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中国科学院昆明植物研究所知识管理系统
Knowledge Management System of Kunming Institute of Botany,CAS
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0.05). For some populations, germination capacity in 12-h photoperiod was significantly higher than that in completed darkness（W-FD: P < 0.01, W-JD: P < 0.05）.Genetic variation within and among six populations was assessed using AFLP markers. Genetic diversity was higher at species level (PPL = 69.19%, HE = 0.221) than at population level (PPL = 26.22%, HE = 0.095, Is =0.140), and populations in southeast Yunnan were strongly differentiated from those in southwest Yunnan (Nei’s GST = 0.575; FST = 0.655). UPGMA analysis demonstrated a clear genetic division between the two populations from DeHong (SW Yunnan; D-JD and D-HG) and the four from WenShan (SE Yunnan; W-FD, W-LH, W-ML, and W-MG). Within-population genetic variation was significantly correlated with population isolation (r(PPL) = -0.94, P = 0.006; r(HE) = -0.85, P = 0.032; r(Is) = -0.87, P = 0.025), but not with population size (r(PPL) = 0.63, P = 0.178; r(HE) = 0.54, P = 0.268; r(Is) = 0.56, P = 0.249).","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.date.issued.year&advanced=false&query1=Time%2BTo%2BGermination&&fq=dc.project.title_filter%3ACraigia%5C+yunnanensis%5C+W.%5C+W.%5C+Smith%5C+%5C%26%5C+W.%5C+E.%5C+Evans%5C+%5C%28Tiliaceae%5C%29%5C+is%5C+an%5C+endangered%5C+deciduous%5C+tree%5C+species%5C+which%5C+has%5C+high%5C+scientific%5C+and%5C+economic%5C+value.%5C+C.%5C+yunnanensis%5C+is%5C+seriously%5C+threatened%5C+and%5C+has%5C+been%5C+pushed%5C+to%5C+the%5C+verge%5C+of%5C+extinction%5C+due%5C+to%5C+vegetation%5C+destruction%5C+in%5C+China%5C+and%5C+consequent%5C+contraction%5C+of%5C+its%5C+distribution.%5C+Hence%2C%5C+it%5C+was%5C+listed%5C+as%5C+a%5C+nationally%5C+rare%5C+and%5C+endangered%5C+plant%5C+in%5C+1999%5C+and%5C+has%5C+also%5C+been%5C+proposed%5C+as%5C+a%5C+second%5C-ranked%5C+plant%5C+for%5C+national%5C+protection%5C+in%5C+China%5C+and%5C+included%5C+in%5C+IUCN%5C+red%5C+list.%5C+As%5C+a%5C+scientifically%5C+important%5C+and%5C+valued%5C+tree%5C+species%5C+with%5C+endangered%5C+status%2C%5C+the%5C+wild%5C+populations%5C+of%5C+C.%5C+yunnanensis%5C+therefore%5C+represent%5C+is%5C+a%5C+genetic%5C+resource%5C+that%5C+must%5C+be%5C+conserved.%5C+To%5C+provide%5C+basic%5C+information%5C+for%5C+its%5C+conservation%2C%5C+the%5C+population%5C+dynamics%5C+and%5C+population%5C+size%5C+structures%2C%5C+pollination%5C+biology%5C+and%5C+breeding%5C+system%2C%5C+eleven%5C+fitness%5C-related%5C+characters%5C+and%5C+the%5C+genetic%5C+variability%5C+based%5C+on%5C+AFLP%5C+were%5C+comprehensively%5C+studied.%5C+The%5C+main%5C+results%5C+are%5C+summarized%5C+as%5C+follows%5C%3A%5C+A%5C+total%5C+of%5C+six%5C+wild%5C+populations%5C+of%5C+C.%5C+yunnanensis%5C+were%5C+found%5C+in%5C+two%5C+disjunct%5C+regions%5C+of%5C+Yunnan%2C%5C+i.e.%5C+WenShan%5C+%5C%28SE%5C+Yunnan%5C%29%5C+and%5C+DeHong%5C+%5C%28SW%5C+Yunnan%5C%29%2C%5C+from%5C+2005%5C+to%5C+2007.%5C+Additionally%2C%5C+in%5C+all%5C+but%5C+one%5C+of%5C+the%5C+populations%5C+we%5C+detected%2C%5C+mature%5C+trees%5C+were%5C+felled%5C+between%5C+2005%5C+and%5C+2007%2C%5C+so%5C+destruction%5C+of%5C+most%5C+of%5C+these%5C+populations%5C+is%5C+ongoing.