899 resultados para High Content Screening
Resumo:
Cubic GaN(c-GaN) films are grown on GaAs(001) substrates by metalorganic chemical vapor deposition (MOCVD). Two GaN samples were grown with different buffer layer, the deposition time of each was 1 and 3 min, respectively. 4-circle X-ray double crystal diffraction (XRDCD) was used to study the secondary crystallographic phases presented in the c-GaN films. The phase composition of the epilayers was determined by X-ray reciprocal space mapping. The intensities of the c-GaN(002) and h-GaN(10 (1) over bar 1) planes detected in the mapping were investigated by omega scans. The content of the hexagonal phase inclusions in the c-GaN films was calculated to about 1.6 and 7.9%, respectively. The thicker buffer layer is not preferable for growing high quality pure c-GaN films. (C) 2000 Elsevier Science S.A. All rights reserved.
Resumo:
A systematic study of electron cyclotron resonance (CR) in two sets of GaAs/Al0.3Ga0.7As modulation-doped quantum-well samples (well widths between 12 and 24 nm) has been carried out in magnetic fields up to 30 T. Polaron CR is the dominant transition in the region of GaAs optical phonons for the set of lightly doped samples, and the results are in good agreement with calculations that include the interaction with interface optical phonons. The results from the heavily doped set are markedly different. At low magnetic fields (below the GaAs reststrahlen region), all three samples exhibit almost identical CR which shows little effect of the polaron interaction due to screening and Pauli-principle effects. Above the GaAs LO-phonon region (B > similar to 23 T), the three samples behave very differently. For the most lightly doped sample (3 x 10(11) cm(-2)) only one transition minimum is observed, which can be explained as screened polaron CR. A sample of intermediate density (6 x 10(11) cm(-2)) shows two lines above 23 T; the higher frequency branch is indistinguishable from the positions of the single line of the low density sample. For the most heavily, doped sample (1.2 x 10(12) cm(-2)) there is no evidence of high frequency resonance, and the strong, single line observed is indistinguishable from the lower branch observed from sample with intermediate doping density. We suggest that the low frequency branch in our experiment is a magnetoplasmon resonance red-shifted by disorder, and the upper branch is single-particle-like screened polaron CR. (C) 1998 Elsevier Science B.V. All rights reserved.
Resumo:
随着工农业的快速发展,土壤重金属尤其Cd和Pb污染日益严重。筛选和培育具有重金属低积累特性的农作物排异品种被认为是当前应对土壤重金属污染最为合理和有效的途径之一。本文通过盆栽试验、大田试验和砂培试验,研究了大白菜品种对Cd和Pb的吸收和积累的品种差异、对Cd、Pb胁迫的响应以及大白菜安全生产的调控技术,得出以下结论: 1) 盆栽梯度试验中,80种大白菜地上部对Cd的吸收存在显著差异(p < 0.05)。在3种Cd处理下(1.0, 2.5和5.0 mg/kg),80种大白菜Cd含量浓度范围分别为(mg/kg) 0.22–2.46, 0.90–14.10和2.03–18.01, 其平均值分别为 0.79, 3.76 和6.79 mg/kg DW。大白菜对Cd胁迫具有较强的耐性。大田试验中,15种大白菜的富集系数和转运系数与盆栽梯度试验的结果基本一致。排异植物的筛选和鉴定标准包括:(1)该植物的地上部和根部的Cd含量都很低或者可食部位低于有关标准;(2) 富集系数(BF) < 1.0;(3) 转运系数(TF) < 1.0;(4)该植物具有较高的Cd耐性,在较高的Cd污染下能够正常生长且生物量无显著下降。采用此标准,结合盆栽梯度试验和大田试验结果,北京新3号、绿星70和丰源新3号可鉴定为Cd排异品种。秋傲和赛新5号具有排异Cd特性,但其对Cd的耐性较差。 2) 盆栽梯度试验中,在Pb投加浓度为500和1500 mg/kg处理下,30种大白菜地上部对Pb的吸收存在显著差异 (p < 0.05),其Pb浓度的范围分别为:0.52–8.68 和1.86–16.20 mg/kg, 其平均值分别为3.01 和6.87 mg/kg DW。并且,随着Pb浓度的增加,白菜地上部Pb含量有随之增加的趋势。大白菜对Pb具有较强的耐性。低浓度的Pb处理对大白菜的生物具有一定的促进作用。结合盆栽试验和大田试验的结果,秋傲、世博秋抗和福星80可鉴定为Pb排异大白菜品种。 3) 砂培试验中,在Cd和Pb胁迫下,大白菜地上部的丙二醛(MDA)含量增加,随着Cd处理浓度的增加,超氧化物歧化酶(SOD)活性呈现先下降后上升接着下降的趋势,酸菜王的SOD活性要高于北京新3号SOD的活性。随着Pb处理浓度的增加,福星80地上部的SOD活性随之增加,而绿星大棵菜地上部的SOD活性先下降后增加。在不同梯度的Cd处理下,大白菜地上部的可溶性蛋白(SP)含量未见显著降低,甚至有所增加,而在不同梯度的Pb处理下,大白菜地上部的SP含量有所降低。 4) 施用改良剂可升高土壤的pH值和降低土壤中的有效态Cd,从而对大白菜的生长具有促进作用。施用改良剂可显著降低大白菜对Cd和Pb的吸收和累积。
Resumo:
