938 resultados para Eubulus, ca. 405-330 B.C.
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En 1991 se estableció un ensayo en época seca (20-30 Abril) en el Centro Experimental del Algodón, Posoltega, Chinandega, con los objetivos de determinar si el efecto de diferentes tiempos de exposición a deshidratación del coyolillo (Cyperus rotundus L.) al arar el suelo en seco con arado de discos para reducir su densidad, se mantiene hasta la época de postrera y asimismo si tiene efecto sobre el crecimiento y rendimiento del cultivo de ajonjolí (Sesamun indicum L.). Para tales efectos se usó un diseño de Bloques Completos al Azar (B.C.A.) con seis tratamientos y seis repeticiones, evaluándose el factor tiempo de exposición a deshidratación del coyolillo. Los tratamientos consistieron en realizar la práctica de arar el suelo en seco en las diferentes parcelas a intervalos de dos días, empezando el 20 de abril con el tratamiento diez días y terminando el 30 de abril con el tratamiento cero días de exposición, aplicándose ese mismo día el riego en todo el área. Los tubérculos quedaron expuestos a la deshidratación durante 1O, 8, 6, 4, 2 y O días. Después de establecer lo tratamientos, se sembró el 2 de mayo el cultivo de soya (Glycine max (L.) Merr.) (Época de primera) y el cultivo de ajonjolí del presente ensayo el 5 de septiembre (época de postrera). Las variables evaluadas consistieron para coyolillo: densidad y para ajonjolí: altura de plantas, número de hojas, diámetro del tallo y rendimiento. El análisis de varianza mostró significancia estadística (P < 0.05) en densidad a los 14, 28 y 42• dds, altura de plantas 42 y 89 dds, número de hojas 28 dds y diámetro del tallo 42 dds. El contraste olinomial planificado presentó significancia estadística para la tendencia lineal (P <0.05) para densidad a los 14, 28.y 42.dds, altura de plantas 14, 42y 89 dds, número de hojas 28 y 42 dds, diámetro del tallo 28 y 42 dds y rendimiento. Los mayores valores de medias para densidad de coyolillo se presentan en el tratamiento cero días y los menores valores en el tratamiento ocho días de exposición a deshidratación del coyolillo para las tres fechas de muestreo. Todas las variables evaluadas en el cultivo de ajonjolí presentan los mayores valores en el tratamiento ocho días y los menores valores en el tratamiento cero días de exposición, excepto altura 14 dds, donde el mayor valor se presenta en el tratamiento cero días y el 'menor valor en el tratamiento seis días de exposición a deshidratación del coyolillo. Los análisis de regresión realizados entre la variable independiente tiempo de exposición y las variables dependientes del coyolillo y el cultivo del ajonjolí, mostraron una respuesta de tendencia lineal negativa en densidad de coyolillo y tendencia lineal positiva en todas las variables del cultivo de ajonjolí, excepto altura 14 dds que presentó una tendencia lineal negativa.
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El presente trabajo se realizó en el Centro Experimental laCompañía, ubicado en el municipio de San Marcos, Carazo, Nicaragua. El objetivo fue evaluar diferentes prácticas de manejo de suelos en el crecimiento, desarrollo y el rendimiento del maíz (Zea mays l.). y la concentración y extracción de nutrientes por el cultivo y las malezas. El estudio consistió en un experimento de campo, realizado en la época de primera; En suelos franco arenosos, con buen drenaje, pH 6.5. los sistemas de labranza estudiados fueron: labranza cero, labranza cero más subsoleo, labranza convencional, labranza convencional mas subsoleo, labranza con bueyes y labranza con bueyes más rastrojos. la fertilización consistió en una sola aplicación, a razón de 100 kg/ha de nitrógeno (urea 46 porciento), al momento de la siembra. El diseno utilizado fue bloques completos al azar (BCA), las poblaciones manejadas fue de 35 757 plantas por hectárea. En la cosecha, las muestras de plantas fueron separadas en raíz, tallo, paja, tuza. olote y grano a las cuales se les realizó análisis de N, P, K, Ca, Mg, S, Mn, Zn, B, Fe, Cu y C. la mayor eficiencia obtenida en rendimiento, altura, diámetro del tallo y extracción de N, P, K, S, Ca, Zn y B en la labranza cero, en comparación con el r&sto de labranza fue aparentemente a que esta labranza garantiza un mayor contenido de humedad, la labranza con bueyes, más rastrojos ejerció un efecto positivo en la absorción de Fe, la labranza con bueyes mas rastrojos en Mn y labranza convencional más subsoleo en Cu. los diferentes sistemas de labranza no ejercieron efectos significativos en la concentración de nutrientes de las malezas, así como en su peso seco, sin embargo, se observó que en el sistema convencional fueron estas favorecidas, la menor extracción de los nuitrientes por las malezas en la labranza con bueyes
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El presente trabajo fue realizado en la estación experimental "La Compañía" Carazo, en época de primera (junio - agosto de 1994), en suelos jóvenes de origen volcánico con altos contenidos de Potasio y deficiente en fósforo. El propósito del experimento fue determinar la variación en la cantidad de Nitrógeno fijado, así como la extracción de nutrientes y acumulación de materia seca, por cuatro variedades de frijol común. Las variedades evaluadas fueron: DOR-364, ESTELÍ 90-A, BRUNCA, CRIOLLA y variedad TESTIGO. El diseño utilizado fue bloques completos al azar (B.C.A) con cuatro tratamientos y cuatro repeticiones. Las variables evaluadas fueron: altura de planta, número de plantas cosechadas por ha, número de vainas por planta, rendimiento de grano, número de nódulos, acumulación de materia seca, concentraciones de nutrientes en la planta y cantidades exportadas por ha. Los datos se procesaron usando análisis de varianza (ANDEVA) y se utilizó la prueba de rangos múltiples de DUNCAN (p 0.05). Los resultados obtenidos se pueden sintetizar de la siguiente forma: La variedad DOR-364 alcanzó una mayor altura así como también una mayor concentración de P y Zn en sus tejidos y acumuló más materia seca, sin embargo disminuyó tanto en el rendimiento de grano como en el número de vainas, además fue deficiente en la población. Esta variedad extrajo la mayor cantidad de N, sin embargo la mayor concentración de este elemento en el follaje la presentó la variedad (testigo. La ESTELÍ 90-A, presentó un mayor número de plantas por ha y un alto rendimiento de grano, fue la variedad más eficiente en la nodulación. Las variedades DOR-364 y ESTELI 90-A exportaron más nutrientes que el resto de las variedades evaluadas. Se desarrolló una tendencia por parte de la variedad TESTIGO a presentar una mayor concentración de K, S, Ca, Mg, Fe, Cu y Mn en sus tejidos en comparación con las otras variedades estudiadas. Se realizaron correlaciones entre cada uno de los elementos y variables en estudio, las cuales mostraron resultados positivos en su mayoría a excepción de la relación entre el número de vainas y los elementos Cu y Fe. El rendimiento se relacionó solamente con el número de nódulos en la segunda evaluación.
