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1.
[目地]研究普通油茶油脂成分形成的调控机制。[方法]通过转录组测序获得2条普通油茶Δ-12脂肪酸脱氢酶基因序列,分别命名为Cofad6和Cofad2-2,并对这两个基因及其编码蛋白的序列特征进行比较,对其基因表达量与脂肪酸成分含量的相关性进行分析。[结果]Cofad6基因c DNA编码区全长1 347 bp,编码448个氨基酸;Cofad2-2基因c DNA编码区全长1 152 bp,编码383个氨基酸。经比对,Co FAD6蛋白与其余物种FAD6蛋白有65.7%83.68%的氨基酸同源,Co FAD2-2与浙江红花油茶FAD2-2蛋白有99.22%的氨基酸同源,与其余物种的蛋白质78.59%81.72%同源。蛋白质二级结构分析表明,Co FAD6和Co FAD2-2均具有一个脂肪酸去饱和酶结构域,属于脂酰-Co A去饱和酶基因家族;两者均为跨膜蛋白,且Co FAD2-2具有定位于内质网的保守模序。定量PCR检测发现,在普通油茶‘长林4号’无性系未成熟种子中,Cofad6表达量随着种子发育先升高后降低,而Cofad2-2基因随着种子发育表达量逐渐降低,与种子油脂中亚油酸、亚麻酸含量变化趋势呈显著正相关,与油酸含量变化呈显著负相关。[结论]推测Cofad2-2基因是调控普通油茶种子油脂中油酸和亚油酸含量的关键基因之一,该研究为普通油茶油脂改良基因工程育种奠定了基础。  相似文献   

2.
竹子捕光叶绿素a/b结合蛋白基因全长的克隆和序列分析   总被引:6,自引:1,他引:5  
采用RT-PCR技术与RACE技术,从绿竹中克隆到捕光叶绿素a/b结合蛋白基因全长cDNA,命名为cab-BO1(GenBank登记号:EF061137).该基因长1 102 bp,从第64~861 bp含有1个开放阅读框,编码265个氨基酸.cab-BO1编码的氨基酸中第64~232位包括典型的捕光叶绿素a/b结合蛋白功能域,第9~11位为蛋白激酶C的磷酸化位点,第27~52位为泛素结构功能域信号,第55~58位为酪氨酸激酶(CK2)磷酸化位点.序列相似性分析结果表明:cab-BO1 DNA序列和编码的氨基酸序列与玉米、小果野蕉、小麦、水稻等的cab基因及其氨基酸序列的相似性分别在85%和89%以上,显示了cab家族基因在进化过程中的保守性.  相似文献   

3.
以油茶‘湘林1号’品种的近成熟种子为材料,采用简并PCR、RACE、交错PCR等技术,获得了油茶乙酰辅酶A羧化酶BC亚基基因的全长cDNA克隆。该基因cDNA序列全长1 901 bp,含有一个1 599 bp的ORF,编码533个氨基酸残基。BC蛋白的等电点pI为6.88,分子量为58 509.3 u,含两个跨膜结构域,是一个不稳定的非分泌蛋白。模体搜索结果表明在BC上具有N-糖基化位点、蛋白激酶C磷酸化位点、ATP结合位点等多个功能结构域。同源建模的模型中发现12个α-螺旋,整个BC蛋白为一个内凹的结构。该基因已登录到GenBank,并被命名为co-bc。  相似文献   

4.
应用RT-PCR和RACE法从麻疯树总RNA中分离出pepc基因全长cDNA,长度为3 142 bp,阅读框2 898 bp,编码965个氨基酸,在GenBank中登录,序列号为EU069413。它的氨基酸序列与蓖麻、陆地棉、橙、大豆、花生、烟草、油菜、拟南芥的氨基酸同源性分别为94.94%、92.46%、90.60%、90.50%、90.50%、88.33%、84.61%、82.44%。该基因编码了pepc基因家族中的pepc1,蛋白属于C3型PEPC。推测了pepc编码蛋白分子量为110.6 kD,并对其稳定性、二级结构、疏水性等特性进行了分析,最后确定了该蛋白的功能位点和结构域。  相似文献   

5.
以油茶近成熟种子为材料,根据油茶EST文库中已知的脂氢过氧化物裂解酶EST序列,利用3’RACE与5’RACE技术,克隆到脂氢过氧化物裂解酶的全长cDNA序列。该基因全长1648bp,,包含一个1476bp开放阅读框长,编码491个氨基酸,5’与3’非编码分别为52bp、121bp。预测该蛋白相对分子量为54.7779KDa,等电点为6.77,是个叶绿体转运肽,N端包含有22个疏水氨基酸残基组成的序列,不仅具有转运肽富含的丝氨酸,还含有大量转运肽中很少存在的脯氨酸。多序列比对发现油茶脂氢过氧化物裂解酶核苷酸序列与茶的相似性达到96%,因此将该基因命名coHPL。  相似文献   

