| Tripterygium wilfordii Hook.f. is the Tripterygium plant of Celastraceae, which is an important toxicnatural medicine resource and a versatile high economic value species. Taining County, Fujian Province isthe largest production base of T.wilfordii Hook.f. in China. Although the production technology of T.wilfordii Hook.f. has become more mature, but there is still some problems in the production, such as heavediseases, nonstandard cultivation methods and fertilization techniques. Therefore, the syudy on GAP keytechnologies of T. wilfordii Hook.f. is very urgent. To achieve T. wilfordii Hook.f. medicine “real, highquality, stable, controllable,” and to promote the need of the standardization, intensivism, modernizationand internationalization of traditional Chinese medicine, GAP key technology of Fujian genuine Chinesemedicinal materials T. wilfordii Hook.f. was reseached.This study started from to evaluate the environmental quality of GAP base for T. wilfordii Hook.f.GAP base; then collect the domestic T. wilfordii Hook.f. resources, to study selective improved variety andgood seedlings breeding technology for T. wilfordii Hook.f. At the same time, fertilization effect,cultivation key technologies, photosynthetic characteristics, resistance to cold damage and other aspects forT. wilfordii Hook.f. were studied. The results were as follows:1. The ranking prior to comprehensive scores of soil nutrients in sixteen T. wilfordii Hook.f. maindistribution areas in Tai Ning county were Dayang Village of Meikou(Jiangkeng), Chongji VillageXialankeng of Shangqing, Changxing Village of Shancheng Town. The comprehensive score was insequence of1.7174,1.0247,0.7638. By comprehensive index method of soil fertility, the prior rankingswere Dayang Village of Meikou, Shengyi Village of Shancheng Town(Zhinongkeng), Guantian Village ofLonghu, Changxing Village of Shancheng Town. The results using the two methods showed that the lowestsoil fertility was Yuxi Work Area in Qiuhong Village of Shancheng Town. In annual dynamic change of soilnutrients, soil pH value, total K, N, available P and organic matter content iapproximately increased year byyear in Changxing Village, Yuxi Work Area, Zhinongkeng and Jiangkeng of T. wilfordii main distributionareas from2009to2011, while others decreased year by year. Soil pH value and total K content ofJiangkeng were the highest in the three years, were5.75,4.22g·kg~-1. The soil total P and organic mattercontent of Zhinongkeng were the highest in the three years, the highest value followed by0.55g·kg-1,103.62g·kg~-1. And hydrolysable N content of Changxing Village was the lowest, the minimum in2011,only32.11mg·kg~-1.2. In2011January to September, the soil pH value in Changxing Village seemed slightly higher,whose changes was in the range of5.24-5.69. The range of total K content in the main distribution areas was little in February to June, relatively gently, which was the lowest in January. The highest was inJiangkeng in April, up to60.35g·kg~-1, and the minimum was in Zhinongkeng, only0.94g·kg~-1. The soil totalP content in Changxing Village, Yuxi Work Area and Jiangkeng changed more mildly than that ofZhinongkeng. The total P content of Zhinongkeng was the highest, reaching0.28g·kg~-1in June. Soilavailable K content in the four main distribution areas approximately increased first and then decreased, thelowest was Yuxi Work Area, only5.37mg·kg~-1in January. The trend of total N content and soil available Pcontent in Changxing Village and Yuxi Work Area was consistent. Jiangkeng’s total N content at theirhighest in February, up to4.29g·kg~-1.3. The environment present situationis of Changxing Village, Jiangkeng and Zhinongkeng in plantingbase of T. wilfordii of Tai Ning county was good. There were no industrial and mining enterprises, withoutany industrial pollution, with good air quality. Soil fertility was lower-middle. Comprehensive pollutionindex of Jiangkeng, Zhinongkeng and Changxing Village was respectively0.33,0.28,0.40, less than0.5.Cultivated soil had no toxic ions and dumping enrichment, no heavy metals pollution. Both single pollutionindex of every pollutants in irrigation water and in air were less than1, of which was suitable forcultivation of T. wilfordii in the overall assessment. Changxing Village of water sources being dynamicmonitored, it found that due to the catastrophic floods in Taining County in2011, the indexes suchas suspended matter, volatile phenol and chemical oxygen demand appeared transient abnormalityin the water, were232.47mg·L~-1、49.7mg·L~-1、276.33mg·L~-1. They have recovered after6-12months,which were in line with national standards. However, copper content in the water