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El presente trabajo fue realizado en la estación experimental "La Compañía" Carazo, en época de postrera 1995, en suelos jóvenes de origen de cenizas volcánicas, el suelo es franco-limoso con altos contenidos de potasio y deficientes en fósforo. De acuerdo a sus propiedades, este suelo puede ser considerado adecuado para la mayoría de los cultivos; pH neutro y buen drenaje. El propósito del experimento fue determinar la capacidad de extracción de macro elementos, elementos secundarios, acumulación de materia seca y la dinámica de plagas, enfermedades y enemigos naturales en el cultivo de frijol, para dos variedades de frijol común (Phaseolus vulgaris L.). Las variedades evaluadas fueron: Dor 364, Revolución 84. El diseño utilizado fue en bloques completamente al azar (B.C.A.) con seis tratamientos y cuatro repeticiones, cada tratamiento era un tipo de labranza. Los datos se sometieron a un análisis de varianza (ANOEVA) y se utilizó la prueba de rangos múltiples según Tukey y SNK al 95 por ciento de confianza. Para la recopilación de información sobre plagas y enfermedades los datos se arreglaron en un diseño de parcelas divididas ubicndose los tipos de labranza en las parcelas grandes y las variedades en las parcelas pequeñas. El análisis de plagas se hizo usando el paquete estadístico (S.A.S.). El análisis estadísticos de datos se realizó a través de ANDEVA y se separaron los tratamientos por medio de la prueba de S.N.K . Los datos obtenidos se pueden sintetizar de la siguiente forma: La variedad Dor 364 alcanzó una mayor altura en el sistema de labranza mínima, las mayores extracciones de macro elementos y elementos secundarios (P, K, Ca, Mg, y S ), se obtuvo en el sistema de labranza cero + mulch + subsoleo y al mismo tiempo acumuló más materia seca. En cuanto a la variedad Revolución 84 la mayor altura se obtuvo en la labranza cero + mulch, las mayores extracciones y acumulación de materia seca se obtuvo en la labranza convencional La labranza con mejores rendimientos fue la convencional, para ambas variedades evaluadas en el ensayo. La planta indicadora (sorgo) mostró mayor extracción de P, K, Ca, Mg, y S en sus tejidos en comparación con las otras variedades estudiadas En los resultados referentes a plagas de suelo se encontró que labranza mínima presentó mayor número de especies de plagas de suelo. En cuanto insecto plagas de follaje, los mas importantes que se presentaron en el ensayo están; Diabrótica spp., Nodonata spp., y Empoasca spp., la mayor presencia fue de Empoasca spp, la cual estuvo en casi todos los muestreos realizados. De los seis sistemas de labranzas evaluados labranza mínima fue la que mejor disminuyó las poblaciones de insectos plagas del follage. La mayor cantidad de benéficos se observó en labranzas mínima mas incorporación, seguida muy de cerca por labranza convencional más subsoleo. En cuanto a la incidencia y severidad de las enfermedades, fue notorio que hubo poca incidencia en las plantas. Sin embargo en las pocas plantas que hubo presencia de agentes patógenos se observó una alta severidad, la cual fue mayor usando, cero labranzas más mulch.
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Three groups of Sarotherodon niloticus fry were fed for 8 weeks on diets either treated with 17- & methyltestosterone (MT), alcohol (CA), or untreated (CO). Growth rate and food utilization in the different groups were compared. Results indicate that the best growth, Feed Conversion Ratio (FCR), Protein Efficiency Ratio (PER) and Mean Growth Rate (MGR) were obtained with the MT diet. There was no significant difference (P 0.05) in growth and food utilization of the CA and CO fry, nor in the mortality rate of the 3 treatments. The androgen, methyltestosterone promotes growth and protein anabolism without producing toxic effects in S. niloticus
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O eixo temático se desenvolveu a partir do questionamento sobre como foi o processo de reconstrução de Sagunto, que foi promovido por Roma. A região de Sagunto configura como o motivo central para o embate entre romanos e cartagineses. Todavia, com o término da Segunda Guerra Púnica (218-202 a.C.), a cidade estava destruída e uma embaixada saguntina foi enviada para Roma, a fim de solicitar ao Senado sua reorganização. O pedido aparece como sendo bem aceito pelos senadores romanos. Contudo, a partir desse momento, começa o silenciamento. A escassez de informações sobre a temática foi o primeiro problema encontrado ao longo da pesquisa. Sendo assim, foi necessário recorrer à documentação arqueológica da cidade, à numismática e à epigrafia para conseguir preencher as lacunas referentes ao tema de pesquisa. Os indícios possibilitaram não somente compreender a cidade de Sagunto e seus vários estatutos jurídicos perante Roma, como também lançar outro olhar sobre as práticas imperialistas. Ao evocar Edward Said como teórico deste trabalho, elemento de inovação da pesquisa, é possível construir a estrutura de atitudes e referências que os romanos, entre os séculos II a.C. e I d.C., aplicaram na região saguntina para consolidar o seu poder. Assim, por meio do estudo das entidades geográficas, compreende-se o espaço físico da cidade e, pelo conceito de entidades culturais, analisam-se o sistema administrativo e os colégios sacerdotais atuantes em Sagunto, no século I d.C. Logo, o imperialismo romano pode ser visto como um mecanismo que se vale de diversos elementos, os quais não se limitam à força no processo de ocupação. Em suma, política e cultura são peças centrais no processo de preservação do poder romano no espaço provincial.