6.
【目的】油茶种仁含油率较低且易遭逆性环境等的影响,这些都严重影响和制约了油茶产业的快速发展。丙二酰单酰Co A:ACP转酰酶是脂肪酸合成酶(FASⅡ)复合体的组成酶之一,在脂肪酸合成中起着装载作用,它的酶促反应产物丙二酰单酰ACP是脂肪酸合成关键组成部分。开展该基因的功能研究,能为油茶的分子遗传育种奠定基础。【方法】以油茶品种‘华硕’种子为材料,借助其种子转录组和表达谱数据库,通过RACE技术,克隆丙二酰单酰Co A:ACP转酰酶基因并对该基因进行生物信息学分析。利用常规的酶切连接技术构建基因的原核表达载体,并利用α酮戊二酸脱氢酶(KDH)偶联系统测定表达产物的酶活性。【结果】丙二酰单酰Co A:ACP转酰酶基因的全长c DNA 1 867 bp,含有1 131 bp的开放读码框,编码376个氨基酸,具有58个氨基酸长度的叶绿体转运肽,该基因被命名为Co MCAT(Gen Bank登录号KJ910337)。在malonyl-Co A结合位点和ACP结合位点上分别存在高度保守的基序"GQGXQ"和"GXSXG"。原核表达载体p ET30a-Co MCAT在BL21(DE3)细胞中被成功地诱导表达,目的重组蛋白分子量约为40 k Da。酶活性分析表明,重组蛋白Co MCAT的最佳催化温度为30℃,最佳p H为7.0,比酶活性约为2.384 U·μg-1。【结论】以本实验室构建的油茶转录组数据库为基础,首次克隆获得油茶MCAT基因的全长c DNA序列,构建其原核表达载体在宿主细胞BL21(DE3)中成功诱导表达目的蛋白,并对重组蛋白进行初步的酶活性分析。研究结果为今后进一步深入开展和揭示油茶MCAT基因在油脂合成的代谢过程中的作用奠定基础。  相似文献   

7.
依据日本落叶松转录组数据库检测到的咖啡酸-O-甲基转移酶(COMT)基因ESTs序列设计引物,分离得到一个编码该酶的新基因。其cDNA克隆全长为1 350 bp,包含长度为1 092 bp的开放阅读框,可编码364个氨基酸残基。序列分析显示,所推导的蛋白质氨基酸序列包含COMT基因5个所特有的保守元件,与海岸松PpCOMT的蛋白质氨基酸序列同源性达94%。在此基础上,利用DnaSP5.0软件对日本落叶松40株基因型个体的LkCOMT序列进行了单核苷酸多样性SNPs分析,共检测到92个SNP位点,SNP发生频率为1/17 bp,多样性指数πT为0.007 80。在这些SNPs中,65个属于转换,27个属于颠换。在外显子区域,共检测到58个SNP位点,其中37个为错义突变,21个为同义突变。研究结果为日本落叶松基因标记辅助育种提供了理论基础。  相似文献   

8.
油茶ACP基因的全长cDNA克隆及序列分析   总被引:7,自引:2,他引:5  
酰基载体蛋白是脂肪酸合成中的关键蛋白质,位于脂肪酸合成酶系的中央,作为脂酰基的载体将脂酰基从一个酶反应转移到另一个酶反应.以油茶近成熟种子cDNA文库和EST文库为基础,采用分子生物学技术分离克隆了油茶酰基载体蛋白基因全长cDNA序列.结果表明:油茶酰基载体蛋白基因的全长cDNA为669bp,包含1个完整的CDS及3′UTR和5′UTR,编码141个氨基酸,该基因被命名为co-acp并提交至GeneBank,登录号是EU717697;油茶酰基载体蛋白基因的推导氨基酸序列与其它15种植物的ACP进行AlignX比较的结果表明,油茶酰基载体蛋白与油橄榄ACP的相似性最高,而且遗传距离最近;预测油茶酰基载体蛋白的二级结构以α螺旋为主,没有跨膜结构域和信号肽,其等电点约为4.995.  相似文献   