%5C+Across%5C+the%5C+six%5C+populations%5C+of%5C+extant%5C+C.%5C+yunnanensis%5C+found%5C+during%5C+our%5C+study%2C%5C+the%5C+total%5C+number%5C+of%5C+mature%5C+%5C%28reproductive%5C%29%5C+individuals%5C+detected%5C+was%5C+584%5C+in%5C+2007%EF%BC%8Cplus%5C+larger%5C+numbers%5C+of%5C+seedling%5C+and%5C+resprouts%5C+from%5C+cut%5C+trunks.%5C+The%5C+result%5C+of%5C+surveying%5C+Population%5C+structure%5C+showed%5C+that%5C+there%5C+are%5C+two%5C+regeneration%5C+types%5C+which%5C+are%5C+seedlings%5C+and%5C+sprouts.%5C+Seedlings%5C+occurred%5C+abundantly%5C+in%5C+gaps%5C+or%5C+open%5C+areas%5C+and%5C+the%5C+size%5C+class%5C+frequency%5C+distributions%5C+were%5C+often%5C+discontinuous%2C%5C+and%5C+the%5C+same%5C+general%5C+pattern%5C+occurred%5C+in%5C+all%5C+the%5C+investigated%5C+populations%5C+for%5C+juveniles%5C+and%5C+adults.%5C+The%5C+numbers%5C+of%5C+seed%5C-origin%5C+individuals%5C+did%5C+however%5C+decline%5C+sharply%5C+with%5C+increasing%5C+size%2C%5C+indicating%5C+a%5C+high%5C+mortality%5C+rate%5C+going%5C+from%5C+seedling%5C+to%5C+sapling%5C+stage%5C+may%5C+be%5C+a%5C+problem%5C+for%5C+this%5C+species.%5C+Additionally%2C%5C+the%5C+cash%5C+crop%5C+cultivation%5C+and%5C+logging%5C+seriously%5C+threaten%5C+the%5C+survival%5C+of%5C+the%5C+species.%5C+We%5C+conducted%5C+field%5C+observations%5C+and%5C+artificial%5C+pollination%5C+experiments%5C+on%5C+the%5C+floral%5C+biology%2C%5C+pollination%5C+process%5C+and%5C+breeding%5C+system%5C+of%5C+Craigia%5C+yunnanensis%5C+in%5C+Fadou%2C%5C+Xichou%5C+county%5C+of%5C+Yunnan%5C+province.%5C+The%5C+lifespan%5C+of%5C+a%5C+single%5C+hermaphrodite%5C+flower%5C+is%5C+approximately%5C+3%5C-4%5C+days.%5C+A%5C+cyme%5C+has%5C+2%5C-9%5C+flowered.%5C+The%5C+flowering%5C+period%5C+of%5C+an%5C+inflorescence%5C+is%5C+usually%5C+5%5C-14%5C+days.%5C+The%5C+flowers%5C+of%5C+C.%5C+yunnanensis%5C+were%5C+protandrous.%5C+The%5C+stamens%5C+were%5C+within%5C+petal%5C-like%5C+staminodes%5C+in%5C+the%5C+opening%5C+flowers%5C+until%5C+the%5C+flower%5C+withered.%5C+Without%5C+touchment%2C%5C+the%5C+bractlike%5C+staminodes%5C+can%E2%80%99t%5C+open.%5C+Self%5C-pollination%5C+was%5C+partially%5C+avoided%5C+by%5C+temporal%5C+and%5C+spatial%5C+isolation%5C+of%5C+male%5C+and%5C+female%5C+organs%5C+within%5C+the%5C+same%5C+flower.%5C+However%2C%5C+autogamous%5C+and%5C+geitonogamous%5C+pollination%5C+is%5C+unavoidable%5C+because%5C+of%5C+the%5C+large%5C+number%5C+of%5C+flowers%5C+on%5C+a%5C+single%5C+tree%5C+and%5C+the%5C+action%5C+of%5C+pollinators.%5C+The%5C+values%5C+of%5C+both%5C+OCI%5C+%5C%28%E2%89%A54%5C%29%5C+and%5C+P%5C%2FO%5C+%5C%281381%5C%29%5C+and%5C+the%5C+results%5C+of%5C+bagging%5C+tests%5C+indicated%5C+there%5C+was%5C+no%5C+apomixes%5C+in%5C+C.%5C+yunnanensis%5C+and%5C+the%5C+breeding%5C+system%5C+of%5C+the%5C+species%5C+was%5C+outcrossing%5C+with%5C+partial%5C+self%5C-compatibili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a field trip at a brule in Shangri-La, a mixed population of Ligularia Cass. was found, which including L. subspicata (Bur. et Franch.) Hand.-Mazz., L. nelumbifolia (Bur. et Franch.) Hand.-Mazz., L. tongolensis (Franch.) Hand.-Mazz., L. cymbulifera (W.W.Smith) Hand.