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A história da pólis de Esparta como, por vezes, nos foi apresentada tomou uma perspectiva historiográfica dotada de pressupostos Atenocêntricos, os quais acabaram apresentando-a como rústica, dotada de uma economia e uma cultura estática e demasiadamente inclinada às atividades militares. No entanto, através de nossa pesquisa verificamos que as práticas político-culturais dos cidadãos de Esparta eram dinâmicas, para sua época. Seguindo por esse viés, verificamos que as representações de Esparta, sobretudo dos esparciatas e dos seus basileus, variaram de acordo com o grupo social e o contexto histórico em que foram empregadas. Com isso, observamos que embora os cidadãos de Esparta tenham sido, em algumas circunstâncias, criticados pelos pensadores antigos, esta não foi uma tendência hegemônica. Sendo assim, mediante os indícios da documentação literária do período Clássico, notamos que os esparciatas e os seus basileus teriam sido homens dotados de um habitus tradicional, o qual valorizava o aprimoramento físico e mental, assim como a responsabilidade com os deveres sagrados. Através da interação entre os vestígios documentais e dos estudos historiográficos mapeamos parte das representações de Esparta que figuraram os diversos discursos no decorrer da história do Ocidente, no intuito de materializarmos as possíveis motivações político-culturais nas apropriações do habitus espartano. Por conseguinte, recorremos à documentação literária para entendermos como parte dos pensadores clássicos concebeu, por meio de uma memória ancestral, a formação da região da Lacedemônia e da pólis de Esparta, a qual teria se dado concomitantemente com a legitimação político-cultural da identidade étnica dos basileus e dos esparciatas. Por fim, analisamos as práticas rituais em honra ao deus Apolo como um mecanismo empregado pelos segmentos sociais hegemônicos da Lacedemônia para ratificar o seu poder político frente a grupos sociais submetidos.
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A bioassay study was conducted using three organic pesticides to determine their comparative toxicity to fingerlings of Hypophthalmichthys molitrix. There was wide variation in the toxicity of different pesticides with 24 hr LC sub(50) values ranging from 0.000403 to 0.294 mg/l. Endosulfan appeared to be the most toxic, whereas BHC was the least.
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比较了云南高黎贡山地区的贡山独龙江和腾冲大蒿坪白颌大角蟾(Megophryslateralis)两个地理种群的核型、C-带和Ag-NORs结果表明,两个地理种群在核型和带型上都有差异两个地理种群的核型均为2n=26,NF=52染色体形态差异不明显,而次缢痕的位置完全不同,贡山独龙江标本的次缢痕位于No.2的长臂上近着丝点处,腾冲标本的次缢痕位于No.5的短臂上近着丝点的部位在腾冲标本中发现一雄性个体中有一条额外的染色体,可能是B染色体两地标本的C-带差异不太显著,贡山独龙江标本的C-带相对较为显著.贡山独龙江标本的Ag-NORs位于No.2长臂近着丝点处,与次缢痕的部位对应,两条同源染色体上大小有显著差异.腾冲标本的Ag-NORs位于No.5短臂上近着丝点处,与次缢痕的部位对应依据核型和带型的比较,对白颌大角蟾的分类和进化问题进行了讨论。
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银额果蝇昆明群体有丝分裂中期核型中存在B染色体, 出现频率为69.1%在已 研究过的来自各个地区的银额果蝇中, 昆明群体的B染色体频率最高。B染色体 数目为1—6条。该群体内单雌系间的B染色体数目不同, 个体间和细胞间的B染色 体数目也不同。在核型中, B染色体最小, 形态稳定, 点状, C-带和G-带呈阳性 。 图版1图2表1参12
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鲤科是鱼类最大的科,在中国淡水鱼类组成中鲤科鱼类的成分占一半以上.鲤科鱼类的演化过程代表了东亚淡水鱼类的整体演化过程.为探讨东亚鲤科鱼类系统发育关系,共分析了包括18种新测序列在内的54种鲤科鱼类细胞色素b基因的全序列.分析的物种涵盖了鲤科鱼类的12个亚科并对问题较多的(鱼丹)亚科(Danioninae)和雅罗鱼亚科(Leuciscinae)进行了广泛的采样.系统发育树的建立使用了多种方法,包括邻接法、最大简约法和最大似然法.亚口鱼科(Catostomidae)的胭脂鱼(Myxocyprinus asia
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本文首次合成了Lar_xSr_xFe_yCor_yO_3(x、y:0.1、0.3、0.5、0.7、0.9)系列化合物和Lar_xBa_xFe_yCor_yO_3(x、y:0.1、0.3、0.5、0.7、0.9)系列化合物,并对其结构,电性及气敏性质进行了研究。关于ABO_3型稀土过渡金属复合氧化物的结构研究,文献上报道比较混乱,不同的人在进行结构研究中采用不同的结构基元得到了不同的晶型和相应的晶胞参数。本文根据上述化合物的X射线谱图,对其进行指标化,得到了上述两系列化合物为主方晶系结构,晶胞参数计算结果表明:在Y = 0.7时,化合物Lar_xSr_xFe_(0.7)Co_(0.3)O_3的晶胞参数随X值的增大而减小。化合物Lar_xBa_xFe_(0.7)Co_(0.3)O_3的晶胞参数随X值的增大而增大。在X = 0.3时,上述两条系列化合物的晶胞参数均随Y值的增大而增大。对以上两系列化合物所表现出的结构变化规律,我们用起决定作用的离子半径大小进行了解释。针对上述两系列化合物,我们研究了它们的电学性质,研究结果表明,上述两系列化合物,在Y = 0.7时,其室温电阻率均随X在 0 < x < 0.5内增大而减小。在0.5 < x < 1内随X值增大而增大。在X = 0.5时的两个化合物具有最小的室温电阻率。在此方面,其电阻率变化规律与LarxMxBO_3(M:Ca Sr、 Ba、 B:Fe Co)化合物的变化规律相似。在X = 0.3时,上述两系列化合物的室温电阻率均随Y值的增大而增大,上述两系列化合物所表现出的室温电阻率变化规律可以运用C. Zener的双交换模型加以解释,同时,低温电阻率测定表明:上述两系列化合物在温度100-263K内,表现半导体性质。液氮温度下它们不具有超导性。除以上研究之外,我们针对Lar_xSr_xFe_yCor_yO_3 (X = 0.3、 Y:0.1、0.5、 0.9, Y = 0.7, X:0.1、 0.9)系列化合物初步地进行了气敏性研究。结果为:化合物La_(0.7)Sr_(0.3)Fe_yCor_yO_3 (Y:0.1、 0.5、 0.9)都不同程度地对乙醇、石油醚、 乙炔、烟具有气敏性,它们对乙醇的气敏响应时间约为1分钟,而对石油醚、乙炔和烟的气敏响应时间要用5分钟左右,在上述化合物中,La_(0.7)Sr_(0.3)Fe_(0.5)Co_(0.5)O_3化合物表现出最强的对乙醇、石油醚、乙炔的气敏性。对化合物La_(0.7)Sr_(0.3)Fe_(0.5)Co_(0.5)O_3的进一步研究表明,在乙醇气氛下,化合物La_(0.7)Sr_(0.3)Fe_(0.5)Co_(0.5)O_3在工作电流I_(Dn) = 0.20A时具有最佳的气敏数应。工作电流高于或低于此值都将导致其气敏效应的大小顺序为La_(0.7)Sr_(0.3)Fe_(0.5)Co_(0.5)O_3 > La_(0.7)Sr_(0.3)Fe_(0.9)Co_(0.1)O_3 > La_(0.7)Sr_(0.3)Fe_(0.1)Co_(0.9)O_3对化合物Lar_xSr_xFe_(0.7)Co_(0.3)O_3 (X:0.1、 0.9)的气敏性研究表明:它们只对乙醇和烟具有气敏性。对上述两种气氛的气敏响应时间为5分钟。上述两种化合物中,La_(0.1)Sr_(0.9)Fe_(0.7)Co_(0.3)O_3气敏效应最强。
