【作者】 杨细明；

【导师】 洪伟；

【作者基本信息】 福建农林大学 ， 森林培育， 2008， 博士

【摘要】 以福建省泰宁县的雷公藤野生植株和人工林为研究对象,开展雷公藤的个体生态学和优良无性系选育技术研究,为藤本植物雷公藤的驯化栽培提供基础研究背景,为藤本植物、药用植物的遗传改良提供理论支撑。根据泰宁县农业区划和土壤普查资料,对泰宁县全境雷公藤自然资源分布状况进行调查;以泰宁县芦下、江坑为观测点,开展雷公藤物候观察;以芦下天然起源雷公藤为研究对象,研究野生雷公藤的种内变异;以泰宁县玉溪的雷公藤人工林为研究对象,研究雷公藤的个体生态学规律,并以之为选择群体,开展优良单株选择技术研究;以选出的优良单株为材料,在福建省林业科学研究院采用根扦插技术进行无性系化,并进行无性系苗期生长测定;采用美国CID公司生产的CI-301 CO₂ GAS ANALYZER研究无性系苗木的光合生理特性;采用ELISA法、高效液相色谱法分别研究无性系苗木的内源植物激素、雷公藤甲素含量变异规律;以扦插时间、扦插材料、扦插基质和生长激素为试验因素,开展雷公藤扦插技术研究。本文的主要研究结果:雷公藤主要分布区的自然环境特点:①气候:＞10℃年积温5000～5500℃,年降雨量1700～1800mm,相对湿度80～86%。②海拔:海拔高度300～500m的高丘地带为雷公藤的适宜分布区;海拔200～300m的河谷地和500～800m的低山为雷公藤的一般分布区;海拔800～1700m的中山为雷公藤的边缘产区。③地形:在300～500m垂直高度上,雷公藤的生长状况,缓坡优于陡坡,南坡优于北坡,下坡优于中、上坡。④土壤:雷公藤自然分布的土壤类型中,红壤占90%,黄壤占5%,紫色土占5%;土壤的PH值为5～6.5,平均为6,显示雷公藤喜弱酸性土壤。⑤植被:雷公藤植物群落中共分布各类植物60科111属135种,其中蕨类植物8科11属11种,裸子植物4科4属4种,被子植物48科96属120种（双子叶植物41科77属98种,单子叶植物7科19属22种）。一年中,雷公藤的生长季约300d,落叶休眠期约65d。生长季为2月中旬～12月中旬,期间藤条生长呈现出“快—慢—快”生长节律,对应于2个藤条生长期,分别称之为春梢生长和秋梢生长。春梢抽梢期约120d,其间约100d为速生期;春梢生长量呈现出下坡＞上坡现象。秋梢抽梢期约70d,其间约50d为速生期;秋梢生长量小于春梢。多年生雷公藤4月中旬现蕾,5月中上旬初花,果实成熟于9月中下旬;上坡和下坡的生殖节律基本一致;多年生雷公藤每年开花结实1次,结实量呈现大小年交替变化规律。野生雷公藤种内存在丰富的变异,冠幅、枝数、地茎、主藤长的变异系数分别为55.73%、59.52%、38.27%、53.2%;野生雷公藤各生长性状存在极显著的相关性,冠幅与地茎、冠幅与主藤长、地茎与主藤长的相关性达到极显著水平,冠幅与枝数的相关性达到显著水平。不同坡位雷公藤根系的分布量都呈现出随土层深度的增加而递减的规律;雷公藤根系生长具有坡位效应,下坡、中坡根量大于上坡;根系的平均含水率为43.88%,呈现出下坡＞中坡＞上坡规律;根系的径级分配在不同坡位上均存在相同的规律,即粗根＞中根＞细根,粗根比例为62.18%;植株根、茎、叶生物量分别占全生物量的52.75%、37.67%、9.58%;植株的地径、平均藤径、枝数等生长性状与全生物量存在显著或极显著相关关系,通过最优方程的拟合,可以建立以枝数、地径、平均藤径、平均藤长为自变量的全生物量估测模型。雷公藤人工林选优的合理对比株数为16株。以枝数为选优目标,其选优标准为1.5倍比值;以地径为选优目标,其选优标准为1.45倍比值;以平均藤长为选优目标,其选优标准为1.45倍比值;以平均藤径为选优目标,其选优标准为1.85倍比值。最后用指数选优法从36株候选株中选出23个优良单株,入选率为1.05%,生长指标为林分平均水平的1.47～1.8倍。23个优良单株的根扦插总成活率为94.4%,生长量均超过对照;扦插苗生长呈现出“快—慢—快—停”年生长节律。苗木的枝数、地径、主藤长的生长差异达到显著性水平;枝数、地径的广义遗传力分别为0.3612、0.4664;系统聚类分析结果,23个无性系分为粗壮型、伸展型、中间型、普通型4个类型;以地径、枝数为主要选择指标,共选出苗期速生无性系10个,与23个无性系的平均数相比,其枝数的平均生长优势为10.17%,地径的平均生长优势为22.75%。雷公藤苗木净光合速率的日变化曲线呈不对称双峰曲线,出现“午休”现象;通过比较上、中、下部叶片的净光合速率,发现下部叶片更能准确反映植株的日变化规律;雷公藤对光的适应性属中等偏下,为中度耐荫植物,其苗木光补偿点为17u mol.m^-2.s^-1,光饱和点为600u mol.m^-2.s^-1;苗木的净光合速率与气温、叶温、光合有效辐射、蒸腾速率呈极显著正相关关系,与大气相对湿度、叶内湿度、胞内CO₂浓度、胞间CO₂浓度呈极显著负相关关系;主藤长伸长生长受净光合速率、蒸腾速率影响显著;雷公藤无性系间光合作用差异明显,编号为17、13、16、10、11、4、6的7个无性系具有较高净光合速率。雷公藤无性系间的IAA、GA、ABA、ZR四种内源植物激素含量具有一定差异性。差异程度:IAA＞GA＞ABA＞ZR。IAA与GA、IAA与ZR、GA与ZR之间呈现出极显著的正相关关系,说明这三种激素含量水平密切相关;ABA的含量与其余三种激素间没有特别的相关性,属独立因素。雷公藤无性系间的内源激素平衡性较差,对其生理活动和生长状态产生了一定影响。雷公藤苗木的根木质部的雷公藤甲素含量是根韧皮部的1.29倍,二者之间的相关性极其显著;无性系间的根韧皮部、根木质部的雷公藤甲素含量差异达到显著性水平;通过联合选择,共选出2个高雷公藤甲素含量无性系,并进一步选出1个兼有速生及高雷公藤甲素含量双优性状的优良无性系。雷公藤根扦插受季节限制不明显,枝条扦插成活率受夏季高温的影响较大,扦插最佳时间为2～3月或9月小阳春;在非适宜季节,根扦插可以选用ABT2号、ABT3号生根粉,枝扦插可以选用ABT1号生根粉进行促根处理;根扦插与枝扦插相比,存在成活率高、根系发达、生长好等优势;根扦插的最适取穗长度为6cm～8cm,枝扦插的最适取穗长度为12cm～15cm,并以枝条中段取穗为佳;根扦插对基质的适应性比枝扦插强,雷公藤扦插最经济适用的基质为沙壤土。本文通过雷公藤生长发育环境调查、物候观察、种内变异、根系分布规律、植株生物量分配规律及全生物量模型等基础研究,开展了雷公藤个体生态学、优良单株选择、苗期生长特性、光合特性、内源植物激素含量、雷公藤甲素含量等研究,选育出雷公藤苗期速生无性系10个,高雷公藤甲素含量无性系2个,兼有速生、高雷公藤甲素含量双优性状无性系1个,同时,探索总结了雷公藤扦插繁殖技术体系,制定了雷公藤育种策略,为雷公藤的遗传改良、良种化、产业化的发展提供了物质基础和技术保障,填补了国内外有关这一研究领域的空白。更多还原