-Mazz., L. lingiana S.W.Liu, and also some individuals morphologically intermediate between L. subspicata and L. nelumbifolia. Hence, these intermediate individuals were preliminarily assumed as natural hybrids of the two Ligularia. According to their morphology, they’re assumed to form hybrids A and B. Through careful comparison of specimens in herbarium and those we collected, the inflorescence of putative hybrid A is close to L. nelumbifolia, but the shape of laminae are intergradation of L. subspicata and L. nelumbifolia; overall morphology of putative hybrids B is similar to L. nelumbifolia, but inflorescence color is as same as L. subspicata. Compared to L. nelumbifolia (39%) and L. subspicata (36.8%), the germination rate of putative hybrid B (45.7%) slightly higher than the two; but that of hybrid A is extraordinarily low (0.3%). One possible interpretation of the low rate is hybridization. 60 individuals were collected, including putative parents, other 4 species of Ligularia nearby, putative hybrid A and B. They were all direct sequenced of four cpDNA fragments, and direct sequenced or cloning sequenced of nrDNA ITS4-5. The results support that L. nelumbifolia and L. subspicata are parents of putative hybrid A, and the majority female parent is L. subspicata, L. vellerea may also be involved in the hybridization in some degree; the nuclear sequences of putative hybrid B have no superposition, and its chloroplast DNA sequences are identical with L. nelumbifolia, so putative hybrid B could not be hybrid; and there are backcross individuals exist among the putative parent L. subspicata. NewHybrids analysis of ISSR markers indicated that, the individuals of putative hybrid A are almost L. nelumbifolia and L. subspicata F1 hybrid generation (10/11), only 1/11 possibly backcross or other forms; all individuals of hybrid B are L. nelumbifolia; except one individual of L. subspicata as backcrossed, the other parent individuals are 100% reliable. This study focused on molecular evidence, complemented by ecological, reproductive and other characteristics, we demonstrated that the morphologically intermediate individuals’ origin, and the probability of belonging to each parental or hybrid class. And concluded that L. nelumbifolia and L. subspicata are the parents of putative hybrid A, L. vellerea may also be involved in the hybridization in some degree, hybrids mainly are the first generation, a few individuals may be involved in backcross, and most probably backcross with L. subspicata according to the anthesis, while the assumption of hybrid B is not 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stresses could limit plant growth, development and propagation. Abiotic stress refers to the negative impact factors to the plants, such as extreme temperature, drought, flood, salinity, irradiation, chemicals and so on. To understand the mechanism of abiotic stress is very important.Membrane is the most sensitive organs in the cell that response to environmental changes. Cells respond and transduct environmental signals by changing content of membrane lipids and membrane proteins. The activity change of membrane phospholipase D (PLD) and the composition and content of membrane lipid molecules is one