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从新几内亚核桃木的树皮中分离得到的吲哚类喹诺里西定生物碱10-Desbromoarborescidine A,因发现其具有阻滞钙离子通道的活性而倍受关注。10-Desbromoarborescidine A由A、B、C、D四个环组成,只有一个手性中心,是吲哚生物碱中结构较简单的一种,常作为此类生物碱全合成方法的模型化合物。但迄今为止,能高效而简便的实现手性10-Desbromoarborescidine A不对称全合成方法线路不多,大多数以不对称诱导的方式建立其手性中心,手性催化的方式仅有一例金属催化。从逆合成分析可知,Desbromoarborescidine A的全合成可以通过亚胺不对称催化还原进行关键的手性中心构建,而本课题组在之前的研究中通过手性有机小分子催化剂的发展,已将三氯硅烷氢转移还原亚胺发展成了一类简便实用、高效、高对映选择性并具有优良底物适应范围的不对称催化反应,我们希望以这一反应作为关键手段,发展一条Desbromoarborescidine A及其类似物不对称合成新路线。 根据我们设计的新路线,首先成功合成了其关键中间体,然后我们进行了关键的不对称催化尝试。用本实验室已有的高性能有机小分子催化剂虽得到了较好的对应选择性,但是产率很低。同时,为了验证整条线路的可行性,我们也用消旋的中间体进行拉通线路的尝试。但不幸的是,在脱除保护基时遇到了很大困难。尝试换不同的保护基,或改变脱保护基的顺序,都未能成功合成目标产物。究其原因可能是由于吲哚的特殊性造成的,吲哚类亚胺与常规的芳香亚胺有较大的差异,其NH基团无论保护还是不保护,对与其2位相联接的C=N双键均有很大的影响,导致其不对称催化还原难以进行。另外,由于所设计的还原产物含有处在吲哚苄位的胺基,稳定性较差,造成保护基脱除困难。 烯胺C-亚磺酰化反应是本课题组最近发现的一个新反应,之前未见文献报道。本研究对该反应进行了反应条件优化和底物扩展,发现带Cbz,Ac,COt-Bu,CO2Et,Bz等保护基的一系列环状和非环状烯胺在亚磺酸钠、DMAc和MeSiCl3的共同作用下能高效高产率生成β-胺基烯基亚砜类新化合物,为合成多官能团化的烯基亚砜新化合物提供了一条简便实用的途径。 The main constituent of Dracontomelum mangiferum B1, indoloquinolizidine alkaloid 10-Desbromoarborescidine A, has drawn great attention due to its calcium channel blocking activity. Its molecular structure is relatively simple compared with the other alkaloids of the same type, which has only one chiral center, albeit with four cycles A, B, C, and D. This compound is often used as a model target for exploring different strategies for the total synthesis of indole alkaloids. Nevertheless, so far there still lack practical and highly efficient methods for the asymmetric total synthesis of 10-Desbromoarborescidine A. Most of the current available methods rely on stoichiometric asymmetric synthesis for the construction of the chiral center. There is only one example reporting utilization of asymmetric catalysis, but with transition metal complex as the catalyst. Our retrosynthetic analysis shows that catalytic asymmetric reduction of imine could be used as the key step for the construction of the chiral center of Desbromoarborescidine A. Since in the previous studies our group has developed the asymmetric reduction of imines by trichlorosilane into a practical and highly efficient and enantioselective method using newly designed chiral organocatalysts, we hope to apply this method to develop a novel synthetic route for the total synthesis of Desbromoarborescidine A and its analogues in this study. According to the newly designed synthetic route, we first accomplished the synthesis of the key intermediates which was then examined for the critical asymmetric catalysis. The asymmetric reduction using the highly efficient organocatalysts, developed in our lab afforded high ee but poor yield. We tried different reaction conditions to improve the yield, but failed to get any good results. Simultaneously, to vertify the feasibility of the synthetic route we designed, we also tired to go through the route toward the racemic synthesis of Desbromoarborescidine A. But unfortunately, protection and deprotection proved to be big hurdles. All the different protection groups and different sequences of protection and deprotection we tried failed to get us through the designed synthetic sequence and furnish the final product. Most likely, the indole part is the culprit behind the failures.The NH group of the indole, no matter protected or not, may impact the catalytic asymmetric reduction of C-N double bond connected with 2-C. Additionally, the reduction product we designed contains an amino group in the β-position of the indole, which may cause problems due to its instability. C-sulfenylation of enamines is a novel reaction discovered recently by our group, which has not been seen before in the literature. In this study, optimization of the reaction conditions and exploration of the substrate scope were further undertaken for this reaction, which reveal that a series of enamines with N-Cbz, Ac, COt-Bu, CO2Et protection groups could all undergo smooth C-sulfinylations with the comined use of sodium benzene sulphinate, DAMc and MeSiCl3, efficiently furnishing the β-amino vinylsulfoxide products in high yield, affording a practical and highly efficient methods for synthesis of functional vinylsulfoxides.