近二十多年来,基于对臭氧层衰减、紫外线B(UV-B)增强的担心,研究者希望了解到紫外线辐射对不同作物的影响情况,增强UV-B辐射条件下是否对作物的生长发育、产量质量构成威胁。在本试验中,我们首先探讨了双子叶作物黄瓜(Cucumis sativus)和大豆(Glycine max)对不同紫外波段的生物效应[分别为B-UVA(315-400 nm),N-UVA(315-340 nm),B-UVB(275-400 nm)和N-UVB(290-340 nm),UV-(>400nm)作对照]。我们观察到所有的UV波段处理都使黄瓜和大豆的生长受到抑制,并且细胞受到不同程度的氧化伤害;UV波段处理的作用效果与不同波段的紫外有效生物辐射剂量有关。处理差异在UV-B波段内部和UV-A波段内部同样存在。植物生长UV辐射公式(BSWF)能很好的预测本试验UV-B波段内的平均植物效应,但不能预测UV-A波段的植物效应。短波UV-A的生物作用强于长波UV-A。光合色素的变化与UV波谱差异和种间差异有关。在高的紫外/可见光背景下,UV-A处理同UV-B同样导致光合色素的降低,但黄瓜类胡萝卜素/叶绿素比例升高。与其他研究者的试验结果比较后,我们认为紫外线B辐射的生物效应一致性很高,但紫外线A波段的生物学效应存在较大争议。因此我们在本试验的基础上仅进行荞麦[苦荞(Fagopyrum tataricum Gaertn.)和甜荞(Fagopyrum esculentum Moench.)]对紫外线B波段的响应研究。 我们对苦荞品种-圆籽荞进行了连续两个生长季节的大田半控制试验以观察UV-B辐射对苦荞生长、发育、产量及叶片色素的影响;试验小区进行降低UV-B、近充足UV-B和增强UV-B辐射处理。我们的试验表明在不同强度UV-B辐射下苦荞的生长、地上部生物量积累及最终产量都有所下降,但苦荞的发育加快;当前条件下的日光紫外线B辐射对植物生长和产量也造成负面影响。植物光合色素被日光及增强UV-B辐射降低;UV化合物及卢丁含量在中低剂量的UV-B辐射强度下显著升高,但在高剂量的增强UV-B辐射下短期升高后迅速下降。我们的试验表明苦荞是一个对UV-B高度敏感的作物。苦荞对UV-B的敏感性与UV-B剂量、外界环境因素及生长季节有关。 单个苦荞品种的试验结果使我们认识到外界UV-B辐射已经对苦荞生长发育构成逆境条件,未来全球气候变化条件下增强紫外线B辐射可能使其处于更不利的生长环境中。因此我们有进行了多个种群进行UV-B响应观察并筛选耐性种群。我们对15个苦荞种群进行增强UV-B辐射处理(6.30 kJ m2 UV-BBE,模拟当地25%的臭氧衰减),我们观察苦荞UV-B辐射效应存在显著的种内差异,UV-B辐射对多数种群具有抑制作用,但对一些种群还有刺激作用。我们采用主成分分析方法与作物UV-B响应指数(RI)来评价苦荞作物UV-B辐射耐性。我们发现作物的UV-B耐性不仅与其原产地背景UV-B强度有关,而且与作物相对生长效率、次生代谢产物含量(如卢丁)及其他因素有关。我们观察到苦荞伸展叶总叶绿素变化与UV-B耐性成正相关;室内苦荞幼苗的UV-B辐射致死试验表明:苦荞种群死亡率与其UV-B耐性成负相关。 此外,我们对甜荞的UV-B辐射响应也进行了初步研究。选取美姑甜荞、巧家甜荞和云龙甜荞进行5个梯度的增强UV-B辐射室外模拟试验。我们观察到UV-B辐射显著降低了甜荞的生长、生物量及产量;并严重影响了甜荞的生殖生长,降低了花序数、种子数和结实率;并且UV-B辐射对甜荞的抑制作用存在显著的剂量效应。三种甜荞品种存在显著的种内差异,其中美姑品种UV-B耐性最强,且膜脂受UV-B辐射氧化伤害最小,这与该品种UV-B辐射下较高的GR酶活性、APX酶活性和PPO酶活性、以及含量更高的抗坏血酸有关。甜荞的次生代谢也受到增强UV-B辐射的影响,其香豆酰类化合物在UV-B辐射下升高显著,而槲皮素含量也在高剂量UV-B辐射下有所增加;卢丁含量依赖UV-B辐射剂量而变化,中低剂量UV-B辐射下其卢丁含量逐渐升高,但在高剂量辐射下逐渐下降。 通过对生长在高海拔地区的荞麦作物(苦荞和甜荞)进行的室外研究,我们认识到作物不同品种存在很大的耐性差异,这就为UV-B耐性育种创造了有利条件。进一步加大荞麦种质资源筛选力度并深入认识荞麦抗性机理,在此基础上通过杂交或其他基因融合手段培育抗性品种,对高剂量UV-B辐射地区的荞麦产量的提高将起到重要推动作用,并使荞麦生产能有效应对未来全球气候变化条件下UV-B辐射可能升高的威胁。 During last few decades, due to concern of ozone layer depletion and enhancement of ultraviolet B radiation(UV-B, 280-315 nm), the agronomist want to know the responses of different crop species to UV-B. In the first experiment of our study, the effect of different UV band [B-UVA(315-400 nm), N-UVA(315-340 nm), B-UVB(275-400 nm), N-UVB(290-340 nm)and UV-(>400nm, as control)] on the cucumber(Cucumis sativus)and soybean(Glycine max)were investigated in growth room. Spectra-dependent differences in growth and oxidation indices existed within UV-A bands as well as UV-B bands. The general biological effects of different band were UV- < B-UVA< N-UVA<N-UVB<B-UVB. The plant growth biologically spectra weighting function(BSWF)matched well with average plant response in UV-B region, but not in UV-A region. Shorter UV-A wavelength imposed more negative impact than longer UV-A wavelength did in both species. The effect on photosynthetic pigment was related to different UV bands and different species. The photosynthetic pigment content was decreased by UV-A spectra as well as UV-B spectra. In comparison with the results of previous studies, we found that the wavelength-dependent biological effect of ultraviolet B radiation has high consistency, but the biological effect of ultraviolet-A radiation was inconsistent. We narrow our following study on the effect of ultraviolet B radiation on the buckwheat(tartary buckwheat and common buckwheat). The tartary buckwheat(Fagopyrum tataricum Gaertn.)cultivars Yuanziqiao was grown in the sheltered field plots for two consecutive seasons under reduced, near-ambient and two supplemental levels of UV-B radiation. The crop growth, photosynthetic pigments, total biomass, final seed yield and thousand-grain weight were decreased by near-ambient and enhanced UV-B radiation, while crop development was promoted by enhanced UV-B radiation. Leaf rutin concentration and UV-B absorbing compound was generally increased by UV-B with the exception of 8.50 kJ m-2 day-1 supplemental levels. Our results showed that tartary buckwheat is a potentially UV-B sensitive species. Study on one cultivars showed that ambient solar radiation had present a stress to tartary buckwheat. This makes it necessary to observe the UV-B response of many cultivars and screen tolerant cultivars. Fifteen populations of tartary buckwheat were experienced enhanced UV-B radiation simulating 25% depletion of the stratospheric ozone layer in Kunming region, and plant responses in growth, morphology and productivity were observed. Principal components analysis(PCA)was used to evaluate overall sensitivity of plant response to UV-B as well as response index. The different populations exhibited significant differences in responses to UV-B. The photosynthetic pigments of young seedlings were also affected significantly under field condition. On the other hand, the healthy seedlings of different populations