of the most anti-stress methods for the plants. It was reported that plants responded to some abiotic stresses such as freezing, thawing, seed aging and dehydration through changing lipid molecules especially the messenger phosphatidic acid (PA) and mutants of PLD were more tolerant to those stresses. It is important to investigate the characteristics and variation of membrane lipids and membrane proteins to understand the streee in plants.Three different kinds of stresses, including alpine scree temperature stress, allelopathy and Gamma irradiation stress, were studied in the present dissertation. And try to understand how plants response to those stresses by changing membrane system and the function of PLD in resistant to those stresses, lipidomic methods were used to profiling the changing of 11 lipids classes (160 lipids molecules) under thoses stresses. Moreover, PLD mutants were also used to study the role of PLD under those stresses. The mechanisms of plants response to stresses were very complicated; PLD and lipid molecules were not the only factors that response to stresses, the metabolism and phytohormones of tested plants under these stresses were also studied.In alpine scree of northwest Yunnan, the temperature was various from 33 °C during the midday to 4 °C at night, and the highest temperature could reach to 35 to 40 °C. Saussurea medusa and Solms-Laubachia linearifolia, which live in this environment, were chosen as studied model. The results showed that membrane lipid of these two plants significantly fluctuated with the temperature, while the double bond index (DBI) that had close relationship to temperature did not change. Furthermore, the the lysolipids which rise significantly under stresses did not change too much either. Laboratory mimic experiments also confermed the characteristics of lipids change to temperature in alpine scree plants. The results suggested that the plants living in such temperature changeable environment had already adapted to this situation and their membrane responded to the temperature was a kind of adaptation instead of stress response.Since the first introduction in Yunnan province of China in 1940s, E. adenophorum has spread very rapidly especially in southwestern China. Without understanding its invasive mechanism, it is impossible to control it. o-Hydroxycinnamic acid (o-HCA), an allelochmeical isolated from leachates of aerial parts of E. adenophorum were studied. o-HCA was abundant in aerial parts of E. adenophorum (1g/10kg fresh weight). The data showed that o-HCA not only had strong allelopathic effect on Arabidopsis seeds germination, but also inhibited seedling growth, and even induced root death of Arabidopsis seedlings. It could be proposed that o-HCA affected seedlings indirectly, through inducing root cell death, and it disturbed the water and ion absorption of plants and finally induced seedling to die. Interestingly, o-HCA could also inhibit E. adenophorum seed germination, while it showed no effect on its seedling growth. E. adenophorum can produce thousands of seeds and has the ability to vegetative reproduction, with which may alleviate the harmful effect of o-HCA on E. adenophorum. Unlike E. adenophorum, its neighbors’ population was inhibited, under this situation, E. adenophorum coule have better condition to live and invade successfully.Arabidopsis were irradiated with gamma rays, and 50-100 Gy gamma irradiation could inhibit seedling growth, and with the dosage above 200 Gy it could inhibit seedling flowering. Treated Arabidopsis wild types and their PLD a and d mutant with gamma ray showed no significant differences among them. The lipid molecules changes of seedlings under stress of gamma ray were also tested, and found that Gamama ray induced lipids degradation, among which, MGDG and DGDG degraded dramatically, while the average carbons in lipids did not changed. The lipids content (nmol per mg dry weight) decreased significantly, while the mol% content (mol% of total) changed slightly. Gamma irradiation also leaded to dramatically change of Arabidopsis seedling metabolomics and the phytohormones (ABA，ZR，JA，IAA).","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.date.issued.year&advanced=false&query1=Time%2BTo%2BGermination&&fq=dc.project.title_filter%3AEnvironmental%5C+stresses%5C+could%5C+limit%5C+plant%5C+growth%2C%5C+development%5C+and%5C+propagation.%5C+Abiotic%5C+stress%5C+refers%5C+to%5C+the%5C+negative%5C+impact%5C+factors%5C+to%5C+the%5C+plants%2C%5C+such%5C+as%5C+extreme%5C+temperature%2C%5C+drought%2C%5C+flood%2C%5C+salinity%2C%5C+irradiation%2C%5C+chemicals%5C+and%5C+so%5C+on.%5C+To%5C+understand%5C+the%5C+mechanism%5C+of%5C+abiotic%5C+stress%5C+is%5C+very%5C+important.Membrane%5C+is%5C+the%5C+most%5C+sensitive%5C+organs%5C+in%5C+the%5C+cell%5C+that%5C+response%5C+to%5C+environmental%5C+changes.%5C+Cells%5C+respond%5C+and%5C+transduct%5C+environmental%5C+signals%5C+by%5C+changing%5C+content%5C+of%5C+membrane%5C+lipids%5C+and%5C+membrane%5C+proteins.%5C+The%5C+activity%5C+change%5C+of%5C+membrane%5C+phospholipase%5C+D%5C+%5C%28PLD%5C%29%5C+and%5C+the%5C+composition%5C+and%5C+content%5C+of%5C+membrane%5C+lipid%5C+molecules%5C+is%5C+one%5C+of%5C+the%5C+most%5C+anti%5C-stress%5C+methods%5C+for%5C+the%5C+plants.%5C+It%5C+was%5C+reported%5C+that%5C+plants%5C+responded%5C+to%5C+some%5C+abiotic%5C+stresses%5C+such%5C+as%5C+freezing%2C%5C+thawing%2C%5C+seed%5C+aging%5C+and%5C+dehydration%5C+through%5C+changing%5C+lipid%5C+molecules%5C+especially%5C+the%5C+messenger%5C+phosphatidic%5C+acid%5C+%5C%28PA%5C%29%5C+and%5C+mutants%5C+of%5C+PLD%5C+were%5C+more%5C+tolerant%5C+to%5C+those%5C+stresses.%5C+It%5C+is%5C+important%5C+to%5C+investigate%5C+the%5C+characteristics%5C+and%5C+variation%5C+of%5C+membrane%5C+lipids%5C+and%5C+membrane%5C+proteins%5C+to%5C+understand%5C+the%5C+streee%5C+in%5C+plants.Three%5C+different%5C+kinds%5C+of%5C+stresses%2C%5C+including%5C+alpine%5C+scree%5C+temperature%5C+stress%2C%5C+allelopathy%5C+and%5C+Gamma%5C+irradiation%5C+stress%2C%5C+were%5C+studied%5C+in%5C+the%5C+present%5C+dissertation.%5C+And%5C+try%5C+to%5C+understand%5C+how%5C+plants%5C+response%5C+to%5C+those%5C+stresses%5C+by%5C+changing%5C+membrane%5C+system%5C+and%5C+the%5C+function%5C+of%5C+PLD%5C+in%5C+resistant%5C+to%5C+those%5C+stresses%2C%5C+lipidomic%5C+methods%5C+were%5C+used%5C+to%5C+profiling%5C+the%5C+changing%5C+of%5C+11%5C+lipids%5C+classes%5C+%5C%28160%5C+lipids%5C+molecules%5C%29%5C+under%5C+thoses%5C+stresses.