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大气臭氧的损耗导致了地球表面具有生物学效应的紫外线-B(UV-B)辐射增强。同时,大气成分变化中除了UV-B辐射增强外,氮沉降是一个新近出现而又令人担忧的环境问题,其来源和分布正在迅速扩展到全球范围,并不断向陆地和水生生态系统沉降。本试验在四川省境内的中国科学院茂县生态站内进行,以云山、冷杉、色木槭和红椋子幼苗为模式植物,从生长形态、光合作用、抗氧化能力和矿质营养等方面研究了青藏高原东缘4种树苗对全球变化-增强UV-B辐射和氮供应(氮沉降)的响应。该试验为室外盆栽试验,包括四个处理:(1)大气UV-B辐射+无额外的氮供应(C);(2)大气UV-B辐射+额外的氮供应(N);(3)增强UV-B辐射+无额外的氮供应(UV-B);(4)增强UV-B辐射+额外的氮供应(UV-B+N)。其目的:一方面有助于丰富我国对全球变化及区域响应研究的全面认识,进一步完善在全球气候变化条件下臭氧层削减和氮沉降对陆地生态系统影响的内容;另一方面,在一定程度上有助于我们更好的理解在全球变化下森林更新的早期过程。具体结果如下: 增强的UV-B辐射在生长形态、光合、抗氧化能力、活性氧和矿质营养方面对4种幼苗都有显著的影响。UV-B辐射增强对幼苗的影响不仅与物种有关,而且,还与氮营养水平相关。总体表现为,高的UV-B辐射导致了色木槭和红椋子幼苗叶片的皱缩和卷曲,并降低了色木槭幼苗的叶片数和叶重,在额外的氮供应下,云杉、冷杉和红椋子的叶重也显著地降低了;色木槭和红椋子幼苗叶片的解剖结构受到了增强的UV-B辐射的影响,增强的UV-B辐射显著地降低了色木槭叶片的栅栏组织厚度,提高了红椋子叶片的厚度;增强的UV-B辐射显著地降低了4种幼苗的单株总生物量、植物地下部分的生长、总叶绿素含量 [Chl (a + b)]、净光合速率和最大量子产量(Fv/Fm),提高了4种幼苗叶片的膜脂过氧化(MDA含量),改变了植物体不同器官中的矿质元素含量;增强的UV-B辐射提高了冷杉、色木槭和红椋子叶片中的过氧化氢含量(H2O2)、超氧负离子(O2-)生成速率,在额外的氮供应下,云杉叶片中的活性氧含量也显著地提高了;在无额外的氮供应条件下,增强的UV-B辐射显著地提高了4种幼苗叶片中的UV-B吸收物质、脯氨酸含量和抗氧化酶的活性(SOD、POD、CAT、GR和APX)。在额外的氮供应条件下,UV-B辐射的增强却显著地降低了冷杉叶片中脯氨酸含量和红椋子叶片中UV-B吸收物质含量,但是,在4种幼苗叶片中,5种抗氧化酶的活性对UV-B辐射的增强没有明显的规律性,增强的UV-B辐射显著地提高了云杉叶片中的POD、SOD和GR的活性,提高了冷杉叶片中的POD和GR活性,提高了色木槭叶片中的POD、SOD和CAT活性和红椋子幼苗叶片中的POD和SOD活性。从这些结果可知,植物在遭受高的UV-B辐射导致的过氧化胁迫时,植物体内形成了一定的保护机制,但是,这种保护不能抵抗高的UV-B辐射对植物的伤害。 额外的氮供应在生长形态、光合、抗氧化能力、活性氧和矿质营养方面对4种幼苗都有一定的影响,不同幼苗对额外的氮供应响应不同,并且受到UV-B辐射水平的影响。在当地现有的UV-B辐射水平下,额外的氮供应显著地提高了幼苗的单株总生物量、植物地下部分的生长、Chl (a + b)、净光合速率(红椋子除外)、UV-B吸收物质(冷杉除外)、脯氨酸含量(红椋子除外)和部分抗氧化酶的活性,降低了H2O2的含量、O2-的生成速率和MDA含量(红椋子除外),改变了植物体内部分矿质元素含量,显著地提高了云杉和冷杉叶片中的Fv/Fm。这些指标总体表明,在当地现有大气UV-B辐射水平下,额外的氮供应对植物的生长和发育是有利的。在增强的UV-B辐射水平下,4种幼苗的生长形态和光合大部分指标都没有受到额外氮供应的影响,额外的氮供应提高了红椋子幼苗的单株总生物量和Chl (a + b)含量,提高了冷杉和色木槭叶片中的活性氧含量和MDA含量,却降低了红椋子叶片中的活性氧含量;额外的氮供应也提高了云杉、色木槭和红椋子叶片中UV-B吸收物质和脯氨酸含量,降低了冷杉叶片中UV-B吸收物质和脯氨酸含量;在抗氧化酶活性方面,额外的氮供应降低了云杉、冷杉叶片中5种抗氧化酶的活性和红椋子叶片中POD和GR的活性,提高了色木槭叶片中的POD和SOD的活性;4种幼苗植物体内的矿质元素含量对额外的氮供应没有显著的规律性。从这些结果可知,在高的UV-B辐射下,额外的氮供应提高了云杉、冷杉和色木槭幼苗对高的UV-B辐射的敏感性,然而,额外的氮供应却促进了红椋子幼苗的生长,原因可能是,在高的UV-B辐射下,额外的氮供应增加了红椋子叶片的厚度、叶重和叶片数,降低了叶片中活性氧含量的结果。表明在高的UV-B辐射水平下,额外的氮供应降低了红椋子幼苗对高的UV-B辐射的敏感性。 在全球变化的趋势下,UV-B辐射增强和氮沉降可能同时存在,我们的研究表明,与大气UV-B辐射+无额外的氮供应处理相比,增强UV-B辐射+额外的氮供应处理显著地降低了幼苗的单株总生物量(红椋子除外)、Chl (a + b)、净光合速率、Fv/Fm(冷杉除外)和MDA含量(红椋子除外),提高了活性氧含量 (云杉除外)、UV-B紫外吸收物质含量(冷杉除外)、脯氨酸含量和部分抗氧化酶的活性,改变了植物体不同器官中的矿质元素含量。结果表明,在当地现有条件下,全球变化(UV-B辐射增强和氮沉降)对云杉、冷杉和色木槭幼苗的生长是不利,尽管植物体内一些抗氧化性指标提高了,然而,却对红椋子幼苗的单株总生物量的累积没有显著的影响。 The depletion of the ozone led to the increase of ultraviolet-B (UV-B) with biological effects in the earth’s surface. At the same time, except for enhanced UV-B radiation, nitrogen deposition was an anxious environmental problem at present, rapidly expanding to the global scope and continuously depositing to land and aquatic ecosystem. The experiment was conducted in Maoxian Ecological Station of Chinese Academy of Sciences, Sichuan province, China. Picea asperata, Abies faxoniana, Acer mono Maxim and Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings were selected as model plants to assess the effects of enhanced UV-B radiation and supplemental nitrogen supply on growth, morphological, photosynthesis, antioxidant and mineral nutrient traits of 4 species seedlings in east Qinghai-Tibetan Plateau. The experiment was potted outdoor, including 4 treatments: (1) ambient UV-B without supplemental nitrogen (control, C); (2) ambient UV-B with supplemental nitrogen (N); (3) enhanced UV-B without supplemental nitrogen (UV-B); (4) enhanced UV-B with supplemental nitrogen (UV-B+N). One hand, it was helpful for enriching our country to comprehensive understanding of the researches in the global change and the region response, further perfecting the effects of the depleted ozone layer and nitrogen deposition on land ecosystem under the global change; the other hand, it was favorable for us to better understanding of the early process of forest renews under the global change. The results were as follows: Enhanced UV-B radiation had significant effects on 4 species seedlings in growth, morphological, photosynthesis, antioxidant and mineral nutrient traits of 4 species seedlings. The effects of enhanced UV-B on plants were not only related with species, but also related with nitrogen nutrient level. Generally, the increase of UV-B radiation led to the shrinkage and curl of leaves in Acer mono Maxim and Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings, and reduced the number of leaf and leaf weight of Acer mono Maxim seedlings, under supplemental nitrogen supply, leaf weight of Picea asperata, Abies faxoniana and Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings significantly also reduced; the anatomical features of leaf in Acer mono Maxim and Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings affected by enhanced UV-B radiation, the increase of UV-B radiation markedly reduced the palisade tissue thickness of Acer mono Maxim leaf and enhanced the leaf thickness of Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings; the enhanced UV-B radiation significantly reduced total biomass per plant of 4 species seedlings, the growth of the underground parts, Chl (a + b), net photosynthetic rate and maximum potential quantum yield of photosynthesis (Fv/Fm), and increased the degree of lipid peroxidation (MDA content) and changed the content of mineral elements in different parts of plants; the enhanced UV-B radiation also increased the rate of superoxide radical (O2-) production and hydrogen peroxide (H2O2) content in leaves of Abies faxoniana, Acer mono