【Abstract】 Based on the wild habitats and plantations of Tripterygium wilfordii（TW） in Taining County（TNC）,the studies focused on both autoecology research and clone selection so as to provide either the basic scientific data for TW’s domestication and the theoretic knowledge for the improvement of canes and medical plants.The overall survey of TW resources was carried out according to the data of TNC agriculture subdivision and soil survey and the phenological observation was implemented in the 2 spots which located on Luxia and Jiangkeng of TNC.The natural generated population of TW in Luxia was applied to explore the variation within the species,while the plantation in Yuxi for the study of TW autoecology and elite individual selection.In Fujina Forestry Academy,the clonalization of the selected plants was achieved using root cuttings,furtherly the growth and photosynthetic physiologic characters of clonal plantlets were tested using CI-301 CO₂ Analyzer,which made in USA.The content of endogenous growth regulators and triptolide in TW seedlings were measured by ELISA and high efficiency liquidphase chromatography（HELC） respectively.Also the clonal propagation by cuttings was experimented using date,cutting, medium and growth regulators as the elements.The following was the achievements.The natural prospects of TW main distribution region were as the following:①Climate:The annual accumulation of temperature higher than 10℃,rainfall and relative humidity is 5000～5500℃,1700～1800 mm and 80～86%respectively;②Altitude:The high hilly area with altitude 300 to 500m is the adequate distribution zone for TW,and the valley at altitude 200 to 300m and mountains with 500 to 800m as the general zone.The mountainous area with altitude 800 to 1700m is TW’s boundary zone.③Topography:In the elevation between 300 and 500m, TW grows better in light and southwide lope than in tiff and northwide. On the same slope,it grows best in the foot.Regarding on the micro-topography,it grows better in the concave than in the convex and platform.④Soil:The soil category of TW natural distribution allocated in the red soil 90%,yellow soil 5%and purple soil 5%.The average PH is about 6,varying from 5 to 6.5 and suggests TW prefers light acid soil.⑤Vegetation:In the TW community there are 135 plant species belong to 111 genus of 60 families,of which 11 fern species of 11 genera in 8 families,4 gymnosperm species of 4 genera in 4 failies,120 angiospermous species of 96 genera in 48 families（98 dicotyledonous species of 77 genera in 41 families and 22 amphibrya species of 19 genera in 7 families）.In a year-round,TW has 2 shoot growing periods which is in the spring and autumn respectively.The spring one lasts about 120d,of which growing peak longests 100d and the growth in the foot part of slope is more than that in the up.The autumn one is about 70d,of which growing peak lasts 50d.Generally the growth of spring shoot is bigger than that of autumn one.TW develops flower buds on middle April,starts flowering on early or middle May,fruits ripen on middle or late September.TW bears flower and fruit once a year and the fruit crop presents the rythum of plenty and poor alternativelyGreat inner-species variance were observed for TW and the variance index of crown width（CW）,branch number（BN）,ground