of ChangxingVillage was always a little high, some even as high as38.997mg·L~-1.4. For safety evaluation of T. wilfordii Hook.f., the results showed that the residue of BHC, DDT,Pentachloronitrobenzene (PCNB), aldrin, total bacteria, coliform bacteria in the roots and leaves of T.wilfordii Hook.f. were all lower than the concentration limits of corresponding national standards. Inaddition to the slightly high copper content in the leaves of T. wilfordii Hook.f., up to25.00mg·kg~-1. The Pb,Cd, Cu, As, Hg residues in its roots were below the concentration limit of corresponding national standards.The test results could ensure the safety of medication for T. wilfordii Hook.f..5. Using quadratic regression orthogonal combination design of four factors, the results showed thatusing the concentration of78.64%methanol, solid-liquid ratio of1:35.96, ultrasonic time of39.1min, theamount of9g chromatography alumina, the maximum extraction of available triptolide was predicted to be0.0192%. According to the three cluster analysis of the leaf biomass, leaf and root triptolide content of T.wilfordii Hook.f. species, elite provenance was selected. The results showed that32kinds of mean leafbiomass was132.0g. Leaf biomass and root lactone alcohol content of Taining of Fujian Sanming were thehighest, respectively200.6g,26.37mg·kg~-1. The highest leaf triptolide content was in Datian of FujianSanming, up to17.25mg·kg~-1. The excellent provenances when these three conditions were met as follows:Taining, Datian, Qingliu of Fujian Sanming, Wuyishan of Fujian Nanping, Jingling of Zhejiang Lishui, the scores was in sequence of2.7733,2.6880,2.2472,2.0626,2.0041.6. Analysising triptolide content of5a,6a,7a,8a,9a in different tissues, the triptolide content of theleaves that would fall off (“defoliate”for short) was much higher than other organizations, the highest levelswas up to74.92mg·kg-1in7a, which was2.72times of average leaves triptolide content. The triptolidecontent of flower was the lowest, only9.19mg·kg-1in7a. The triptolide content of T. wilfordii Hook.f. mainbranch was higher than lateral branch in the same year. The highest was in7a, main branch was22.78mg·kg~-1, lateral branch was17.05mg·kg~-1. The order of triptolide content in each year roots was the>fibrous root> root core, the highest was45.09mg·kg-1in main root of7a. The triptolide content in根皮washigher than main root in5a,6a,7a, but was opposite in8a and9a, the root bark7a. was the highest, up to46.87mg·kg~-1.7. The cuttage method helped to improve cutting survival rate during November to January, with0.5-0.7cm cutting coarseness, using yellow soil or1:2perlite and yellow soil as substrates, under conditionswith concentration100mg·L-1available rooting powder soaking1h, whose cutting survival rate was up to88.0%. The correlation of cutting survival rate and temperature was max, whose correlation coefficient was-0.98, indicating that the higher the temperature, the survival rate of cutting for T. wilfordii Hook.f. was thelower.8. Using L9（43）orthogonal design, the effect of precursor substance on triptolide contents for T.wilfordii Hook.f. tissue culture seedlings was studied. Under lighting conditions, the triptolide content andof treatment2tissue culture seedling was the highest, up to59.44mg·kg~-1, whose comprehensive score wasthe highest, up to0.8536. Under shading conditions, the comprehensive score of treatment4was thehighest, up to0.8542. When it was shade, the best combination effect was the treatment of0.5mg·L~-1lysineand1.0g·L~-1yeast extract. But no matter the light or not light, adding1.0g·L~-1yeast extract was beneficialto secondary metabolism products of tissue culture. In the light and shade conditions, the average triptolidecontent of tissue culture seedlings with precursor substance were respectively33.22mg·kg~-1,31.40mg·kg~-1,and the average whole plant biomass was2.4506g,2.2504g, respectively, higher than that of CK1.36times,1.24times.9. N, K, Ca, Mg content in roots was low in January to Months. P content in roots was generally lowin every month, N, K, Ca content in which was less than10g·kg~-1, P and Mg content in which was lessthan1g·kg~-1. N, K, Mg content in leaves was higher than in roots. P content in leaves was slightly higherthan in roots. The N content of9a in roots and leaves were the highest in April, respectively, up to22.31g·kg~-1,31.69g·kg~-1. The root and leaf Ca content of1a in September were the highest, respectively15.01g·kg~-1,26.70g·kg~-1; Root P content of5a in February was the highest, up to1.3g·kg~-1. Root K contentof2a was the highest in June, up to24.03g·kg~-1. Root Mg content of5a was the highest in April, up to7.63g·kg~-1. Leaf P content of2a was the highest in September, up to1.31g·kg~-1. Leaf K content of5a inApril, up to61.27g·kg~-1. Leaf Mg conten of9a was the highest in April, up to6.90g·kg~-1. 