Maxim, Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings, under supplemental nitrogen supply, the reactive oxygen species in leaves of Picea asperata seedlings also significantly increased by enhanced UV-B radiation; under without supplemental nitrogen supply, enhanced UV-B radiation evidently induced an increase in UV-B absorbing compounds, proline content and the activities of antioxidant enzymes (SOD, POD, CAT, GR and APX) of leaves in 4 species seedlings. Under supplemental nitrogen supply, enhanced UV-B radiation induced a decrease in proline content of leaves in Abies faxoniana seedlings and UV-B absorbing compounds of leaves in Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings, but, there were no obvious rules in the activities of five antioxidant enzymes of 4 species seedling leaves to enhanced UV-B radiation, enhanced UV-B radiation significantly increased the activities of POD, SOD and GR in Picea asperata leaves, the activities of POD and GR in Abies faxoniana leaves and the activities of POD, SOD and CAT in Acer mono Maxim leaves. The results indicated that some protective mechanism was formed when plants were exposed to enhanced UV-B radiation, but the protection could not counteract the harm of high UV-B radiation on plants. Supplemental nitrogen supply had some effects on 4 species seedlings in growth, morphological, photosynthesis, antioxidant and mineral nutrient traits. The response of 4 species seedlings was different to supplemental nitrogen supply, and was affected by UV-B levels. Under local ambient UV-B radiation, supplemental nitrogen supply significantly increased the total biomass per plant, the growth of underground parts, Chl (a + b), net photosynthetic rate (except for Acer mono Maxim seedlings), UV-B absorbing compounds (except for Abies faxoniana seedlings), proline content (except for Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings) and the activities of some antioxidant enzymes, and reduced H2O2 content, the rate of O2- production and MDA content (except for Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings) and changed the content of mineral elemental in different parts; supplemental nitrogen supply also evidently increased Fv/Fm in Picea asperata and Abies faxoniana seedlings. These results indicated that supplemental nitrogen supply was favorable for the growth of plants under local ambient UV-B radiation. Under enhanced UV-B radiation, mostly parameters in growth and morphology of 4 species seedlings were not affected by supplemental nitrogen supply. Supplemental nitrogen supply increased the total biomass per plant and Chl (a + b) of Swida hemsleyi (Schneid. et Wanger.) Sojak seedling, increased the reactive oxygen species and MDA content in Abies faxoniana and Acer mono Maxim leaves, and reduced the reactive oxygen species in Swida hemsleyi (Schneid. et Wanger.) Sojak leaves; supplemental nitrogen supply also increased UV-B absorbing compounds and proline content in Picea asperata, Acer mono Maxim and Swida hemsleyi (Schneid. et Wanger.) Sojak leaves, decreased UV-B absorbing compounds and proline content in Abies faxoniana leaves; in the activities of antioxidant enzymes, supplemental nitrogen supply significantly reduced the activities of antioxidant enzymes in Picea asperata and Abies faxoniana leaves and the activities of POD and GR in Swida hemsleyi (Schneid. et Wanger.) Sojak leaves, and increased the activities of POD and SOD in Acer mono Maxim leaves; the content of mineral elements in 4 species seedlings was no significantly rule to supplemental nitrogen supply. We knew from the results, under enhanced UV-B radiation, supplemental nitrogen supply made Picea asperata, Acer faxoniana and Acer mono Maxim seedlings more sensitivity to enhanced UV-B radiation, however, accelerated the growth of Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings. The reason was probably that supplemental nitrogen supply increased the leaf thickness, leaf weight and leaf number, reduced the reactive oxygen content of leaf in Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings grown under high UV-B radiation. This showed that supplemental nitrogen supply reduced the sensitivity of Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings to high UV-B radiation. Under the tendency of the global change, enhanced UV-B radiation and nitrogen deposition may probably coexist. The results showed, compared with the treatment of ambient UV-B radiation without supplemental nitrogen supply, the treatment of enhanced UV-B radiation with supplemental nitrogen supply significantly reduced the total biomass per plants (except for Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings), Chl (a + b), net photosynthetic rate, Fv/Fm and MDA content (except for Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings), and increased reactive oxygen content (except for Picea asperata seedlings), UV-B absorbing compounds (except for Abies faxoniana seedlings), proline content and part antioxidant enzymes, and changed the content of mineral elements of different parts. The results indicated that the global change (enhanced UV-B and nitrogen deposition) were not favorable for the growth of plants under local ambient UV-B radiation and nitrogen nutrient level,, though increased some antioxidant indexes, however, the treatment of enhanced UV-B with supplement nitrogen supply did not significantly affect on the biomass accumulation of Swida hemsleyi (Schneid. et Wanger.) Sojak seedlings.
