和17.1%,差异达显著水平（P<0.05）,三个海拔间CAT活性降低幅度随海拔升高而增加,依次为13.6%和14.6%;POD活性随海拔升高呈现先升高后降低的变化趋势,海拔2780m处理当归体内POD活性最低,比海拔2360m和2570m处理分别低26.1%和31.5%,差异达显著水平（P<0.05）,三个海拔间POD活性降低幅度随海拔升高而增加,在海拔2360m基础上,海拔2570m处理浮动4.27%,而在海拔2570m基础上,海拔2780m处理浮动24.0%。三种保护酶活性与MDA含量间相关分析表明, CAT与MDA呈显著的正相关关系（R²=0.3068）,且三种保护酶间均呈极显著的正相关关系（R²>0.96）。5.当归根中阿魏酸含量随海拔升高而升高,海拔2780m处理比海拔2360m处理增加14.5%,差异达显著水平（P<0.05）,而海拔2570m和其他两个海拔间差异均不显著;阿魏酸含量在海拔梯度间的变化幅度随海拔升高而降低,依次为8.6%和5.5%,但差异不显著（P>0.05）。6.当归根中挥发油收率、挥发油含量和藁本内酯相对含量均随海拔升高而增加。当归挥发油收率海拔2360m处理最低,为0.86%,海拔2780m和2570m处理分别比低海拔2360m处理显著高于14.0%和11.6%;三个海拔处理当归挥发油含量分别为85.55%、95.16%和96.67%,且GC-MS成分分析表明,挥发油主要化学成分为藁本内酯,三个海拔处理相对含量分别为58.72%、60.91%和61.59,海拔间差异不显著（P>0.05）,但对藁本内酯和其异构物含量的综合分析表明,海拔2570m（64.28%）和2780m（65.29%）处理比海拔2360m处理（58.99%）分别高出8.97%%和10.68%,海拔2780m与2360m处理间差异显著（P<0.05）。7.通过相关分析表明,影响当归产量和品质的关键生态因子均为温度和降雨量,相关程度为0.7417～0.9714,其次是日照时数;影响当归产量的关键生理生化因子为MDA （R²=-0.9118）,其次为可溶性糖（R²=-0.8831）,而影响阿魏酸的关键生理生化因子为可溶性糖（R²=0.9749）,其次为SOD（R²=0.8408）,影响挥发油的关键生理生化因子为可溶性糖（R²=-0.8990）和MDA（R²=-0.8958）,其次为SOD（R²=-0.7226）;影响当归产量的关键光合参数为胞间CO₂浓度（R²=-0.7393）和净光合速率（R²=-0.7332）,其次是空气湿度（R²=-0.5955）、气孔导度（R²=-0.5553）和光合有效辐射（R²=-0.5277）,而影响阿魏酸的关键光合参数为空气相对湿度（R²=0.9699） ,其次为光合有效辐射（R²=0.9467）、Chlb（R²=0.9380）和气温（R²=-0.9050）,影响挥发油的关键光合参数为胞间CO₂浓度（R²=-0.9706）和净光合速率（R²=-0.96849）,其次为空气相对湿度（R²=0.9060）、气孔导度（R²=0.8839）、光合有效辐射（R²=0.8682）和空气CO₂浓度（R²=0.8598）。综合分析,影响当归产量和品质的关键光合参数为胞间CO₂浓度、净光合速率、空气相对湿度以及光合有效辐射和空气CO₂浓度。综合以上研究结果认为,在试验设计范围内,随海拔升高,当归产量增加、品质改善。从生态因子角度分析,降低温度、增加降雨量有利于当归产量和品质形成;从生理生化因子分析认为,降低膜质过氧化产物MDA、渗透调节物质可溶性糖含量以及保护酶SOD活性,有利于当归产量和品质形成;从光合参数角度分析,大气相对湿度增加、有效光辐射增强和空气CO₂浓度增加均有利于产量形成和品质改善。因此,在2300m～2800m范围内,建议扩大高海拔当归栽培面积。更多还原

【Abstract】 Angelic sinensis is one of the genuine medicinal materials of Min County in Gansu province,and Min Angelic sinensis renowned at home and abroad because of its high quality. However,the limited appropriate ecotopes and traditional customs of planting Angelic sinensis on relatively smooth and low altitude land led to the lack of planting area,and further led to the situation that the supply is inadequate to meet the demand. In order to explore the appropriate planting area of Angelic sinensis from existing ecological resources,ecological adaptability tests （field and pot experiment） were conducted in Chabu country of Min County in Gansu province during 2009-2010. In this study,we discussed the difference of yield and quality of Angelic sinensis and revealed the influencing mechanism of altitude on yield and quality of Angelica sinensis from protective enzyme,osmotic regulation substances and photosynthetic characteristics,to look for the key factors and its influence size to effect yield and quality of Angelic sinensis. The main results are as follows:1. Altitudes effected the Angelic sinensis yield formation through changing sink-source relationships and dry matter accumulation rate. The partitioning of photosynthates to root arrived ahead of time and dry matter accumulation rate increased with the increase of altitude. Partitioning of photosynthates in Angelic sinensis of 2780m altitude to root began 10 days ahead than the others and the percentage of photosynthates distribute to root was much more than the latter in later growth stage,which a solid base for the yield formation of Angelica sinensis in harvest period. Biological yield increased by 33.1%,43.7% and 46.7%,respectively,with the increase of altitude,compared with than in flat land. The sequence of Angelic sinensis root yield was 2570m（28.4g/plant）,2780m（26.6g/plant） and 2360m （21.8g/plant）,and the root yield of 2360m treatment significantly decreased by 22.6% and 20.8% compared to the other altitudes（P<0.05）.The variance of root yield and dry matter accumulation rate reduced with the increase of altitude.2. Altitudinal gradients significantly influenced the photosynthetic pigment content and photosynthetic parameters of Angelic sinensis. The carotenoids,net photosynthetic rate（Pn）,stomatal conductance（Gs） and intercellular CO₂ concentration（Ci） were decreased,but the chlorophyll content was increased and