were exposed to the high level of UV-B radiation in growth chambers to determine the plant lethality rate. The plant tolerance evaluated by multivariate analysis was positively related to total plant chlorophyll change, but negatively related to lethality rate. In other hand, the UV-B responses of the other important cultivated buckwheat species, common buckwheat(Fagopyrum esculentum Moench.), were also studied preliminarily. Three widespread cultivated variety(Meigu, Qiaojia and Yunlong cultivars)were provided with five level of enhanced UV-B radiation outdoors. We observed that the crop growth, development and production were significantly decreased, and reproductive production, like anthotaxy number, seed number and seed setting ratio, was also decreased. Dose-dependent inhibition effect caused by enhanced UV-B radiation also existed in common buckwheat. Significant intraspecific difference existed in those three cultivars. The Meigu cultivars with dwarfed growth and lower production have highest UV-B tolerance as well as lowest damage in cell membrane, this could be associated with profound enhancements of glutathione reductase(GR)activity, ascorbate peroxidase activity and polyphenol oxidase activity as well as higher ascorbic acid concentration. The secondary metabolism was also affected by UV-B radiation, with profound elevation of coumarin compound and moderate increase of quercetin concentration. Rutin concentration was peaked in 5kJ m-2 UV-B. The contrasting effect of UV-B radiation on different populations indicated that there existed abundant genetic resources for selecting tolerant populations of common and tartary buckwheat. Much effort needed be pose on screening of buckwheat germplasm and clarification of mechanism of buckwheat tolerance to UV-B. On this base the tolerant cultivars could be bred by hybridization and other gene transfusion method, this would help increase buckwheat yield in high ambient UV-B region and counteract the effect of possible enhanced UV-B radiation in future.
Resumo:
小麦条锈病(Puccinia striiformis f. sp. tritici)是世界性小麦病害,可导致受害小麦减产30%以上,甚至绝收。小麦条锈病在我国西南、华北麦区危害严重,四川麦区是小麦条锈病发病最重的地区之一,每年因条锈病流行造成小麦产量损失巨大。利用抗条锈病品种是控制该病害最安全、经济的有效途径,因此挖掘利用抗病新基因,开展抗病遗传基础研究是当前育种工作中面临的重要任务。 偏凸山羊草(Aegilops ventricosa,DDMvMv,2n=28)是一年生草本植物,起源于地中海西部沿岸地区,具有对小麦白粉病、锈病等高抗或免疫、耐盐、抗寒、蛋白质含量高等优良性状,是小麦遗传育种很好的种质资源。本研究以高抗条锈病的小麦—偏凸山羊草6Mv/6B代换系(Moisson 6Mv/6B)为材料,对其含有的带条锈病抗性基因的偏凸山羊草6Mv染色体在四川小麦背景中的传递情况、与小麦—簇毛麦双端体附加系所具有的白粉病抗性的聚合以及对Moisson 6Mv/6B进行电离辐射诱变筛选抗条锈病的小麦—偏凸山羊草易位系三个方面进行了研究。取得的主要研究结果如下: 1. Moisson 6Mv/6B与高感条锈病的四川地区普通小麦品种绵阳26、绵阳93-124和SW3243的杂种F1与其普通小麦亲本分别作为父、母本回交,通过对其BC1和F2的结实率、根尖细胞有丝分裂中期染色体的观察以及对条锈病抗性的鉴定,发现含6Mv染色体的F1植株作母本时的回交结实率(83.10%)普遍高于含6Mv染色体的F1植株作父本(48.61%),结实率与普通小麦基因型密切相关(χ2=34.15>>χ20.05=5.99(df=2));6Mv染色体在三种四川小麦中通过雌、雄配子传递的传递方式与其传递率间没有显著相关性,其传递率与普通小麦基因型呈显著相关性(χ2=6.42>χ20.05=5.99(df=2))。 2. Moisson 6Mv/6B与高抗白粉病的小麦—簇毛麦双端体附加系Pana(2n=42+2t)正反杂交,希望在聚合两者抗性的同时观察不同受体背景下的抗性反应。对Moisson 6Mv/6B和Pana正反杂交的结实率、杂交后代的农艺性状进行观察,并对杂交后代进行基因组荧光原位杂交(GISH)分析及条锈病和白粉病的抗性鉴定。结果表明Moisson 6Mv/6B作母本时杂交结实率(80.56%)高于Pana作母本时(58.33%),结实率与杂交方式间紧密相关(χ2=4.96>χ20.05=3.84(df=1));Moisson 6Mv/6B和Pana杂交后代株高比最高亲本高约10cm,成熟期也较两亲本提前两个星期左右;正反杂交后代中具有偏凸山羊草6Mv染色体的植株具有条锈病抗性,具有簇毛麦端体的植株具有白粉病抗性,同时筛选到4株含有偏凸山羊草和簇毛麦遗传物质并对条锈病和白粉病兼抗的材料,证明来自偏凸山羊草6Mv染色体的条锈病抗性与来自簇毛麦端体的白粉病抗性已经聚合在一起,且没有产生相互抑制的作用,暗示通过这两个抗性基因的聚合是完全能获得兼抗条锈病和白粉病的小麦新种质。 3. 对Moisson 6Mv/6B在减数分裂时期的成株进行总剂量为6Gy、辐射频率为120rad/min的60Co-γ射线辐射,对辐射植株自交后代进行农艺性状及根尖细胞有丝分裂中期染色体形态观察和条锈病抗性鉴定。结果为辐射植株自交结实率为2.22%,根尖细胞有丝分裂中期的染色体存在明显碎片,辐射自交后代植株对条锈病具有成株期抗性。 小麦—偏凸山羊草6Mv/6B代换系对条锈病抗性稳定,是培育条锈病抗性品种的良好供体。本研究证明在四川小麦背景中要利用该品种抗性,在结实数满足需要时,可将其作父本,亦可作母本,但关键是要选择好一个优良的受体基因型;同时其条锈病抗性与来自簇毛麦的白粉病抗性没有相互抑制作用,可将两者抗性有效聚合用于小麦育种中。 