diameter（GD） and main cane length（MCL） was 55.73%,59.52%,38.27%and 53.2% respectively.The growth traits were closely correlated with each other and very obvious correlation happened between CW and GD,CW and MCL,GD and MCL,while visible between CW and BN.The distribution of TW root system in different slope position decreases along the increment of soil depth.The slope position effect is obviously presented in TW root growth,which more roots in the bottom and mille than that in the up.The water content in TW root increases gradually from the bottom to the top of slope with an average at 43.88% and the distribution of roots at different diameter grades was simillar in different position on slope,i.e the roots with big diameter is more than that mediate and small and the big ones accounts for 62.18%of the total. The biomass of TW root,stem and leaf accounts for 52.75%,37.67%and 9.58%respectively.The correlation between biomass and GD,average cane diameter and BN is visible or very visible and a model for estimation of biomass was successfully formatted using BN,GD,ACD and ACL as the self elements by best formula.The reasonable select ratio for plus tree selection in TW plantation is 16.For BN as selection objective the reasonable select ratio is 1.5 times of the standard.And that for GD,ACL,and ACD is 1.45,1.45 and 1.85 respectively.In the end 23 elite trees were selected by exponant from 36 candidate trees at select ratio of 1.05%,of which the growth is 1.47 to 1.8 times of the stand average.The average survival rate of the cuttings from the 23 plus trees was 94.4%and no disease or insect damage was found.The TW cuttings grow rhythm is typical as "rapid- slow-speed -stop".The variance of BN,GD, ACL at 1.5-years-old plantlets is visible and the generalized heritibility of BN,GD is 0.3612,0.4664.The growth of the 23 clones is better than the checked and the 23 clones can be classified into 4 groups by hierachical cluster analysis,ie.the trong,stretch,mdiate and normal.A general correlation was found among the 3 growth traits in the seedling stage,of which GD is the central trait correlated with the others.Using GD and BN as the select criteria,10 elite clones were selected with rapid growth at nursery stage and their average growth gain of BN and GD was 10.17%and 22.75%respectively,compared to the average of the 23 clones.The daily variance of net photosynthetic rate in TW plantlets tends to be a two-peak curve with a pause at noon.Also the leaves in the down part were found to present the daily variance trend of net photosynthesis more precisely than that of both the up and middle and the results suggested TW is a mediate shade-resistant plant as its general or poor adaptability to sunlight with a photo compensation point at 17u mol.m^-2.s^-1 and saturation at 600u mol.m^-2.s^-1.The net photosynthesis rate was found correlated positively with air temperature,leaf temperature,efficient radiation of photosynthesis and vapouring rate,negatively with air relative humidity,leaf water content,concentration of CO₂ inner cell or between cell and invisibly with air CO₂ concentration and.The growth