transpiration rate（Tr） increased in 2570m treatment then decreased Along with altitude elevating,Carotenoids content decreased by 14.3% and 55.4%,mean net photosynthetic rate decreased by 10.0% and 18.4%,stomatal conductivity decreased by 5.3% and 16.0%,mean intercellular CO₂ concentration decreased by 7.6% and 14.0%,chlorophyll-a increased by 7.9% and 70.5% and chlorophyll-b increased by 21.1% and 75.8%,compared 2570m and 2780m treatments with 2360m treatment. Mean transpiration rate significantly increased by 13.3% in 2570m treatment and significantly decreased by 13.3% in 2780m treatment compared to that in 2360m treatment.Analysis of environment factor influencing the net photosynthetic rate showed that the air relative humidity was the key factor,followed by photosynthetic available radiation （PAR） and air CO₂ concentration.3. The excessive accumulation of active oxygen metabolism caused by high temperature in low altitude area led to the formation of MDA which is the membranous peroxide and can produce a toxic effect on cells in Angelica sinensis. Angelic sinensis reduced the toxicity of MDA by increasing the content of cell osmotic regulation substances and this was consistent with Angelic sinensis’properties of being fond of cool ecological conditions. MDA content decreased with the increase of altitude that in 2360m treatment was significantly increased by 52.4% and 38.1% compared with that in 2570m treatment and 2780m treatment （P<0.05）;The reducing range between treatments decreased with altitude elevating,it was 52.4% and 30.0%. Free proline content showed the altitude effect of "high—low—high" and there was no significant difference between the treatments in 2360m and 2780m,but free proline content in the two treatment above was significantly higher than that in 2570m,and increased by 12.4% and 13.6%,respectively. The variation ranges of free proline content between treatments increased by 12.4% and 13.6%with the increase of altitude. Soluble sugar content of 2360m and 2570m treatments were significantly higher than that of 2780m by 19.7 % and 14.7% （P<0.05）,while the difference between the treatments of 2360m and 2570m was not significant（P>0.05）,the variance range in three altitudes were increased by 7.7% and 14.7% with altitude elevatsing. The analysis of the relationship between osmotic regulation substances and membranous peroxidation product showed that the change of osmotic regulation substances content was in accordance with the change of MDA content,and there was a complementary relationship between the content of osmotic regulation substances.4. Low altitude caused active oxygen metabolism disordere,active oxygen accumulation and finally the formation of MDA in Angelic sinensis. In order to sheltered cells from the suffering of MDA,Angelic sinensis’cells increased the antioxidant enzymes activity to decompose peroxide. The studies showed that SOD activity reduced with the increase of altitude,and that in 2780m treatment was decreased by 40.8% and 39.3%,relatively,compared with that in 2360m and 2570m treatments （P<0.05）. The reducing range of SOD activity in three altitudes were increased with altitude elevating,and that in 2570m treatment was significantly decreased by 2.5% compared to that in 2360m treatment and that in 2780m treatment was significantly decreased by 39.3% compared to that in 2570m（P<0.05）. CAT activity reduced with the increase of altitude and that in 2780m treatment significantly decreased by 13.6% and 17.1% compared to that in 2360m and 2570m treatments （P<0.05）. The reducing range of CAT activity in three altitudes were increased with the increase of altitude,it was 13.6% and 14.6%. POD activity showed the tendency of rising at first and then decreased with the increase of altitude. POD activity in 2780m treatment was lowest and lowered by 26.1% and 31.5% compared to that in 2360m and 2570m treatments and the difference between them were significant （P<0.05）. The reducing ranges of POD activity in three altitudes were