10. Using the quadratic regression current rotation compose design, the quantities of nitrogen,phosphorus and potassium fertilizer with leave biomass and triptolide contents for planting six years’ T.wilfordii Hook.f. leaf and root were obtained. The results showed that the optimal combination for urea wasfound to be51.4-68.6kg·hm~-2, calcium superphosphate70.8-97.9kg·hm~-2and potassium sulfate111.0-154.1kg·hm~-2, biomass for T. wilfordii Hook.f. leaf was437.3-456.1g, which was1.7-1.8times of the biomass ofcontrol group. The fertilizations affected triptolide content in the order, N>P>K. The effect of nitrogenfertilizer on increasing triptolide content of leaf was relatively large. The optimal combination by computersimulation for urea was found to be66.3-86.4kg·hm~-2, calcium superphosphate93.9-119.1kg·hm-2andpotassium sulfate124.0-164.1kg·hm~-2, triptolide content for T. wilfordii Hook.f. leaf was90.462-110.541mg·kg~-1, which was4.2-5.1times of the triptolide of control group. The optimal combination for urea wasfound to be55.2-77.6kg·hm~-2, calcium superphosphate60.6-78.5kg·hm-2and potassium sulfate69.9-87.8kg·hm~-2, triptolide content for T. wilfordii Hook.f. roots was47.474-53.661mg·kg~-1, which was1.73-1.96times of the triptolide content of control group.11. The results showed that1a T. wilfordii Hook.f. seedlings after spraying fertilizer, the average plantheight and the longest length of vine were40.62cm,28.09cm, higher than control group. The root biomasswith spraying compound microbial agents leaching solution (P7) was the highest,600times Guoguangamino acid leaf fertilizer (P11) followed by43.70g,39.83g, that of CK (P16)4.22times,3.84times. Theaverage root, branch, leave triptolide content with chemical treatments were higher than CK, respectively52.167mg·kg~-1,16.677mg·kg~-1,51.281mg·kg~-1. The order that spraying pesticides impacting triptolidecontent was leaf>root> branch. The triptolide content of treatment11was the highest, up to164.84mg·kg~-1.Spraying with0.2%potassium dihydrogen phosphate (P2) the four kind of roughness of0.1-0.2cm,0.2-0.4cm,0.4-0.6cm and4.0-6.0cm in the root distribution were the most, respectively accumulated to461.583cm,134.345cm,53.907cm,9.479cm, were that of CK3.96,3.82,3.22,4.55times. Spraying600times Guoguang amino acid leaf fertilizer (P11) processing the roughness of0-0.1cm,0.6-0.8cm,0.8-1.0cm,1.0-2.0cm,2.0-4.0cm and greater than6.0cm in the root distribution were the most, respectivelyaccumulated up to4.407cm,48.884cm,37.078cm,90.644cm65.002cm and2.014cm, were that of CK3.79,2.80,2.66,2.77,3.55and21.84times. It can be seen that spraying600times Guoguang amino acids leaffertilizer and0.2%potassium dihydrogen phosphate (P2) wass the most beneficial to root growth, whoseroot was the most developed.12. The planting spacing of1.5m×2m was the best planting density, which leaf triptolide content wasthe highest, up to21.43mg·kg~-1, promoting the improvement of yield and quality of T. wilfordii Hook.f.. Itis recommended that afforestation mode of T. wilfordii Hook.f. mainly used mixed mode of T. wilfordii×Magnolia officinalis and pure forest model. The average triptolide content in the roots and leaves of themixed mode of T. wilfordii×Magnolia officinalis reached the maximum, respectively38.86mg·kg~-1,35.61mg·kg~-1. The average diameter of mixed Magnolia and Chinese Fir were5.36cm,16.01cm, higher than the control18.92%、35.79%. The average tree height of Magnolia is higher than pure forest22.35%.13. Using the L16(45) orthogonal experiment, irrigation situation of1-year T. wilfordii Hook.f. wasstudied. The results showed that the size of soil temperature was June>May>November>December. Thesize of plant water content with irrigation treatment was roughly May>June> July>August, respectively77.39%,72.07%,67.08%,65.22%. The size of plant water potential with irrigation treatment was roughlyOctober>September>August>June, the average water potential was respectively-1.874MPa,-1.875MPa,-2.147MPa,-2.906MPa. Irrigation treatment can improve the triptolide content of roots and leaves. Theroots triptolide content of treatment16was up to28.20mg·kg~-1, and leaves of treatment13was the highest,reaching14.51mg·kg~-1. The average root heave, number and longth of each irrigation treatment were9.53g,7.5,15.5cm. Ranking prior was treatment3(irrigation amount of20mm from Month to June, amount of60mm from July to October, amount of30mm from November to December) which score was1.8722.14. The soil sterilization with quick lime could