9.
为得到高质量的油茶总RNA及稳定的内参基因,以油茶良种‘华硕’为试验材料,采用ambion试剂盒结合CTAB法进行总RNA提取。获得的总RNA完整性好,纯度高;从前期构建的油茶转录组数据中选取ACT(肌动蛋白基因)等14个基因作为候选内参基因,共设计22对引物,通过RT-PCR反应体系的优化及琼脂糖凝胶电泳检测,快速筛选出了适合油茶不同组织(器官)的内参基因。ALB(清蛋白基因),EF1α(延伸因子基因),ETIF3H(启动因子基因),UBC2(泛素结合酶基因)可以作为油茶果实初选内参基因,EF1α可以作为油茶根、茎、花及果实内参基因,ETIF3H可以作为油茶根、茎、叶、花瓣的内参基因。  相似文献   

10.
以杜仲叶片cDNA为模板,采用反转录RCR及RACE技术分离出DXR基因cDNA全长。序列分析结果表明该基因序列全长1 814 bp,共编码478个氨基酸,推导的蛋白质分子量为51.71 kD,理论等电点5.79,命名为EuDXR。推导的EuDXR蛋白质序列具有植物DXR酶的5个典型基序,并预测出26个潜在的功能位点。系统进化树分析表明EuDXR蛋白与玉米、水稻亲缘关系最近,其次为橡胶、拟南芥、烟草。  相似文献   

11.
Five aromatic plants, Carum carvi (caraway), Apium graveolens (celery), Foeniculum vulgare (fennel), Zanthoxylum limonella (mullilam) and Curcuma zedoaria (zedoary) were selected for investigating larvicidal potential against mosquito vectors. Two laboratory-reared mosquito species, Anopheles dirus, the major malaria vector in Thailand, and Aedes aegypti, the main vector of dengue and dengue hemorrhagic fever in urban areas, were used. All of the volatile oils exerted significant larvicidal activity against the two mosquito species after 24-h exposure. Essential oil from mullilam was the most effective against the larvae of A. aegypti, while A. dirus larvae showed the highest susceptibility to zedoary oil.  相似文献   

12.
An understanding of growth and photosynthetic potential of subtropical rainforest species to variations in light environment can be useful for determining the sequence of species introductions in rainforest restoration projects and mixed species plantations. We examined the growth and physiology of six Australian subtropical rainforest tree species in a greenhouse consisting of three artificial light environments (10%, 30%, and 60% full sunlight). Morphological responses followed the typical sun-shade dichotomy, with early and late secondary species (Elaeocarpus grandis, Flindersia brayleyana, Flindersia schottiana, and Gmelina leichhardtii) displaying higher relative growth rate (RGR) compared to mature stage species (Cryptocarya erythroxylon and Heritiera trifoliolatum). Growth and photosynthetic performance of most species reached a maximum in 30–60% full sunlight. Physiological responses provided limited evidence of a distinct dichotomy between early and late successional species. E. grandis and F. brayleyana, provided a clear representation of early successional species, with marked increase in Amax in high light and an ability to down regulate photosynthetic machinery in low light conditions. The remaining species (F. schottiana, G. leichhardtii, and H. trifoliolatum) were better represented as falling along a shade-tolerant continuum, with limited ability to adjust physiologically to an increase or decrease in light, maintaining similar Amax across all light environments. Results show that most species belong to a shade-tolerant constituency, with an ability to grow and persist across a wide range of light environments. The species offer a wide range of potential planting scenarios and silvicultural options, with ample potential to achieve rapid canopy closure and rainforest restoration goals.  相似文献   

13.
Buruk K  Sokmen A  Aydin F  Erturk M 《Fitoterapia》2006,77(5):388-391
The Eastern Black Sea Region has an extensive flora because of ample rainfall lasting all year. In this study, antimicrobial effects of 74 crude extracts of 22 endemic plants were investigated. Among the 30 active crude extracts, water-insoluble crude extracts from Betula medwediewii, Heracleum platytaenium, Primula longipes, Anthemis cretica ssp. argaea and Centaurea helenioides were the prominent ones with their MIC values.  相似文献   

14.
从落叶松梢栖真菌筛选出对落叶松枯梢病病原菌有抑制作用的菌株,对落叶松枯梢病进行生物防治。将自落叶松梢部分离到的11种梢栖真菌与落叶松枯梢病病原菌进行对峙培养,根据被覆盖程度、抑菌率、拮抗系数和防治预试验的结果,综合评价后发现:粪生粪壳、深绿木霉和球毛壳3种梢栖真菌对落叶松枯梢病有良好的拮抗作用。利用这3个菌株在黑龙江省勃利县通天一林场国富沟进行林间防治试验,结果表明:防治效果为粪生粪壳>代森锰锌400倍液>深绿木霉>球毛壳;深绿木霉和粪生粪壳喷洒浓度50%和球毛壳喷洒浓度75%时,真菌多样性最多,梢栖真菌系统较其他浓度更为稳定,与化学防治相比,既达到了防治目的又具有不污染环境的优势。  相似文献   