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高等植物种子胚乳贮藏蛋白是种子发芽时的主要氮源,也是人类和动物食用植物蛋白的主要来源。大麦种子胚乳贮藏蛋白主要是醇溶蛋白(hordeins),占大麦胚乳总蛋白的50–60%。根据大麦醇溶蛋白的大小和组成特点,大麦醇溶蛋白被划分为三种类型:富硫蛋白亚类(B,γ-hordeins)、贫硫蛋白亚类(C-hordeins)以及高分子量蛋白亚类(D-hordeins)。B组和C组醇溶蛋白是大麦胚乳的两类主要贮藏蛋白,它们分别占大麦总醇溶蛋白成分的70–80%和10–12%。遗传分析表明,大麦B、C、D和γ-组醇溶蛋白分别是由位于大麦第五染色体1H(5)上的Hor2、Hor1、Hor3和Hor5位点编码。Hor2位点编码大量分子量相同但组成不同的B组醇溶蛋白(B-hordein)。B-hordein的种类、数量和分布是影响大麦酿造、食用及饲养品质的重要因素之一。为深入了解B-hordein基因家族的结构和染色体组织,探明Hor2位点基因表达的发育调控机制,最终达到改良禾谷类作物籽粒品质的目的,本研究以青藏高原青稞为材料,采用同源克隆法,分别克隆B-hordein基因和启动子,通过原核生物表达验证B-hordein基因功能,并利用实时定量PCR探索B-hordein基因表达时空关系,取得如下研究结果: 1. 以具有特殊B组醇溶蛋白亚基组成的9份青藏高原青稞为材料,根据GenBank中三个B-hordein基因序列(GenBank No. X03103, X53690和X53691)设计一对引物,通过PCR扩增,获得23个B-hordein基因克隆并对其进行了序列分析。核苷酸序列分析表明,所有克隆均包含完整的开放阅读框。有11个克隆都存在一个框内终止密码子,推测这11个克隆可能是假基因。推测的氨基酸序列分析表明,所有大麦B-hordein具有相似的蛋白质基本结构,均包括一个高度保守的信号肽、中间重复区以及C-端结构域。不同大麦种重复区内重复基元的数目有较大差异。青稞材料Z07–2和Z26的B-hordeins仅具有12个重复基元结构,更接近于野生大麦。这些重复基元数目的差异导致了重复区序列长度和结构的变异。这种现象极可能是由于醇溶谷蛋白基因在进化过程中染色体的不平衡交换或复制滑动所造成的。对所克隆基因和禾本科代表性醇溶谷蛋白基因进行聚类分析,结果表明所有来自栽培大麦的B-hordeins聚类成一个亚家族,来自野生大麦的B-hordeins以及普通小麦的LMW-GS聚类成另外一个亚家族,表明这两个亚家族的成员存在显著差异。此外,我们发现B-hordein基因推测的C-末端序列具有一些有规律的特征:即具有相同C-末端序列的B-hordein基因在系统发生树中聚类为同一个亚组(除BXQ053,BZ09-1,BZ26-5分别单独聚为一类外)。这个特征将有助于我们对所有B组醇溶蛋白基因家族成员进行分类,避免了在SDS-PAGE电泳图谱上仅依靠大小分类的局限性。 2. 根据上述克隆的青稞B-hordein基因的5’端序列设计三条基因特异的反向引物,以青稞Z09和Z26的基因组DNA为模板,采用SON-PCR和TAIL-PCR技术分离克隆出8个B-hordein基因的上游调控序列(命名为Z09P和Z26P)。序列分析表明,推测的TATA box位于–80 bp,CAAT–like box位于–140 bp处。此外,Z09P和Z26P中有六个序列在–300 bp处均存在一个由高度保守的EM基序和类GCN4基序构成的胚乳盒(Endosperm Box,EB),在约–560 bp处存在一个胚乳盒类似结构。而Z09P-2和Z26P-3不存在保守的胚乳盒或其类似结构,预示着这两个启动子所调控的基因表达可能受不同类型反式作用因子的调节,推测该启动子对基因的表达调控具有多样性。 3. 将B-hordein基因的开放阅读框定向克隆到表达载体pET-30a中,将其导入大肠杆菌表达菌株BL21中进行外源基因的诱导表达以验证所克隆基因的功能。结果表明仅含重组子pET-BZ07-2和pET-BZ26-5的BL21细菌有目的表达蛋白产生。在诱导3 h时的蛋白表达量最高;3 mM IPTG诱导的蛋白表达量要高于1 mM IPTG诱导的表达量。这为分离纯化B-hordein蛋白以及进一步研究其对大麦籽粒品质的影响奠定基础。 4. 根据从青稞Z09和Z26中分离克隆的B-hordein基因序列设计一对基因特异的引物,同时,选择大麦α-微管蛋白基因(GenBank no. U40042)为看家基因并设计特异引物,利用实时荧光定量PCR检测了青稞籽粒4个胚乳发育时间段的B-hordein基因表达,荧光定量结果显示:两份材料中B-hordein基因的表达量均随发育过程的进行而逐渐升高。Z09中B-hordein基因在开花后7天开始转录,而Z26开花4天后就有低水平B-hordein的表达,这表明Z26中B-hordein基因可能比Z09表达的较早或者Z09中B-hordein基因表达水平较低以致于不能被检测到。此外,在4个不同的胚乳发育时期中,Z26中B-hordein基因的表达量均高于Z09材料。在开花12天到18天的过程中,Z09和Z26中B-hordein基因的表达水平有一个急剧性的升高。这说明在不同胚乳发育时期,Hor2位点的B-hordein等位基因变异体存在mRNA的差异表达。 Seed endosperm storage proteins in higher plants are the main resources of nitrogen for germinating and plant proteins for human and animals. Barley prolamins (also called hordeins) are the major storage proteins in the endosperm and account for 50–60% of total proteins. Hordeins are classically divided into three groups: sulphur-rich (B, γ-hordeins), sulphur-poor (C-hordeins) and high molecular weight (HMW, D-hordeins) hordeins based on the size and composition. B-hordeins and C-hordeins are two major groups and each respectively account for about 70-80% and 10-12% of the total hordein fraction in barley endosperm. Genetic analysis showed that B-, C-, C-, γ-hordeins are encoded by Hor2, Hor1, Hor3 and Hor5 locus on the chromosome 1H (5). Hor2 locus is rich in alleles that encode numerous heterogeneous B-hordein polypeptides. It is reported that B-hordein species, quantity and distribution are significant factors affecting malting, food and feed quality of barley. To understand comprehensively the structure and organization of B-hordein gene family in hull-less barley and explore the developmental control mechanisms of Hor2 locus gene expression and eventually to better exploitation in crop grain quality improvement, we isolated and cloned B-hordein genes and promotors of hull-less barley from Qinghai-Tibet Plateau by PCR, and testified their expression founction in bacteria expression system and explore their spatial and temporal expression pattern by quantitative real time PCR. Our results are as followed, 1. Twenty-three copies of B-hordein gene were cloned from nine hull-less barley cultivars of Qinghai-Tibet Plateau with special B-hordein subunits and molecularly characterized by PCR, based on three B-hordein genes published previously (GenBank No. X03103, X53690 and X53691). DNA sequences analyses