of TW main cane was influenced obviously by net photosynthesis rate and vaporescence rate but non-obviously by BN,GD growth and plant photosynthesis.Finally the photosynthesis variance among the TW clones was significant,of which clone 17,13,16,10,11,4 and 6 had higher net photosynthesis rate.Significant variance of endogenous hormone content was found among the TW clones regarding on IAA,GA,ABA and ZR and obvious correlation was found among the 4 endogenous hormones,of which very positive among IAA,GA and ZR.It suggested the content of 3 hormones is closely correlated with each other and the variance trend similar among different clones.By the way the ABA content is independent with the others.The hormones should play influential effect on the growth and development of TW plantlets by interaction or.The content of triptolide in the root xylem of TW clone plantlets was 1.29 times as that in the phloem and its variance between the 2 parts was visible,which was 2.09 and 3.1 times as those of the checked respectively. The triptolide contents of root xylem and phloem were correlated with each other very obviously,but the correlation was invisible between the triptolide contents of root and growth traits,photosynthesis factors and content of endogenous hormones.Fatherly 2 elite clones with high triptolide content were selected by index screening,of which one with double advantages,i.e.rapid growth and high triptolide content.Although seasoning was not very restricted for TW cutting propagation,the high temperature in the summer did affect the cutting survival rate.As a result,the best season for cutting is on February to March or in the ’little spring’ of September.No root promotor needed on February to March.Otherwise ABT2 or ABT3 was suitable for root cutting and ABT1 for branch.The plantlets from root cuttings seemed to grow more branches,better roots and growth than that from stems.Of the branch cuttings the highest survival rate was achieved using cuttings form meddle section,which followed by the bottom and the poorest from the upper.The most suitable section length for root cutting was 6 cm to 8 cm and that of branch 12 cm to 15 cm.More substrates could be used for root cutting than that for branch cutting and the sandy soil was the best economically.Based on the studies concerned with development environment survey,phonological observation,inner-species variance,root distribution, plant biomass allocation pattern and biomass model,the research covered an area including autoecology,plus tree selection,growth characters, photosynthesis traits and contents of endogenous hormone and triptolide at nursery stage.As a result,10 elite clones with rapid growth at nursery stage,2 clones with high content of triptolide and 1 clone with both rapid growth and high triptolide content were selected and the TW cutting propagation procedures and breeding policy were successfully developed. The results could provide the material base and technical assurance for TW breeding,improved varity deployment and industry development and filled the gap of research concerned in the domestic and abroad.更多还原