15.
The potential benefits of species mixture were investigated using pair-wise comparisons of four timber tree species in northern Viet Nam. Chukrasia tabularis, Canarium album, Michelia mediocris and Eucalyptus urophylla were grown in monocultures and in pair-wise mixtures. The trial was established as a randomized block design with each treatment replicated four times. Volume production gain or loss in mixtures was assessed by calculating a mixture index, which is defined as Relative Yield Total (RYT). At age 48 months, the trial indicated mixed performance with both positive and negative impacts of growing some species in mixtures compared to monocultures. The largest gain was shown in the mixture of a shade-intolerant species, Chukrasia with a more shade-tolerant species, Michelia (47% gain in relative yield), and Eucalyptus with Michelia (45% gain in relative yield) at the first 38 months. The other three mixtures tested, and which were not successful (i.e. had lower relative yields) at 38 months, was the mixture of all shade-intolerant species, including Chukrasia with Canarium, Canarium with Eucalyptus and Chukrasia with Eucalyptus. This suggests that species with different shade tolerances can form complementary pair-wise mixtures, but this changed significantly (P < 0.05) over the following 10 months. At age 48 months the RYT of Chukrasia with Michelia increased by 12%, those of Michelia with Eucalyptus decreased by 20% at 48 months compared to 38 months, suggesting that Eucalyptus should be thinned at around year 5 years. Likewise, the RYT of Michelia with Canarium declined significantly by 23% at 48 months. The RYT of other tested mixtures remained almost unchanged over time.  相似文献   

16.
Ungulate browsing greatly influences regeneration dynamics of some forest ecosystems, yet the relationship between browse susceptibility and foliar chemistry of forest tree seedlings is not well understood. We applied field fertilization (15N-9P-10K controlled-release fertilizer at 0, 20, 40, and 60 g per seedling) and investigated how subsequent changes in terpenoid production and foliar nutrition influence ungulate browse preference for Douglas-fir (Pseudotsugamenziesii (Mirb.) Franco), western hemlock (Tsuga heterophylla Raf. Sarg.), and western redcedar (Thujaplicata Donn ex D. Don) seedlings across four sites in northwestern Oregon, USA. Fertilization increased foliar N concentration of all three species, but above-ground growth of only Douglas-fir and western hemlock. Foliar monoterpene concentrations for western hemlock and western redcedar also increased at higher fertilization rates, while Douglas-fir monoterpene production was not affected by fertilization. Regardless of monoterpene levels, ungulate browse preference was greater for fertilized western hemlock seedlings. The opposite response, however, was observed for western redcedar at two of the four study sites where the likelihood of browse was greater for non-fertilized than fertilized seedlings. Differences in browse preference among species may depend on the type and amount of individual monoterpenes manufactured in response to fertilization. Western redcedar produce α- and β-thujone, oxygenated monoterpenes known to promote gastroenteritis and possibly inhibit microbial rumen activity, which were absent from Douglas-fir or western hemlock. Higher concentrations of α- and β-thujone associated with increasing fertilizer rate provide a plausible explanation as to why ungulates preferred non-fertilized western redcedar. Our results illustrate species-specific adaptation in browse avoidance and selective ungulate browsing behavior of individual trees as linked to foliar chemistry.  相似文献   