Wheat stripe rust (Puccinia striiformis f. sp. Tritici) is a worldwide disease of wheat, and could lead to victims of 30 percent or even total destruction of wheat production. Wheat stripe rust harms badly in China's southwest and North China. Sichuan province is one of the regions damaged by wheat stripe rust heavily. The use of resistant varieties is the most secure and economical way to control the wheat stripe rust. Therefore, it is essential to identify new disease-resistant genes and genetically research of disease resistance. Aegilops ventricosa (DDMvMv, 2n = 28) is an annual herbaceous plant, originating in the coastal areas of the western Mediterranean, with good characters such as resistance of wheat powdery, rust, salt, cold and high protein content. It is a good germplasm resource. In this study, the wheat- Aegilops ventricosa 6Mv/6B substitution line Moisson 6Mv/6B (highly resistant to the wheat stripe rust) was used to study on the transmission of chromosome 6Mv of Aegilops ventricosa in different genetic background of Sichuan wheat varieties, hybridization with wheat- Haynaldia villosa ditelosomic addition line Pana (highly resistant to the powdery mildew) and screening of wheat- Aegilops ventricosa translocation line by exposuring Moisson 6Mv/6B under ionizing radiation. The main results are as following: 1. Moisson 6Mv/6B was crossed with Sichuan wheat varieties mianyang26, mianyang93-124 and SW3243 (highly susceptible to stripe rust), respectively. Their F1 hybrids were further backcrossed as male and female to corresponding wheat varieties. The seed-setting rate, chromosomes confirmation in the mitotic metaphase of root tip cells, and resistance to stripe rust of the subsequent BC1 and F2 plants were investigated. The average seed-setting rate of backcross via 6Mv as female donor (83.10%) was higher than that of backcross via 6Mv as male donor (48.61%), suggesting that the seed-setting rate was associated with the wheat genotypes(χ2=34.15>>χ20.05=5.99(df=2)). In all analyzed populations, transmission frequencies of chromosome 6Mv were not correlated with the ways of 6Mv through male or through female. However, transmission frequencies of chromosome 6Mv were significantly correlated with Sichuan wheat genotypes(χ2=6.42>χ20.05=5.99(df=2)). 2. To aggregating the resistances to stripe rust and powdery mildew, as well as research on the resistance reactions in different genetic background, Moisson 6Mv/6B was reciprocally hybrided with the wheat- Haynaldia villosa ditelosomic addition line Pana (highly resistant to the powdery mildew). The seed-setting rate, agronomic characters, genomic in situ hybridization (GISH) of hybrid progenies,and resistances to stripe rust and powdery mildew were investigated. The results showed that the seed-setting rate of hybridization via Moisson 6Mv/6B as female donor (80.56%) was significant higher than that via Pana as female donor (58.33%). The seed-setting rate was associated with the hybrid methods (χ2 = 4.96> χ20.05 = 3.84 (df = 1)). The plant height of hybrid progenies was about 10 cm higher than Pana, the parent with maximum height. And the maturity of hybrid progenies was about two weeks earlier than that of the parents. In the hybrid progenies, the plants with the 6Mv chromosome have the resistance to stripe rust and the plants with the telosome from Haynaldia villosa have the resistance to powdery mildew. It was found that four plants with both the 6Mv chromosome and the telosome from Haynaldia villosa were resistant to stripe rust and powdery mildew. It indicated that the resistance to stripe rust and powdery mildew aggregated, and no mutual inhibition was found. It implied that the aggregation of the two resistance genes was able to provide the new wheat germplasm with the resistances to stripe rust and powdery mildew. 3. Moisson 6Mv/6B was irradiated with 60Co-γ rays of 6Gy (120rad/min) during meiosis. The agronomic characters and chromosomes confirmation in the mitotic metaphase of root tip cells,as well as resistance to stripe rust were investigated. The seed-setting rate of irradiated plants was only 2.22%. The chromosomes in mitotic metaphase had clear fragments. The resistance to stripe rust of progeny of irradiated plants was the adult-plant resistance. The wheat- Aegilops ventricosa 6Mv/6B substitution line is a good stripe rust resistance donor for its stabile resistance. Our study demonstrated that the key for use the resistance is to choose a good receptor. There is no difference between Moisson 6Mv/6B be the female and be the male if the seed number meets the requirement. At the same time, the stripe rust resistance of Moisson 6Mv/6B did not have the mutual inhibition with the powdery mildew resistance from Haynaldia villosa. It is able to aggregate the two resistances for wheat breeding.