increased with the increase of altitude and it was 4.27% and 24.0%,respectively. The correlation analysis between protective enzyme and MDA showed that the content of MDA and CAT had a significantly positive correlation （R²=0.3068） and the POD,SOD and MDA are significantly positively correlated with each other （R²>0.96）.5. Ferulic acid content in Angelic sinensis root was increased with the increase of altitude,Ferulic acid content of Angelic sinensis in 2780m treatment was significantly increased by 14.5%（P<0.05） compaed with 2360m treatment,while there were no significant difference between 2570m treatment and the other two altitudes. The variation scope of ferulic acid content decrease with the increase of altitude and there were no significant difference between them.6. The yield and amount of essential oil,relative ligustilide content in Angelic sinensis increased with the increase of altitude. Yield of essential oil in Angelic sinensis in 2360m treatment,was 0.86% and it was the lowest in all treatments and was decreased by 11.6% and 14.0% compared to that in 2570m and 2780m treatments,respectively. Essential oil content under three altitudes were 85.5%,95.16% and 96.67%. GS-MS analysis showed that ligustilide was the main chemical compositing of essential oil and the relative content in Angelic sinensis of three altitudes were 58.72%,60.91% and 61.59%,respectively,and there were no significant difference between them. Synthetic analysis of ligustilide and its isomer showed that the content in 2570m treatment （64.28%） and 2780m treatment （65.29%） were increased by 8.97% and 10.68% compared with that in 2360m treatment,respectively and the difference between that in 2780m treatment and 2360m treatment was significant（P<0.05）.7. Correlation analysis showed that the key ecological factors effecting the yield and quality of Angelic sinensis was temperature and rainfall which has the correlation degree from 0.7417 to 0.9714,sunshine hours. MDA and soluble sugar were the key physiological and biochemical factors effecting yield of Angelic sinensis （R²=-0.9118） and the determination coefficients were -0.9118 and -0.8831,respectively. The physiological and biochemical factor effecting ferulic acid of Angelic sinensis,the soluble sugar was of first importance （R²=0.9749） and then followed by SOD （R²=0.8408）. The physiological and biochemical factors effecting essential oil of Angelic sinensis,soluble sugar （R²=-0.8990） and MDA （R²=-0.8958） were of first importance and then followed by SOD （R²=-0.7226）. In the photosynthesis parameters,that effecting the yield of Angelic sinensis,Ci（R²=-0.7393） and Pn（R²=-0.7332） were the key factors and followed by RH （R²=-0.5955）,Gs（R²=-0.5553） and PAR（R²=-0.5277）,in that effecting ferulic acid,RH（R²=0.9699） was the key factor and followed by PAR（R²=0.9467）,Chlb （R²=0.9380）and air temperature（R²=-0.9050）,and in that effecting effect essential oil of Angelic sinensis,Ci（R²=-0.9706） and Pn（R²=-0.96849） were the first important,followed by RH,Gs,PAR and air CO₂ contentration. Comprehensive analysis showed that the key photosynthesis parameters effecting the yield and quality of Angelic sinensis were Ci, Pn, RH, PAR and air CO₂ contentration.Based on the results described above, the yield and quality of Angelic sinensis increased with the increase of altitude. From the angle of ecological factors, the reduce of temperature and the the increase of rainfall are beneficial to the yield and quality of Angelic sinensis. From the angle of physiological and biochemical factors,the reduce of MDA content,soluble sugar content and protective enzyme activity are beneficial to the formation of the yield and quality of Angelic sinensis. From the angle of photosynthesis parameters,the increase of air relative humidity,photosynthetically active radiation （PAR） and air CO₂ density are beneficial to the yield increasing and quality improving. So,in the altitude range from 2300m to 2800m,we suggested expanding the cultivation areas of Angelic sinensis.更多还原