promote the growth of T. wilfordii Hook.f., which rootsand leaves triptolide content was respectively14.62mg·kg-1,12.77mg·kg~-1. Leaf disease index sprayingchemical treatment was lower than the control, of which spraying400times of65%amobam WP was thelowest, only14.56%. The average triptolide content of roots and leaves spraying chemical treatment washigher than the control. The highest average root triptolide content with spraying65%Zineb wettablepowder was the highest, up to17.18mg·kg~-1.15. The triptolide content in roots and leaves first increased and then decreased with ages for T.wilfordii Hook.f.. The root triptolide content of8years old was the highest, up to35.48mg·kg~-1, and6-year-old leaves triptolide content was the highest, up to21.60mg·kg~-1. Therefore, the root of T. wilfordiiHook.f. was determined to dig after being planted seven years. T. wilfordii Hook.f. leaves can be harvestedafter3-8years, but six years later to pick was the best. The root triptolide content of7years old plantsincreased significantly on the end of October, which was the highest in December, up to36.8mg·kg~-1.Therefore, it is suggested that the best period excavation of the roots of T. wilfordii Hook.f. was in the lateautumn to winter first. T. wilfordii Hook.f. leaf was determined to pick in July to August each year.16. As the drying temperature increases, the triptolide content of roots showed an increased first andthen downward trend, the triptolide content of which using60℃drying was the highest, up to30.26mg·kg-1. The effect of three kinds of drying methods on the quality of T. wilfordii Hook.f. medicines waslittle. Difference in moisture content only reached an extremely significant level, and difference oftriptolide content in the roots reached a significant level. It is recommended that drying shoud use dryingbox or mixed cross-drying method.17. The light saturation point of T. wilfordii Hook.f. with seasons showed an “increase-decrease-increase” trend. In June, the LSP of1-3a T. wilfordii Hook.f. was large, whose order was1a<3a<2a, the2aof which was the highest, reaching4650.757μmol·m~-2·s~-1. In general, light compensation point of T.wilfordii Hook.f. showed an increased and then decreased trend with seasons. In June, the LCP of1-3a T. wilfordii Hook.f. was large, the size of the order:1a>2a>3a,1a of which was the highest, reaching61.001μmol·m~-2·s~-1. The LCP of1a and3a were the lowest in October,3a of which was the lowest, only13.608μmol·m~-2·s~-1. The size of the order between1a Pnwas generally August>June>April>October. Thesize of order between Ci and Ls of1a and2a T. wilfordii Hook.f. each month was generally October>August>June>April. The size of the order between1a Trand2a WUE was generally August>June>October>April.18. One year-old T. wilfordii Hook.f. cutting seedlings leaves were sprayed with0,5,10,15,20,25mg·L~-1exogenous ABA under low temperature stress. The results showed that spraying15-20mg·L~-1ABA improved chilling resistance, effectively reducing relative electrical conductivity and theaccumulation of MDA, which increased free proline content of T. wilfordii Hook.f. cuttings, weakened theeffect of low temperature stress on SOD、POD、CAT enzyme activities. Relative electrical conductivity andMDA content of15mg·L~-1ABA were the lowest, which were45.74%and153.28nmol·g~-1respectively.Proline content raised by72.99μg·g-1compared with the control. When ABA content was20mg·L~-1,chilling injury degree was significantly lower than other treatments. SOD activity with ABA treatmentshowed obvious double peak tendency with the prolongation of stress time. POD activity showed the singlepeak tendency first increasing then decreasing. When ABA content was15and20mg·L~-1, CAT activityshowed “decreasing-increasing-decreasing” pattern.19. Spraying20mg·L~-1ABA slowed down the decrease amplitude of Pn, Tr, Gs, Ciin low temperature,increasing photosynthetic capacity of T. wilfordii seedling leaves. After6days exposure to low temperaturestress, Fodecreased, Fmand Fv/Fmshowed an upward trend with the rise of ABA concentration as a whole.ΦPSand qPshowed a downward and then upward trend, qNshowed “decreasing-increasing-decreasing”pattern. At20mg·L~-1ABA, Pn, Gs, qP, Fmand Fv/Fmreached their peak values. RETR showed thetendency firstly increasing and then decreasing as PAR increased. When PAR was395μmol·m~-2s~-1, rETRof every treatment reached the highest value.25mg·L~-1ABA was the highest,20mg·L~-1ABA took thesecond place, which raised by17.1%and5.2%compared with the control respectively. The lightresponse curves of ΦPSⅡdecreased as PAR increased, which of qNperformed reserved. |
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