17.
18.
Bark beetles are largely known for their ability to undergo intermittent population eruptions that transform entire landscapes and pose significant economic hardships. However, most species do not undergo outbreaks, and eruptive species usually exert only minor disturbances. Understanding the dynamics of tree-killing noneruptive species can provide insights into how beetles persist at low densities, and how some spatiotemporal patterns of host predisposition may more likely favor breaching eruptive thresholds than others. Elucidating mechanisms behind low-density populations is challenging, however, due to the requirement of long-term monitoring and high degrees of spatial and temporal covariance. We censused more than 2700 trees annually over 7 years, and at the end of 17 years, in a mature red pine plantation. Trees were measured for the presence of bark beetles and wood borers that breed within the primary stem, root weevils that breed in root collars, and bark beetles that breed in basal stems. We quantify the sequence of events that drive this decline syndrome, with the primary emergent pattern being an interaction between below- and above-ground herbivores and their fungal symbionts. This interaction results in an expanding forest gap, with subsequent colonization by early-successional vegetation. Spatial position strongly affects the likelihood of tree mortality. A red pine is initially very likely to avoid attack by tree-killing Ips beetles, but attack becomes increasingly likely as the belowground complex spreads to neighboring trees and eventually make trees susceptible. This system is largely internally driven, as there are strong gap edge, but not stand-edge, effects. Additional stressors, such as drought, can provide an intermittent source of susceptible trees to Ips beetles, and elevated temperature slightly accentuates this effect. New gaps can arise from such trees as they subsequently become epicenters for the full complex of organisms associated with this decline, but this is not common. As Ips populations rise, there is some element of positive feedback, in that the proportion of killed trees that were not first colonized by root organisms increases. This positive feedback is very weak, however, and we propose the slope between beetle population density and reliance on host stress as a quantitative distinction along a gradient from noneruptive through eruptive species. Almost all trees colonized by Ips were subsequently colonized by wood borers, likely a source of negative feedback. We discuss implications to our overall understanding of cross-scale interactions, between-guild interactions, forest declines, and eruptive thresholds.  相似文献   

19.
One of the arguments against using prescribed fire to regenerate oak (Quercus spp.) forests is that the improvement in species composition of the hardwood regeneration pool is temporary and multiple burns are necessary to achieve and maintain oak dominance. To explore this concern, I re-inventoried a prescribed fire study conducted in the mid-1990s to determine the longevity of the effects of a single prescribed fire on hardwood regeneration. The initial study was conducted in three oak shelterwood stands in central Virginia, USA. In 1994, each stand was divided into four treatments (spring, summer, and winter burns and a control) and the hardwood regeneration was inventoried before the fires. During the burns, fire intensity was measured and categorized in each regeneration sampling plot. Second-year postfire data showed marked differences in species mortality rates, depending on season-of-burn and fire intensity: oak and hickory (Carya spp.) regeneration dominated areas burned by medium- to high-intensity fire during the spring and summer while yellow-poplar (Liriodendron tulipifera) and red maple (Acer rubrum) seedlings dominated unburned areas and all areas treated with low-intensity fire regardless of season-of-burn. The treatments were re-inventoried in 2006 and 2007 to determine whether these fire effects were still present. The new data show that the species distributions by season-of-burn and fire intensity found in 1996 still existed 11 years after the treatments. The fact that fire effects in oak shelterwood stands can last at least a decade has important management implications for resource professionals interested in sustaining oak forests in the eastern United States.  相似文献   

20.
Efforts in Europe to convert Norway spruce (Picea abies) plantations to broadleaf or mixed broadleaf-conifer forests could be bolstered by an increased understanding of how artificial regeneration acclimates and functions under a range of Norway spruce stand conditions. We studied foliage characteristics and leaf-level photosynthesis on 7-year-old European beech (Fagus sylvatica) and pedunculate oak (Quercus robur) regeneration established in open patches and shelterwoods of a partially harvested Norway spruce plantation in southwestern Sweden. Both species exhibited morphological plasticity at the leaf level by developing leaf blades in patches with an average mass per unit area (LMA) 54% greater than of those in shelterwoods, and at the plant level by maintaining a leaf area ratio (LAR) in shelterwoods that was 78% greater than in patches. However, we observed interspecific differences in photosynthetic capacity relative to spruce canopy openness. Photosynthetic capacity (A1600, net photosynthesis at a photosynthetic photon flux density of 1600 μmol photons m−2 s−1) of beech in respect to the canopy gradient was best related to leaf mass, and declined substantially with increasing canopy openness primarily because leaf nitrogen (N) in this species decreased about 0.9 mg g−1 with each 10% rise in canopy openness. In contrast, A1600 of oak showed a weak response to mass-based N, and furthermore the percentage of N remained constant in oak leaf tissues across the canopy gradient. Therefore, oak photosynthetic capacity along the canopy gradient was best related to leaf area, and increased as the spruce canopy thinned primarily because LMA rose 8.6 g m−2 for each 10% increase in canopy openness. These findings support the premise that spruce stand structure regulates photosynthetic capacity of beech through processes that determine N status of this species; leaf N (mass basis) was greatest under relatively closed spruce canopies where leaves apparently acclimate by enhancing light harvesting mechanisms. Spruce stand structure regulates photosynthetic capacity of oak through processes that control LMA; LMA was greatest under open spruce canopies of high light availability where leaves apparently acclimate by enhancing CO2 fixation mechanisms.  相似文献   

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