Resumo:
作物的抗旱性是一个多基因控制的、极为复杂的数量性状,植物对干旱在分子水平上的差异反应通过植物组织生理和细胞生物学水平,最终表现为植物抗旱性的不同。在我国,旱地农业超过耕地面积的50%,但水资源短缺,因此培育和选育抗旱高产作物是发展节水型农业最有效的途径。 青藏高原气候恶劣、年均降雨量少,也是世界大麦初生起源中心,因而蕴藏了十分丰富的与抗逆相关的种质资源材料,从这些特殊的资源材料克隆抗旱基因,不仅对培育抗旱、优质、高产大麦新品种具有重要理论意义和经济价值,而且对整个作物抗旱基础和育种应用研究都具重大促进作用。 为了筛选青稞(裸大麦,Hordeum vulgare ssp. vulgare)抗旱性材料,本研究选用来自青藏高原不同地区的84份青稞为材料,在叶片失水率(water loss rate, WLR)检测分析的基础上,选择失水率值差异显著的12个品种,通过相对含水量(relative water content, RWC)和反复干旱法评价其抗旱性,并通过植株对干旱胁迫下的丙二醛(MDA)含量和游离脯氨酸(free-proline)含量变化,了解不同抗旱性材料的生理反应特性。选择抗旱性强弱不同的品种各两份进行LEA2蛋白基因(Dhn6基因)、LEA3蛋白基因(HVA1基因)的克隆,比较LEA蛋白结构差异与作物抗旱性之间的关系。同时,对抗旱性不同的青稞品种受到干旱时间不同的失水变化率(dynamics water loss rate, DWLR)进行了检测;对抗旱性不同的青稞对照材料进行2 h、4 h、8 h和12 h的快速干旱处理,通过SYBR Green实时荧光定量RT-PCR技术对Dhn6基因、Dhn11基因、Dhn13基因和HVA1基因在不同抗旱性材料受到不同干旱时间处理后的相对表达水平进行了检测。本研究对LEA蛋白基因在抗旱性不同的青稞材料中的干旱胁迫分子水平上的差异反应进行了研究,也对植物的抗旱机理进行了初步探讨。主要研究结果如下: 1. 青稞苗期进行离体叶片失水率测定结果表明,来自青藏高原的84份青稞材料的WLR在0.086~0.205gh-1g-1DW之间。选择WLR低于0.1gh-1g-1DW和WLR高于0.18gh-1g-1DW的品种各6份,并对苗期分别进行未干旱及干旱12小时的处理。相对含水量检测结果表明,低失水率青稞材料干旱后的具有更高的相对含水量,盆栽缺水试验也显示叶片失水率低的材料耐旱能力强于失水率高的材料。通过水合茚三酮法测定离体叶片游离脯氨酸的含量,结果表明,所有品种未干旱处理时,游离脯氨酸含量差异不大(17.10~25.74 µgg-1FW);干旱12小时后,低失水率的品种游离脯氨酸含量明显增高(32.99~53.45µgg-1FW),高失水率品种的游离脯氨酸含量与干旱前变化不明显(P<0.05)。硫代巴比妥酸法测定离体叶片丙二醛(MDA)含量,结果显示,12份所选对照品种中,丙二醛的含量在0.97~2.74nmolg-1FW,干旱12小时后丙二醛的含量显著上升(1.46~4.74nmolg-1FW),高失水率的6个品种的丙二醛含量在未干旱和干旱处理时都明显高于低WLR品种。本研究结果表明青稞的低失水率、低丙二醛含量、高相对含水量和高脯氨酸含量具相关性(P<0.05)。综上研究,我们认为作物失水率的测定可以作为快速检测作物抗旱性的指标之一,因此,强抗旱品种喜玛拉10号(TR1)、品比14号(TR2)和弱抗旱品种冬青8号(TS1)、QB24 (TS2)被选作抗旱基因克隆和表达分析的研究材料。 2. 高等植物胚胎发育晚期丰富蛋白(late embryogenesis abundant proteins, LEA proteins)与植物耐脱水性密切相关,为了探讨青稞LEA蛋白结构差异性与植物抗旱性的关系,本研究以强抗旱品种(喜玛拉10号、品比14号)和弱抗旱品种(冬青8号、QB24)为材料,利用同源克隆法,通过RT-PCR,分别克隆了与抗旱性密切相关的Dhn6基因和HVA1基因。Dhn6基因序列分析结果表明,强抗旱品种品比14号和弱抗旱品种冬青8号Dhn6基因所克隆到的序列为1026bp,它们之间只有5个碱基的差异;喜玛拉10号和QB24克隆到的序列长963bp。在强弱不同的抗旱品种中有22个核苷酸易突变位点,相应的脱水素氨基酸序列推导结果表明,22个核苷酸突变位点中,仅有8个位点导致相应的氨基酸残基的改变,其余的位点系同义突变,另外,21个富含甘氨酸序列的缺失并没有联系作物抗旱性特征。推测这些同义突变位点的氨基酸残基对维持青稞DHN6蛋白的正常结构和功能起着非常重要的作用,也可能DHN6蛋白对青稞长期适应逆境胁迫和遗传进化的结果。对HVA1基因的序列分析结果表明,冬青8号、QB24、品比14号和喜玛拉10号的目的基因核苷酸序列全长分别为661bp、697bp、694bp和691bp,它们都包含1个完整的开放阅读框。相应的LEA3蛋白氨基酸序列结果表明,11个高度保守的氨基酸残基组成基元重复序列的拷贝数与青稞抗旱性之间没有必然关系,在强抗旱品种(喜玛拉10号、品比14号)中三个共同的氨基酸突变位点Gln32、Arg33和Ala195可能对抗旱蛋白的结构和功能有影响;另外,强抗旱青稞品种LEA3蛋白质中11-氨基酸保守基元序列拷贝数和极性氨基酸占蛋白的比例更高,推测LEA3蛋白中基元序列拷贝数和极性氨基酸占蛋白的比例对该蛋白的结构和功能影响更大。 3. LEA蛋白基因的表达水平的上调与植物的耐脱水性密切相关,我们对强抗旱性材料(喜玛拉10号、品比14号)和弱抗旱材料(冬青8号、QB24)进行干旱处理2 h、4 h、6 h、8 h和10 h的失水变化率进行测定,结果表明弱抗旱品种在2~4小时之间失水率变化最明显,而四个对照品种的失水率在8小时后和24小时的失水率值变化不大。进一步提取青稞苗期进行2 h、4 h、8 h和12 h的干旱处理后的总RNA,通过SYBR Green实时荧光定量RT-PCR技术对青稞脱水素基因(Dhn6、Dhn11和Dhn13)和LEA3蛋白基因(HVA1)的相对表达水平受干旱时间和作物抗旱性的影响进行了检测。研究发现,抗旱性不同的青稞品种随干旱处理的时间延长,Dhn6、Dhn11、Dhn13和HVA1基因的相对表达水平不同。 Dhn6基因的相对表达水平在强抗旱青稞品种干旱8小时后快速上升,但在弱抗旱青稞品种干旱处理12小时后检测到更高表达量;Dhn11基因在对照青稞抗旱品种的表达累积水平随干旱时间的延长持续下降;整个干旱过程中,Dhn13基因的相对表达水平在弱抗旱品种持续上升,在强抗旱品种中干旱处理8小时快速上升并达到最高,干旱12小时后降低。与脱水素基因相比较,强抗旱青稞品种在干旱2小时后HVA1基因的相对表达水平显著升高,相对表达量随干旱处理的时间持续上升,在干旱12小时后达到最高;与之相比较,在整个干旱过程中,弱抗旱品种的相对表达水平显著低于强抗旱品种,在干旱8小时之前弱抗旱品种的相对表达水平变化不明显;在干旱8~12小时后却显著上升。上述结果表明,不同的LEA蛋白在植物耐脱水过程中的干旱表达累积水平不同;干旱不是诱导高等植物Dhn11基因表达的主要因素;植物的抗旱性不同,不同LEA蛋白基因对干旱的反应有差异。推测某些LEA蛋白基因的干旱胁迫早期表达累积程度与植物的抗旱性直接相关;其中,Dhn11基因和Dhn12基因不同的表达模式可能与干旱调控表达顺式作用成分(dehydration responsive element, DRE)的有无或结构上的差异有关。 本研究结果认为,(1)失水率和相对含水量可作为植物抗旱性检测的指标之一;(2) DHN6同义突变位点的氨基酸残基对维持该蛋白的正常结构和功能起着重要作用;(3) 11-氨基酸保守基元序列拷贝数和极性氨基酸的比例对LEA3蛋白结构和功能有重要影响;(4)LEA蛋白表达随着干旱胁迫程度而增加,但Dhn11基因并不受干旱诱导表达;(5)作物的抗旱性不同,LEA蛋白对干旱的累积反应并不相同,干旱早期LEA蛋白的累积程度可能会影响植物的抗旱性。 Drought resistance was a complex trait which involved multiple physiological and biochemical mechanisms and regulation of numerous genes. Because its complex traits, it is difficult to understand the mechanisms of drought resistance in plants. Plants respond to water stress through multiple physiological mechanisms at the cellular, tissue, and whole-plant levels. Tibetan hulless barley, a pure line, is a selfing annual plant that has predominantly penetrated into the Qinghai-Tibetan Plateau and remains stable populations there. The wide ecological range of Tibetan hulless barley differs in water availability, temperature, soil type and vegetation, which makes it possess a high potential of adaptive diversity to abiotic stresses. This adaptive genetic diversity indicates that the potential of Tibetan hulless barley serves as a good source for drought resistance alleles for breeding purposes. 