%5C+Moreover%2C%5C+PLD%5C+mutants%5C+were%5C+also%5C+used%5C+to%5C+study%5C+the%5C+role%5C+of%5C+PLD%5C+under%5C+those%5C+stresses.%5C+The%5C+mechanisms%5C+of%5C+plants%5C+response%5C+to%5C+stresses%5C+were%5C+very%5C+complicated%5C%3B%5C+PLD%5C+and%5C+lipid%5C+molecules%5C+were%5C+not%5C+the%5C+only%5C+factors%5C+that%5C+response%5C+to%5C+stresses%2C%5C+the%5C+metabolism%5C+and%5C+phytohormones%5C+of%5C+tested%5C+plants%5C+under%5C+these%5C+stresses%5C+were%5C+also%5C+studied.In%5C+alpine%5C+scree%5C+of%5C+northwest%5C+Yunnan%2C%5C+the%5C+temperature%5C+was%5C+various%5C+from%5C+33%5C+%C2%B0C%5C+during%5C+the%5C+midday%5C+to%5C+4%5C+%C2%B0C%5C+at%5C+night%2C%5C+and%5C+the%5C+highest%5C+temperature%5C+could%5C+reach%5C+to%5C+35%5C+to%5C+40%5C+%C2%B0C.%5C+Saussurea%5C+medusa%5C+and%5C+Solms%5C-Laubachia%5C+linearifolia%2C%5C+which%5C+live%5C+in%5C+this%5C+environment%2C%5C+were%5C+chosen%5C+as%5C+studied%5C+model.%5C+The%5C+results%5C+showed%5C+that%5C+membrane%5C+lipid%5C+of%5C+these%5C+two%5C+plants%5C+significantly%5C+fluctuated%5C+with%5C+the%5C+temperature%2C%5C+while%5C+the%5C+double%5C+bond%5C+index%5C+%5C%28DBI%5C%29%5C+that%5C+had%5C+close%5C+relationship%5C+to%5C+temperature%5C+did%5C+not%5C+change.%5C+Furthermore%2C%5C+the%5C+the%5C+lysolipids%5C+which%5C+rise%5C+significantly%5C+under%5C+stresses%5C+did%5C+not%5C+change%5C+too%5C+much%5C+either.%5C+Laboratory%5C+mimic%5C+experiments%5C+also%5C+confermed%5C+the%5C+characteristics%5C+of%5C+lipids%5C+change%5C+to%5C+temperature%5C+in%5C+alpine%5C+scree%5C+plants.%5C+The%5C+results%5C+suggested%5C+that%5C+the%5C+plants%5C+living%5C+in%5C+such%5C+temperature%5C+changeable%5C+environment%5C+had%5C+already%5C+adapted%5C+to%5C+this%5C+situation%5C+and%5C+their%5C+membrane%5C+responded%5C+to%5C+the%5C+temperature%5C+was%5C+a%5C+kind%5C+of%5C+adaptation%5C+instead%5C+of%5C+stress%5C+response.Since%5C+the%5C+first%5C+introduction%5C+in%5C+Yunnan%5C+province%5C+of%5C+China%5C+in%5C+1940s%2C%5C+E.%5C+adenophorum%5C+has%5C+spread%5C+very%5C+rapidly%5C+especially%5C+in%5C+southwestern%5C+China.%5C+Without%5C+understanding%5C+its%5C+invasive%5C+mechanism%2C%5C+it%5C+is%5C+impossible%5C+to%5C+control%5C+it.%5C+o%5C-Hydroxycinnamic%5C+acid%5C+%5C%28o%5C-HCA%5C%29%2C%5C+an%5C+allelochmeical%5C+isolated%5C+from%5C+leachates%5C+of%5C+aerial%5C+parts%5C+of%5C+E.%5C+adenophorum%5C+were%5C+studied.%5C+o%5C-HCA%5C+was%5C+abundant%5C+in%5C+aerial%5C+parts%5C+of%5C+E.%5C+adenophorum%5C+%5C%281g%5C%2F10kg%5C+fresh%5C+weight%5C%29.%5C+The%5C+data%5C+showed%5C+that%5C+o%5C-HCA%5C+not%5C+only%5C+had%5C+strong%5C+allelopathic%5C+effect%5C+on%5C+Arabidop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Chapter 1, we isolated a flavonoid prenyltransferase-like gene from traditional Chinese medicinal herb, Epimedium L. (berberidaceae). Epimedium species have a high content of the prenylated flavonol glycosides. Icariin and epimedin A, B and C are frequently used as marker compounds for the quality control of Epimedium. Here we speculate prenyl flavonoids biosynthesis pathway in Epimedium: The flavonoid prenyltransferase is responsible for the prenylation of flavonoids (naringenin 、kaempferol or apigenin) at the 8-position or 3'' or 