confirmed that the six clones all contained a full-length coding region of the barley B-hordein genes. Eleven clones all contain an in-frame stop codon and they are probably pseudogenes. The analysis of deduced amino acid sequences of the genes shows that they have similar structures including signal peptide domain, central repetitive domain, and C-terminal domain. The number of the repeats was largerly variable and resulted in polypeptides in different sizes or structures among the genes. Twelve such repeated motifs were found in Z07–2 and Z26, and they are close to those of the wild barleys, and it is most probably caused by unequal crossing-over and/or slippage during replication as suggested for the evolution of other prolamins. The relatedness of prolamin genes of barley and wheat was assessed in the phylogenetic tree based on their polypeptides comparison. Our phylogenetic analysis suggested that the predicted B-hordeins of cultivated barley formed a subfamily, while the B-hordeins of wild barleys and the two most similar sequences of LMW-GS of T. aestivum formed another subfamily. This result indicated that the members of the two subfamilys have a distinctive difference. In addition, we found the B-hordeins with identical C-terminal end sequences were clustered into a same subgroup (except BXQ053,BZ09-1 and BZ26-5 as a sole group, respectively), so we believe that B-hordein gene subfamilies possibly can be classified on the basis of the conserved C-terminal end sequences of predicted polypeptide and without the limit of SDS-PAGE protein banding patterns. 2. The specific primers were designed according to the published sequences of barley B-hordein genes from Z09 and Z26. Using total DNA isolated from them as the templates, eight clones (designated Z09Pand Z26P) of upstream sequences of the known B-hordein genes was obtained by TAIL-PCR and SON-PCR. Sequences analysis shows that the putative TATA box was present at position –80 bp and CAAT-like box at position –140 bp. Besides, a putative Endosperm Box including an Endosperm Motif (EM) and a GCN4-Like Motif was found at position –300 bp in six clones, and another Endosperm-like box was found at positon –560 bp. While the Endosperm Box or Endosperm-like box was not found in Z09P-2 and Z26P-3. This may indicate that gene expression drived by the two promtors was probably controlled by different trans-acting factors and the genetic control mechanism of corresponding gene expression may be diverse. 3. The B-hordein genic region coding for the mature peptide was cloned into expression vector pET-30a and transformed into bacterial strain BL21 for identifying gene expression fountion. Protein SDS–PAGE analysis showed that only the transformed lysate with the pET-BZ07-2 and pET-BZ26-5 constructs produced proteins related to B-group hordeins of barley, and the mounts of proteins induced by 3 mM IPTG and 3 h were higher than other conditions. This established a base for isolating and putifying B-hordein and further exploring their effects on barley grain quality. 4. The gene-specific primers of B-hordein genes from Z09 and Z26 were used for the quantification of B-hordein gene expression. The α-tubulin gene from Hordeum vulgare subsp. vulgare (GenBank accession number U40042) was used as a control gene. The result shows the transcription of the B-hordein genes in Z09 was found 7 days after flowering, while the transcription of the B-hordein genes in Z26 was found 4 days after flowering, but at a very low level, and it suggested that the B-hordein genes in Z26 probably expressed earlier than those in Z09, or the B-hordein genes in Z09 expressed at so a lower level than Z26 that it can not detected. In addition, B-hordein genes in Z26 accession showed higher expression levels than those in Z09 in four developing stages. Furthermore, a progressive increase in the expression levels of the B-hordein genes between 12 and 18 days after anthesis was observed in both Z09 and Z26. It implies that the B-hordein allelic variants encoded by Hor2 locus exist the differential expression in mRNA levels of during barley endosperm development.