12 contrasting drought-tolerant genotypes were selected to measure relative water content (RWC), maldondialdehyde (MDA) and proline content, based on values of water loss rate (WLR) and repeated drought methods from Tibetan populations of cultivated hulless barley. As a result of the screening, sensitive and tolerant genotypes were identified to clarify relationships between characteristics of LEA2/LEA3 genes sequences and expression and drought-tolerant genotypes, associated with resistance to water deficit. In addition, dynamics water loss rate (DWLR) was measured to observe the changes on diffrential drought-tolerant genotypes. Real-time quantitative RT-PCR was applied to detect relative expression levels of Dhn6, Dhn11, Dhn13 and HVA1 genes in sensitive and tolerant genotypes with 2 h, 4 h, 8h and 12 h of dehydration. In the present study, differential sequences and expression of LEA2/LEA3 genes were explored in Tibetan hulless barley, associated with phenotypically diverse drought-tolerant genotypes. 1. The assessments of WLR and RWC were considered as an alternative measure of plant water statues reflecting the metabolic activity in plants, and the parameters of MDA and proline contents were usually consistent with the resistance to water stress. The values of detached leaf WLR of the tested genotypes were highly variable among 84 genotypes, ranging from 0.086 to 0.205 g/h.g DW. The 12 most contrasting genotypes (6 genotypes with the lowest values of WLR and 6 genotypes with the highest values of WLR) were further validated by measuring RWC, MDA and free-proline contents, which were well watered and dehydrated for 12 h. Results of RWC indicated that the values of 12 contrasting genotypes RWC ranged from 89.94% to 93.38% under condition of well water, without significant differences, but 6 genotypes with lower WLR had higher RWC suffered from 12 h dehydration. The results indicated that lower MDA contents, lower scores of WLR and higher proline contents were associated with drought-tolerant genotypes in hulless barley. Remarkably, proline amounts were increased more notable in 6 tolerant genotypes than 6 sensitive genotypes after excised leaves were dehydrated for 12 h, with control to slight changes under condition of well water. Results of MDA contents showed that six 6 tolerant genotypes had lower MDA contents than the 6 sensitive genotypes under both stressed and non-stressed conditions. As a result of that screening, drought- resistant genotypes (Ximala 10 and Pinbi 14) and drought-sensitive genotypes (Dongqing 8 and QB 24) were chosen for comparing the differential characteristics of LEA2/LEA3 genes and their expression analysis. It was conclusion that measurements of WLR could be considered an alternative index as screening of drought-tolerant genotypes in crops. 2. Late embryogenesis abundant (LEA) proteins were thought to protect against water stress in plants. To explore the relationships between configuration of LEA proteins and phenotypically diverse drought-tolerant genotypes, sequences of LEA genes and their deduced proteins were compared in Tibetan hulless barley. Results of comparing Dhn6 gene in Ximala 10 and QB24 indicated that absence of 63bp was found, except that only 5 mutant nucleotides were found. While 22 mutant sites were taken place in Dhn6 gene between sensitive and tolerant lines, 14 synonymous mutation sites appeared in the contrasting genotypes. The additional/absent polypeptide of 21 polar amino acid residues was not consistent with phenotypically drought-tolerant genotypes in hulless barley. It was deduced that synonymous mutation sites would play important roles in holding out right configurations and functions on DHN6 protein. The sequencing analysis results indicated that each cloned HVA1 gene from four selected genotypes contained an entire open reading frame. The whole sequence of HVA1 gene from Dongqing 8, QB24, Pinbi 14 and Ximala 10 was respectively 661bp, 697bp, 694bp and 691bp. Results of DNA sequence analyses showed that the differences in nucleotides of HVA1 gene in sensitive genotypes were not consistent with that of tolerant genotypes, except for absence of 33 nucleotides from +154 to +186 (numbering from ATG) in QB24. Database searches using deduced amino acid sequences showed a high homology in LEA3 proteins in the selected genotypes. Multiple sequence alignments revealed that LEA3 protein from Dongqing 8 was composed of 8 repeats of an 11 amino acid motif, less the fourth motif than Pinbi 14, Ximala 10 and QB24. Consistent mutant amino acid residues appeared in contrasting genotypes by aligning and comparing the coding sequence region, including Gln32, Arg33 and Ala195 in tolerant genotypes as compared to Asp32, Glu33 and Thr195 (Thr184 in Dongqing 8) in sensitive lines. It was concluded that consistent appearance of Gln32, Arg33 and Ala195 would contributed to functions of LEA3 protein in crops, as well as higher proportion of 11-amino-repeating motifs and polar amino acid residues. 