5''-position. Leaves of Epimedium acuminatum Franch in the nursery were collected every month, and then detected the icariin content. The results show that leaves in March have the highest icariin content. Total RNA was extracted from leaves in March as template. A similarity-based cloning strategy yielded a flavonoid prenyltransferase-like gene, named EaPT1. In E. coli. expression system, pET32a(+) was chosen as the expression vector for use in Rosetta-gamiB(DE3)、RosettaTM 2（DE3）plysS、BL21(DE3)plysE、BL21(DE3)gold and BL21(DE3) cells. The full length ORF and truncated sequence were ligated with pET32a(+). We did not detect the target protein in SDS-PAGE. In Saccharomyces cerevisiae expression system, the full length ORF was ligated with pYES2. In this expression system, we still could not detect the protein in SDS-PAGE. LC/MS did not detect the activity of prenyltransferase, with naringenin as substrate. Chapter 2 describes functional expression and characterization of two copalyl pyrophosphate synthase gene from Isodon ericalyx (Dunn) Kudo, named IeCPS7 and IeCPS11. Their full length ORF and truncated sequence were ligated into pET32a(+). These vectors were used to transform E.coli BL21(DE)3. The truncated IeCPS7 sequence expressed a soluble His-tag recombinant protein, 104699.41D, pI5.87, 924aa. The recombinant protein was characterized for diterpene synthase activity by using geranylgeranyl diphosphate(GGPP) as substrates and subsequent GC/MS analysis of products. The purified recombinant IeCPS showed optimum activity at pH7.1. In addition, IeCPS showed maximum activity at 30℃. The enzymatic activity was increased by addition of MgCl2 to the reaction mixture. Unexpectedly, MnCl2 actually inhibited the enzyme activity. In addition,only insoluble recombinant proteins were expressed for IeCPS11 in BL21(DE)3, Rosetta-gamiB(DE3) and RosettaTM 2（DE3）plysS. The last part reviews the advances in molecular studies of aromatic prenyltransferase in plants and fungi, focusing on membrane-bound homogentisate prenyltransferses, flavonoid prenyltransferases as well as soluble indole prenyltransferases.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.date.issued.year&advanced=false&query1=Time%2BTo%2BGermination&&fq=dc.project.title_filter%3AIn%5C+Chapter%5C+1%2C%5C+we%5C+isolated%5C+a%5C+flavonoid%5C+prenyltransferase%5C-like%5C+gene%5C+from%5C+traditional%5C+Chinese%5C+medicinal%5C+herb%2C%5C+Epimedium%5C+L.%5C+%5C%28berberidaceae%5C%29.%5C+Epimedium%5C+species%5C+have%5C+a%5C+high%5C+content%5C+of%5C+the%5C+prenylated%5C+flavonol%5C+glycosides.%5C+Icariin%5C+and%5C+epimedin%5C+A%2C%5C+B%5C+and%5C+C%5C+are%5C+frequently%5C+used%5C+as%5C+marker%5C+compounds%5C+for%5C+the%5C+quality%5C+control%5C+of%5C+Epimedium.%5C+Here%5C+we%5C+speculate%5C+prenyl%5C+flavonoids%5C+biosynthesis%5C+pathway%5C+in%5C+Epimedium%5C%3A%5C+The%5C+flavonoid%5C+prenyltransferase%5C+is%5C+responsible%5C+for%5C+the%5C+prenylation%5C+of%5C+flavonoids%5C+%5C%28naringenin%5C+%E3%80%81kaempferol%5C+or%5C+apigenin%5C%29%5C+at%5C+the%5C+8%5C-position%5C+or%5C+3%27%27%5C+or%5C+5%27%27%5C-position.%5C+Leaves%5C+of%5C+Epimedium%5C+acuminatum%5C+Franch%5C+in%5C+the%5C+nursery%5C+were%5C+collected%5C+every%5C+month%2C%5C+and%5C+then%5C+detected%5C+the%5C+icariin%5C+content.