3. Most of the LEA genes are up-regulated by dehydration, salinity, or low temperature, are also induced by application of exogenous ABA, which increases in concentration in plants under various stress conditions and acts as a mobile stress signal. Higher levels of proteins of LEA group 3 accumulated was correlated well with high level of desiccation tolerance in severely dehydrated plant seedlings. Dehydrins (DHNs), members of LEA2 protein, are an immunologically distinct protein family, and Dhn genes expression is associated with plant response to dehydration. Dynamic water loss rate was measured between sensitive genotypes and tolerant genotypes after they were dehydrated for 2 h, 4 h, 6h and 8 h. Detailed measurements of WLR at the early stage of dehydration (2, 4, 6, and 8 h) showed that WLR was stabilizing after 8 h, and there were no significant changes between these values and WLR after 24 h. Drought stress was applied to 10-day-old seedlings by draining the solution from the container for defined dehydration periods. Leaf tissues of the selected genotypes were harvested from control plants (time 0); and after 2, 4, 8, and 12 h of dehydration. Differential expression trends of Dhn6, Dhn11, Dhn13 and HVA1 genes were detected in phenotypically diverse drought-tolerant hulless barleys, related to different time of dehydration. Results of quantitative real-time PCR indicated that relative level of HVA1 expression was always higher in tolerant genotypes, rapidly increasing at the earlier stages (after 2-4 h of dehydration). However, HVA1 expressions of sensitive genotypes had a fast increase from 8 h to 12 h of stress. Significant differences in expression trends of dehydrin genes between tolerant genotypes and sensitive lines were detected, mainly in Dhn6 and Dhn13 gene, depending on the duration of the dehydration stress. The relative expression levels of Dhn6 gene were significantly higher in tolerant genotypes after 8 h dehydration, by control with notable higher expression levels after 12 h water stress in sensitive ones. The relative expression levels of Dhn13 gene tended to ascend during exposure to dehydration in drought-sensitive genotypes. However, fluctuate trends of Dhn13 expression level were detected in drought-resistant lines, including in lower expression levels of 12 h dehydration as compared to 8 h water stress. It was conclusion that (1) diverse LEA proteins would play variable roles in resisting water stress in plants; (2) expression of Dhn11 gene was not induced by dehydrated signals because of the trends of expression descended in contrasting genotypes suffered from water deficit and (3) variable accumulations on LEA proteins would be appear in diverse drought-tolerant genotypes during dehydrations. It is deduced that higher accumulations of Dhn6 and Dhn13 expression in 8 h dehydration are related to diverse drought-tolerant lines in crops. The present results indicated that different dehydrin genes would play variable functional roles in resisting water stress when plants were suffered from water deficit. The authors suggest physiologically different reactions between resistant and sensitive genotypes may be the results of differential expression of drought-resistant genes and related signal genes in plants. In addition, contrarily induced expression of Dhn11 and Dhn12 was related to dehydration responsive element (DRE) in barleys. The present study indicated that (1) measurements of WLR and RWC could be considered as one index of drought-tolerant screenings; (2) synonymous mutation sites would play important roles in holding out right configurations and functions on DHN6 protein, (3) higher proportion of 11-amino-repeating motifs and polar amino acid residues would contribute to functions on LEA3 protein, (4) the longer drought, the more accumulation on LEA proteins, except for Dhn11 gene in crops and (5) differential responses on expression of LEA protein genes would result in physiological traits of drought tolerance in plants.