%5C+The%5C+results%5C+show%5C+that%5C+leaves%5C+in%5C+March%5C+have%5C+the%5C+highest%5C+icariin%5C+content.%5C+Total%5C+RNA%5C+was%5C+extracted%5C+from%5C+leaves%5C+in%5C+March%5C+as%5C+template.%5C+A%5C+similarity%5C-based%5C+cloning%5C+strategy%5C+yielded%5C+a%5C+flavonoid%5C+prenyltransferase%5C-like%5C+gene%2C%5C+named%5C+EaPT1.%5C+In%5C+E.%5C+coli.%5C+expression%5C+system%2C%5C+pET32a%5C%28%5C%2B%5C%29%5C+was%5C+chosen%5C+as%5C+the%5C+expression%5C+vector%5C+for%5C+use%5C+in%5C+Rosetta%5C-gamiB%5C%28DE3%5C%29%E3%80%81RosettaTM%5C+2%EF%BC%88DE3%EF%BC%89plysS%E3%80%81BL21%5C%28DE3%5C%29plysE%E3%80%81BL21%5C%28DE3%5C%29gold%5C+and%5C+BL21%5C%28DE3%5C%29%5C+cells.%5C+The%5C+full%5C+length%5C+ORF%5C+and%5C+truncated%5C+sequence%5C+were%5C+ligated%5C+with%5C+pET32a%5C%28%5C%2B%5C%29.%5C+We%5C+did%5C+not%5C+detect%5C+the%5C+target%5C+protein%5C+in%5C+SDS%5C-PAGE.%5C+In%5C+Saccharomyces%5C+cerevisiae%5C+expression%5C+system%2C%5C+the%5C+full%5C+length%5C+ORF%5C+was%5C+ligated%5C+with%5C+pYES2.%5C+In%5C+this%5C+expression%5C+system%2C%5C+we%5C+still%5C+could%5C+not%5C+detect%5C+the%5C+protein%5C+in%5C+SDS%5C-PAGE.%5C+LC%5C%2FMS%5C+did%5C+not%5C+detect%5C+the%5C+activity%5C+of%5C+prenyltransferase%2C%5C+with%5C+naringenin%5C+as%5C+substrate.%5C+Chapter%5C+2%5C+describes%5C+functional%5C+expression%5C+and%5C+characterization%5C+of%5C+two%5C+copalyl%5C+pyrophosphate%5C+synthase%5C+gene%5C+from%5C+Isodon%5C+ericalyx%5C+%5C%28Dunn%5C%29%5C+Kudo%2C%5C+named%5C+IeCPS7%5C+and%5C+IeCPS11.%5C+Their%5C+full%5C+length%5C+ORF%5C+and%5C+truncated%5C+sequence%5C+were%5C+ligated%5C+into%5C+pET32a%5C%28%5C%2B%5C%29.%5C+These%5C+vectors%5C+were%5C+used%5C+to%5C+transform%5C+E.coli%5C+BL21%5C%28DE%5C%293.%5C+The%5C+truncated%5C+IeCPS7%5C+sequence%5C+expressed%5C+a%5C+soluble%5C+His%5C-tag%5C+recombinant%5C+protein%2C%5C+104699.41D%2C%5C+pI5.87%2C%5C+924aa.%5C+The%5C+recombinant%5C+protein%5C+was%5C+characterized%5C+for%5C+diterpene%5C+synthase%5C+activity%5C+by%5C+using%5C+geranylgeranyl%5C+diphosphate%5C%28GGPP%5C%29%5C+as%5C+substrates%5C+and%5C+subsequent%5C+GC%5C%2FMS%5C+analysis%5C+of%5C+products.%5C+The%5C+purified%5C+recombinant%5C+IeCPS%5C+showed%5C+optimum%5C+activity%5C+at%5C+pH7.1.%5C+In%5C+addition%2C%5C+IeCPS%5C+showed%5C+maximum%5C+activity%5C+at%5C+30%E2%84%83.%5C+The%5C+enzymatic%5C+activity%5C+was%5C+increased%5C+by%5C+addition%5C+of%5C+MgCl2%5C+to%5C+the%5C+reaction%5C+mixture.%5C+Unexpectedly%2C%5C+MnCl2%5C+actually%5C+inhibited%5C+the%5C+enzyme%5C+activity.%5C+In%5C+addition%2Conly%5C+insoluble%5C+recombinant%5C+proteins%5C+were%5C+expressed%5C+for%5C+IeCPS11%5C+in%5C+BL21%5C%28DE%5C%293%2C%5C+Rosetta%5C-gamiB%5C%28DE3%5C%29%5C+and%5C+RosettaTM%5C+2%EF%BC%88DE3%EF%BC%89plysS.%5C+The%5C+last%5C+part%5C+reviews%5C+the%5C+advances%5C+in%5C+molecular%5C+studies%5C+of%5C+aromatic%5C+prenyltransferase%5C+in%5C+plants%5C+and%5C+fungi%2C%5C+focusing%5C+on%5C+membrane%5C-bound%5C+homogentisate%5C+prenyltransferses%2C%5C+flavonoid%5C+prenyltransferases%5C+as%5C+well%5C+as%5C+soluble%5C+indole%5C+prenyltransferases."},{"jsname":"lastIndexed","jscount":"2024-04-15"}],"资助项目","dc.project.title_filter")'>
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