Resumo:
Hydrogen peroxide (H2O2)/kerosene is a prospective bipropellant due to its high-energy content, high storage density, and environmentally benign properties. The possibility of making it hypergolic renders this option even more attracting. Self-ignitable H2O2/kerosene bipropellants were prepared by combining different candidate catalysts and promoters. Preliminary screening evaluations were conducted by using a dropping-test method. Propulsive performances of the combinations having passed satisfying dropping-test requirements were then investigated on a specially designed thrust engine. The results revealed that short ignition delay and reliable propulsion performances could be acquired in both steady-state and pulse-mode operations, and the combination of kerosene with additives and H2O2 of 90% concentration could still have good performances after 3 months storage time. It is expected that the combination of H2O2 and kerosene can be an efficacious alternative for storable toxic propellants used currently.
Resumo:
A multistream reactor for high-throughput examining the surface acidity by NH3-TPD method by application of multistream mass spectrometer screening (MSMSS) technique has been developed. This method allows for examining the surface acidity of 10 catalyst samples in about 6 h, which is an improvement over the traditional process. The demonstration of the feasibility of high-throughput TPD can be significant in convincing the hardened traditionalists in the heterogeneous catalysis community that, combinatorial methods indeed should have an important place in scientific catalyst research and development. The developed method could also be used for almost all the temperature-programmed analysis theoretically with careful designed multistream reactors. (C) 2003 Elsevier B.V. All rights reserved.
Resumo:
High melt strength polypropylene (HMSPP) was synthesized by in situ heat induction reaction, in which pure polypropylene (PP) powders without any additives were used as a basic resin and vinyl trimethoxysilane (VTMS) as a grafting and crosslinking agent. The grafting reaction of VTMS with PP was confirmed by FTIR. The structure and properties of HMSPP were characterized by means of various measurements. The content of grafted silane played a key role on the melt strength and melt flow rate (MFR) of HMSPP. With increasing the content of grafted silane, the melt strength of HMSPP increased, and the MFR reduced. In addition, due to the existence of cross-linking structure, the thermal stability and tensile strength of HMSPP were improved compared with PP.
Resumo:
Cobalt porphyrin complex ((TPPCoX)-X-III) (TPP = 5, 10, 15, 20-Tetraphenylporphyrin; X = halide) in combination with ionic organic ammonium salt was used for the regio-specific copolymerization of propylene oxide and carbon dioxide. A turnover frequency of 188 h(-1) was achieved after 5 h, and the byproduct propylene carbonate was successfully controlled to below 1%, where the obtained poly(propylene carbonate) (PPC) showed number average molecular weight (M-n) of 48 kg/mol, head-to-tail content of 93%, and carbonate linkage of over 99%.
Resumo:
In vitro a-glucosidase inhibition assays and ultrafiltration liquid chromatography with photodiode array detection coupled to electrospray ionization tandem mass spectrometry (ultrafiltration LC-DAD-ESI-MSn) were combined to screen a-glucosidase inhibitors from hawthorn leaf flavonoids extract (HLFE). As a result, four compounds were identified as alpha-glucosidase inhibitors in the HLFE, and their structures were confirmed to be quercetin-3-O-rha-(1-4)-glc-rha and C-glycosylflavones (vitexin-2 ''-O-glucoside, vitexin-2 ''-O-rhamnoside and vitexin) by high-resolution sustained off resonance irradiation collision-induced dissociation (SORI-CID) data obtained by Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS).
Resumo:
A new strategy for preparing ammonium-type ionic liquid (IL) by acid/base neutralization reaction was proposed. The method contributed to preparing hydroxide-based ammonium IL and resulting task specific ionic liquid (TSIL) with high purity using a low-costly and environment-friendly synthetic. route. Halide contamination in the prepared ILs could be markedly decreased than those prepared by well-established anion metathesis method. Moreover, some novel TSILs composed of cations and anions with big steric hindrances could be prepared by this method. Physicochemical properties of the bifunctional TSILs, i.e., density, water content, decomposition temperature, and munal solubility, were also studied in this article.
Resumo:
A series of red-light emitting electrophosphorescent polyfluorenes (PFs) with varying content of a quinoline-based iridium complex, (PPQ)(2)Ir(acac) (bis(2,4-diphenylquinolyl-N,C-2') iridium(acetylacetonate)), in the side chain are synthesized by Suzuki polycondensation. Because of the efficient Forster energy transfer from the PF main chain to (PPQ)(2)Ir(acac) and direct charge trapping on the complex, the electroluminescent emission from PF is nearly completely quenched, even though the amount of iridium complex I incorporated into the polymers is as low as 1 mol %. Based on a single-layer device configuration, a luminous efficiency of up to 5.0 cd A(-1) with a luminance of 2000 cd m(-2) and Commission Internationale de L'Eclairage coordinates of (0.63, 0.35) (x, y) is realized, which is far superior to that of previously reported red-light emitting PFs containing benzothiazole- and isoquinoline-based iridium complexes.
Resumo:
Composite membranes based on Sulfonated poly(ether ether ketone) (SPEEK) and sulfonated organically modified Si-SBA-15 (S-SBA-15) were investigated with the purpose of increasing the proton conductivity. The novelty of the composite membranes was attributed to two special structures and different ion exchange capacities (IEC) of S-SBA-15 fillers, which were embedded in membranes. The typical hexagonal channels array of S-SBA-15 was confirmed by XRD and TEM. The regular vermiculate and amorphous structures of the inorganic fillers were proved by SEM. Composite membranes were prepared through common solvent casting method. SEM images indicated that the inorganic filler with regular structure dispersed homogeneously in the composite membranes, but the amorphous filler caused an agglomeration phenomenon at the same loading content.
Resumo:
A novel hard material of (W0.25Al75)C has been successfully prepared by the high-pressure sintering process without the addition of any binder phase. The high-pressure is a suitable and powerful technique for sintering the binderless hard material, the relative density of the hard material can reach 99.6% under high-pressure sintering. The density of the novel light hard material is only 6.2371 g cm(-3), which is much lighter than the normal hard material. The hardness of the light hard material can reach 18.89 GPa even the aluminum content get the astonished 75%.